V Danube Symposium on Chromatography Yalta, November 11-16, 1985

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V DANUBE SYMPOSIUM ON CHROMATOGRAPHY YALTA, NOVEMBER 11-16, 1985

ABSTRACTS

NAUKA" ACADEMY OF SCIENCES OF THE USSR GENERAL AND TECHNICAL CHEMISTRY DiVISION

THE SCIENTIFIC COUNCIL ON CHROMATOGRAPHY INSTITUTE OF PHYSICAL CHEMISTRY

V DANUBE SYMPOSIUM ON CHROMATOGRAPHY

YALTA, NOVEMBER 11-16, .1985

ABSTRACTS

к "NAUKA" troc 5t3.5H

V Danube Symposium on Chromatography. ABSTRACTS. И.: "Nauka", 1985.

The volume contains short abstracts of plenary lectures,section papers and poster contributions presented at the V Danube Symposium on Chromatography, Proceedings of the Symposium reflect recent advances and prospects of further development in all branches of modern chromatographic techniques, including column liquid chromatography, gas chromatography, ion exchan ge, thin-layer chromatography, flow electrophoresis as well as equipment development. PROGRAM COMMITTEE K.I.Sakodynskii (chairman), B.G.Belenkii, V.G.Berezkin, G.A.Chikin, V.P.Chizhkov, V.A.Davankov, O.G.Larionov, K.H.Saldadze, G.V.Samsonov, A.A.Zhukhovitskii, P.P.Zolotarev

(g) Institute of Physical Chemistry of the USSR Academy of Sciences, 1985 GENERAL PROBLEMS OF CHROMATOGRAPHY

001 QUANTIFICATION OF THE INFORMATION OF CHROMATOGRAPHIC RETENTION DATA FOR THE IDENTIFICATION OF ANALVTES КОЛИЧЕСТВЕННАЯ ОБРАБОТКА ПАРАМЕТРОВ ХРОКАТОГРАФИЧЕСКОГО УДЕРЖИВАНИЯ С ЦЕЛЬЮ ИДЕНТИФИКАЦИИ АНАЛИЗИРУЕМЫХ ВЕЩЕСТВ J.F.K.Huber, G.Reich., E.Kenndler, Vienna, Austria

In qualitative analysis the nature of the components of a sample have to be determined. In principle two approaches can be usedt Identification by means of characteristic data and elucidation of the chemical structure. In the first approach the identity of species is determined by charac terizing the species by means of data. Comparison of the data allows to decide if two species are identical or not. Due to the limited precision and accuracy of the data such a decision can be made only with a certain probability. The capacity of an identification method depends on its discrimination power and can be described quantitatively by applying information theory. In chromatography the components are discriminated by the separation process and the retention data can be used for the characterization of the species. The identifica tion capacity of this method depends on the efficiency and selectivity of the separation process as well as on the probability distribution of the species occurring wi thin a given retention range. Quantification of the information content of the different modes of chroraato- graphy leads to the conclusion that single stage opera tion gives less than 9 bit and this means not a very high identification capacity.

The identification capacity can be increased significan tly if multidimensional chromatography is applied. In this technique the sample is separated on a cascade of columns by transfering the effluent of a given column in frac tions to the columns of the next separation stage of the system. It is crucial that columns of different retention characteristics are used. Such columns, which are little correlated in retention, can be selected by pattern recog nition. If carried out in a proper way a high identifica tion capacity can be achieved by multidimensional chroma tography. 002. PL COMPUTER CHROMATOGRAPHY POSSIBILITIES AND APPLICATIONS: TWO EXAMPLES TO CAPILLARY GC AND HIGH PERFORMANCE TLC ВОЗМОЖНОСТИ ШИРОКОГО ПРИМЕНЕНИЯ КОМПЬЮТЕРОВ В ХРОМАТОГРАФИИ: ДВА ПРИМЕРА В КАПИЛЛЯРНОЙ ГХ И В ВЫСОКОЭФФЕКТИВНОЙ тех R.E.Kaiser, R.I.Rieder, Bad Duerkheira.FRG

ON-line using a 10 megabyte small laboratory Personal Computer (Apple lie) one can improve gas and liquid chro matographic techniques remarkably. In the case of two series coupled capillary gas chroma tography system the laboratory computer changes the gas pressure between the two series coupled capillaries, digi tizes, stores, integrates and evaluates the chromatogram series of an unknown sample mixture. The computer commands 5 (or more) consecutive injections of the same sample and after the series of runs it can do either adjusting the chromatographic system to optimum polarity conditions for further routine analysis or offers multilevel qualitative and quantitative analysis of the GC sample. As a result at least four sets of retention index increments and in formation about peaks under peaks are given, thus offering a new chromatographic evaluation principle: Multi Chroma tography, MC.

The pressure adjusting system between the capillaries is a digital controlled piezo electronic fluidix system and adjusts hydrogen pressure reproducible at the millibar level thus controlling flow at the microliter per minute level. This determines the residence time in polare and apolare capillaries and therefore changes retention index data for polare compounds for several hundred unites in the pressure ranges applicable.

In the case of planar chromatography the computer con- trolls sampling, separation and quantitation on high per formance thin-layer plates. One of the surprising results is that HPTLC analysis now reaches the top of accuracy of all existing chromatographic techniques, although many chromatographers consider TLC as semiquantitative only. By total area quantitation in one mm wide tracks touching each other the lab computer offers a complete change of the quantitation techniques including automatic wide range calibration, which leeds to HPTLC in the sense of High Precision. The computer chromatography principles of the two examples mentioned and further possibilities are shown and will be discussed.

4 003 PL STATE AND TRENDS IN OPLC СОСТОЯНИЕ И ТЕНДЕНЦИИ В ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ ПРИ ПОВЫШЕННОМ ДАВЛЕНИИ E.Tyihak, Budapest, Hungary The special possibilities of totally closed planar liquid chromatographic system were first realized by deve lopment of over-pressured-layer chromatography (OPLC) using a pressurized ultra-micro chamber. OPLC is a logical extension of the theory-and practice developed in HPLC to the field of planar liquid chromato graphy. However there are important differences between OPLC and HPLC. It is obvious that the development trends in OPLC are based upon these facts. The outstanding properties of chemically bonded phases on various chromatoplates were used in OPLC for the effi cient separation of different substance groups. Because of the complexity of the retention mechanism, it is possible to separate simultaneously compounds of a wide polarity range with excellent resolution and short reten tion times. Using a two-dimensional separation in OPLC the effi ciency increases quadratically. The versions of OPLC employ constant flow rate which has special advantages in the case of chromatoplates with concentrating zone. The extremely narrow line of substan ce that is formed at the interface provides an optimal start for chromatographic separation'. This separation mo de is especially attractive when larger sample amounts are applied. It is important that in this system very dilute solutions can be used, too. The overpressured multi-layer chromatography is a very attractive version of OPLC because a large number of samples can be separated at the same time. One- and two-dimensional on-line separation and detec tion in OPLC increase the efficiency of this technique and the application possibilities, alike. Preparative OPLC separation modes are more suitable for isolation of substances of different structure than the classical TLC. OPLC opens a new way in this field as the separation is many times faster in a pressurized chamber than in normal TLC chambers and so the diffusion of substances in the sorbent layer decreases dramatically.

004. MICROCOLUMN CHROMATOGRAPHY OF SYNTHETIC POLYMERS, PROTEINS AND THEIR FRAGMENTS МИКРОКОЛОНОЧНАЯ ХРОМАТОГРАФИЯ СИНТЕТИЧЕСКИХ ПОЛИМЕРОВ, БЕЛКОВ И ИХ ФРАГМЕНТОВ B.G.Belenkii, E.S.Gankina, V.G.Maltsev, Leningrad,USSR The application of exclusion and hydrophobic micro column chromatography to the analysis of synthetic poly mers, proteins, peptides and amino acids has been consl-

5 dered. Since a small amount of the eluent is used in columns 0,5 mm in diam. ( V » 50/ul) for exclusion chroma tography, it is possible to employ 'any solvents as eluents including expensive and toxic solvents. Spectrofotometric, fluorometric and refractometric detectors with the cell volume of O.l-l.O ml were used for the detection of the eluate in microcolumn chromatography. Steel and fluoroplastic microcolumns 0.5-1.0 mm in diam. with the efficiency of 40-70,10,J theoretical plates/m were used. Examples are presented of microcolumn exclusion chroma tography used for analysis of molecular-weight distribu tion of synthetic oligomers and polymers Including poly esters and polyamides and for the molecular-weight deter mination of peptides and proteins in the random coil con formation. The method of gradient microchromatographic analysis of DMS-amino acids with the aid of.a laser fluorimetric detector with a sensitivity of 10 mol was developed. 005. MICROCOLUMN SEPARATION METHODS I PAST, PRESENT AND FUTURE МЕТОДЫ МИКРОКОЛОНОЧНОГО РАЗДЕЛЕНИЯ: НАСТОЯЩЕЕ, ПРОШЛОЕ И БУДУЩЕЕ M.Novotny, Bloomington, Indiana, USA Whereas in the field of gas chromatography, the column miniaturization already started with the development of open tubular columns by M.J.E.Golay /1/, similar efforts in liquid chromatography (LC) can only be traced to the late 1970s. Encouraged by the theoretical considerations of Giddings /2/, as well as the successful efforts to decrease particle size in LC /3-5/, our laboratory pursued various investigations, aiming at a drastic decrease of the column inner diameter /6,7/. Initially, for somewhat different reasons, Scott and Kucera /e,9/ and Ishii et.al. /10/ Independently followed their LC equipment miniaturi zation ideas. An increasing number of laboratories have subsequently joined these efforts. Today, microcolumn LC belongs to the most popular areas of investigation in analytical separations, as evidenced by a rapidly increasing number of communications and the three book's /11-13/ edited within a period of just one year. The principal reasons for this vivid interest in the subject stem from: (a.) the high-performance potential of microcolumn techniques; (b.) possibilities of using "exotic" mobile-phases and column materials; (c.) the emergence of novel detection techniques due to' extremely low flow-rates associated with microcolumns; and, (d.) compatibility with various micromanipulation techniques of modern biology and medicine. Recently developed methods of capillary supercritical fluid chromatography /14/ and capillary electrophoresis /15/ share certain common features with microcolumn LC..

6 Various column technologies have rapidly advanced during the last several years. Mills open tubular columns remain somewhat Illusive in LC (but not in supercritical fluid chromatography and electrophoresis), nueerou» unique applications of microbore colusn», semipermeable packed capillaries, and slurry-packed capillary columns have already bean demonstrated. In addition, miniaturized instrumentation is being rapidly developed, including sampling devices, small-volume (nanoliter level) detectors, gradient elution systems, etc. Similar advances are being continuously made in capillary supercritical fluid chroma tography. s fi Column efficiencies in the range of 10 -lO theoretical plates were already achieved with all three abovementioned methods, providing the basis for effective analyses of complex nonvolatile mixtures. During the last year, our laboratory has demonstrated analyses of several environ mentally and biochemically important mixtures obtained at very high resolution. In the area of detection and ancil lary techniques, significant advances have also been made in association with microcolumns; these Include primarily devices that combine extremely well with very small detec tion volumes and low flow-rates: flame- and laser-based detectors, microelectrodes, and mass spectrometers. Minia turized optical concentration- sensitive detectors that work remarkably well Include Fourier transform infrared spectroscopy, a laser-based fluorescence detector, and a fluorescence imaging (intensified photodiode array) device; their unique applications from our laboratory will be discussed. Further improvements in the separation performance and systems versatility appear feasible. New avenues have also been opened for extremely high sensitivities (Femtogram levels and below) due to the development of miniaturized systems. With the recent revolutionary advances in optical devices and microsensor systems, the overall area is likely to grow in importance. Related chemical advances will be equally important to ascertain further expansion of the powerful microcolumn techniques into various bran ches of science and technology. 1. H.J E.Golay, in D.B.Desty (Editor), Gas Chromatography 1958, Butterworths, London, 1958, p. 36. 2. J.C.Giddings, Anal.Chem., 36, 1890 (1964). 3. J.H.Knox , M.Saleem, J.Chromatogr. Sci.,7, 614 (1969). 4. J.F.K.Buber, in E.Kovats (Editor), Column Chromatogra phy, Chimia Supplement, Sauerlander, Aarau, 1970, p.24. 5. J.J.Kirkland, in S.G.Perry (Editor), Gas Chromato graphy 1972, Applied Science Publishers, Barking, U.K., 1973, p. 39. 6. T.Tsuda, M.Novotny, Anal.Chem., 50, 271 (1978). 7. T.Tsuda, M.Novotny, Anal.Chem.j50, 632 (1978). 8. R.P.H.Scott, P.Kucera, J.Chromatogr.,125, 251 (1976). 9. R.P.W.Scott, P.Kucera, J.Chromatogr.,Тб7, 51 (1979). 10. D.Ishii, K.Asai, K.Hibi, T.Jonokuchl, M.Nagaya, J.Chromatogr., 144, 157 (1977). 7 11. P.Kucera (Editor), Microcolumn High-Performance Liquid Chromatography, Elsevier, Amsterdam, 1984. 12. R.P.W.Scott (Editor), Small Bore Liquid Chromatography Columns, Wiley-Interscience, New York, 1984. 13. M.V.Novotny, D.Ishii (Editors), Microcolumn Separa tions, Elsevier, Amsterdam, 1985. 14. M.Novotny, S.R.Springeton, P.A.Peaden, J.C.Fjeldsted, M.L.Lee, Anal.Chem.,53, 407,(1981). 15. J.W.Jorgenson, K.D.LuTcacs, Anal.Chem., 53, 1298,(1981). 006 CHROMATOGRAPHY OF VAPOURS AND CHROMADISTILLATION ХРОМАТОГРАФИЯ ПАРОВ И ХРОМАДИСТИЛЛЯВДЯ А.А.zhukhovltskii, Moscow, USSR Solution of some problems (introduction of large doses for analysis of admixtures, in order to prepare pure sub stances and to determine their physico-chemical characte ristics, establishing the distribution of components according to their boiling points, especially for high boiling components, continuous separation, etc) by gas chromatographic methods is rather difficult. In chromadistillation (CD) one makes use of the featu res of chromatography at vapour pressures close or nearly equal to saturation (existence of "restrictive regime", efficiency of the Immobile temperature gradient). Application of CD allows to introduce doses of mixtures to be analysed which may be as much as Ю citr of liquid, to attain the sensitivity of trace determination about 1»10"6, to carry out the distillation of substances with boiling points near 600°C, to measure the vapour pressu res for small doses (microgramms), ensuring the purity of components, as well as to carry out continuous separation of two-component mixtures. 007. GENERAL SEPARATION THEORY AND CHROMATOGRAPHY ОБЩ! ТЕОРИЯ РАЗДЕЛЕНИЯ И ХРОМАТОГРАФИЯ V.P.Chlzhlcov, M.P.Zabokritsky, N.V.Sterkhov.E.F.Litvin, Moscow, USSR In thermodynamics of equilibrium processes it is consi dered idealized gas separation (mixing) models based on the assumption concerning the existence of ideal semiper meable membranes and the possibility of obtaining of absolutely цо'е substances in final energy loss. The con cept of ideal cascade is often used with respect to real processes of fine separation. Concepts of final separation factor, single separation step and separation potential are introduced into thermodynamic theory of the ideal cascade. These a priori concepts limit the completeness of binary mixture separation in cascade of final dimen sions and in final energy.loss; energy and time necessary for obtaining the set mixture resolution increase with decreasing the differences between the components. The 8 general separation theory has been spread to the aspect of chromatography dealing with the principle of ideal cascade, The model of ideal chromatographic cascade has been proposed. The ideas concerning the ideal (non-equi librium) chromatography and the least time for binary mixture separation in chromatographic ana-lysis are dis cussed. The thermodynamic ground of the work on cascade step separation for particular case of partition chroma tography has been given.

008 PRINCIPLE OF NON-ADDITIVITY FOR SORPTION ENERGY AND ITS APPLICATION IN CHROMATOGRAPHY ПРИНЦИП НЕАДДИТИВНОСТИ СВОБОДНОЙ ЭНЕРГИИ И ЕГО ПРИМЕНЕНИЕ В ХРОМАТОГРАФ»! R.V.Golovnya, Moscow, USSR Therwodynamic study of the gas chromatographic behavi our of homologous series of organic compounds has been car ried out. As a conclusion of our study we can postulate the reason for the violation of the addltivity principle for the energy contribution of members of a homologous series to the free energy sorption: the energy of Van der Waale interactions between the stationary phase and the sample molecules having the functional groups bound directly to

the methyl (CH3) radical is lower than the same energy of interactions between the stationary phase and sample

molecules having more than 3-4 methylene (CH2) units. Based on the thermodynamic representation of the GC separa-lon process we obtained the uniform equation for the accurate description of the dependence of AG, log V„,

I and log tR on sequential number (m) of a homologue (including the first members) of any homologous series

RmX. The high accuracy of retention index calculations by uniform equations obtained on polar and nonpolar capilla ry columns are demonstrated on five homologous series. The possibility of the application of uniform equations for calculation of retention parameters in GLC, TLC and gas-adsorption chromatography was established. The uniform equation can be used also for the calcula tion of the distribution coefficients К for homologues in liquid-gas, liquid-liquid, solid-gas systems with a good accuracy. It was illustrated on 15 examples for different organic series.

009 MOLECULAR STRUCTURE AND SOLUTE-SOLVENT INTERACTION OF CHEMICALLY BONDED PHASES МОЛЕКУЛЯРНАЯ СТРУКТУРА И ВЗАИМОДЕЙСТВИЯ СОРБАТА С РАСТЕОРИТЕЛЕМ ПРИ ХР0МАТ0ГРА9ЙИ НА ХИМИЧЕСКИ СВЯЗАННЫХ ФАЗАХ E.Bayer, Tubingen, FRG Summary not arrived on time for. printing.

9 010 S.l-5 MODERN TRENDS OF DEVELOPMENT OF THE GAS CHROMATOGRAPHIC HEAD SPACE ANALYSIS СОВРЕМЕННЫЕ ТЕНДЕНЦИИ РАЗВИТИЯ ГА30ХР0МАТ0ГРАФИЧЕСК0Г0 ПАР0ФАЗН0Г0 АНАЛИЗА B.V.Ioffe, Leningrad, USSR

Abstract not arrived on time for printing.

Oil PL CHROMATOGRAPHIC PEAK OVERLAP ПЕРЕКРЫВАНИЕ ХРОМАТОГРАФИЧЕСКИХ ПИКОВ J.C.Giddings, Utah, USA Abstract not arrived on time for printing 012 S.7-S ' THE DEVELOPMENT OF THE METHODS OF QUANTITATIVE INTERPRETATION OF CHROMATOGRAMS РАЗВИТИЕ МЕТОДОВ КОЛИЧЕСТВЕННОЙ ИНТЕРПРЕТАЦИИ ХРОМАТОГРАММ M.S.Vigdergauz, Kuibyshev, USSR Some new versions of carrying out analyses and interpre ting the chromatograms obtained are suggested here to realize the system approach to both qualitative and quan titative chromatographic analysis: 1. Interpolation methods requiring the addition of two or more standards to the sample with the. chromatographic separation to follow. 2. Methods requiring asynchronous injections of samples and standards. 3. Methods concerned with the use of marker-standard system. 4. Methods concerned with the use of addition-standard system. 5. Methods requiring the extrapolation of the data ob tained (the adjustment to the zero quantity of the standard and the consideration of the nonlinearitxes of the calib ration factor) . 6. The interpolation methods of determining the calib ration factors for the peak hights and peak areas. The application of the suggested versions of interpre tation to the chromato-extraction methods (liquid extrac tion, head space analysis) is of special interest. The use of correlations between the solute retention and the corresponding characteristics of the detector response is considered to be an aspect of the system approach to the qualitative 'and quantitative analysis. The approach discussed above is used in working out the chromatographic fragments of the universal system of chemical analysis. 10 013 . И- STATE-OF-THE-ART REPORT OF THEORY OF MULTICOMPONENT CHROMATOGRAPHY WITH SPECIAL EMPHASIS ON PREPARATIVE SEPARATIONS СОВРЕМЕННОЕ СОСТОЯНИЕ ТЕОРИИ МУЛЬТИКОМПОНЕНТНОЙ ХРОМАТОГРАФИИ И ПРЕПАРАТИВНЫХ РАЗДЕЛЕНИЙ F.G.Helfferich, Pennsylvania, USA Abstract not arrived on time for printing.

014 S.6-4 POTENTIALITIES OF FOCUSING FIELD-FLOW FRACTIONATION ПРИНЦИПИАЛЬНЫЕ ВОЗМОЖНОСТИ ПРОТОЧНОГО ФРАКЦИОНИРОВАНИЯ В ПОПЕРЕЧНОМ ПОЛЕ S.N.Semionov, A.A.Kuznetsov, P.p.Zolotarev, Moscow, USSR

.Focusing field-flow fractionation (FFFF) is a modifica tion of field-flow fractionation (FFF). In the technique proposed, to separate the particles introduced as a thin layer into a laminar flow a transversal force is used. It does not accumulate the particles to the channel's wall as it is in the classical FFF, but focuses the par ticles with different physical and cheaical properties in central regions of 'the channel where the velocities of the?'- notion in the flow are different. The lack of inte raction between the particles and the channel's walls ma kes it possible to obtain better stability of separated particles, to decrease the peak dispersion and to reduce the separation time. The processes of perticle density separation in centrifugal force field and in medium den sity gradient, magnetic susceptibility separation in a high-gradient magnetic field, isoelectric points (pi) sepa ration in electric field and in the pH gradient are theoreti cally analysed. It has been shown that the particles with the same pi aay be additionally separated according to correlation slope of their effective charge on pH, and particles with the same densities - in their volume. Due to its advantages, FFFF may become an effective tool in chemical and biological research.

11 015 S.6-3 CHROMATOGRAPHIC REACTORS. PERSPECTIVES FOR PRODUCTION-SCALE PROCESSES ПЕРСПЕКТИВЫ ИСПОЛЬЗОВАНИЯ ХРОМАТОГРАФИЧЕСКИХ РЕАКТОРОВ . В ПРОМЫШЛЕННОМ МАСШТАБЕ S.A.Volkov, G.V.Eroshenkova, S.M.Zamanskii, K.F.Khalilov, Moscow, USSR Reversible reactions can be forced to completion in a chromatographic reactor. The conversion in such reactor may be higher than equilibrium value and temperature may be reduced. The experimental values of conversion and productivity are obtained in fixed bed reactor for cyclohexane dehydrogenation reaction. Effect of sample volume on coversion and effect of catalyst particle size reactor length and diameter on productivity are investigated. Experimental data are compared with the theory based on assumption that sorption isotherm is linear. The productivity can be increased if the moving or fluidized beds of catalyst instead of fixed bed are used. The behaviour of counter-current moving bed chromatographic reactor has been studied theoretically. For the first- order reaction A •* В + С with kinetics and sorption iso therms following linear equation the expressions for cal culation of component concentrations and conversion distri butions were obtained. In addition to a one-section, chro matographic reactor with second exhausting section was examined. The conversion in this two-section reactor is substantially higher than in one section reactor. The effect of various parameters on the conversion was studied too. The possibility of the chromatographic separation in fluidized bed was investigated. The poor separation was obtained in non-organized bed whereas in the bed organi zed by horizontal wire-nets separation achieved did not differ from separation in fixed bed. Reactions in this organized bed may be performed. Some industrially impor tant reactions which may be performed in chromatographic reactor are discussed.

016 S.7-4 ISOMER-SPECIFIC DETERMINATION OF POLYCHLORODIBENZO- DIOXTNS (PCDD) AND -FURANS (PCDF) IN EFFLUENTS FROM INCINERATION PROCESSES СПЕЩШЧЕСКОЕ ОПРЕДЕЛЕНИЕ ИЗОМЕРОВ ПОЛИХЛОРБЕНЗО- ДИОКСИНОВ И ФУРАНОВ В ПРОДУКТАХ ПРОЦЕССОВ СЖИГАНИЯ K.Ballschmiter, Ulm/Donau, FRG The formation of PCDD and PCDF in incineration proces ses is part of the fundamental possibilities of flame

12 chemistry. The formation of soot as a final product from Co-precursors has its counterpart in the formation of oligocycllc arenes and oxyarenee. The pattern of PCDD and PCDP formed by flame chemistry suggests successive reac tions of chlorobenzenes to chlorobiphenyls and chloro- phenole which finally react to PCDD/PCDF as a thermodyna mic sink. Depending on the type and the technology of the incine ration and the physico-chemical properties of the molecu les formed, the pollutants will be found distributed bet ween gas phase, particle phase and water phase of the total effluents. The isomer-specific determination of PCDD/PCDF by high-resolution gas chromatography and mass spectrometric detection will be reported.

017 PL THE COMPARISON OF DIFFERENT TYPE COVALENTLY BONDED CHIRAL PHASES USED IN THE ENANTIOMERIC ANALYSIS OF OPTICALLY ACTIVE COMPOUNDS BY HIGH-PERFORMANCE LIGAND-EXCHANGE CHROMATOGRAPHY СРАВНЕНИЕ РАЗЛИЧНЫХ ТИПОВ ПРИВИТЫХ ХИРАЛЬНЫХ СТАЦИОНАРНЫХ ФАЗ В ЭНАНТИОМЕРНОМ АНАЛИЗЕ ОПТИЧЕСКИ АКТИВНЫХ СОЕДИНЕНИЙ МЕТОДОМ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ V.A.Davankov, A.A.Kurganov, T.M.Ponomareva, Moscow, USSR

The optical purity analysis of chiral compounds acqui red greater importance during the last few years due to greater restrictions on the impurites content in the pre parations used in medicine, pharmacology, biochemistry. One of the most successful techniques developed for this purpose is the ligand-exchange chromatography of optical isomers which permits to achieve high enantiomer resolu tion selectivities.

Among the various modes of the ligand-exchange chroma tography method the most attractive one is using of covalently bounded chiral phases. Such sorbents synthesis includes two principal moments: the selection of an opti mal silica matrix to be modified, and the selection of chiral ligand type to be attached on to silica surface. In this work the results of examination in each of these fields are given together with the data on proper ties of sorbents synthesized. The eorbent evaluation was carried out on the base of Knox-dependences which 'were

13 obtained at different temperatures. The role of some other parameters of the chromatographic process (mobile phase composition, type of organic modifiers of the eluent et cetera) has been investigated too. The analysis of these regularities enabled to single out a number of speci fic features in the ligand-exchange chromatographic reso lution which must be taken into account. The examples of efficient resolution of the amino acid optical isomers on the synthesized sorbents are given.

018 PL NEW ASPECTS OF APPLICATION OF HPLC IN BIOLOGY AND MEDICINE НОВЫЕ АСПЕКТЫ ПРИМЕНЕНИЯ ХРОМАТОГРАФИИ В БИОЛОГИИ И МЕДИЦИНЕ R.T.Toguzov, Yu.V.Tikhonov, Moscow, USSR The modern level of development of chromatography has given rise, with adeguate obviousness, to new fields of application of chromatography in biology and medicine. Chromatography, particularly HPLC, not only greatly broa dens the range of methods used by researchers but also enables a qualitatively new interpretation of some tradi tional views existing in practical medicine. It becomes possible, on the basis of chromatographic methods, to make use of fundamentally new approaches in medical biochemistry in analyzing the most informative biochemical parameters for automatic computerized diagnos tics of diseases, predicting their course, and checking the effectiveness of medical treatment.

The progress in solving the urgent problems of practi cal medicines the identification of the main pathogenesis links of some diseases and risk groups, the supply of laboratory instruments for mass prophylactic medical exa mination of population, the search for new groups of medi cal preparations and new effective methods of medical treatment - is directly related to achievements and success in chromatography. Moot promising in this respect seem to be the development and batch production of versa tile automatic analyzers on the basis of a high-performan ce liquid chromatograph. The identification and study of laws governing the operation of the natural chromatographic systems in biolo gical objects and the determination of their biological importance are the new fields of science; working in this new field, researchers have-"* right to expect results of significance both in the theoretical and practical sense. The data obtained by the authors and illustrating the theses set forth above are discussed in the paper. 14 019 P.1-01 CONTROL BY THE MOLECULAR TRANSFER PROCESSES THROUGHOUT NARROW CHANNELS IN THE RESONANCE LASER RADIATION FIELD УПРАВЛЕНИЕ ПРОЦЕССАМИ ПЕРЕНОСА МОЛЕКУЛ ЧЕРЕЗ УЗКИЕ КАНАЛЫ,ПОЛЕМ РЕЗОНАНСНОГО ЛАЗЕРНОГО ИЗЛУЧЕНИЯ N.V.Karlov, A.S.Laguchev, A.N.Orlov, Yu.N.Petrov, A.M.Prokhorov, M.A.Yakubova, Moscow, USSR 1. The flows of resonance molecules throughout capilla ries and porous structures under the effect of laser radiation can both increase and decrease. This effect could not be explained by the converting processes of laser radiation in heat. 2.The presence of laser radiation near the surface leads to the change of the adsorption potential for the resonance molecules. The executed estimation of this adsorption potential change have shown it being more or on the order of kT. 3. When surface diffusion prevailed, an accurate solu tion shows the adsorption potential depth growth inside the channel to cause degreasing the. molecular flow; just on the contrary, the adsorption potential depth growth on the external barrier surface to cause increasing the mole cular flow. 4. The diffusion equation solution follows to control of resonance molecules throughout capillaries by laser radiation being possible rather for the case of prevailing bulk diffusion, but surface diffusion making a small cont ribution. 020 P.1-02 THE PACKED BED STRUCTURE AND MOBILE PHASE VELOCITY FIELD AND ITS EFFECT ON LONGITUDINAL DISPERSION IN GAS AND LIQUID CHROMATOGRAPHY ПРОДОЛЬНОЕ РАЗМЫВАНИЕ В ГАЗОВОЙ И ЖИДКОСТНОЙ ХРОМАТОГРАФИИ И ЕГО СВЯЗЬ СО СТРУКТУРОЙ ЗЕРНИСТОГО СЛОЯ И ПОЛЕМ СКОРОСТЕЙ ПОДВИЖНОЙ ФАЗЫ S.A.Volkov, V.I.Reznikov, K.F.Khalilov, Moscow, USSR The longitudinal spreading in packed bed, conditioned by nonuniformity of this bed was investigated. The gas and liquid velocity fields at the bed outlet for different parameters was studied by laser anemometry. Experiments were carried out in 4C-140 an tubes filled with diatomite supports usually used in gas chromatography. The velocity varies widely from point to point of cross section. These velocity variations indicate that the par ticles probably cluster into close-packed aggregates, which are "bridges" by loose particles. The velocity varia tions are random and may be characterized by certain sta tistical moments. The experimantal data were processed by computer. This statistical processing quantitatively characterizes the nonuniformity in the velocity field and packed bed structure. Influence of packed bed parameters

IS (size and shape of the support particle, size distribu tion, diameter of tube) on the nonuniformity of velocity field was studied and empiric equations for the statisti cal moments were obtained. A relation between the velocity variations and longi tudinal dispersion in the packed bed was discussed. Auxiliary terms in the expression for the effective longitudinal diffusion coefficient was included.

021 p.1-03 AN ITERATIVE METHOD FOR COMPUTING THE GAS HOLD-UP TIME ON THE BASE OF NON-ADJUSTED RETENTION TIMES OF HOMOLOGUES МЕТОД ИТТЕРАЦИИ ДЛЯ РАСЧЕТА ВРЕМЕН УДЕРЖИВАНИЯ НА ОСНОВЕ НЕПРИВЕДЕННЫХ ВРЕМЕН УДЕРЖИВАНИЯ ГОМОЛОГОВ B.Wronski, eialystok, Poland Z.Witkiewicz, Warsaw, Poland This paper represents an iterative method (IM) for the calculation of the gas -hold-up time (tM) on a base of non- -djusted retention time values (tjy,) of homologues. In the IM it is assumed, that the exact linear depen dence between the logarythmus of the adjusted retention time (lg tjy.) and the number of carbon atoms in a molecu le "i" (ni) is obligatory, like lg th± * an^ + b. The value t(4 is calculated according to the following algo rithm. At first, it is admitted, that tM(pj - 0 ("0" and successive natural numbers in braces are iterative steps) and the system of k-equations is solved by:

2S fcRi * a(o)ni + b(o)' where i • 1» 2, 3, ... , k, concerning a(0i and b/0\. Secondly the values of the gas hold-up times for individual homologues tMi(o) = tRi - 10a'°> ni + b(o> and the arithmetic mean tJJ(i) = j" jjr tj, are calculated. Thirdly the

) a + b equation lg(tRi -"tM(i) * (l) (l)' where i = 1,2,,.k, is solved. Concerning to a^) and b(D the values 10а<1,П 1 +Ь Ш W>= *«i - »* *«(2Г k g 4(1(1) are computed^ At last the equation

a n + b wnere lg(tRl - tM(2)) - (2) i (2) i = 1» 2,... k, is solved and the values a(2), b(2)/ ^M/ii a*e calculated. This algorithm is repeated p-times, as long as tM. .= t„ = const by the established accuracy of the numerical value t„ (e.g. t~. - 0,00 min.).

The calculatedvalues or t4 with IM or other methods for such homologues like n-alkanes and fatty acid methyl esters were compared. Conformable results were obtained. Packed columns with squalane and liquid crystalline sta tionary phases were employed. Conclusions for the calcu lation of tH with reference to the optimal selection for homologues were made. 16 022 P.1-14 MEASUREMENT OF MASS TttANSFER COEFFICIENTS BY THE REVERSED-FLOW GAS CHROMATOGRAPHY TECHNIQUE ИЗМЕРЕНИЕ КОЭФФИЦИЕНТА МАССОПЕРЕНОСА С ПОМОЩЬЮ ГАЗОВОЙ ХРОМАТОГРАФИИ В ОБРАЩЕННОМ ПОТОКЕ G.Karaiskakis, P.Aghathonos, A.tUotis, N.A.Katsanos, Patras, Greece Reversed-flow gas chromatography, a new method for studlng heterogeneous catalysis and other related pheno mena, as diffusion, adsorption etc., is applied here to simultaneous determination of mass transfer coefficients for the evaporation of pure liquids or liquid mixtures and the diffusion coefficients of vapours from these liquids into the carrier gas. The necessary mathematical equations were derived and used to calculate the above parameters for the pure liquids n-pentane, n-hexane, n-heptane, n-octane, metha nol, ethanol, propan-1-ol, butan-1-ol and pentan-1-ol, as well as for the mixtures methanol/water, ethanol/water and propan-1-ol/water, evaporating into carrier gas helium. In the case of mixtures the variation of the mass transfer coefficient for the evaporation of alcohols with their mole fraction, the working temperature and the carrier gas flow-rate was studied. The mass transfer coefficients measured depend, not only on the nature of the alcohol and the other component in the mixture, but also on their mole fraction. Moreover, it increases with increasing temperature, but it is inde pendent of the carrier gas flow-rate. The values of diffusion coefficients found are in good agreement with those calculated theoretically or reported in the literature. 023 P. 1-15 SOLUTION OF INVERSE PROBLEM AND CALCULATION OF SORPTION ISOTHERMS IN CHROMATOGRAPHY РЕШЕНИЕ ОБРАТНОЙ ЗАДАЧИ И РАСЧЕТ ИЗОТЕРМ СОРБЦИИ В ХРОМАТОГРАФИИ A.V.Larin, Moscow, USSR The theory of nonideal chromatography has been inade quately developed, in this context the methods of solu tion of the reverse problem has not been worked out for arbitrary sorption isotherms. In the present report the possibilities of the new variant of plate theory - the model of layer of the equilibrium adsorption -are exami ned. In contrast to the layer-by-layer method the model proposed uses only a length scale for a characterization of the kinetic constants. The equilibrium adsorption layer Le, for which the outlet concentration is numeri cally equal to the concentration in equilibrium with the average adsorption of whole layer, is determined as the effective kinetic constant. This assumption allows us ко

2.3uc. 872 17 obtain the quite total and reliable solutions in real chromatography. On the basis of the model proposed it has been demonst rated that the shape of the elution outlet curves is de fined by the related length of the chromatographic column

(1/Le) and by the sorption isotherm, At l/£e > lOOf150 chromatographic systems both with concave and with convex isotherms are written by the equation of ideal chromato graphy and sorption isotherms are calculated by de Vault's method. Isolength line, calculated with the ideal chromatography equation connects the maxima of peaks for different sample sizes only at a certain length of the column. For example, for anysol adsorption on silica SCP under the given conditions a calculation of the sorption isotherm by Huber's method becomes possible only with the column length 1/Le>8. The algorithms of a solution of the inverse problem and a calculation of sorption isotherms are worked out. Practical examples of definition of sorption isotherms on microporous adsorbents in elution and frontal chromato graphy are included in this report. Isotherms calculated are in a good agreement with the experimental isotherms

024 P.1-17 CONCENTRATION WAVES AND THEIR INTERFERENCE IN A MULTICOMPONENT NONLINEAR CHROMATOGRAPHIC SYSTEMS КОНЦЕНТРАЦИОННЫЕ ВОЛНЫ И ИХ ВЗАИМОДЕЙСТВИЕ В МНОГОКОМПОНЕНТНЫХ НЕЛИНЕЙНЫХ ХРОМАТОГРАЧИЧЕСКИХ СИСТЕМАХ A.I.Kalinitchev, Moscow, USSR Exact analytical solutions describing the steady stage of concentration waves with equal dispercing factors for systems with multicomponent Langmuir sorption isotherms or with linear isotherms of components and with concentra tion dependence of the mobile-phase velocity are presen ted. It is shown that the relations between the concentra tions in this waves are described by straight lines in multicomponent composition space. It follows from this solutions that the interference of species produces the additional broadening of concentration waves. The interference {"crossover") of two concentration waves in two component chromatographic system with the concentration dependence of separation factors is studied on the ground of ideal equilibrium theory. It is shown that the interference of widening and selfsharpening con centration waves may be calculated only by the use of individual isotherms if the initial and input compositions of species are on the boundaries of concentration space. The results of this theoretical analysis are used for the elaboration of mathematical model and calculation of industry preparative process of chromatographic separa tion.

18 025 P.5-14 APPLICATION OF PERSONAL COMPUTER IN PROCESSING OF CHROMATOGRAPHIC DATA ПРИМЕНЕНИЕ ПЕРСОНАЛЬНОГО КОМПЬЮТЕРА ПРИ ОБРАБОТКЕ ХРОМАТОГРАЧЙЧЕСКИХ ДАННЫХ I.Raisz, Hungary Recently we reported about off-line evalution of chromatograms. Using our experience we developed our on-line chromatograph - personal computer system. Chromatron GCHF 18.4 type chromatograph was connected to VIDEOTON PERSONAL COMPUTER, so we created an on-line data evaluation system on the basis of equipment produced in socialist countries. Using Commodore VC-64 personal computer we can plot the chromatogram evaluate the data, perform common chemical analysis and other calculations. We studied the effect of data sampling frequency on accuracy of evaluation. The optimum data sampling frequ ency was determined as function of half value width, so we can make accurate evaluation with optimum usage of data storage capacity. Kith the help of multi-channel interface we made possible storage of chromatographic parameters in addi tion to chromatogram data acquisition and evaluation. He demonstrate on some example examination of effect of some parameters in developed chromatograph - personal computer system.

026 PL TRACE ANALYSIS BY CHROMATOGRAPHY ХРОМАТОГРАЖЧЕСКИЙ АНАЛИЗ МИКРСПРИМЕСЕЙ J.Janak, Brno, CSSR Abstract not arrived on time for printing.

19 ION-EXCHANGE CHROMATOGRAPHY

027 S.3-1 ION-SELECTIVE CHROMATOGRAPHY IN CHEMICAL TECHNOLOGY ИОНОСЕЛЕКТИВНАЯ ХРОМАТОГРАЗИЯ В ХИМИЧЕСКОЙ ТЕХНОЛОГИИ K.M.Saldadze, Moscow, USSR The discovery of synthetic ion-exchange resins in 40's gave rise to a new brancb.-ion-exchange chromatography based on ion-exchange heterogeneous chemical processes taking place at contact of electrolyte solutions with ionites. As the mentioned reactions do not change the agregate condition of separated substances, such chroma tography, as a method of separation, finds its direct application in purification, concertration and separation of substances. In 50's ion-exchange technology, based on ion-exchange chromatography, was developed, which helped to solve seri ous problems, such as separation of rare earths elements aminoacids, concentration and separation oi rare and scattered elements, etc. The application of ion-exchange chromatography in water preparation for getting highly desalinated water, free of silicic acid, was a true revolution in thermo- and atomic energetics, because it became possible now to use atomic reactors and vessels of high and superhigh parameters. It was found out, that if ion-exchange chromatography is indispensable in making water highly desalinated, high- purity and high-ohmic, it is non-profitable when used for freshening and demineralizing of salt waters, because of high consumption of chemical reagents for ionites regene ration. For demineralization of salt waters, including sea water, it is more reasonable to use electro-membrane chromatography with ion-exchange membranes in electric field. During the last twenty years, based on electro-membrane chromatography, there was developed a reagent-free electro- membrane technology of purification, separation and con centration of substances. This method is profitable only for purification and demineralization of waters rich in salt and doesn't provide highly desalinated water. As is shown in our research, for high desalination of water from waters rich in salt it is more practical to use a mixed sorption-membrane technology, based on ion-selective chro matography with application of polymeric ion-selective membranes and ion-exchange resins.

30 028 S.3-2 AMERICIUM SEPARATION AND PURIFICATION BY CHROMATOGRAPHY ВЫДЕЛЕНИЕ И ОЧИСТКА АМЕРИЦИЯ С ПОМОЩЬЮ ХРОМАТОГРАФИИ J.D.Navratil, Colorado, USA Americlum-241, one of the most useful actlnide isotopes, is produced as a byproduct of plutomiun scrap recovery operations. Americium is separated from chloride and mono valent cations by cation exchange displacement chromato graphy. The americium is further purified by an extraction chromatography process. The process chemistry and evalua tion of separation materials for both processes will be described.

029 S.3-3 APPLICATION OF ION-EXCHANGE FOR DEEP PURIFICATION OF SUBSTANCES ПРИМЕНЕНИЕ ИОННОГО ОБМЕНА ДЛЯ ГЛУБОКОЙ ОЧИСТКИ ВЕЩЕСТВ V.I.Gorshkov, Moscow, USSR When we deal with deep purifying of substances we encounter with two groups of problems possible to be sol ved by using ion-exchange. 1. Purification of non electrolytes or week electroly tes from admixtures of strong electrolytes. 2. Purification of strong electrolytes from admixtures of other strong electrolytes. The solution of the first group of problems is based on substitution of admixtures ions by suitable ions of ion- exchanger (usually H+ or OH"), while the substances to be purified does not interact with ion-exchanger. As a rule the conditions are chosen so that to guarantee the preferable sorption of removed ions. In this case for deep purifica tion we do not need very strong multiplying of equilib rium separation effect. Such processes are realised in a fixed bed ionyte or in countercurrent columns. As examples, the deaineralization of water, the purification of organic solvents, the purification of some aminoacids and liquid crystal substances are considered. The problems of second group are more difficult. For the solution it is necessary to conduct a considerable multiplying of single separation effect, which is charac terized by equilibrium separating coefficient of substan ce to be purified and admixtures, Л,. In order to solve such a problem only effective columns (that is columns with low value of HETS) must be used. While choosing con ditions of purification a maximal different at from 1 must be guaranteed and such a scheme of process in which the expenditures on auxiliary operations should be mini mal must be worked out. Moreover, it is desirable that the columns were not only effective, but more productive. In this report the schemes of continuous processes of deep purification (with refluxes of mixture and alternation of the direc- 21 tion of transfer between phases) and some aspects of equipment construction for countercurrent ion-exchange are discussed. Example of using ion-exchange for purifi cation of Jf from Tb, Dy, Gd, Bo, purification of doublecharged ions from onecharged ones, purification of Kb from Cs and Cs from Kb are regarded. In conclusion the requirements to purity of auxiliary substances - ion,exchangers, solvents and auxiliary elec trolytes are considered.

030 S.3-4 SEPARATION OF MIXTURES OF UNI- AND BI-VALENT IONS WITH CHARGE-SELECTIVE IONIC MEMBRANES РАЗДЕЛЕНИЕ СМЕСИ ОДНО- И ДВУХЗАРЯДНЫХ ИОНОВ С ПРИМЕНЕНИЕМ ЗАРЯД-СЕЛЕКТИВНЫХ ИОНИТОВЫХ МЕМБРАН V.D.Grebenyuk, Kiev, USSR Separation of a mixture of mono- and bi-charged ions is necessary not only for analytical but for practical pur poses for example for complex processing of saline water aimed to produce fresh water and important mineral sub stances. It is shown in the paper that selectivity of the membranes in respect to singlecharged ions may be achieved by different transport mechanisms. Firstly, it may be achieved by change of relation of mobility between single- and doublecharged ions in the membrane. This mechanism was realised as the result of treatment of mem brane MA-40 by active bright yellow dye 5 "B". Secondly it is possible to put neutral or charged films on the membrane, which have high permeability in relation to co-ions but substantially different permeability in rela tion to single- and doublecharged counterions. This mecha nism was realised in three different ways. Firstly, it was realised by combining of sulphocationlte membrane MK-40 with acetate cellulose membrane МГА-95, secondly, by putting a layer of positively charged polyelectrolyte on the sulphocationit* membrane MK-40, thirdly, by amination of chlorosulphated matrix of membrane MF-100 by ethylenediamine. Selectivity of membranes in relation to singlecharged ions may be achieved by partial using of a current to produce H+ and OH" - ions. These ions precipitate the ions of hardness and remove them from realm of transport.

22 031 S.3-5 USE OF COMPLEX-FORMING ION-EXCHANGE CHROMATOGRAPHY FOR SELECTIVE ISOLATION OF NICKEL IONS FROM THE SEWAGE OF GALVANIC PRODUCTION ИСПОЛЬЗОВАНИЕ КОМПЛЕКСООБРАЗОВАТЕЛЬНОЙ ИОНООБМЕННОЙ ХРСМАТОГРАЗЙИ ДЛЯ СЕЛЕКТИВНОГО ВЫДЕЛЕНИЯ ИОНОВ НИКЕЛЯ ИЗ СТОКОВ ГАЛЬВАНИЧЕСКОГО ПРОИЗВОДСТВА V.B.Voitovich, M.Yu.Khazel, Voronezh, USSR The use of complex-forming ion exchangers for the iso lation of nickel ions from the sewage of galvanic produc tion is expedient. The investigation of the processes of complex-formation of nickel ions with carboxylic and &!~L.~io carboxylic ion exchangers from aqueous and model solutions, containing the lemon-acid complex of nickel, was carried out. Conditions of the least stability of the complex in the studied media were determined spectrophotometrically depending on pH. The structure and composition of comp lexes were established. The possible composition of nickel complexes with the fixed groups of ion exchangers were examined on the basis of the results of IR-spectroscopy and potentiometric tit ration. The degree of covalence of COO-Ni bond was deter mined from the lR-»pectro«copy data according to the fre quency difference of asymmetric and symmetric stretching vibrations. The strength of COO-Ni bond was evaluated also with the help of coefficients of- self-diffusion of nickel ions in the phase of the exchangers, calculated according to the results of electric conduction measure ments. Curves of nickel ions sorption by ion exchangers from the examined solutions were obtained. It was shown that the type of nickel-ionogene group complex and its strength determine the shape of the cur ves and the formation of sorption front. Conditions and the choice of ion exchanger for the iso lation of nickel from the sewage of galvanic production were formulated on the basis of the joint analysis of the obtained physico-chemical characteristics.

032 S.9-2 ION-EXCHANGE CHROMATOGRAPHY OF AMINO ACIDS ON MACROPOROUS ION EXCHANGERS ИОНООБМЕННАЯ ХРОМАТОГРАЗИЯ АМИНОКИСЛОТ НА МАКРОПОРИСТЫХ ИОНИТАХ G.A.Chikin, V.F.Selemenev, G.Yu.Oros, Voronezh, USSR The object of the work was to study the peculiarities of interaction between insoluble AA (glutamic - Q, tyro sine - V, tryptophan - И) and high-base anion!te (HBA-17- 2P (porous) and strong-acid cationic exchanger (SACE-23 (30/100) and also the chromatographic isolation of the above-mentioned acids in the form of crystals from the

23 production solutions. It was shown that the adsorption of Q in the form of cation and zwitterion on the sulphocatio- nic exchanger in the H-form goes on practically without complications. In the case of sorption Q2~ the maximum observed on the curves indicates that the absorption of the glutamic acid is accompanied by side effects, the skipping of an amino acid preceding the emergence of sodium ions. It was shown that along the height of the column there goes on the redistribution of the mineral cation and various ionic forms of amino acid: (Q2~ + Q")< Na+<(Q+ + Q*!. The concentration cf the end product ex ceeds the solubility by 2,6-3,2 tim-s. The sorption of glutamic acid in the presence of the coloured organic substances (melanoidines - M+) and mineral ions on the anionite ВДА-17-2Р in OH-form showed that Q2" is selecti vely absorbed by anionite, the front of sorption being sharp at small concentrations of amino" acid in the exter nal solution. For Q* and Q" smooth curvet are observed which shows a small selectivity of absorption of these ions by anionite. At pH 1,2 the maximum is observed on the curves and the corresponding concentration of the acid in the ion exchanger layer twice exceeds its solubi lity. Using the IRS method it was shown that in the maxi mum band of the glutamic acid absorption the resin phase contains cation and zwitterion forme Q. The sorption order of the substances under consideration on the anio nite HBA-17-2P iss (Q+ + Q±)

033 S.9-4 APPLICATION OF PRECIPITATE-FORMATION AND COMPLEX- FORMATION REACTIONS IN LIQUID CHROMATOGRAPHY ПРИМЕНЕНИЕ РЕАКЦИИ ОСАДКООБРАЗОВАНИЯ И К0МШ1ЕКС00БРА30ВАНИЯ В ЖИДКОСТНОЙ ХРОМАТОГРАШИ V.D.Eristavi, Tbilisi, USSR The use of modified ion exchange resins by various precipitate and complex formers enables to expand the abilities of chromatographic methods, and particularly to apply the anion exchange resins for concentrating and separating of metal cations. The x-ray, IR-spectral and thermographic research of modified anionite samples before and after their contact

24 with metal-containing solutions enabled to make clear the mechanism of metal sorption on the investigated sorbents and also the nature of compounds being formed. There have been considered kinetics and dynamics of these processes. There have been also established the kinetics area of sorption dynamics, the distribution of the sorbed substance concentration between the solution and the sorbent, and the moving rate of sorption fronts. The paper presents the experimental results obtained for the sorption and desorption of number of metals (Be, U, Th, Zr, In, Ga, Tl, Zn, Cd, Pb, Sn, Bi, Cr, Mn, Ni, Co, Fe, Al, Cu, Ti) on anion exchange resin (AB-16, AB-17, ЭДЭ-10, АН-2Ф) modified by carbonate- and oxalate ions. On these grounds the methods of separation and extraction of one of these ions out of complex mixtures have been worked out.

034 S.9-5 PHYSICO-CHEMICAL PROCESSES DURING CHROMATOGRAPHIC SEPARATION ON COMPLisX-FORMING ION EXCHANGERS ФИЗИКО-ХИМИЧЕСКИЕ ПРОЦЕССЫ В ХРОМАТОГРАФИЧЕСЧИХ РАЗДЕЛЕНИЯХ НА КОКПЛЕКСООБРАЗУЩИХ ИОНИТАХ N.N.Matorina, L.V.Shepetyuk, T.I.Bakaeva, L.V.Karlina, O.v.Kryuchkova, Moscow, USSR The development of industrial chromatographic separa tion on selective complex-forming ion exchangers requires consideration of separate stages of the chromatographic process and investigation of their influence on the be haviour of the complex-forcing sorbents under the condi tions of chromatographic separation. At present in most cases the complex character of the regularities governing the complex-forming processes in such packings cannot be predicted even qualitatively. The present report uses an example of absorption of the ions of rare earths and transition metals by the amphoteric vinyl pyridine and imino diacetate resins to represent the results of investigation of the equilibrium sorption, kinetics and dynamics of sorption, which may be used to intensity the processes of chromatographic separa tion on the complex-forming ion exchangers. A method has been suggested to characterize the selec tivity of sorption and describe the sorption isotherms. The method is based on using the concentration constants of the Ion-exchange equilibrium on weakly dissociating ionites and on the dependence of these constants on the ionite filling. The relationship between those constants and the shape of chromatographic zones has been demonstra ted. The influence of the nonuniformity of functional groups on the equilibrium and kinetic characteristics of the complex-forming ion e changers has been established. The kinetics of sorption on the complex-forming ion exchangers has been investigated. The possibility of applying ultra-

25 sound to intensity the diffusion processes in these sys tems has been demonstrated. An example of separation of rare earths and transition metals was used to show the advantages of the complex-for ming ion exchangers as compared with the use of strongly dissociating ones.

035 P.1-27 CALCULATION THEORY FOR SUCCESSIVE LAYERS OF MULTICOMPONENT ION-EXCHANGE CHROMATOGRAPHY ТЕОРИЯ ПОСЛОЙНОГО РАСЧЕТА В МНОГОКОМПОНЕНТНОЙ ИОНООБМЕННОЙ ХРОМАТОГРАФИИ ПРИ СМЕШАННОМ МЕХАНИЗМЕ ДИФФУЗИИ И НЕЛИНЕЙНЫХ ИЗОТЕРМАХ G.A.Chikin, V.A.Kusminykh, Yu.D.Chuprakov, Voronezh, USSR Basic results obtained in the field of phenomenological theory of sorption kinetics and dynamics and ion exchange for mixed diffusion mechanism and non-linear isotherms have been presented. Due to versatile experimental data comparison it has become possible to describe selective ion-exchange sorption with constant effective coefficients of interdiffusion in solid and liquid phases and to expand this approach for multicomponent solutions.

The calculation theory for successive layers for diffe rent variants of non-equilibrium ion-exchange chromatogra phy with arbitrary component number in the solution, mixed mechanism of ion diffusion and non-linear isotherms of ion-exchange equilibrium has been developed. The proposed method is mathematically valid and makes it possible to calculate with any required precision and a small number of "layers" in ion-exchange column which saves computer time in solving invert and optimization problems. The average calculation times of one frontal chromatography variant for five-component solution is 20-30 minutes for M-222 computer. Calculation experiment has shown the range of applica tion of chromatography equilibrium theory., its practical results and derived calculation formulas. Distribution of the components to be separated in ion-exchange column and in filtrate output is defined not only by their rela tive sorption affinity for ion-exchange but it also greatly depends upon kinetic of chromatography process and in particular upon kinetic properties of ion-exchange material used. Sifficient effective ion separation even with essentially different individual exchange constants is possible only for ion-exchangers with high kinetic indices.

26 036 P.1-28 THE THEORY OF THE INFLUENCE OF CHEMICAL REACTIONS AND OTHER NONLINEAR FACTORS ON THE KINETICS OF THE ELEMENTARY STAGE OF ION-EXCHANGE CHROMATOGRAPHY ТЕОРИЯ ВЛИЯНИЯ ХИМИЧЕСКИХ РЕАКЦИЙ И ДРУГИХ НЕЛИНЕЙНЫХ ФАКТОРОВ НА КИНЕТИКУ ЭЛЕМЕНТАРНОЙ СТАДИИ ИОНООБМЕННОЙ ХРОМАТОГРАФИИ Yu.S.Ilnitsky , Novosibirsk, USSR

The interactions of an ion-exchange element with the surrounding solution, which form a basis for ion-exchange chromatography, have been insufficiently studied. Actual ion-exchange systems have features which cannot be ex plained by idealized models. Experiments show that the representative physical and chemical parameters of a sys tem depend on the properties and state of a process. This means that such systems are nonlinear and must be analyzed carefully.

We study the nonsquilibrium stage of ion exchange in a particular heterogeneous system, "ion-exchanger - water solution", to determine in what manner the selectivity of ion-exchangers and chemical reactions between ions in ao- lution modify the kinetics of such processes. The work was performed by means of computational experiments with the use of a previously described theoretical model. As a basis for the work, we used our previously derived equa tions of ion interdiffusion accompanied by fast chemical reactions between ions. They are in the form of time-de pendent Ginzburg-Landau equations.

ut= M(U,V)UXX + N

It has been found that there are some peculiar regimes of behaviour in the modelling of nonequiiibrium systems, caused by the influence of the chemical reactions and the selectivity of the ion-exchanger. Apart from the appea rance of a reaction underlayer in the diffusion layer near the interface boundary, under certain values of the representative parameters of the system, time- and space- dependent dissipative structures have been observed in the form of mass and change density waves and a correspon ding pulsing of the diffusional fluxed. This indicates that oscillations of the concentrations of the analyzed substances and spreading of chromatography fronts in some regimes of ion-exchange chromatography pannot, in princip le, be eliminated, i.e. that there exist definite limits to the analysis of ralcroquantities of various substances.

27 037 P.1-29 ON THE RELATION OF THE MASS EXCHANGE RATE IN IONITE AND SOLUTION FOR SELECTIVE SYSTEMS IN THE CASE OF A CONCAVE EXCHANGE ISOTHERM О СООТНОШЕНИИ СКОРОСТЕЙ МАССООБМЕНА В ИОНИТЕ И РАСТВОРЕ ДЛЯ СЕЛЕКТИВНЫХ СИСТЕМ В СЛУЧАЕ ВОГНУТОЙ ИЗОТЕРМЫ ОБМЕНА A.I.Kali.:itchev, E.V.Kolotinskaya, T.D.Semenovskaya, Moscow, USSR The results of computer analysis of ion-exchange kine tics in selective systems presented in the paper show that the widely used method of determination of the limiting stages (particle diffusion control - p.d.c. or film diffu sion control - f.d.c.) is not applicable to the kinetic processes in selective systems. This conclusion is due to the dependence of the ion-exchange rate on the separation factor and solute concentration. The dependence is typical not only for the mass transfer in the solution but also for the particle diffusion in the case of concave exchange isotherm. Previously the effect of selectivity and solute concentration for the p.d.c. kinetics was obtained by a computer analysis and confirmed by experiments on complex forming ionites. For the selective systems the Helfferich-Tunitsky crite rion gives the inadequate estimate of f.d.c. or p.d.c. contribution since the interdiffusion coefficients in the criterion may decrease greatly when the direct exchange is replaced by the reverse one. Usually this fact is not ta ken into consideration for the unselective exchange. The disagreement with ideal ion exchange kinetics cau sed by the influence of selectivity is possible of comp lex forming, weak dissociating, chemically and structuraly heterogenuous ionites, where the presence of the free and bound ions is not excluded. The arguments of the paper are confirmed by the kinetic experiments on various ionites for different solution con centrations. It is shown that very slow exchange rate in the grains of the macroporous ionites may be explained only by the influence of the equilibrium factors on the rate of ion transfer.

038 P.l-60 KINETIC CHARACTERISTICS OF COUNTER IONS WITH DIFFERENT MOBILITIES, WHEN AS INTRAPARTICLE AS FILM DIFFUSION CONTROL OF PREPARATIVE ION EXCHANGE CHROMATOGRAPHY TAKES PLACE ОПРЕДЕЛЕНИЕ КИНЕТИЧЕСКИХ ХАРАКТЕРИСТИК ПРОТИВОИОНОВ С РАЗЛИЧНОЙ ПОДВИЖНОСТЬЮ ПРИ ПРЕПАРАТИВНОЙ ИОНООБМЕННОЙ ХРОМАТОГРАФИИ В УСЛОВИЯХ СМЕШАННОЙ ДИФФУЗИИ G.E.El'kin, T.A.Atabekyan, G.V.Samsonov, Leningrad,USSR The non-linear problem of ion exchange kinetics nas been solved by the Storm's, linearisation method. The model has been investigated with the concentration dependence of 28 the sa intraparticle as film diffusion coefficients and with the rectangular isotherm for the description of the sorption equilibrium. We have got the solution for a semi- infinite body as a relation between the F°-value and Biot's diffusion criterion, Bi. The F°-value is equal to the extent of the process completeness., F(t), at the moment t°/ when the initial linear relation between_F and t is destorted. The effective coefficients D and D have been introduced for film and intraparticle diffusion respecti vely . The relations between effective diffusion coeffi cients and counter ions selfdiffusion coefficients D,, D,, D., D, have been determinated. The values of 6 and Bi have been estimated from the oxytetracycline and chlortetracycline desorption data. Using the asimpthotic formulae:

5 a (4/.C)\TD\D2, DJ^D,;

D =• (16/ЗГ )D2{ln(51/JT52f- (3/2)1п1п\П^7ГБ2|, D^D, one can estimate D. (or D,), if D and D, (or D,) are known. 03Э P.1-30 ELUTION CHROMATOGRAPHY FOR THE CASE OF INTRAPARTICLE DIFFUSION WITH REGARD TO VARIOUS BROADENING FACTORS ЭЛЩИОННАЯ ХРОМАТОГРАФИЯ ПРИ ВНУТРИДИФФУЗИОННОИ КИНЕТИКЕ С УЧЕТОМ РАЗЛИЧНЫХ ФАКТОРОВ РАЗМЫВАНИЯ ЗОН ВЕЩЕСТВ G.E.El'kin, A.T.Melenevsky, E.B.Chizova, G.V.Samsonov, Leningrad, USSR A theoretical analysis was made for elution of twocom- ponent reacting zone in equilibria with regard to axial dispersion in absense of adsorption of one of the compo nents. The influence of the axial effects on elution of the infinitely narrow zone for the case of intraparticle diffusion at equilibria and nonequilibria was discussed as a part of the former problem. A number of generelized variables and parameters was introduced. The solution of the problem was given by: F

G = J°G1(x1f ).G2(f , FQ -f )df , (1) where the functions G. and G, - the influence functions of nonequilibrium factors/axial dispersion and chemical reac tion - for the former problem; axial dispersion and intra particle diffusion - for the latter/. The influence of nonequilibri-um factors on the form and location of the elution curve maxima was estimated. The extent of declinationAfrom equilibria of chromatographic process may be estimated by the criterion , Using the Laplas asympthotic method, that is substitu ting one of the influence functions by Dirack's Ъ-tunc tion of the respective variable, we have got the relation between the output time of the elution curve maxima (the bend point of the boundary) and the geometric parameters of the column and broadening factors. 29 040 P.1-4 3 DETERMINATION OF Cl" IN WATER SOLUTIONS BY ION CHROMATOGRAPHY ОПРЕДЕЛЕНИЕ 01" В ВОДНЫХ РАСТВОРАХ МЕТОДОМ ИОННОЙ ХРОМАТОГРАФИИ К.Punning, T.Somer, J.Veisserik, Tallinn, USSR The ion chromatograph IVK-11 (designed and produced by Special Design Office, Academy of Sciences of Estonian SSR) was applied to the analysis of Cl" in water solutions. In separation column (250 x 4 mm) the anion exchange resin VARS-2 (capacity 12,7 meq/cm , synthesized by the Institu te of Chemistry, Academy of Sciences of Estonian SSR) and in suppression column KY-2(8) were used. The eluent was

0,003 M NaHC03 and 0,0025 Na2C03. The number of theoreti cal plates varies from 380 to 970, depending on the eluent flow-rate, sample volume and concentration. The number of effective theoretical plates varies from 11 to 57. At con stant column pressure the eluent flow-rate increase causes the decrease of column efficiency and the increase of limit of detection (LOD). At constant eluent flow-rate-the column pressure increase results in lower LODs due to the decrease of detector noise level caused by the concrete pump pulsation, LOD varies from 0,6 to 11 ppb, in units of amount from 70 to 220 pg, depending on the analysis condi tions. The plot of Cl" amount versus peak height is linear up to 4 jug injected Cl". 041 P.1-32 THE THEORETICAL BASIS OF DESCRIPTION OF ION-EXCHANGE CHROMATOGRAPHIC PROCESSES UNDER HIGH PRESSURE ТЕОРЕТИЧЕСКИЕ ОСНОВЫ ОПИСАНИЯ ИОНООБМЕННЫХ ХРОМАТОГРАФИЧЕСКИХ ПРОЦЕССОВ ПРИ ВЫСОКИХ ДАВЛЕНИЯХ V.L.Bogatyryov, Novosibirsk, USSR The analysis of chromatography processes with ion-ex change mechanisms and their classification from the point of view of high-pressure influence on specific components is given. On the basis of thermodynamical Planck equation the changes of partial molar volume for weak-ionized sorbents and solution phase components under sorption of ions with different possible mechanism are calculated'. The calcula tions are based on the theoretical analysis of volumetric effects of partial parallel-consecutive reactions and their contributions into the total volumetric effect of sorption. The calculated results are compared with experi mental values, obtained by graphic differentiation of the curve, representing the dependence of logarithm of ion- exchange equilibrium constant on pressure. In this way the corresponding mechanisms of sorption for certain sys tems representing the weak electrolyte solution and anion- exchanger of агЛпо type, were established. In the systems with strongly ionized components the

30 degree of changing of such parameter as "swelling pressu re" from Gregor's osmotic theory of ion exchange is the factor determining the change of the exchange constant with pressure. The values of swelling pressure in sorbent particles can be calculated from the data of direct measu rement of specific and equivalent volumes of ion-exchan gers under high hydrostatic pressure. Such method of description of ion-exchange systems has been tested experimentally with sulfonated polystyrene cation exchangers. It has been shown that the methodology proposed allows to predict selectivity changes for ion exchangers under high pressures.

042 P.1-33 SCIENTIFIC BASIS WORKING OUT AND UTILIZATION OF THE METHOD DISPLACEMENT CHELATION CHROMATOGRAPHY FOR REE AND THE EXTRACTION AND SEPARATION РАЗРАБОТКА НАУЧНЫХ ОСНОВ И ПРИМЕНЕНИЕ МЕТОДА ВЫТЕСНИТЕЛЬНОИ КОМПЛЕКСООБРАЗОВАТЕЛЬНОИ ХРОМАТОГРАФИИ ДЛЯ ВЫДЕЛЕНИЯ И РАЗДЕЛЕНИЯ РЗЭ И ТПЭ P.P.Nazarov, E.A.Chuveleva, G.B.Maslova, O.V.Haritonov, V.v.Belov, L.A.Firsova, S.P.Kudryavtseva, Moscow, USSR The method of displacement chelation chromatography was used to prepare great number of high purity radioactive ra re earth elements (REE), transplutonium (TRE) and alka- liearth (AEE). To intesify the research, the calculation method for optimal conditions of displacement and separa tion of elements was developed. It was shown theoretically that displacement mode is possible if isotherm of displa ces on the elements to be displaced has S-like shape. The method of calculation of equilibrium concentrations in chromatographic bands is proposed taking into account the penetration of the displacement ion into the zone of re taining ion. The calculations were confirmed by the expe riment of REE separation in presence of diethylenetriamin- pentaacetic acid (DEPA). The influence of ionizing radiation of high-power (2>lO6 Gr) at the chromatographic process of REE and TPE exstraction and separation has been studied. It was shown that in fields with high radiation it's better to use eluents, containing nitrilotriacetic acid. REE separation at high mobile phase velocity (up to 300 cm/min) at 290-350 К has been studied. The producti vity of such mode is approximately lOO times higher, whereas the loss of efficiency is not significant. The work gives the examples of radioizotopes prepara tion of high purity for 147pm, 244Cm, 241^ and 90Sr.

31 043 P.1-34 DETERMINATION OF GLYCINE AND HIS BYPRODUCTS BY LOW-PRESSURE ION-EXCHANGE CHROMATOGRAPHY ОПРЕДЕЛЕНИЕ ГЛИЦИНА И ЕГО АНАЛОГОВ ИОНООБМЕННОЙ ХРОМАТОГРАФИЕЙ НИЗКОГО ДАВЛЕНИЯ L.Balogh, Debrecen, Hungary A method is described for the separation of glycine and his pollutions. The components were eluted with Na-citrate/ citric acid buffer-solutions and buffer/methanol mixed solvent on Dowex ion-exchanger resins. Conductivity was measured which allows to quantify the organic and inorganic components in the same run. The method advantageous to determine the purity of glycine.

044 P. 1-36 COMPLEXITES IN CHROMATOGRAPHY КОМПЛЕКСИТЫ В ХРОМАТОГРАФИИ V.D.Kopylova, K.M.Saldadze, V.B.Kargman, Moscow, USSR Functional groups of a great number of ion-exchangers in deprotonized condition contain atoms possessing unsha red pairs of electrons; in contact with molecules and ions having vacant orbits they sorb the latter as a result cf donor-accept interaction whereby ion-exchanger exercise the function of peculiar ligands (complexites). In the process of sorption with the participation of complexites in dynamic condition, separation of components of the solu tion takes place, in case energy of interaction of the latter with the functional groups of the polymer is varied. Experiments prove the theoretic conclusion that the order and place of zones at the primary chromatogram cor relate to diminition of stability constants forming in the complexites phase in coordination centers. Quantitative separation of components in the system, at the primary chromatogram takes place under the condition whereby sta bility constants of forming complexes differ in 3 or more orders. Efficiency of using complexites in chromatography grows in case of bathing the premary chromatogram in a set of developers, such as an acid solution of a definite concen tration gradient or solutions of ligands forming complexes of various stability with the components of the primary chromatogram. The paper gives concretion instances of separation of a number of cations of transient metals in a macroporous anion-exchanger, a copolymer of 2,5-methylvinylpyridin and divinylbenzen (AH-251).

32 045 P.1-37 SELECTIVE SEPARATION OF COMPONENTS (COPPER, NICKEL, ZINC, CHROMIUM(VI) IN THE PROCESS OF ION EXCHANGE PURIFICATION OF SAWAGE СЕЛЕКТИВНОЕ РАЗДЕЛЕНИЕ КОМПОНЕНТОВ /МЕДЬ, НИКЕЛЬ,ЦИНК, ХР0М(У1) В ПРОЦЕССЕ ИОНООБМЕННОЙ ОЧИСТКИ СТОЧНЫХ ВОД L.P.Sokolova, V.V.Skornyakov, V.B.Kargman, K'.M.Saldadze, Moscow, USSR

The main direction in the purification of sawage at pre sent is the development of methods securing the possibi lity of utilization of the components which have been ex tracted from the sawage as well as circulation of purified water. In order to render harmless acid sawage washing water of etching departments of factories manufacturing the non- ferrous metals containing (milligramm/litre); Cu - 50-150; Zn - 70-250; Ni - 3-25; Cr(Vl) - 50-150; H2SO4 - O,3-0,25 gramme/litre; pH - 1,5-3,5 the method is elaborated allow ing tc separate above-mentioned components during the pro cess of ion-exchange purification. On the using of home selective sorbents with complex forming compounds. First chromium (VI) on the weak-alkaline anionite, con taining the piridin with the macroporous structure AN-251. Then the copper on the aminocarboxyl polyampholit ANKB-50 in the hydrogen form is selective extracted from the solu tion. The conditions for the most complete selection of chromium (VI) and copper from the other components of solu tion by applied selective sorbents are piched out.

After chromium (VI) and copper have been selected zinc and nickel remained in the solution are simultaneously sorbated on the cathionite KU-2-8 in the hydrogen form. In order to select this two components the well-known proper ty of these ions to form anion complexes in the hydrochlo ric acid-solution of the different concentrations is used. Zinc and nickel are eluated from the cathionite 2-N by the hydrochloric acid-solution. During the passing of hydro chloric acid eluate through the anionite AB-17-8 in the chlorine form the anion complexes of zinc are sorbated on the anionite while the nickel chloride passes through the anionite without being sorbed. The elaborated method of the purification with the use of selective sorbents allows to get during the regenera tion single-component concentrated solutions and all the " components of sawage as commodity products. The purified water returns into the washing bathes of the etch departments of the factory and is used for washing of non-ferrous metals and their alloyes after the etching.

3. Зак. 872 33 046 P.1-38 LARGE ORGANIC MOLECULES SORPTION BY ION-EXCHANGE FIBERS СОРБЦИЯ БОЛЬШИХ ОРГАНИЧЕСКИХ МОЛЕКУЛ ИОНООБМЕННЫМИ ВОЛОКНАМИ L.A.Volf, L.V.Emets, I.l.Paptsova, Leningrad, USSR Under the investigation were the peculiarities of adre naline and insulin binding with carboxyl-containing fib- rious ion-exchangers. The latter were obtained by way of polyacrylonitrile (PAN) and polyvinylalcohol (PVA) fibres treatment with various modifiers. Alonoside as a sorbent we used a fibre based on the natural alginic acid. It was estimated that the nature of polymeric matrix and the structure of fibres essentially influenced the character of the above mentioned binding. It was found that insulin sorption value (up to 1350 mg/g of sorbent) increased greatly during the transition of the carboxyl-containing PAN-fibre from hydrogen to salt form (Na+); as for the other fibres the increase of their selectivity was noticed during the transition to the H- form. The kind of fibre .also influenced the values of me dium acidity which corresponded the maximum value of insu lin sorption. The nature of polymeric matrix also influenced the strength of the formed complexes fibre-insulin. Thus comp lex PAN fibre-insulin could be destroyed only by alkaline, agents, complex alginate fibre-insulin by 0.1 M solution of hydrocarbonic acid. As for PVA fibra-insulin was desorbed by water for 80%. The feature of fibrous ion-exchangers was the high ki netics of ion-exchange processes (especially the processes of desorption) . Adrenaline sorption from its water suspensions was car ried out with the help of carboxyl-containing PVA fibres in their hydrogenous form. Under these conditions the fib re sorbed only pure adrenaline found in the raw-adrenaline and the coloured products of its oxidation either remained in the solution or were easily desorbed by washing with distilled water. The degree of the extracted pure adrena line was 98-100%. Adrenaline desorption was carried out with the help of hydrochloric acid. We managed to obtain 0,1% solutions of adrenaline hydrochloride that met all the requirements of State Pharmacopy X. 047 P.l-40 STATIONARY ION-EXCHANGE ELECTROCHROMATOGRAPHY СТАЦИОНАРНАЯ ИОНООБМЕННАЯ ЭЛЕКТРОХРОМАТОГРАФИЯ V.A.Shaposhnik, Voronezh, USSR Method of frontal electrochromatography, based on elec- trodialysis with the use of ion-exchange membranes and intermembrane diluting cells filling with the mixed-bed of cation- and anion-exchange resin is studied. Intermem- brane filling plays the role of chromatographic column, where ion separation according to their electric mobility 34 and selectivity coefficient is fulfilled. Potential gradi ent is applied in the direction perpendicular to the direc tion of the solution supply. Stationary state is positioned as a result of uninterrupted electrochemical regeneration of ion exchanger, when the iones under the influence of the electrical potential gradiant migrate into the adjacent concentrating cells along the chain of equipolar granules. First front in the intermemrane filling is created by the least selective uni-changed iones, followed by the front of iones in the increasing order of their selectivity in the ion exchanger. Experimental dependence of concentra tion distribution along the entire length of the intermembrane filling is exponentially reducing, corresponding to the found solution of the stationary sorption dynamic equa tion. Electrolytic mixtures are separated by the method of fraction selection along the different length of the cell. Primary field of this method application is the ever lasting ultrahigh water desalting process. As an initial material the natural water, containing small quantity of mineral impurities, obtained by reverse osmosis, which allows to remove electrolytes up to electric resistivity of 0,03 MOhm cm and also a considerable part of non-elec trolytes is used. The process of ultrahigh water desalting is fulfilled in two steps at the linear velocity of water supply of 0,3 cm/s. On the first step current density is 0,5 mA/cm2 and resistivity of water is 0,3-0,5 MOhm cm. On the second step optimal desalted water with resistivity of 18-25 MOhm cm is produced with current density of 0,05 mA/cm2. Particularities of this method are continuity of the process, no need for regeneration of ion-exchangers and maximum possible ecologic expendiency due to the rea gent mass increment absence in sewage as compared with the mass of impurities removed from the water.

04B P.1-41 LIQUID CHROMATOGRAPHY OF HERBICIDE COMPOUNDS ON CHEMICALLY BONDED ION-EXCHANGE MATERIALS • ЖИДКОСТНАЯ ХРОМАТОГРАФИЯ ГЕРБИЦИДОВ НА ХИМИЧЕСКИ МОДИФИЦИРОВАННЫХ ИОНООБМЕННЫХ МАТЕРИАЛАХ P.Jandera, J.Churacek, P.Butzke, Pardubice, CSSR M.Smrz, Brno, £SSR Possibilities of using new chemically bonded anion-exchange (Silasorb N - Et,) and cation-exchange (Silasorb KAT) materials for HPLC of various herbicide compounds was investigated. Separations of certain phenoxyacid her bicides that are difficult to achieve on aminosilica or alkylsilica chemically bonded phases, can be accomplished on Silasorb N - Et, columns in aqueous - methanolic mobile phases containing phosphate buffers-. Under these condi tions, nitrophenols and chlorophenols can be also sepa rated. The columns packed with Silasorb N - Et, or with Sila sorb KAT ion-exchange materials can be used for efficient

35 separations in normal-phase systems using mobile phases composed of n-propanol and n-hexane. Here, specific selec tivity effects for urea - type herbicides can be achieved in comparison to chromatography on non-modified silica or on chemically bonded aminosllica or cyanosilica, especial ly with Silasorb KAT columns. The influence of the composition of mobile phase and of the structure of urea herbicides on chromatographic behaviour on Silasorb KAT and Silasorb N - Et, columns was investigated in detail and the possibilities of separation are illustrated by several examples.

049 P.1-42 ONE-DIMENSIONAL MATHEMATICAL MODEL'S APPLICATION FOR ION-EXCHANGE COLUMN•DYNAMIC PARAMETERS DETERMINATION ПРИМЕНЕНИЕ ОДНОМЕРНЫХ МАТЕМАТИЧЕСКИХ МОДЕЛЕЙ ДЛЯ ОПРЕДЕЛЕНИЯ ДИНАМИЧЕСКИХ ХАРАКТЕРИСТИК ИОНООБМЕННЫХ КОЛОНН M.S.Safonov, Ya.N.Malykh, V.A.Ivanov, V.I.Gorshkov, V.K.Bel'nov, N.M,Voskresenskii, Moscow, USSR The determination of the dynamic characteristics of ion- exchange column ie one of thi- n.ost important issues in de fining the conditions for substances purification and mix tures separation. There exist various mathematical models (equilibrium diffusion model, plug flow model, etc),where ПЕТР, HTU or other characteristic dimensions are contained. The method of pulse perturbation on concentration of exchangeable substance /1/ is preferably to be used for column dynamic parameters determination. In present paper the comparison of usage of a number of models for experimental data description was considered. These experiments were carried out in packed-bed columns (120 cm height) of fixed resin layer and in countercur rent column (218 cm height) with continuos ion-exchanger motion under gravity for Na+ - H+ exchange on cation resin (KU-2x8) from 0,2 N chlorides solutions. The hydrogen ion "label" in the form of H+- ion-exchanger was injected into the packed column, equilibrious with the mixture solution of 0,199 N NaCl and 0,001 N HC1. Then the solution and the resin equilibrious with it were continiously fed into the column. The 0,1 ml solution samples were extracted with the help of a syringe from different column levels through rubber seals. The concentration of hydrogen ions in these samples was measured by using'microelectrodes technique. The experimental results were treated while using the direct-task analytical solution and random search method for reverse-task, thus defining parameter sets of corres ponding mathematical models. The suggested technique gave simultaneous determination of such model parameters as characteristic dimension, capacity of the column volume

36 unit, the impulse velocity along the column and ion-solu tion/resin phase mixture flow. The experimental conditions region were obtained where the models well agree with the experiment that is the calculated dynamic system parame ters obtained from models are in accordance with the para meters resulted from the independent experiments, and the calculated curves are adequate to the experimentally ob tained ones. 1. M.S.Safonov, A.A.Larikov, S.V.Yevdokimov, V.K.Bel'nov, N.I.Afonina. Teor. osnovy chim. technol., 1982, v. 16, N 5, p. 604. LIQUID CHROMATOGRAPHY

050 PL CURRENT DEVELOPMENTS IN COMPUTER-AIDED RAPID-SCANNING UV-VISIBLE DETECTION IN HPLC РАЗВИТИЕ КОМПЬЮТЕРИЗОВАННОГО СКАНИРУЮЩЕГО УФ-ДЕТЕКТОРА В ВЭЖХ A.F.Fell, Bradford, VK In this lecture recent advances in the field of rapid- scanning detectors in HPLC are surveyed. The remarkable impact of the new technology of linear photodiode array multichannel detection on the practice of IJPLC is clearly apparent in the fields of pharmaceutical, industrial, cli nical and environmental analysis (1-3). Computer-aided manipulation of the spectral data permits the homogeneity of chromatographic peaks to be systematically tested (4-6). Furthermore, rapid identification of eluting compounds can be achieved using archive retrieval techniques recently development (7) . This paper will critically review the use of 3-dimensional projections of chromatograms and of 2-dimensional contour maps, with and without colour graphics (8). The application of new techniques for sensitivity enhancement in HPLC using photodiode array detectors (7), will also be discussed and future perspectives for multichannel detection will be outlined. 1. A.F.Fell, Anal Proc.,.17 (1980) 512-519. 2. C.J.Warwick, D.A.Bagon, Chromatographia,15 (1982) 433-436. 3. A.F.Fell, H.P.Scott et al, Chromatographic,16 (1982) 69-78. 4. A.F.Fell, H.P.Scott et al, J.Chromatogr.,282 (1983) 123-140. 5. B.J.Clark, A.F.Fell et al, J.Chromatogr.,286 (1984) 261-273. 6. G.T.Carter, R.E.Schiesswohl et al, J.Pharm. Sci.,71 (1982) 317-321. 7. A.F.Fell, B.J.Clark, H.P.Scott, J.Chromatogr.,316 (1984) 423-440. 051 S.2-2 INVESTIGATION OF ADDITIVES IN LUBRICATING OILS ВУ. HPLC TECHNIQUES ИССЛЕДОВАНИЕ ПРИСАДОК В СМАЗОЧНЫХ МАСЛАХ МЕТОДОМ ВЭЖХ L.Szepesy, M.Horvath, J.Szanto, T.Veress, Budapest, Hungary Lubricating oils may contain a large number of additi ves of different molecular weights and different polari ties and the list of the possible additives is continually changing. An analysis scheme was developed for the separation 38 and identification of some generally used additive types in lubricating oils. First the high molecular weight poly mers were separated by size exclusion chromatography on a C2 reversed phase column by using THF mobile phase. The low molecular weight fraction was further separated on SiCb into an unpolar and a polar fraction. These fractions were analyzed on a normal phase РАС column as well as on a reversed-phase C18 column, by using OV detection at two different wave lengths (254 and 280 run) . From these analy ses hindered phenols and some polar additive types could be determined. Identification of some additives was sup ported by using reference standards as well as by TLC using specific reagents.

052 S.2-3 OPTIMIZATION OF RESOLUTION IN LIQUID-SOLID CHROMATOGRAPHY ОПТИМИЗАЦИЯ СТЕПЕНИ РАЗЛЕЛЕНИЯ В ЖИДКОСТЬЮ- АДСОРБЦИОННОЙ ХРОМАТОГРАФИИ Ya.I.Yashin, Moscow, USSR High performance liquid chromatography exhibits high separation power enabling successful separation and analysis of complex multicomponent mixtures. However, being mainly empirical the selection of optimum separation conditions is generally laborious. Besides, the empirical methods usually do not provide the most optimum separation modes. These problems may be solved using computer-aided opti mization provided that reliable data on efficiency, selec tivity and capacity of separating systems are available. Columns of guaranteed efficiency are relatively easy to obtain. The reliable data on selectivity and capacity of separating systems are rarely found in literature. The paper offers an estimation system for selectivity and capacity of both most common adsorbents and promising silica, polymer and carbon ones (including those with donors and acceptors on the surface) using a certain set of benzene derivatives with different polar substituents and widely accepted eluents. This system permits to esti mate the separation selectivity for homologues, isomers and compounds of various classes differing in polar group nature. Data on selectivity and capacity of some standard sepa rating systems (adsorbent-eluent) are presented. The possi bility to optimize the separation of certain mixtures is shown. The contributions of substance-adsorbent and substance-eluent interactions to retention are estimated. The potentialities of dynamic modification methods of various adsorbents in different modes are considered, crown ethers, different compounds with long alkyl chains etc. being used as modifiers.

i 053 S.2-4 SELECTIVITY OF SEPARATION IN REVERSED-PHASE LIQUID CHROMATOGRAPHY - CALIBRATION, PREDICTION AND OPTIMIZATION СЕЛЕКТИВНОСТЬ РАЗДЕЛЕНИЯ В ОБРАЩЕННО-ФАЗОВОИ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ: КАЛИБРОВКА, ПРОГНОЗИРОВАНИЕ И ОПТИМИЗАЦИЯ P.Jandera, Pardubice, 6SSR Selectivity in liquid chromatography is usually charac terized by ratios of capacity factors or net retention times of a sample and a reference compound. If the compo- siton of mobile phase is changed, the selectivity usually changes more or less, too, and often even a reversal of the order of elution with changing composition of mobile phase is observed, which makes difficult a quantitative characterization and prediction of selectivity. Using the model of interaction indices developped recently, it is possible to characterize quantitatively polarities of solutes and to predict their chromatographic behaviour in reversed-phase liquid chromatography, however, this approach does not allow differentiation of selective inte ractions between the solutes and various organic solvents used as the components of mobile phases. This inconvenience is overcome by calibration of reten tion scale over a wide range of mobile phase compositions using a calibration homologous series, which makes possible distinguishing between non-specific (hydrophobic) and specific polar contributions to retention and selec tivity. These contributions can be characterized quantita tively for each solute and organic solvent used as a com ponent of mobile phase. Furthermore, if the constants characterizing the specific polar conributions to reten tion of various solutes for one solvent are plotted against similar constants obtained for these solutes in other solvent

054 S.2-5 DETERMINATION OF SOLUTE-SOLVENT ASSOCIATION EFFECTS BY USING LIQUID CHROMATOGRAPHY DATA ОПРЕДЕЛЕНИЕ ЭФФЕКТОВ АССОЦИАЦИИ РАСТВОРЕННОЕ ВЕЩЕСТВО- РАСТВОРИТЕЛЬ С ПОМОЩЬЮ ДАННЫХ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ J.K.Rozylo, J.A.Jaroniec, Lublin, Poland The competitive solute and solvent adsorption is one of the roost important phenomena determining the liquid/so lid chromatography process. This phenomenon is represented 40 by the phase exchange reaction describing the displacement of solvent molecules from stationary (surface) phase by solute molecules contained in the mobile phase. Two simple equations, involving formation of the multimolecular solvates in the mobile phase, are discussed. These equations define the dependence of "-^t capacity ratio upon mobile phase composition. In tuis paper the effects due to the specific solute-solvent interactions in the mobile phase are discussed. The LC data for different alcohols chromatographed in binary mixtures of n-heptane with alkyl acetate and tetrahydrofuran on Partisil 10 at 29 8 К are applied to examine two simple equations involv ing formation of the multimolecular solvates in the mobile phase. The models considered are based on the following assump tions : a) only unassociated solute and solvent molecules participate in the displacement process, b) solute and solvent molecules form solvates in the mobile phase, which are destroyed in the surface phase because of strong solute-adsorbent interactions, c) solute and solvent mole cules have spherical shapes of different molecular sizes, d) the molecular sizes of all solvent molecules in the surface phase is constant and independent of the presence of solute molecules because of their infinitely low con centration, f) the adsorbent surface is energetically homogeneous.

The linear dependence (ksxx) vs. xx, where xx is mole fraction of 1-st solvent in the mobile phase, shows positive slope for the chromatographic systems with strong solute-solvent interactions in the mobile phase, however,

the dependence ksx± vs. x^ has negative slope for the chromatographic systems in question. The above facts leads to the conclusion that in case of phenyl methyl carbinol ch smatographed in alkyl acetate/ n-heptane eluents the model of multimolecular solvates is more relative. Perhaps, the molecules of phenyl carbinol, having greater molecular sizes than the molecules of normal alcohols studied, may annex more than one solvent molecule.

055 S.4-1 HPLC OF INSULIN AND INSULIN DERIVATIVES ВЭЖХ ИНСУЛИНА И ЕГО ПРОИЗВОДНЫХ B.Mehlis, U.Kertscher, J.Oehlke, K.-D.Kaufmann, Berlin, GDR KPLC separation of human, porcine and bovine insulin and some of its derivatives was carried out on various reversed stationary phases. Packed columns were tested for their performance parameters by the BRISTOW-KNOX method (1). It has occasionally been assumed that a "good" column for small-molecule separations (large N value) may often be of less efficiency when applied to large molecular. Therefore, the separation of a mixture of insulins was used as a proof for a good column performance within the

41 range of the relative molecular masses of the insulins

(Mr ~ 6 000). In this study glass columns (CGC, SIX C18) were included and found to be suitable for these difficult separations, The recovery from the columns was studied at different insulin concentrations using tritium labelled insulin at the lower concentrations. In contrast to some smaller peptides (which were quantitatively recovered) the reco very was found to be as low as 70% in some cases. 1. P.A.Bristow, J.H.Knox, Chromatographic»., 10, 279 (1977).

056 S.4-4 APPLICATION OF HPLC IN DIAGNOSTICS C? PATHOLOGIC STATES IN EXPERIMENTS AND CLINIC ПРИМЕНЕНИЕ ВЭЖХ В ДИАГНОСТИКЕ ПАТОЛОГИЧЕСКИХ СОСТОЯНИИ В ЭКСПЕРИМЕНТЕ И КЛИНИКЕ Yu.V.Tikhonov, R.T.Toguzov, P.A.Kol'tsov, Vu.S.Butov, T.E.Novikova, A.M.Pimenov, Moscow, USSR

The development of analytical methods of study of bio- molecules in the tissues of living organisms acquires a decisive importance in investigating the mechanisms of origin and progress of metabolic perturbations in experi mental and clinical pathology. A particular pathologic process, perturbing tissue homeostasis, brings about many biochemical changes, first touching purine and pyrimidine metabolism. Cell destruction and reparative proliferation occurring in an organ activate the enzymic systems of anabolism and catabolism of nucleotides and their inter mediate metabolites which leads to the formation of a certain nucleotide pool peculiar to the tissue, stage, and gravity of the pathologic process.

The only method making it possible to analyze the whole class of purine and pyrimidine derivatives contained in biological samples is HPLC enabling the analysis of the pathogenesis and the development of the diagnostic crite ria of various diseases. From this standpoint, different HPLC method were emp loyed for the analysis of the biopsy material and blood of patients with cardiovascular (myocardial infarction of various gravity, angina pectoris) and autoimmune diseases (systemic and discoid lupus erythematosus, der matitis) as well as with diseases of the gastro-intesti- nal tract, liver and others. The criteria of evaluation of pathologic status were elaborated on the basis of qualitative and quantitative characteristics of the pool of purine and pyrimidine compounds.

42 057 S.ll-1 SIMULTANEOUS ANALYSIS OF PURINE AND PYRIMIDINE NUCLEOTIDES, NUCLEOSIDES AND BASES BY MIXED-MODE HPLC VIA COLUMN SWITCHING ОДНОВРЕМЕННЫЙ АНАЛИЗ ПУРИНОВЫХ И ПИРИМИДИНОВЫХ НУКЛЕОТИДОВ, НУКЛЕ03ИД0В И ОСНОВАНИЙ С ПОМОЩЬЮ МНОГОМЕРНОЙ ВЭЖХ ПУТЕМ ПЕРЕКЛЮЧЕНИЯ КОЛОНОК P.R.Brown, A.P.Halfpenny, Kingston, USA A method is presented for the separation of purines and pyrimidines, their nucleosides and nucleotides in one analysis using column switching. This method uses both the ion-exchange and reversed-phase modes for the analysis of these compounds. The modes are coupled through automatic switching valves. The use of these valves permits t.he iso lation of either column during the analysis. The nucleoti des are separated on an anion-exchange column and the nucleosides and bases on a Cig column. Total analysis time is approximately one hour. The technique is flexible in that it permits optimization of separation of both the ionic nucleotides and the non-ionic nucleosides and bases. In addition, smaller sample volumes are required than if two separate analyses had to be performed. The method is also applicable to various eample matrices and to diffe rent classes of compounds. Application of the method to biological samples is reported.

058 S.ll-2 SEPARATION OF METHYLATED ARGININE AND LYSINE DERIVATIVES ON VARIOUS SILICAS BY SALT SOLUTIONS РАЗДЕЛЕНИЕ МЕТИЛИРОВАННЫХ АРГИНИНА И ЛИЗИНА НА РАЗЛИЧНЫХ СИЛИКАГЕЛЯХ В СОЛЕВЫХ РАСТВОРАХ T.Cserhati, B.Bordas, E.Tyihak, Budapest, Hungary Retention behaviour of tri-, di-, monomethylarginine, arginine, formyllysine, tri-, di-, monoroethy1lysine and lysine has been studied on I. DC-Alufolien Kieselgel 60 (Merck), II. Polygram Sil G (Macherey-Nagel), III. Silufol (Kavalier, Czechoslovakia) and on IV. Silpress (Labor MIM, Hungary) plates. Hand made Kieselgel G plates impregnated with 5% paraffin oil in hexane (V) and without impregna tion (VI) served as controls. Hater, 0,05; 0,1; 0,25; 0,5 M NaCl and Na acetate solutions were applied as eluents. Plates I., II. and IV were suitable for the separation of amino acid derivatives at low salt concentrations.

Principal component analysis (PCA) of Rf values showed that the first PC explained about 90% or total variance. Nonlinear mapping of PC loadings proved that supports I. and II. differed the most strongly on the basis of their retention behaviour, supports III. and IV. were similar, the impregnation modified only slightly the retention of support VI. The application of sodium acetate instead of sodium chloride changed the retention behaviour of support IV. to a little extent. Two dimensional nonlinear mapping

43 of PC variables showed that the lipophilicity of amino acid derivatives and the salt concentration of eluent determined the retention. 059 S.ll-3 INVESTIGATION OF STRUCTURAL RELATION OF PHARMACOLOGICALLY ACTIVE COMPOUNDS BY RP-HPLC ИЗУЧЕНИЕ СТРУКТУРНЫХ ОСОБЕННОСТИ? ФАРМАКОЛОГИЧЕСКИ АКТИВНЫХ СОЕДИНЕНИИ С ПОМОЩЬЮ 0БРАШЕНН0-ФА30В0И ВЭЖХ K.Valko, Budapest, Hungary Drug molecules acting on the same receptor sites should be structurally related although their structural formulae are not obviously similar. Structural similarity is impor tant for a good correlation of the biological activity and physicochemical parameters used in quantitative structure- activity relationship (QSAR) studies, however, one does not usually know in advance whether or not compounds are congeneric. Therefore a quantitative measure has been developed for characterizing the structural similarity of a compound set. Reveraed-phase retention data (expressed by log k' values) of 50 drug molecules have been investigated as a function of the organic modifier concentration (

44 considerable variations from one brand of silica to another /3/. Possible measures to avoid the problems for both types of separation will be discussed. Reversed-phase separa tions may be performed on dynamically modified silica. This approach has shown to exhibit only minor brand to brand variations in selectivity /4,5/. To ensure a more uniform selectivity of straight-phase separation methods it maybe useful to control the pH of the silica surface. This may be carried out by treating the bulk material with a suitable buffer or by an appropriate of the silica column in-situ. 1. A.P.Goldberg, Anal. Chem., 5_£ (1982) 342. 2. I.Wouters, S.Hendrickx, E.Roets, J.Hoogmartens, H.V&iiderhaeghe, J.Chromatogr., 29_1 (1984) 59« 3. H.Engelhardt, H.Mu'ller, J.Chromatogr., 2_18 (1981) 395. 4. S.H.Hansen, P.Helboe, M.Thomsen, U.Lund, J.Chromatogr., £±2 (1981) 453. 5. P.Helboe, J.Chromatogr., 1245 (1982) 229. 081 S.8-1 SOME APPLICATIONS OF MICRO-COLUMN LIQUID CHB0MAT0G!?APHV AND MASS-SPECTROMETRY WITH DIRECT INLET OF LIQUIDS TO PROTEIN CHEMISTRY НЕКОТОРЫЕ ПРИМЕНЕНИЯ МИКР0К0Л0Н0ЧН0И ЖИДКОСТНОЙ ХРОМАТОГРАОИИ И МАСС-СПЕКТРОМЕТРИИ С ПРЯМЫМ ВВОДОМ ЖИДКОСТИ В ХИМИИ БЕЛКА M.L.Alexandrov, O.A.Mirgorodskaya, Leningrad, USSR G.I.Baram, M.A.Grachev, Novosibirsk, USSR Combination of the micro-column liquid chromatograph OB-4 (Milichrom) /1/ and a mass-spectrometer having a device for direct extraction of ions from solutions /2/ may have many applications to protein chemistry. The chromatograph may be used to perform HPLC with typical volumes of eluent ca. 2000/il, on 200/il (2x62 mm) columns having efficiencies up to 6000 theoretical plates. The detector of the instrument - a programmable double- beam microspectrophotometer - enables parallel monitoring of the absorbance at several wavelengths over the range 19Q-360 ran. The latter facility is especially useful in chemical modification studies helping to identify non protein chromophores. The standard reversed-phase micro columns give resolution sufficient for the separation of complex mixtures of peptides by gradient elution, and pure peptides may be obtained from protein enzymatic digests on the nanomole and sub-nanomole scale. Compared with chromatographs having columns of greater volume, Milichrom puts less rigorous demands to purity of solvents and pro vides better detectability of peptides. An essential feature of the new method of direct extrac tion of ions from solution into mass-spectrometer /2/ is that depending on the operation conditions it may give either mass-spectra of fragments of these ions, or mass

es i spectra containing exclusively the quasi-molecular ion lines of even high molecular weght compounds like amino acids and peptides consisting of up to ten amino acid residues. This feature facilitates studies of the chemical and enzymatic conversions of peptides in solutions. To record a mass-spectrum up to lOOO a.m.u. it is sufficient to have some 100 pmoles of a peptide. The rates of the flow of liquids in the above mentioned device for the direct extraction of ions into mass-spectrometer (a few microliters per minute) and in Milichrom are of the same order of magnitude, and it is possible to couple the two instruments "on-line" /3/. Examples will be given of the application of the instru ments to protein chemistry, particularly, to studies of the affinity modification of RNase and histones, to investigation of the probiin of tick-borne encephalitis virus, to sequencing of peptides, to studies of the kine tics of chemical modification and enzymatic hydrolysis of bradykinin /4/. 1. G.I.Baram et al., J.Chromatogr., 264 (1983) 60-90. 2. M.L.Alexandrov et.al., Dokl.Akad.nauk SSSR, 277 (1984) 379-383. 3. M.L.Alexandrov et al., Bioorg.khymia, Ю (1984) 710-712. 4. M.L.Alexandrov et al., Bioorg.khymia, IT (1985) 724-726. 062 S.8-2 SEPARATION OF ENANTI0HERIC AMINES BY SILICA BONDED o^-ACID GLYCOPROTEIN РАЗДЕЛЕНИЕ ЭНАНТИОМЕРНЫХ AMHhOB НА СИЛИКАГЕЛЕ,

МОДИФИЦИРОВАННОМ о11-ГЛИК0ПР0ТЕИН0М G.Schill, Uppsala, Sweden I.W.Wainer, Washington, USA A new chiral stationary phase, crosslinked mi-.-acid glycoprotein ionically bonded to diethylamino ethyl silica (EnantioPac, LKB, cf. /1/), has been applied to resolution of enantiomeric ammonium compounds of physiological impor tance. Cationic compounds of widely different structure can be separated by varying mobile phase composition and other chromatographic conditions, Chiral resolution has been obtained if the molecule, in addition to the ammonium function, contains a hydrogen bonding function and fairly large cyclic structures in the vicinity of the cationic and/or the hydrogen bonding groups. The distance between the bonding* groups does not appear to be critical and enantiomers with widely diffe rent lengths between the groups have been separated. Retention and selectivity* are often strongly affected by temperature. Charged and uncharged modifiers, inorganic salts and pH can also' be used to alter retention and chi ral selectivity. The magnitude of these changes is often strongly dependent on the structure of the analyte and even enantiomers of a single compound can be affected differently.

46 The separating efficiency is fairly low and highly dependent on the mobile phase speed. The peak symmetry is also due to a variety of factors such as structure and concentration of the analyte, pH, mobile phase additives, etc. However, the separation factors are often larger than 1,2-1,3 and resolution can be obtained even though H values are about 0,2-0,3 mm at a flow, of 0,3 ral/min. Separation conditions will be reported for chiral drugs of widely different hadrophobicity including terbutaline, doxylamine, metoprolol, labetalol, methadone, homatropine, propranolol, methorphan, propoxyphene and cocaine. 1. J.Hermansson, J.Chromatogr., 298 (1984) 67. 063 S.8-3 MECHANISMS OF ENANTIOMER SEPARATION ON CHIRAL HPLC COLUMNS МЕХАНИЗМ РАЗДЕЛЕНИЯ ЭНАНТИОМЕРОВ МЕТОДОМ ВЭЖХ НА КОЛОНКАХ С ХИРАЛЬНЫМИ ФАЗАМИ W.H.Pirkle, Urbana, USA Abstract not arrived on the time for printing. 064 S.8-4 TRACE ENRICHMENT AND REACTION DETECTION IN PACKED CAPILLARY LIQUID CHROMATOGRAPHY КОНЦЕНТРИРОВАНИЕ И РЕАКЦИОННОЕ ДЕТЕКТИРОВАНИЕ В МИКРОКОЛОНОЧНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ D.Westerlund, D.Nilsson, Uppsala, Sweden The continuing development within modern liquid chroma tography has demonstrated the advantages of using low dis persion columns. Decreasing the diameter of the columns gives significant .improvements in mass sensitivity. How ever, this feature is in prectice only useful when the sample volume is limited., like in certain bioanalytical applications; thorough pharmacokinetic studies generating numerous of samples, and especially for determinations in small animal cerebrospinal fluid or interstitial cell fluid. Different neurotransmitters, like catecholamines and peptides, were studied in ion-pair reversed-phase liquid chromatographic systems based on packed fused sili ca tubings (I.D. 2OO-5O0 urn). UV-absorbance and fluores cence were used for detection of the analytes. A high concentration sensitivity can only be obtained in systems of this kind if comparably large volumes can be injected. Trace encrichment effects are obtained when the sample is injected in a solvent which is weaker than the mobile phase. The Influence of kind and concentration of ion-pair reagent as well as the content of organic modi fier was studied in this respect. Special attention was paid to the system (anomalous) peaks which may be obtained when the analytes are injected in a solvent which deviates in composition from the mobile phase. Post-column reactions with reagents specific for amino- 47 groups, like o-phtaldialdehyde and fluorescaraine, provide possibilities for significant improvements of detection limits. The design of mixing cells and reaction tubings is critical in order to keep the extra-column bandbroade nlng as low as possible. 1 065 S.12-1 COMPUTER SIMULATION OF INDIRECT UV DETECTION IN HPLC КОМПЬЮТЕРНОЕ МОДЕЛИРОВАНИЕ ПРИ НЕПРЯМОМ УФ-ДЕТЕКТИРОВАНИИ В ВЭЖХ Gy.Vigh, A.Leitold, Veszprem, Hungary Previously, we reported on the use of nonionic UV active eluent additives for the detection of non-UV-absor- blng solutes, both ionic and nonionic, in reversed-phase HPLC /1/. Equations were proposed relating solute reten tion to detection agent concentration and detection agent retention to solute concentration /2/. Based on these results a program package has been developed to simulate, by an EMG 777 computer, the entire chromatographic process, the development of induced peaks, both positive and nega tive. The program, written in BASC, permits the observa tion of the motion, through the column, of the excess and vacancy peaks of the detection agent and the calculation of the chromatogram, including the integration of the area of the induced peaks. Variation of the parameters of the model over the experimentally realistic ranges indicated that often the area of the main positive induced peak did not agree with the area of the negative induced peak (system peak). The main positive induced peak is frequent ly followed by partially separated other positive induced peak (shoulder peaks). The results indicate that quatita- tion based on the induced peaks must be approached with great care and a full understanding of the behaviour of the system is desirable. 1. Gy.Vigh, A.Leitold, J.Chromatogr..312 (1984) 345. 2. A.Leitold, Gy.Vigh, Magy. Kern. Lapja, in press. 066 S.14-4 PROBING SEPARATION MECHANISMS OF IONIC COMPOUNDS IN 'PLC ИССЛЕДОВАНИЕ МЕХАНИЗМА РАЗДЕЛЕНИЯ ИОННЫХ СОЕДИНЕНИИ В ВЭЖХ S.Ahuja, New York, USA Ionic compounds can be chromatographed by several modes in high pressure liquid chromatography (HPLC). These compounds can be chromatographed as a cation or anion on cationic or anionic exchange columns respectively, or as an ion-pair with the oppositely changed counterion by the reversed-phase liquid chromatography (RPLC). Ionization suppression techniques have also been used in RPLC. The separation mechanisms involved in chromatography of baclofen /4-amino-3-(p-chlorphenyl)butyric acid/ from its

48 transformation product were investigated by both ion- exchange and ion-pair reversed-phase HPLC, These investi gations were primarily aimed at the amino group, which was protonated for the cation exchange chromatography or "ion-paired" with pentane sulfonate anion for the'reversed- phase chromatography. Model compounds were used to postu late separation mechanisms for ionic and nonionic com pounds. Data will also be presented to show the importance of investigations on stationary phase dynamics in separa tions by HPLC. 067 S.14-1 ADVANCES IN THE CHROMATOGRAPHIC ANALYSIS OF POLYMERS ДОСТИЖЕНИЯ В ХРОМАТОГРАФИЧЕСКОМ АНАЛИЗЕ ПОЛИМЕРОВ G.Gl'dckner, Dresden, GDR J.H.M. van den Berg, Geleen, The Netherlands Size exclusion chromatography (SEC) is the most widely used technique for the chromatographic characterization of polymers. The method is highly efficient and provides the satisfactory solution of the problem of evaluating the molar mass distribution (MMD), but it has only limited power for detecting chemical differences in the macromolecules of the sample under investigation. On the other hand, the measurement of chemical composition distribution (CCD) is an important topic in the characterization of copoly mers. Thin-layer chromatography is used in several labora tories for this purpose. A few papers report on the sepa ration of chemically different polymers and copolymers by means of column adsorption liquid chromatography (LC). We have found that HPLC analyses using gradient elution with solvent/nonsolvent mixtures are suitable for the measurement of CCD of copolymers. Using tetrahydrofuran as a solvent and alkane hydrocarbons as nonsolvents we have separated poly (styrene-co-acrylonitrile) or poly( ol-methyl- styrene-co-acrylonitrileTsamples with good results. With these copolymers, the thermodynamic quality of the eluent mixture is the most important variable. The contribution of adsorption phenomena or similar interactions between the solute and the surface of column packing material is small. The separation is caused by precipitation and re'dissolution of the polymer specimen (HP precipitation LC, HPPLC). Copolymers from styrene and methyl methacrylate behave differently. Here we observed a pronounced influence of the stationary phase. This influence is to be expected according to the general understanding of liquid chromato graphy. With poly(styrene-co-methyl methacrylate) samples, separation by means of HPPLC using reversed-phase packings is possible but does not reach the excellent level of resolution that can be obtained by using bare silica packing materials.

4. 3lK. 872 49 068 S.14-2 CHROMATOGRAPHY OF MACROMOLECULES UNDER "CRITICAL CONDITIONS" AND PROBLEMS OF MOLECULAR HETEROGENEITIES OF POLYMERS ХРОМАТОГРАФИЯ МАКРОМОЛЕКУЛ В "КРИТИЧЕСКИХ УСЛОВИЯХ И ПРОБЛЕМ МОЛЕКУЛЯРНОЙ НЕОДНОРОДНОСТИ ПОЛИМЕРОВ A.V.Gorshkov, V.V.Evreinov, • Moscow, USSR A new method of defining the molecule heterogeneity of polymers chromatography under critical conditions at the interface between exclusion ( ДЕ«£ТД S) and adsorp tion (ЛЕ>ТДБ) modes of separation - has been first realized and grounded experimentally. Virtually complete disappearance of size separation of the linear homoge neous macromolecules over a wide range of MM (200 to 100.000) is characteristic of the "critical region". This enables a fundamentally new approach (compared to the traditional exclusion and adsorption chromatography) to study of the elementary heterogeneities of polymers, such as functionality, cyclicity, branching, etc. The chromatograms obtained with the separation in the "critical region" (SCR) mode provide information solely on the types of polymer heterogeneities uncomplicated by size separation. The molecular heterogeneity of polymers is treated in terms of elementary heterogeneities of the local and global macromolecular structures. The theoretical ideas of the SCR method are discussed together with the prin ciples underlying its experimental realization for sepa ration in functional and topological types of telechelic polymers (separation of linear from cyclic macromolecules). The possibilities opened up by the method for study of the molecular heterogeneity of block-copolymers, of branched and other more complicated spatial structures of macromolecules are discussed in brief, as well as the physical limitations of the SCR chromatography mode.

069 S.14-3 THEORY OF CHROMATOGRAPHY OF POLYMER MACROCYCLES ТЕОРИЯ ХР0МАТ0ГРА4ЙИ КОЛЬЦЕВЫХ МАКРОМОЛЕКУЛ A.A.Gorbunov, A.M.Skvortsov, Leningrad, USSR A theory of chromatography of ring polymers has been developed. The model under consideration was that of a flexible Gaussian cyclic chain interacting with the walls of a slit-like pore with arbitrary energy. Analyti cal expressions for a distribution coefficient К and for some other equilibrium characteristics (monomer density in pore,' degree of chain bonding) have been derived. In the exclusion range, when the adsorption interaction is shown to be absent, the molecular-mass dependences of К for ring and linear polymers are of similar type. On the other hand, sharp, qualitative differences in chromatographic behaviour of ring and linear polymers

SO appear in the case of weak polymer-adsorbent interactions. In this range the К value decreases with the growth of molecular mass for linear polymers, but increases for cyclic ones. These differences may be used for effective chromatographic fractionation of polymers according to their structure. For the separation of linear and cyclic chains one can use "the critical chromatography" as well. In the critical conditions entropy losses of the linear chain in the pore are completely compensated by the gain in energy, caused by the adsorption of chain units. Therefore К = 1 for linear chain irrespective of its radius R and of pore width D, while for ring molecule at К > 1 К = 1 + V1^ R/D. The theoretical results have been compared with experi mental data on chromatography of macrocycles. 070 P.1-44 THEORETICAL ASPECTS OF THE PORE DISTRIBUTION PROBLEM BY SIZE EXCLUSION CHROMATOGRAPHY ТЕОРЕТИЧЕСКИЕ АСПЕКТЫ РАСПРЕДЕЛЕНИЯ ПОР ПО РАЗМЕРАМ В ЭКСКЛЮЗИОННОЙ ХРОМАТОГРАФИИ R.N.Nikolo'v, Sofia, Bulgaria Usually, the experimentally determined pore distribu tion of dispersive materials is presented graphically by the differential change of the relative pore volume with respect to the pore radius or diameter. In fact, this is a typical pore volume distribution that can be very useful in studying the material porosity. Practically, it is also interesting to know the number of pores having some defi nite diameter regarding all pores of the material studied. Such an information would be substantial to investigating the pore structure of solids and could be obtained from another type of distribution, called here actual pore size distribution. However, there is no any technique for its direct determination. A generalized treatment of the pore distribution prob lem is proposed with no reference to the geometrical form of the pores. Actual pore size distribution is clearly difined and a relationship between it and the pore volume distribution is derived. The possibilities of obtaining both distributions from size exclusion chromatography data are demonstrated on silica gels intended for HPLC. All important numerical characteristics of the distributions (e.g. mean value, standard deviation and variance) have been evaluated by applying the method of moments; Obtained results and plotted distributions of different silica gels are presented and thoroughly discussed.

SI 071 P.1-46 CONCERNING THE PECULIARITIES OF THE INTERPRETATION OF THE DATA OF THE SCREEN CHROMATOGRAPHY OF POLYMERS IN TERMS OF RATE CONSTANTS OF CHEMICAL REACTIONS ОБ ОСОБЕННОСТЯХ ИНТЕРПРЕТАЦИИ ДАННЫХ СИТОИЯ ХРОМАТОГРАФИИ ПОЛИМЕРОВ В ТЕРМИНАХ КОНСТАНТ СКОРОСТЕЙ ХИМИЧЕСКИХ РЕАКЦИЙ N.G.Podosyenova, V.P.Budtov, E.L.Ponomareva, E.G.Zotikov, Leningrad, USSR This presentation considers the original methods for the solution of the reverse task developed by us based on the information of SC. The methods are simple because for the determination of the values of rate constants and their dependence on the conversion the analytical results of the investigation of samples produced in the laborato ries and commercial plants are used. The peculiar feature of the methods is differentiation of the corresponding pa rameters of the experimental MMD instead of the integra tion of the differential equations describing the kinetics and MMD of the polymer and the comparison of the calcula tion results and experimental data. As far as the application of "gradient" methods of MMD transfers the tasks to "mathematically non-correct" class this presentation pays significant attention to the dis cussion of the ways to increase its reliability.lt is shown that check of the results by using the approximate method by decomposition into a series by conversion and by the method of "Difference" chromatograms increases the relia bility. The application of the first of these methods is limited by the range of small conversions and that of the second of these methods - by possible significant experi mental error. The application of the method of "averaging" of derivatives calculated by the experimental dependence of the moments of MMD on the conversion increases the reliability. Similar requirements are used to SC depending on the type of the task. Thus while considering the kinetics of the process special attention should be paid only to the rate of the reproducibility of the shape of chromatograms, its low and high molecular ranges. At the same time while using investigation results for checking models high reliability of absolute values of parameters of MMD is required first of all, it means high accuracy of calibra tion as well as possible correction for imperfect chroma tograms. While determining rate constants by moment derivative method the significant accuracy of the calibra tion dependence does not exceed 5%. Examples were considered and it was shown that deve loped methods for the interpretation of SC of polymers based on the computer programs allow ;1) study of the pecu liar features of the radical polymerization (to clear out the role of different elementary reactions while changing temperature and time of the process and its reagents);

52 2 ) scale up of processes that allow to calculate process kinetics and MWD of the formed product,- 3) determination of the rate constants of elementary reactions at different stages of the monomer conversion; 4) determination of the catalytic activity distribution.

072 P.1-45 APPLICATION OF GEL- AND AFFINITY CHROMATOGRAPHY FOR IMMUNOASSAYS ПРИМЕНЕНИЕ АФФИНОИ И ГЕЛЬ-ХРОМАТОГРАФИИ ДЛЯ ИММУНОЛОГИЧЕСКИХ АНАЛИЗОВ W.Hubl, Dresden, GDR Gel- and affinity chromatography are used to prepare pure, active antibodies and tracers for immunoassays. Steroidhormone enzyme conjugates for enzyme immunoassays prepared by the mixed anhydride reaction were purified from unconjugated haptens by gel-chromatography. A radio active labelled steroid is used as marker for the chroma tography . For special purposes, e.g. high sensitive non-isotopic immunoassays, the antibodies were isolated by affinity chromatography on hormones, coupled to activated Sepharose. Using purified tracers and antibodies, non-isotopic immunoassays for the determination of aldosterone, 17-hydroxyprogesterone, 11-deoxycortisol,,digoxin and angiotensin-1 were developed and used for the clinical diagnosis of patinets with various endocrine diseases. 073 P.3-47 PECULIARITIES OF fi-\,3-GLUCAN BEHAVIOUR IN COMPARISON WITH DEXTRANS AT GEL PERMEATION CHROMATOGRAPHY ОСОБЕННОСТИ ПОВЕДЕНИЯ /4-1,3-ГЛЮКАНОВ В СРАВНЕНИИ С ДЕКСТРАНАМИ ПРИ ГЕЛЬ-ПРОНИКАЩЕИ ХРОМАТОГРАФИИ A.A.Dementyev, T.N.Zvyagintseva, L.A.Elyakova, Vladivostok, USSR At present dextrans are widely used as standards for determination of molecular masses (MM) of different poly- sacharldes by gel chromatography. However their uniform use for these purposes is not always justified. The aim of this investigation is to elucidate the possibility for determination of fi -1,'3-glucan MM by gel filtration. The series of fi -1,3-glucans having narrow MM distribution have been obtained by various methods: laminarin - ,/3-1,3; 1,6-D- glucan from Laminaria cichorioides; ,/3-1,3-glucan 1 isolated from laminarin digest with endo- /}-l,3-glucana- se L0 from Chlamys albidus and an alkaline soluble & -1,3-glucanII obtained by acid hydrolysis of pachyman from Poria cocos. Exact molecular masses of these glucans have been defined by end analysis methods. To determine the Л-1,3-glucan MM by gel filtration the inulin (MM 5000) and the following dextrans from "Farmacia" were used; T-80 (KM 80000), T-40 (MM 40000), T-20 (MM 20000), T-10 (MM lOOOO). The results are given in the tables

S3 ^s. Method End Gel filtration analy sis Sephadex G-50, Sepharose Sepharose 0,05 M CH3COOH GL-4B, (GL-4B, Glucan ^4^ 0,05M NaOH 0,3M NaOH j8-l,3-glucan I 2O0O 4000 Laminarin 50O0 11000 11500 11500 £ -1,3-glucan II 6500 - 15700 15700

That values of the ^-1,3-glucan MM determined by liquid chromatography on a column containing different gels at various elution conditions well coincide while MM of j8-l,3-glucans obtained by end analysis are twice as small. Consequently, it is necessary to have a set of related polymer standards for exact determination of fi -1,3-glucan MM by gel permeation chromatography. The anomalous behaviour can be explained by different confor mational states of these polysacharides in the solutions. 074 S.8-6 CHROMATOGRAPHY OF BIOPOLYMERS ON MACROPOROUS SILICA ХРОМАТОГРАФИЯ БИОПОЛИМЕРОВ НА МАКРОПОРИСТЫХ • КРЕМНЕЗЕМАХ Yu.S.Nikitin, T.D.Khokhlova, Moscow, USSR Chromatography of proteins and viruses on macroporous silica has become widely used for analysis and purifica tion of these biopolymers, in particular for preparation of vaccines. For this reason, investigation of the mechanism of retention of proteins and viruses by silica is not only important for theory, but necessary for prac tice. Silica gels, silochromes, and macroporous glass with surface that was either unmodified, or coated with various organic and inorganic agents were used in the investigation. Retention was proved to be determined by a combination of adsorption, ionic and steric exclusion factors. On silica with unmodified surfaces, strong adsorption of alkaline proteins was observed. Adsorption decreased with increase in the eluent pH. At pH over 8, the acidic and neutral biopolymers - thyroglobulin, urease, ferritin, serum albumin, ovalbumin, tobacco mosaic and influenza viruses (viruses with acidic protein shells) - eluted in the order which is characteristic of size exclusion chromatography. The chromatography on unmodified silica was used for the purification of influ enza virus suspension and enzyme preparations. Since biopolymer elution demands high eluent pH, which promotes silica solubility, silica with lower solubility, whose surface was modified with aluminum salts, was also used for biopolymer purification. Silica modified with aminopropyl groups via correspon-

54 ding silane exhibits anion-exchange properties. At low pH and ionic strength of the eluent, a series of proteins exhibited the phenomenon of ionic exclusion, when proteins with 30-50 A globules would not penetrate into the pores with the diameter of 500 A due to electrostatic repulsion from the positively changed aminopropyl silica surface. The treatment of aminopropylsilica with carbohydrates resulted in preparation of the molecular sieves characte rized by the reduced adsorption activity as regards proteins and viruses. These silicas were used for chroma tographic purification of proteins and viruses. 075 P.2-1 A METHOD FOR CALCULATING CAPACITY FACTORS FOR A LINEAR GRADIENT ELUTIOM IN REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY МЕТОД РАСЧЕТА ФАКТОРОВ ЕМКОСТИ В 0БРАЩЕНН0-ФА30В0Й ВЭЖХ С ЛИНЕЙНЫМ ГРАДИЕНТОМ P.Dufek, Praque, CSSR The dependence of In k' on In (methanol concentration) in isocratic high-performance liquid chromatography has been proved to be linear In k- = a In «Methanol + b' (1)

In (k'C2 A'C1)

where a = , Ь - In k'cl - a-In c,, x In (c2/Cl) k'c and k' are respective capacity factors of a compound when eluted with mobile phase of methanol concen tration сj and c,. Supposing a linear concentration gradient xt methanol

s~l, methanol concentration at time t equals c.= c0 + x>t с being methanol concentration at the start of the gra dient run. Thus

In k' = a.In (c0 + x t) + b (2) Inserting egn. 2 into the definition of capacity factor we obtain a b In tR = In tM + In /(c0 + xt ) e + 1 J and retention time equals a b tR = tM /"(c0 + x t ) . e + 1 J. (3) The linear velocity v is in chromatography given by expression

= L L V (4) ~ tfc_ - t /(c(c + x t)A eb + V R VM o0 and hence the length of a column L is /R vdt = /»- -**- ° ° 1м'/,со + х tr hence t ** = /* r -* a D о 1 + (cQ + x-t ) . e (5) 55 Because primitive function of this term is not known there are two possible waye of solving this integral for calculating tp: 1) neglecting 1 relative to the term (c_ + x-t)a-eb, or 2) using a numerical method of computation. The accuracy of the latter method Is much better but it is more complicated. The calculated values of retention time have been compared with those measured for N-alkylphthalimides.

076 P.2-02 APPLICATION OF A UNIVERSAL EQUATION FOR RETEHTION PARAMETERS CALCULATION OF 2,4-DNPH CARBONYL COMPOUNDS IN LIQUID CHROMATOGRAPHY ИСПОЛЬЗОВАНИЕ УНИВЕРСАЛЬНОГО УРАВНЕНИЯ В ЖИДКОСТНОЙ ХРОМАТОГРАФИИ ДЛЯ РАСЧЕТА ПАРАМЕТРОВ УДЕРЖИВАНИЯ ?,4-ДНФГ КАРБОНИЛЬНЫХ СОЕДИНЕНИЙ L.I.Sokoiova N.I.Svetlova, D.N.Grigoryeva, R.V.Golovny Moscow, USSR GC analysis Oi carbonyl compounds in free form is undesirable due to their lability. Usually they are analy sed by reversed-phase liquid chromatography (RPLC) in form of 2,4-dinltrophenylhydrozone (DNPH) derivatives. Much work has been done to develop approach allowing prediction and justification of retention data in RPLC. But the models that have been proposed up to now are not conve nient for practical application or for accurate prediction of the retention. Recently a universal equation has been suggested for calculation of GC retention indices for homologous seria of organic compounds: I - oC + fi m + J"lgm/m + J / [(m-2)2 + 0,l] , (1) where V is the homologue's sequential number. The egn.l is based on thermodynamical model describing the deviation from additivity principle for the energy of van-der-Waals Interaction of homologues with a stationary phase in GLC conditions. In order to check the validity of the egn.l in RPLC, we employed it for retention parameters calculation of two homologous seria of DNPH derivatives of C,-C„ n-aldehydes and alk-2-ene aldehydes. A "Yanako" liquid cnromatograph, "Yanopak" ODS (50 cm x 5 mm I.D.) column and UV detector operated at 254 ran were used. The eluent was 80:20% iv/v) methanol-water. The inlet pressure was 170-180 kg/cm . The adjusted retention times were used as retention para meters. The values of the coefficients for egn.2 and 3 and standard deviation (s) were cal-ulated by LSM: lg t'= 1.18 + 0.16m - 0.437 lgm/m + O.0O5/ [(m-2)2+ 0.ll(2) for n-aldehydes; s « 0.007, lg t'= 1.1572+0.168m-0.248 lgm/m+O.OOl/[(m-2)2+0. l] (3) for alk-2-ene aldehydes; s = O.O06. 56 The results showed high accuracy of retention parameters calculation for all homologues including the first members. Hince eqn. 1 can be recommended for identification of ..lulticoraponent mixtures of carbonyl compounds by RPLC method.

077 P.2-03 QUANTITATIVE COMPOSITION OF THE MOBILE PHASE AND RETENTION IN ADSORPTION CHROMATOGRAPHY КОЛИЧЕСТВЕННЫЙ СОСТАВ ПОДВИЖНОЙ ФАЗЫ И УДЕРЖИВАНИЕ В АДСОРБЦИОННОЙ ХРОМАТОГРАФИИ V.D.Shatz, O.V.Sahartova, V.D.Grigoryeva, Riga, USSR The use of binary mobile phases is a common feature of normal phase chromatography on silica and reversed phase chromatography on alkyl silicas. Generally one of the mobile phase components has a negligible elution strength towards the class of compounds under study. The presence of the stronger component in the mobile phase usually leads to decreased retention values. Some typical relationships between capacity factors k" and the stronger solvent concentration С are discussed. When compounds of moderate polarity are chromatographed on silica, retention data usually follow the equation

lg k< = a0 + a! lg С (1) and a^ value is approximately constant for different mem bers of a given variety of compounds, so that the lg k' - lg С plots are parallel. The fanwise pattern of lg k' - lg С plots is characte

ristic of reversed phase chromatography, because aQ and

a, are correlated. The relationship between a0 and a^ is linear, so that lg k" - lg С plots tend to intercept at one point.. When basic compounds are chromatographed in a reversed phase mode, the lg k' - lg С relationship is parabolic:

2 lg k' = a0 + ax lg С + a2 lg C. (2) The different types of к'- С relationships in different modes of HPLC can be explained by differences in solvation equilibria existing in the chromatographic system and by the fact that sorption properties of the system components can be altered as a result of solvation.

57 078 P.2-04 STERIC EFFECTS OF SUBSTITUENTS IN NORMAL PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY СТЕРИЧЕСКИЕ ЭФФЕКТЫ ЗАМЕСТИТЕЛЕЙ В ПРЯМОФАЭНОЙ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ A.E.Ermoshkin, N.P.Makarenko, Gorky, USSR K.I.Sakodynskii, Moscow, USSR The influence of alkyl (Alk) substituents on the retention of the silicon-containing compounds OH OSiMe2OAlk

SiMe2OAlk t-Bu-

t-Bu t-Bu II

III

(where Alk = CH3, CHjCHj, CH(CH3>2, C(CH3)3; Me = CH3, t-Bu = C(CHj)- ) on Silasorb 600 ("Lachema", Brno, CSSR) when using n-nexane as an eluent has been investigated. The increase of the Alk-substituents dimensions (going over from CH„ to C(CH3)3 ) results in the retention decrease of the compounds investigated owing to an increase of the steric hindrance of the oxygen atom in dimethylalkoxysilyl substituent making a decisive contri bution to the retention. It is found that there is a correlation between the net retention times logarithms of the compounds investiga ted and the Palm's and Charton's steric constants of the Alk-substituents. 079 P.2-05 THE EFFECT OF ORGANIC SOLVENT MODIFIER AND ION-PAIRING REAGENTS (THE TETRABUTILAMMOKIUM SALTS) ON THE CHROMATOGRAPHIC BEHAVIOUR OF PROTECTED PEPTIDES IN REVERSED-PHASE HPLC ВЛИЯНИЕ ОРГАНИЧЕСКОГО РАСТВОРИТЕЛЯ И ИОН-ПАРНОГО РЕАГЕНТА /СОЛИ ТЕТРАБУТИЛАММОНИЯ/ НА ХРОМАТОГРАФИЧЕСКОЕ ПОВЕДЕНИЕ ЗАЩИЩЕННЫХ ПЕПТИДОВ В УСЛОВИЯХ 0БРАЩЕНН0-ФА30В0И ВЭЖХ E.V.Titova, M.A.Chlenov, L.I.Kudryashov, Moscow, USSR The adsorption, size-exclusion and reversed-phase HPLC are used for analysis of synthetic peptides with protec ted groups. However the behaviour of these peptides in 58 case of reversed-phase HPLC (RP HPLC) is studied unsuffi ciently. The peculiarities of the application of № HPLC for analysis of synthetic peptides with protected groups are described. The retention dependence on zorbax C8 column (Du Pont, USA) of several peptides with free and protected N-ter minus on organic solvent (acetonitrile or methanol) content has been investigated in the range of solvents concentrations Ю-90 % (v/v). It has been shown that in case of acetonitrile the parabolic-like dependence of k' of peptides with free aminogroup on acetonitrile concent ration takes place with the minimum at ca. 80 % of aceto nitrile. This is not the case for analysis of fully pro tected peptides: their retention is reduced when aceto nitrile content is increased up to 90 %. It was also found that in case of methanol the anomalous behaviour of peptides investigated was not observed. When studying the effect of ion-pairing reagents: tetrabutylammonium bromide (TBAB) or tetrabutylammonium hydrogensulphate (TBAHSO.)- on chromatographic behaviour of protected peptides it has been shown that together with already revealed effects of sorbent modification by-TBA+- ions the influence of Br" and HSO." anions takes place. The data obtained show that in the presence of TBAHSO4 retention times of peptides are less than those in the presence of TBAB. It can be concluded that the classic regularities of RP HPLC are valid for analysis of synthetic fully protec ted peptides whereas in case of peptides having free aminogroups some deviations are observed due to additional effects. The counter ions effects were found in the presence of tetrabutylammonium salts in the eluent.

080 P.2-06 LIGAND-EXCHANGE HPLC OF NOCLEOBASES AND NUCLEOSIDES ON 8-HYDROXTOUINOLINE - SILICA - POLYOL ЛИГАНДООБМЕННАЯ ВЭЖХ НУКЛЕИНОВЫХ ОСНОВАНИЙ И НУКЛЕ03ИД0В hA ДИОЛ-СИЛИКАГЕЛЕ, МОДИФИЦИРОВАННОМ 8-0КСИХИН0ЛИН0М G.-J.Krauss, Halle, GDS Immobilized chelating agents are gaining increasing importance for the ligand-exchange chromatography of nucleic acid constituents. In the present study, a new immobilized chelate, 8-hydroxyquinoline-silica-polyol (Serva, Heidelberg, FRG) has been used. Copper binds to this matrix and retains the ability to coordinate ligands so that completing agents can be sorbed on the stationary phase. This new bonded phase was used to investigate the chromatographic behaviour of nucleobases, nucleosides and nucleotides. The mechanism for retention must be conside red as a composite mechanism (ligand-exchange and adsorp tion) . Using water as mobile phase the mixed mode support 5» is a particularly useful system for high speed separation of uracil derivatives. Nucleotides and more basic purine bases and nucleosides are retained strongly on the column. The system reported here allows the quantitative measu rement of nucleoside and nucleobase level in biological samples and illustrates possibilities for the application of these separations in the biochemical field.

081 P.2-07 LIGAND-EXCHANGE CHROMATOGRAPHY OF NUCLEASES ЛИГАНДООБМЕННАЯ ХРОМАТОГРАФИЯ НУКЛЕАЗ V.P.Varlamov, S.A.Lopatin, G.E.Bannikova, I.A.Andryushina, S.V.Rogozhin, Moscow, USSR The ligand-exchange chromatography (metal chelate affi nity chromatography) is one of the effective purification techniques for enzymes. This technique is based on the ability of proteins tc bind ions of metals immobilized on chelate gels. This chromatographic technique has the following advantages: high selectivity, which allows to separate proteins with similar properties, a possibility to use sorbents many times, a simplicity of preparation and regeneration of gels. - Ligand-exchange supports were synthesized based on inorganic material "Silochrome" and organic polymers "Spheron" and "TSK-Gel Toyopearl" with iminodiacetic, iminodimethylphosphonic and ethylendiamintetraacetic acids as stationary ligands. These sorbents were loaded by ions Cu2+, Ni2+, Znz , Mnz+, Coz+, Fe3+. Concentrations of ligands changed from Ю to ICO micromol per ml of gels. The exonuclease A5 from actinomyces was separated from the phosphatase and the 5'-nucleotidase by using tandem columns packed with the above

083 P.2-09 APPLICATION OF .(.-, j8 -CYCLODEXTRIN AND HEPTAKIS- (2,6-DI-O-METHYL)-jS-CYCLODEXTRIN AS THE MOBILE PHASE COMPONENTS FOR SEPARATION OF SOME CHIRAL BARBITURATES INTO ENANTIOMERS BY REVERSED-PHASE HPLC ПРИМЕНЕНИЕ oL, £ -ЦИКЛ0ДЕКСТРАН0В И ГЕПТАКИС/2,6-ДИ- -0-МЕТИЛ/- jS-ЦИКЛОДЕКСТРИНОВ В КАЧЕСТВЕ КОМПОНЕНТОВ ПОДВИЖНОЙ ФАЗЫ ДЛЯ РАЗДЕЛЕНИЯ НЕКОТОРЫХ ХИРАЛЬНЫХ БАРБИТУРАТОВ ПУТЕМ 0БРАЩЕНН0-ФА30В0Й ВЭЖХ D.Sybilska, Warsaw, Poland J.Bojarski, Krakow, Poland The remarkable ability of cyclodextrins (CDs) to form stereoselective inclusion compounds with various molecules and ions is well known and has been exploited in many separation techniques. We have recently found that very effective separation including those of enantiomers may be achieved in chromatographic systems if appropriate CD receptor molecu les are placed either in liquid stationary phase (gas- liquid chromatography) or mobile phase (RP HPLC). This communication reports a systematic study on reso lution of a series of imidazol and pyrimidine derivatives (including some therapeutically useful drugs) into enan tiomers on LiChrosorb RP-18 or RP-8 columns with the mobile phase containing

61 complexes as well as their stability constants in the mobile phase solution were calculated. It has been found that the presence of Js-CD and heptakis-(2,6-di-0-methyl)-3-CD in the RP-systems results in a distinct enantioselectivity in regard to mephenytoin and the barbiturates containing a chiral center in the pyrimidine ring. The elaborated method will be exemplified by the racemic mixture separations and the obtained data will be discussed with consideration of Dalgiesh "three point attachment concept".

084 P.2-10 HPLC OF PHENOLS. A NEW APPROACH FOR IDENTIFICATION ВЭЖХ ФЕНОЛОВ. НОВЫЙ ПОДХОД К ИДЕНТИФИКАЦИИ V.P.Dmitrikov, Dneprodzerzhinsk, USSR HFLC is a convenient method for analysis of coal tar especially of phenols. Silica gel is an excellent choice for analytical sepa ration of solutes having a middle polarity and acidity. In the presence of steric and electronic effects solute retention is not directly proportional to number and type of substitution groups. The most convenient for predict retention is method, of usage constants acidity of phenols in a various solvents. Measurements were performed by a "Hilikhrom" (USSR) on a column, packed with Sylasorb-600 (7,5 jam) , and using the mixture of hexane - i-propanol (100:2 = v/v).

085 P.2-11 THE STUDIES ON OPTIMIZATION OF PARAMETERS OF PREPARATIVE LC COLUMNS FOR PRODUCTION OF CARDIAL GLYCOSIDES ИЗУЧЕНИЕ ПАРАМЕТРОВ ОПТИМИЗАЦИИ КОЛОНОК ДЛЯ ЖХ ПРИ ПРОИЗВОДСТВЕ СЕРДЕЧНЫХ ГЛИКОЗИДОВ B.Sledzinska, Gdansk, Poland The paper presents the results of investigations aiming at maximization of output of the process of production of cardial Ianatosides from their mixtures using chromato graphy. The investigations dealt with the dependence of the yield of the process on column length, particle size of the packing and linear velocity of the mobile phase. It has been established that, among others, due to occa sional limited solubility of the separated substances in the mobile phase it is disadvantageous to use modern HPLC packings with the particle size < 10 jum. On the other hand, simultaneous increase of particle size, column 62 length and the mobile phase flow rate resulted in improve ment of the yield of the process. However, such procedure .is limited due to technological reasons. Technological limitations and optimization criteria which should be taken into account when optimizing indust rial application of preparative chromatography have been formulated. On this basis optimum production conditions have been characterized.

086 P.2-12 ELECTROKINETIC DETECTION IN ION-EXCLUSION HIGH PERFORMANCE LIQUID CHROMATOGRAPHY ЭЛЕКТРОКИНЕТИЧЕСКОЕ ДЕТЕКТИРОВАНИЕ В ИОНООБМЕННОЙ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ W.Kemula, Warsaw, Poland The results of the electrokinetic detection of acidic substances separated using ion-exclusion mechanism in HPLC has been elaborated. It enabled the determination of mixtures of carboxylic acids. The benzoic acid in the mustard - as example- is presented. Detectability of these substances is inverse propor tional to their pKa values.

Strong acids - until pKa« 2.5 - are c'uted unseparated at the beginning of the elution , the weak acids - pKa> 6,5 - similarly are not separated and eluted at the end of elution.

The retention volumes of the substances having pKa values from 2,5 to 6.5 are proportional to their pKa values. From this dependence dead and inner volume of the chromatographic column can be determined. Equations describing suitable dependencies are proposed. 087 P.2-13 STREAMING CURRENT DETECTION OF STRONG ACIDS IN NORMAL PHASE LIQUID CHROMATOGRAPHY ПРОТОЧНЫЙ ДЕТЕКТОР СИЛЬНЫХ КИСЛОТ В НОРМАЛЬНО-ФАЗОВОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ R.Vespalec, M.Cigankova, Brno, CSSR Streaming current detection of strong acids in a normal phase polarity chromatographic system was studied by a model of hydrolyzed extraction solution of tributyl phosphate. In addition to tributyl phosphate (TBP), hydro carbon solvent and water, the model solution contains hydrolytic decomposition products: butanol, dibutylphospho- ric (DBP), monobutylphosphoric (MBP) and orthophosphoric acids. All the components can be separated on silica gel perfectly provided thr.t a mixture of n*-alkane and alcohol, partly saturated with /„ter, serves as a mobile phase. Phosphoric acid zone tailing can be suppressed by trace concentrations of HC10.. At the streaming current detection the chromatographic column served as a detection element. By comparison with refractometer response no record of responses of hydro- «3

I carbon solvent, TBP and butanol was found. Symmetry of acid zones was better in the refractometric record than at the streaming current detection despite that the appa ratus zone broadening is less at the streaming current signal sensing from the column than at the refractometric detection. The sensitivity of the streaming current detec tion of acids, relative to their refractometric responses,

increased in the series DBP - MBP - H3PO. in the ratio 1:4:15. Retentions of these compounds were in the ratio 1:2:4. The minimum detectable concentration of H,PO. in the eluate was 2.10 mol/1.

088 P.2-14 EVALUATION OF SOME VOLUMETERS FOR LIQUID CHROMATOGRAPHY ХАРАКТЕРИСТИКА НЕКОТОРЫХ ИЗМЕРИТЕЛЕЙ ОБЪЕМА ДЛЯ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ T.Macko, O.Berek, Bratislava, CSSR O.Chiantore, Torino, Italy The precise knowledge of the volume of effluent leaving the column is important for quantitation of measurements in various modes of liquid chromatography. He have evalua ted the reliability, precision and repeatibility of three different commercially available volumeters for conven tional and high performance liquid chromatography, namely siphon systems, drop counter and two types of thermopulse capillary flowmeters. The actual volume of testing liquid that has flowed through the volumeter was determined by weighing while the measures were taken to prevent the system from evaporation. The correlations were determined between the particular volumeter response (one dumping of the siphon, one hundred of droplets and ten thermopulses) and the operational variables: nature or composition of liquid, its flow rate and temperature as well as position of volumeter. It was found that the response of all volumeters fairly depended on temperature, nature and flow rate of liquid. Siphon and drop counter responses were sensitive also to the position of the device and to the presence of small amounts of additives in the measured liquid. In the case of drop counter, the additional errors may be caused by evaporation of liquid during measurement. In any case, all volumeters tested yielded only relative values of volume of passed liquid and the corresponding calibrations were necessary to obtain absolute data. The repeability of measurements decreased in the series:Thermopulse volu meter (Knauer Electronic Volumeter Type 68.OO, cell volume lOjul), drop counter (Model DC lOOl, Laboratory Instruments Works, Prague, .Czechoslovakia), siphon systems (home made devices), thermopulse volumeter (Knauer Electro nic Volumeter, cell volume about lOO ^il) and was + 0.14 %, + 0.24 %, + 0.45 %, and + 1.16 % vol. resp. ~

64 089 P.2-15 SEPARATION OF COLLAGEN HYDROLYSATE AMINO ACIDS BY ION-PAIR REVERSED -PHASE HPLC РАЗДЕЛЕНИЕ АШНОКИСЛОТ.ГИДРОЛИЗАТОВ КОЛЛАГЕНА МЕТОДОМ ИОН-ПАРНОИ ОБРАЩЕНН0-ФА30В0И ВЭЖХ J.Macefc, L.Miterovii, M.Adam, Prague, CSSR For characterization of different collagen types and/or collagen fragments the estimation of amino acid composi - tion is of prime importance. Ion-exchange chromatography of amino acids entails problems with high cost of columns, possible citrate buffers contamination and/or column efficiency. As an alternative we have developed a chromatographic system, which is capable to completely resolve all collagen hydrolysate amino acids in 70 minutes. The glass column used in our procedure was SEPARON SI CJQ (15 cm x 3.3 mm I.D.) and the mobile phase A: 0.01 M aqueous sodium dodecylsulfate (SDS) adjusted with trichlo roacetic acid to pH 2.6; mobile phase B: 0.01 M SDS in mixture n-propanol-water (80:20 v/v). Gradient elution was applied as follows: 0 to 8 min 961 A 4% B; 8 to 12 ' rain 96% A 4% В to 86% A 14% B; 12 to 44 min 86% A 14% B; 44 to 64 min 86% A 14% В to 60% A 40% B; 64 to 70 min 60% A 40% B, The mobile phase flow rate was 0.5 ml/min, column temperature 30°C. The detection was accomplished by post-column reaction with ninhydrin and monitoring absorbance at 440 nm and 570 nm, respectively. The problems of separation of aspartic acid/serine and threonine/glycine pairs reported by some authors has been overcome by initial isocratic step. For the separation of hydroxyproline/aspartic acid the temperature was important not to exceed 30°C. For isoleucine/phenylalanine resolu tion it was necessary to introduce the second isocratic step. Ninhydrin detection allowed to obtain quantitative results with collagen amount as low as 20O да. 090 P.2-16 RAPID ISOCRATIC HPLC OF PORPHYRIN ESTERS ON THE AMINOPROPYLSILICA БЫСТРАЯ ИЗОКРАТИЧЕСКАЯ ВЭЖХ ПОРФИРИНОВЫХ ЭФИРОВ НА АМИНОПРОПИЛ-СИЛИКАГЕЛЕ P.Kotal, B.Porsch, M.Jirsa, V.KordaS, Prague, CSSR For the separation of porphyrin esters according to the number of carboxyl groups (usually 2-8) silica-based HPLC columns are usually used; frequently gradient-elution is necessary. Apparently, the reproducibility and/or the use of isocratic mode strongly depends on the whole history of silica materials Application of the aminopropyl-bonded phase of the different origin results in simple rapid isocratic separation of porphyrin esters. By means of che mical modification of the surface of NH2-silica the mecha nism of partition together with the influence of the amino

5. Зак. 872 group was elucidated. Tailor-made silicas with different surface concentration of silanol groups were prepared and the influence of the concentration of these groups on the separation of porphyrin esters was studied. It is shown that the primary amino groups determine the selectivity of isocratic separation of porphyrin ester?: on the plain silica it is possible to obtain isocratic separation if the surface concentration of silanol groups is decreased. Suggested optimum isocratic procedure allows the base-line separation of the porphyrin esters (2-8) together with their Cu complexes. Moreover, synthetic tetraphenylporphyrin can be .conveniently used as the inter nal standard.

091 P.2-17 MICBO-HPLC OF PHYT0H0RM0NES МИКРОКОЛОНОЧНАЯ ВЭЖХ ФИТ0Г0РМ0Н0В G.Schneider, Halle, GDR In plant hormone analysis as In any other trace analy sis the detection limit of the chromatographic system is playing an outstanding role. For HPLC the reduction of the column diameter and the cell volume of the detector was shown to improve the detecting sensitivity. Thus, micro column technology attracts increasing attention. He UBed a microcolumn chromatograph 0B4 "Milichrom" fitted with a 2 x 62 mm stainless steel column. Samples of gibberellins, gibberelliu glucosides, gibberellin gluco- syl esters, abscisic acid, abscisic acid 'jlucosyl ester etc, were isocratically run on silica RP 18 using mixtures of methanol and 0.1 % phosphoric acid. Besides retention times and column performance, the detection limits in both qualitative and quantitative ana lysis are discussed and compared with results obtained with "normal" HPLC. 092 P.2-18 ION-PAIR - HPLC OF XANTHURENIC ACID AND OTHER TRYPTOPHAN METABOLITES ИОН-ПАРНАЯ ВЭЖХ КСАНТУРЕНОВОЙ КИСЛОТЫ И ДРУГИХ МЕТАБОЛИТОВ ТРИПТОФАНА Th.Jira, Th.Beyrich, K.Reinhardt, U.Grimm, Greifswald, GDR The tryptophan metabolism of men can be disturbed through a kynureninase-defect, which has its origin in a genetic defect. This disturbance is dependent on vitamin В 6 and presented in a strong secretion of xanthurenic acid (XA), kynurenic acid and other metabolites in urine (described as xanthurenic acid - urea). Under application of ion-pair reversed-phase - HPLC a new method for the identification of XA in the presence of other metabolites in biological fluids is described improving the diagnosis of this metabolism-defect in this way. Detectable amounts: 12 pmol XA (2.5 ng/ 5 ,ul). 66 A suitable counter ion is the surface-active cetyltri- roethylammoniurabromide in 1/15 mol kaliumdihydrogenphos- phate-buffer with pH 3. Methanol is better than acetonitrile as organic modifier. In connection with examinations of further aromatic aminoacids and carbonacids we received results about the influence of various factors, as pH, concentration of pairing ion, type and concentration of buffer, type and concentration of organic modifier a.o, on the chromato graphic parameters of anallzed compounds.

09 3 P.2-19 ON-LINE SAMPLE CLEANING AND ENRICHMENT TECHNIQUE FOR CATECHOLAMINES IN MICROBORE COLUMN LC ПРИГОТОВЛЕНИЕ ПРОБ ДЛЯ АНАЛИЗА КАТЕХОЛАМИНОВ МИКРОКОЛОНОЧНОЙ ВЭЖХ У V у* * D.Kourilova, K.Slais, M.Krejci, Brno, CSSR Catecholamines from the sample are sorbed on alumina at pH about 8,5. They are then desorbed to the sampling loop (0,2 ml) of a six-port valve. The desorbing solution having pH about 2 contains a suitable counter ion. The counter ion in the eluent from alumina bed causes peak- focusing /1,2/ of the solute during the sampling on the analytical microbore column. The alumina (Boehringer, Mannheim, d «*» lOO um) sorp tion bed (2 mm long, 2 mm in diameter) is placed in a little plastic column (4 mm long) one end of which is adapted for setting injection syringe with the sample and, later, with the eluent. The other end is attached to the sampling valve. A six-way valve is attached to the column (150 mm long, I.D. - 0.7 mm) packed with Separon SIC 18 (Laboratory Instruments, Prague). An EMD 10 electrochemi cal detector (Laboratory Instruments, Prague) with a cell volume of 20 nl is used. Recoveries were measured on the alumina bed down to the concentrations on the order of 10"° mol/1 of adrenaline and noradrenaline, 1. M.Krejci, K.Slais, D.Kourilova, M.Vespalcova, J.Pharm. Biomed. Anal.,2 (1984) 197. 2. D.Koufilova, K.Slais, M.Krejci, Chromatographia (in press). 094 P.2-20 DETERMINATION OF THE OXIDATION DECOMPOSITION OF CH-105 ОПРЕДЕЛЕНИЕ ОКИСЛИТЕЛЬНОГО РАЗЛОЖЕНИЯ CH-105 A.Toth, E.Kotsis, I.Ratonyi, K.Kaloy, Budapest, Hungary CH-105 (1.6-dimethyl-4-oxo-l,6,7,8-tetrahydro-4H-pyrido-/l,2-0/pyrimidine-3-carbamide) is a newly developed original antiphlogistic and inhibitor of platelet aggrega tion drug of Chinoin Pharmaceutical Works. To study the speed of its oxidation is very important for evaluating the stability under storage.

67 It was determined the speed of oxidation at different pH-values, The oxidation products were separated on TLC plate and extracted. The structure of dissolved products was identified by MS. Knowing the structure of decomposi tion products a HPLC method was developed for the quanti tative determination of decomposition products and the intact compound as well.

095 P.2-21 CHROMATOGRAPHY OF GLYCOLIPIDS ХРОМАТОГРАФИЯ ГЛИК0ЛИПИД0В J.Pick, J.Vajda, N.Anh-Tuan, L.Leisztner, S.R.Hollan, Budapest, Hungary The glycolipids comprise minor components of the cell membranes and they are present in extremely small amounts on the outer leaflet of the plasma membranes. The two classes of glycosphingolipids in the focus of our inves tigations, the neutral and the acidic glycolipids, play very important biological roles. Of the many different biological functions attributed to these species we are interested in their antigeneity which is associated with the oligosaccharide moiety of the glycolipid molecules. Different variants of chromatography are commonly used techniques for the separation and Identification of lipids. Low pressure column chromatography is used for preparative work and prefractionation (e.g. florisil, Sephadex 6 and ion-exchange etc.). HPLC is applied to separation of glycolipids with UV detection on the basis of differences in fatty acid composition for non deriva- tlzed samples and in oligosaccharide composition with derivatization. Layer chromatographic methods are commonly used for analytical purposes with in situ staining. The OPLC technique has been introduced to lipid analy sis in our previous publications and in the present paper we report new methodological developments and summarize our experiences in the development and application of chromatographic methods in glycolipid analysis. 096 P.2-22 HPLC STUDY OF DITHIOCARBAMATES DEGRADATION IN NATURAL SAMPLES ВЭЖХ ДЕГРАДАЦИИ ДИТИОКАРБАМАТОВ В ПРИРОДНЫХ ИСТОЧНИКАХ v / E.Brandsteterova, J.Lehotay, J.Garaj, Bratislava, CSSR HPLC analysis of metal complexes of N-disubstituted_ dithiocarbamic acids confirmed that dithiocarbamates (DTC) are not stable and decompose very quickly. As DTC complexes are known to have toxicological and mutational effects, it was necessary to develop a rapid and accurate method for the separation, identification and determination of DTC fungicides and their degradation products. 68 The aim of this work was to use HPLC for the determina tion of very low concentrations of iron (III) dimethyldi- thiocarbamate, iron (Ill)dibutyldithiocarbamate and tetra ethylthiuram disulphide in some agricultural products and the soil. Optimal conditions for the simultaneous HPLC separation of DTCs and their possibile degradation products with minimum separation time and with maximum resolution have been recommended. Adsorption and reversed-phase chromatography were inves tigated. Results of HPLC studies were verified by mass spectrometry, The limits of detection for dithiocarbamate residues and their degradation products correspond with the crite ria of the FAO concerning methods for the determination of DTC in food. High performance liquid chromatography and mass spectro metry provide the possibility of determining of DTC resi due concentrations in natural samples. This paper deals with the identification and determina tion of DTCs and their degradation products in some agricul tural products grown on plastic materials with certain con tents of iron (III) dibutyldithiocarbamate as an antioxidant The stability of DTCs and conditions of their degrada tion were studied by HPLC and the identification of degra dation products was veryfled by elemental analysis, gas chromatography, mass spectrometry. This paper presents a suitable method for the rapid analytical control of trace concentrations of DTCs.

097 P. 2-23 MEASURING THE RELEASE OF DOPAMINE AND 3,4-DIHYDROXY- PHENYLACETIC ACID FROM ISOLATED STRIATAL PREPARATIONS USING HPLC-ECD ИЗМЕРЕНИЕ ОСВОБОЖДЕНИЯ ДОПАМИНА И ЗН-ДИОКСИФЕНИЛ- УКСУСНОЙ КИСЛОТЫ В ПРЕПАРАТАХ СТРИТИАЛА С ПОМОЩЬЮ ВЭЖХ С ПРИМЕНЕНИЕМ ДЕТЕКТОРА ПО ЭЛЕКТРОННОМУ ЗАХВАТУ L.Kerecsen, H.Kala'sz, J.Knoll, Budapest, Hungary The measurement of the release of endogenous dopamine and its main metabolite 3,4-dihydroxyphenylacetic acid serves direct information about the mechanism of action of drugs used in the therapy of Parkinson's disease. The effect of various MAO inhibitors including (-) diprenyl has been studied using HPLC-ECD method to determine the release of dopamine and DOPAC from isolated striatal preparations of the rat. Organ baths were collected and spiked with 40 ng of 3,4-dihydroxynorephedrine or with the same amount of 3,4-dihydroxybenzylamine as internal standard. After a prepurification step on alumina microcolumns samples were injected onto a HPLC system (Liquopump 312, injector fitted with 20 /il loop, 4 x 150 mm i.d. column packed with LiChrosorb RP-18 7 jam particle size, thin-layer cell electrochemical detector with paraffin oil based carbon paste working electrode). A pH 3 triethylamine-phosphate

69 buffer containing 18 mg/1 Na-EDTA, 150 mg/1 octanesulfonic acid sodium salt and 8 % acetonitrile proved to be optimal for the separation of the internal standard, dopamine and DOPAC. One run was accomplished within 9 minutes.

098 P.2-24 HPLC OF SUBSTANCE P AND ITS DERIVATIVES ВЭЖХ ВЕЩЕСТВА Р И ЕГО ПРОИЗВОДНЫХ U.Kertscher, H.Bienert, J.Stein, B.Mehlis, Berlin, GDR

The methionine residue in peptides is both susceptible to alkylation and spontaneous oxidation but results on the physico-chemical properties of these derivatives are relatively rare. Therefore, a chromatographic study of series of compounds related to this problem has been undertaken with the naturally occurring undecapeptide substance P (X • S) and some of its derivatives•

Arg-Pro-Lys-Pro-Gln-Gln-Fhe-Phe-Gly-Leu-NH-CH-CO-NH2

1 (CH2)-X-CH3 substance P (SP) Met - analogs + X - CH2, S, SO, S02, S -CH3 In addition, the corresponding acyl methionine amides were prepared for comparison. HPLC separation of these compounds was achieved on RP stationary phases using phosphate buffer + NaClO^/acetonitrile and phosphate buffer + alkylsulfate/methanol, both at acid pH, as mobile phases. The analogs were + eluted in the order CH2» S >S -CH3 > S02 > SO (k*) (first 3 system) and CHo> S -CH3> s >S0> SO2 (second system). ll For the Nle analog (X » CH2) a remarkably high retention time was observed which indicates strongly increased lipophilicity in comparison to SP (X - S). This was also predicted by the increment method /1,2/ and is supported by the measurement of the distribution coefficients. In the first system the sulfoniura derivative was eluted before SP but, surprisingly, exhibits higher retention times than the S-oxide and the sulfone. Because our data of the peptide analogs show a correla tion between the lipophilicity (calculated or measured) and the observed retention times on the RP phases, these results may be useful in general for the prediction of the HPLC retention and lipophxlicity of peptides modified at methionine residues. 1. J.-L.Fauchere, V.Pliska, Eur. J. Med.. Chem., 18, 369 (1983). 2. T.Sasagawa, T.Okuyama, D.C.Teller, J.Chromatogr., 240, 329 (1982).

70 099 P.2-25 ANALYTICAL CONTROL OF THE SYNTHESIS OF CYTOSINE ARABINOSIDE BY HPLC METHOD АНАЛИТИЧЕСКИЙ КОНТРОЛЬ СИНТЕЗА АРАБИНОЗИДА ЦИТОЗИНА МЕТОДОМ ВЭЖХ K.Butkiewicz, Warsaw, Poland

Cytosine arabinoside {4-amino-l-;8-I5-arabinofuranosyl- -2(lH)-pyrimidone = Ara-c), an important anticancer drug, was prepared in three-step synthesis from cytidine. Syn theses bf acetylsalicyloyl chloride and 2,2'-anhydro-l- -/-D-arabinofurynosylcytosine hydrochloride as well as hydrolysis of this last to Ara-C were controlled. The quality of the final product was analyzed on a 250 x 5 mm stainless column packed with LiChrosorb-NH2 (grain size 10/am). The mobile phase was a mixture of acetonitrile + water (75 + 25 by vol.) and a UV-detector was used. Following compounds were found and separated: cytidine 5,29, Ara-C 4.70, uracil arabinoside 3.2B, 2,2'-anhydro- -Ara-C 2.72, 2,2'-anhydro-10-acetyl-Ara-C 1.95, and one unidentified compound 1.55 min. Determination of the compounds separated was carried out by the external calibration method. 100 P.2-26 HPLC MONITORING THE GOSSYPOL CONTENT OF COTTONSEED FLOUR DURING MICROWAVE TREATMENT ВЭЖХ КОНТРОЛЬ ЗА СОДЕРЖАНИЕМ ГОССИПОЛА В МУКЕ ИЗ ХЛОПКОВЫХ СЕМЯН ПОСЛЕ МИКРОВОЛНОВОЙ ОБРАБОТКИ K.Ganzler, Budapest, Hungary Defatted cottonseed flour contains approximately 30 % protein and has a protein efficiency ratio (PER) similar to that of casein /1/. Most of the commercially available cottonseed contains a toxic terpenoid aldehyde, the gossypol. Because of the toxicity of gossypol, cottonseed flour has limited use either in human consumption or in animal feeding. Several time-, and energy-consuming processes have been reported, producing degossypolyzed cottonseed flour /2,3/. Our aim in this work was to find a new quick, and energy-saving method for minimizing the gossypol content in different cottonseed samples. Due to 15 minutes microwave irradiation the gossypol content decreased as much as 70-75 % of the total gossy pol. For the gossypol analysis a new extraction method has been developed reducing the extraction time to 2-4 minutes /4/.The analytical conditions developed for the monitoring were the following: The stationary phase was Dimesil С 18 (10 fi) packed in a 250 x 4,6 mm column. The mobile phase consisted of methanol-water-glacial acetic acid (80:13:7). The flow rate was 1,0 ml/min. Gossypol was detected at 254 nm, using an ultraviolet absorbance monitor. 71

i Due to the gossypol degradation caused by the microwave treatment different breakdown products of the gossypol could be observed in the chromatograms, Identification and examination of the toxic-, and antinutritive effects of these components are in progress. 1.H.H.Martinez, D.T.Hopkins: Cottonseed protein products. In: Protein Nutritional Quality of Foods and Feeds, ed. by Mendel Friedman., 1975, 355-373. 2. H.L.Vix et al.: Degossypolized cottonseed flour, J. Am. Oil Chem. Soc.,48 (1971) 611-615. 3.L.C.Berardi et al.: Cottonseed orotein isolates. Food Technol.,2_3 (1969) 1305-1312. 4.K.Ganzler et al.: A new method for extraction and determination of vicine and ccnvicine in fababeans by HPLC. In. Proceedings of the IV. American Easters Europian Chromatography Symposium (in press) ed. by H.Halasz.

101 P.2-51 THE SPECIFIC SORPTION OF ATHEROGENIC LIPOPROTEINS OUT OF BLOOD PLASMA СПЕЦИФИЧЕСКАЯ СОРБЦИЯ АТЕРОГЕННЫХ ЛИПОПРОТЕИНОВ ИЗ ПЛАЗМЫ КРОВИ I.P.Andrianova, Moscow, USSR The development of sorption methods of organism detoxi- cation. requires specific sorbents which selectively extract toxins or decrease the level of metabolites whose pathological accumulation leads to functional dicordered state. It is recognized at the present time that the increase of the concentration of low and very low density lipoproteins (LDL and VLDL) and the decrease of the concentration of high density lipoproteins promotes athero sclerosis. In order to correct the changed level of lipo proteins the specific and nonspecific sorbents on the basis of silica carries, carbon fibres and hydrogels have been obtained. The correlation was determined between the pore sizes of macroporous sorbents and the selectivity to the absorption of lipoproteins of different classes. The modification methods of solid carriers which allow to carry out immobilization of ligands on their surface were elaborated. It has been demonstrated, that the immobiliza tion of ligands to cholesterol increases the binding of atherogenic lipoproteins and reduces the nonspecific protein sorption on the surface of the carrier. The action of the size of a charge and the presence of hydrophobic groups and polyanions on lipoproteins was studied. The effectiveness of the LDL and VLDL removal depends on the initial cholesterol level in the blood plasma. The haemo- compatibility of the obtained sorbents was estimated.

72 102 P.2-52 SEPARATION AND DETERMINATION OF THE SPECIES PATTERN 0Г PHOSPHATIDYLETHANOLAMINE BY HPLC АНАЛИЗ И ОПРЕДЕЛЕНИЕ ПУТЕЙ МЕТАБОЛИЗМА ФОСФАТИДИЛЭТАНОЛАМИНА С ПОМОЩЬЮ ВЭЖХ H.Rabe, G.Reichmann, B.Riistow, Berlin, GDR Intramicrosomal diacylglycerols are the immediate pre cursors in the biosynthesis of glycerolipids like phospha tidylcholine, phosphatidylethanolamine and triacylgly- cerols. A precondition explaining the differences of the species pattern of these three lipid classes in relation to that of their diacylglycerol substrate, is a method by which the species pattern of these glycerolipids can exactly be analysed. We have already described a method for analysing the species pattern of phosphatidylcholine and diacylglycerol. The aim of this work is io show a method for the deter mination of the species pattern of diacyl-phosphatidylethanolamine. The difficulty of analysing the phosphatidylethanolamine pattern is its content of plasmalogens which have to be separa J in a primar chromatographic step in our experimental approach. The method is based on the conversion of phospholipase С generated diacylglycerols to fluorescent naphthyl-ure- thanes with cC-naphthylisocyanate. After separation of the plasmalogens the diacyl-species were separated on a rever se phase column (RP 18, 5 yu). 103 P.2-53 PHYTOANALYTICAL APPLICATION OF HPLC ON SOME BIOLOGICALLY АСТЦГЕ PLANT CONSTITUENTS ФИТОАНАЛИТИЧЕСКИЕ ПРИМЕНЕНИЯ ВЭЖХ НЕКОТОРЫХ БИОЛОГИЧЕСКИ АКТИВНЫХ ВЕЩЕСТВ РАСТИТЕЛЬНОГО ПРОИСХОЖДЕНИЯ E.Csapo, G.Verza'r-rPetri, Budapest, Hungary HPLC method has an extensive importance in the analysis of medicinal plants containing various groups of consti tuents . This work reports the qualitative and quantitative exar •.. tion of characteristic and anly partially known cc. : ,.winds in three plants, frequently used in folk medicine. 1. In Corydalis cava the following alkaloids were separa ted, quantitatively determined and identified: bulbo- capnine, tetrahydropalmatine, corydaline etc. 2. The qualitative and quantitative composition of the characteristic iridoid 'type constituents (aucubine etc.) in different Veronica species were studied. 3. The third problem involved in this work was the sepa ration of the multicomponent flavonoid mixture in Lysimachia nummularia, which resulted in their identi fication. 73 Foi* HPLC separation Liquochrom 307 instrument and RP-18 250 x -.',6 mm i.d. column was used. In e.xch group of constituents (alkaloids, iridoid- and flavonold glycosides) several eluents were used systemati cally and the optimal ones selected for solving the sepa ration problem. The separated compounds were identified by comparison with authentic samples using standard addition (HPLC, TLC) and/or spectroscopic techniques.

Ю4 P.2-54 GPC CHARACTERIZATION OF EPOXY RESINS BASED ON POLYETHYLENE GLYCOLS ИЗУЧЕНИЕ ЭПОКСИДНЫХ СМОЛ НА ОСНОВЕ ПОЛИЭТИЛЕНГЛИКОЛЕЙ МЕТОДОМ ПК T.Spychaj, A.Bledzki, Szczecin, Poland Two aliphatic epoxy resins based on diethylene and triethylene glycols were investigated. The resin and frac^ tions from their molecular distillation were analysed by means of conventional and high performance GPC. The values of epoxy. numbers of the fractions were compared too. The following resins were studied: -industrial resin EDG-2 based on diethylene glycol, epoxy numberi 0.559 g-equiv./lOO g (Chemical Works, Sarzyna, Poland)i - pilot resin RG-207 based on triethylene glycol, epoxy number! 0.578 g-equiv./lOO g (Institute of Industrial Chemistry, Warsaw, Poland). Both resins were separated into two fractions in each case: a distillate (EDG-2md, RG-207md, respectively) and a residue (EDG-2rd,• RG-207rd, respectively). GPC studies of epoxy resins and fractions were perfor med by use of two GPC systems: a conventional one - filled with Sephadex LH-20 in methanol (two columns 2004-120 cm, ф 0.9 cm) and high performance one filled with Ultrastyra- gel 100 A (two columns), Ultrastyragel 500 A (two columns) and Styragel 10* A (one column) in THF (5 x 30 cm, 4 0.8cm). Some conclusions may be drawn from the chromatographic analysis: - EDG-2 resin is broader in molecular mass distribution than RG-207; - The contents of the lowest diglycidyl ether oligomer (n » 0) are higher in the case of triethylene glycol resin (И « 262) than in the diethylene glycol resin (И - 218); - The fraction RG-207 md contains mainly oligomer n « О and trace amounts of products of incomplete epoxidation and oligomer n « 1; - The fraction EDG-2md contains four main components in appreciable amounts. There are: the chlorohydrin of mono-

74 glycidyl ether of diethylene glycol and diglycidyl ether oligomerE n « 0-2f The better results of GPC separation of RG-207 were obtained with the Sephadex LH-20 column system than with Ultrastyragel/styragel one. The EDG-2 resin was better separated with Ultrastyragel/styragel columns.

105 P.2-55 SEPARATION OF THE at AND fi ISOMERS OF MONOFUNCTIONAL NAPHTHALENE DERIVATIVES BY MEANS OF GAS CHROMATOGRAPHY РАЗДЕЛЕНИЕ <*И^ -ИЗОМЕРОВ МОНОФУНКЦИОНАЛЬНЫХ ПРОИЗВОДНЫХ НАФТАЛИНА МЕТОДОМ ГАЗОВОЙ ХРОМАТОГРАФИИ M.Sajewicz, J.Sliwiok, J.Rzepa, Katowice, Poland The following aL and fi isomeric monofunctional naphtha lene derivatives were investigated: aC and A methylnaph- thalene, ei and fi naphthyl aldehyde, el and A naphthol, ot and fi naphthylamine , , methyl and ethyl esters of «<• and fi naphthoic acid, and A acetylnaphthalene. Separations were executed by means of gas chromatogra phy, applying.various stationary phases, column temperatu res and carrier gas flow rates. On the basis of the obtained separations a dependence was established between the chromatographic behavior of the investigated isomers and their structure. A scheme was suggested, enabling a good qualitative interpretation of the above mentioned dependence.

106 P.2-56 ANALYSIS OF SOME ADDITIVES USED IN PLASTIC INDUSTRY BY HIGH PRESSURE LIQUID CHROMATOGRAPHY (HPLC) АНАЛИЗ НЕКОТОРЫХ ПРИСАДОК, ИСПОЛЬЗУЕМЫХ В ПРОИЗВОДСТВЕ ПОЛИМЕРОВ, МЕТОДОМ ВЭЖХ M.Krajmer, Veszprem, Hungary F.Barla, Gyor, Hungary In plastics industries a great number of additives are applied, which form the final characteristics of the poly mers. So the quality of the plastic is affected not only by the type, but in particular by the quality of the added materials, too. The quality testing of different additives is important from this point of view. In this paper HPLC separation methods are introduced, which are found suitable for quality control of different additives of PVC-processing. During development of the testing methods in several cases we had to employ spectrophotometrical methods for

75 getting more information about the chemical structures of the additives. Testing methods of some additives with different chemi cal structure are introduced for plasticizers (affecting the rigidity, cold resistance, the makeup characteristics of polymere), polymer plasticizers 'modifying the proces sing viscosity characteristics) and flame resistance increasing additives. 107 P.4-39 COMPUTER SIMULATION OF PROTEI?' ADSORPTION AND CHROMATOGRAPHY МОДЕЛИРОВАНИЕ НА ЭВМ АДСОРБЦИИ И ХРОМАТОГРАФИИ БЕЛКОВ A.A.Gorbunov, A.E.Lukyanov, V.A.Pasechnik, Leningrad, USSR Adsorption behaviour of protein molecules in chromato graphy has been theoretically studied. A silt of width D has served as a model of the adsorbent pore, a spherical particle with radius RQ has modelled the protein molecule. Adsorption-active sites were supposed to be located in a certain way on the surfaces of adsorbent and protein mole cule. Analytical expressions for the distribution coeffi cient Kd have been obtained for a number of cases with regular arrangement of active sites. In general, Monte- Carlo simulation procedure has been used to calculate the dependences of Xd on D, Ro, adsorption energy £ , on the surface density of active sites and on the mode of their arrangement.- The principal result is the possibility of chromatogra phic separation of proteins according to their surface hydrophobicity. Conditions required for the good separa tion have been analyzed. As it is shown, one can expect the fine separation using adsorbents with the hydrophobic patches of optimal size. The influence of an active site arrangement on the protein molecular surface upon the chromatographic retention has been specifically studied. The proteins differing only in active site disposition proved to have different Kg values. Therefore such pro teins can be chromatographically separated. Some experimen tal features of protein chromatography can be explained by means of proposed theoretical model. 108 P.1-58 HPLC AND TLC INVESTIGATIONS ON RESIDUES AND METABOLIZATION OF THE GROWTH PROMOTING QUINOXALINE-DI-N-OXIDE DERIVATIVE OLAQUINDOX ИЗУЧЕНИЕ С ПОМОЩЬЮ ВЭЖХ И ТСХ ПРОМОТОРА РОСТА ЖИВОТНЫХ - ХИНОКСАЛИН-ДИ-Н-ОКСИДА И ЕГО МЕТАБОЛИТОВ H.Much, w.Netsch, P.Dietrich, H.-J. Niclas, Berlin, GDR HPLC and TLC procedures were developed to determine residues of Olaquindox and their metabolites in slaughter products of farm animals in order to exclude any consumers 76 risk resulting from feeding with such growth promoting (•rgotropic) substances. The TLC method subsequent to reduction with SnCl2 (detection limit 20 ppb) gives the overall amount of original residue and metabolized compounds. For the separation of these substances, the HPLC pro cedures were developed using gradient elution of lyophi- lized organ material, identification by on-line regictra- tion and confirmation by selective detection. The metabo lites were identified-by means of on-line UV-peak scanning and by chromatographic data. Heat, liver, kidney and fat of pigs taken from feeding trials with Olaquindox were analyzed to establish a withdrawal time before slaughte ring. No detectable residues were found in the same slaugh ter products from calves after application of effective doses with milk replacer feed. A preliminary metabolization scheme is given on basis of the HPLC results. 109 P.1-59 RAPID RECOVERY OF DABTC - MODIFIED PROTEINS FROM GEL SLICES FOLLOWED BY N-TERMINAL SEQUENCE ANALYSIS СКОРОСТНОЕ ВОССТАНОВЛЕНИЙ ДАБТК - МОДИФИЦИРОВАННЫХ БЕЛКОВ ПРИ N-ТЕРМИНАЛЬНОМ СТУПЕНЧАТОМ АНАЛИЗЕ R.Kraft, Berlln-Buch, GDR То prove the uniformity of single protein bands separa ted on polyacrylamide gels the N-terminal amlnoacld analy sis is a reliable method. He propose a rapid and simple procedure especially suitable for not too complex protein mixtures. The method involves the following steps: 1. Application of the powerful resolving SDS-PAGE system for rod gels as described by Weber and Osborn using DABTC-(dimethylaminoazobenzenethiocarbamyl)-prelabelled peptides. The migration of the yellow coloured peptides can be followed up visually by the DABTC-aodified samples. 2. Slicing of coloured gel bands of interest. Transferring of the DABTC-modified proteins fro* the rod gel slices into a minor volume of buffer (100 - 150/ll) by rever sed electrophoresis (duration: 15 - ЗО aln} using dis continuous conductivity gradient as has been suggested by Otto and Snejdarkova. 3. Isolation of the peptides suitable for N-terainal sequence analysis after acetone-precipitation and removal from residual salts from the precipitate by distribution In a two-phase water - pyridine (1:1) ternary system. The proposed procedure is demonstrated on six peptides (range of molecular weight 6.7 to 36 kDa) recovered from one to two reeluted gel slices followed by manual N-termi nal sequence analysis using the DABITC/PITC-technique as described by Chang. Approximately six to eight steps of the' N-terminal part of the peptides were obtained.

77 110 P.3-01 BACKFLUSHING METHOD FOR TESTING HPLC COLUMNS МЕТОД ОБРАТНОГО ИМПУЛЬСА ДЛЯ ТЕСТИРОВАНИЯ КОЛОНН В ВЭЖХ V Т.Маоко, D.Berek, Bratislava, CSSR Recently, it was observed /1,2/ that pressure in the system containing a sorbent and a nixed liquid influences the extent of preferential sorption i.e. the composition of liquid layer on the sorbent surface. This means that pressure variations in a HPLC system with mixed eluent cause the local change in the effluent composition detect able by a non specific detector. The volume of ef f .uent with a changed composition that we call "eigenzone", corresponds to the total volume of liquid within the column, while its size i.e. the extent of compositional change of the effluent is a function of nature of both sorbent and liquid mixture and depends also on the extent of pressure variation. The shape of the eigenzone, however, reflects the course of pressure changes within the column and, consequently, also the sorbent bed geometry which can be applied in the column diagnostics /3/. The sudden changes in pressure, ДР, within HPLC columns can be performed in various ways /2/, the most convenient method is the reversing the effluent flow through the column ("backflushing"). In the present study we have tested the above method by investigating numerous columns packed with different sorbents. Both the UP values and the eluent compositions for sorbents of different nature were optimized. The data obtained were correlated with the conventional values like heiqht equivalent to the theoretical plate, peak symmetri- city, column permeability and column stability. The latter , parameter was obtained by a multiple eluent changes and pressure strokes. It was proved that the backflushing diagnostics yield the valuable information on the HPLC column stability. Moreover, the method can be used for estimating the column dead volume and the density of the sorbent bed and presents an appropriate supplement to the existing column test pro cedures. 1. D.Berek, M.Chalanyova, T.Macko, J.Chromatogr., 286 (1984) ,185. , 2. M.Chalanyova, T.Macko, J.Kandrac, D.Berek, Chromatogra- phia,18 (1984), in press; 3. T.Macko, D.Berek, New diagnostic method for HPLC column packing procedures, 15th Int. Symp. on Chromato graphy, Nurnberg, FRG, October 1984, Abstracts, pp.216.

78 111 P.3-02 "ARMSORB" COLUMNS FOR HIGH-EFFICIENCY LIQUID CHROMATOGRAPHY. PACKING AND PROPERTIES ПЛОЙКИ "АРМСОРБ" ДДЯ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ. УПАКОВКА И СВОЙСТВА A.M.Arutunyan, S.G.Chaylyan, V.K.Aliev, A.A.Isackov, A.A.Karapetyan, S.G.Babayan, Yerevan, USSR Grinding and fractioning processes for "ARMSORB" type eorbents has been studied,including the jet-grinding by MC-40 mill, followed by separation on"Alpina MZR lOO" type classifier and sedimentation from acqueous suspensions carried out by УФС 01 plant. HPLC coluan fill process by suspensions has been deve loped, using low-viscosity liquids and viscous packing method. Fill process optimization has been carried out by means of experimental planning method, in which the full process and roughness of column inner surface in case "ARMSORB-Sil 10" (10 jam) silica gel have been investigated. A set of HPLC columns has been developed, including the columns 150 x 4 mm and 250 x 4 mm, filled by sorbents based on silica gels with average pore diameters 6, 10 and 30 nm, as well as by sorbents modified by Cg, Сде and CN phases with particle size 5 and 10 yum. Chromatographic characteristics of the filled columns have been investi gated. 112 P.4-01 PRECHROMATOGRAPHIC CHEMICAL DERIVATIZATION OF ENDOGENOUS AND EXOGENOUS COMPOUNDS OF PLASMA IN HPLC ANALYSIS ПРЕДКОЛОНОЧНАЯ МОДИФИКАЦИЯ ПРИ ИССЛЕДОВАНИИ ЭКЗОГЕННЫХ И ЭНДОГЕННЫХ ВЕЩЕСТВ ПЛАЗМЫ КРОВИ МЕТОДОМ ВЭЖХ S.P.Katrukha, V.G.Kukes, Moscow, USSR Within the last years, the HPLC method was widely used in medicine, for the analysis of endogenous and exogenous compounds of biological fluids and tissues. The basic difficulties in the analysis of these compounds are asso ciated with the hydrophilic character of most drugs which are poorly extracted from plasma by organic solvents. Due to the absence of suitable detectors it is more preferable to employ the derivatization of the analysed compounds before their injection on the column. In our studies the pre-column derivatization of carbonyl and amino groups were used. For the modification of the carbonyl group, 2,4-dinitrophenylhydrazine which forms with keto-compounds strongly dyed hydrazones with absorp tion at 360 nm in the acidic medium was chosen. The 2,4- DNP-hydrazons of the ketoacids were extracted from plasma by benzene and the extract was injected on the column with silica gel. 2,4-DNP-hydrazones were eluted from the column by chloroform with 2,1 t formic acid. The developed method allows the determination of Ю"8 moles/ml of pyruvate and

79 acetoacetate in only 0,1 m3 of plasma without its prelimi nary deproteinization. All analysis were carried out within 30 min. With small changes, the method can be used for analysis of acetone and «t-ketoglut^ric acid. For the selective modification of the amino compound propranolol, used in the treatment of hypertension, dansyl chloride at basic pH was applied. N-DNS-propranolol was also extracted from the reaction medium with benzene and the extract was chromatographed on the silica column, using the mixture of acetone and chloroform as a mobile phase. From 1 ng to 1 g of DNS-propranclol was determined by fluorometric detector.

113 P.4-02 AN APPLICATION OF REVERSED-PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH OPTIMIZED CAPACITY FACTOR FOP. CORTICOID HORMONES AND CATECHOLAMINES ANALYSIS ПРИМЕНЕНИЕ МЕТОДА 0БРАЩЕНН0-ФА30В0Й ЖИДКОСТНОЙ ХРОМАТОГРАФИИ ВЫСОКОГО РАЗРЕШЕНИЯ С РЕГУЛИРУЕМЫМ КОЭФФИЦИЕНТОМ ЕМКОСТИ К АНАЛИЗУ КАТЕХОЛАМИНОВ И КОРТИКОСТЕРОИДОВ A.S.Kanev, G.S.Katychin, A.M.Vorontsov, Leningrad, USSR A method for controlled change of capacity factor by mixing of two buffers for optimal separation in reversed- phase high performance liquid chromatography is realized. The programmed selection of mobile phase modifiers not only determines the separation, but also affects resolu tion and selectivity. These effects are considered and discussed. The identification of catecholamines as well as corti- coids is processed by examination of retention in compa rison with retention of specially added labels. The quantitative analysis is performed by means of double internal standard method to ensure the results in condi tions of mobile phase programming. The chromatographic experiments were carried out using RP-18 with a particle size of Ю jam, current eluents (methanol-water, acetonitrile-water) and buffers, (citrate, phosphate, glycine, ace tate) , subsequent ion-pairing reagents and modifiers. There was used the gradient chromatograph "Okhta-A-101" with spectrophotometric and electrochemical detectors, UV-monitor, two binary gradient modules, data control system. The probes of 5 and 20/jl were injected'by a valve connected with titanium alloy columns packed using a pump and packing device of the chromatograph.

80 99m_ r',UJ SEPARATION OF MYOCARDAFEINE ""Тс-DMPE COMPLEXES BY HPLC РАЗДЕЛЕНИЕ КОМПЛЕКСОВ МИОКАРДАФФИНА С ПОМОЩЬЮ ВЭЖХ K.H.Heise, Dresden, GDR Lipophilic complexes of the short lived isotope ""тс (t]/2 = 6.1 h) and Bis(dimethylphosphino)ethane (DMPE), formed by reduction of TcO^" by DMPE, are among the new reagents for myocardial imaging in nuclear medicine. Best clinical results are given by the technetium (I) and tech netium (III) complexes, whereas technetium (II) and tech netium (V) complexes are inefficient. Consequently the characterization of formed products is required in any case before application. We have analysed the "raTc complexes by HPLC with a BECKMAN chromatograph, Model 342, in connection with an effective self-made flow-trough scintillation counter for radiodetection. The chromatographic system is formed by a RP-8 column (SEPARON SIX) from Laboratorni Pristroje Praha and a solvent of buffered methanole, pH 7...S The retention of complexes is strong-.- depending on the buffer concentration in the solvent. The complete separation of all complexes in substoicho- metric concentration has been obtained with methanole, buffered by 0,05 m KH2P04 and 0,01 m triethylamine in water. Through blocking the free siloxyl groups in the silica matrix by aliphatic amines we exclude trapping of micro- components at the sorbent, 115 P.4-04 ANALYSIS OF BIOGENIC AMINES AND THEIR METABOLITES BY MICROBORE HPLC WITH ELECTROCHEMICAL DETECTION БИОГЕННЫЕ АМИНЫ И ИХ МЕТАБОЛИТЫ: ОПРЕДЕЛЕНИЕ МИКРОКОЛОНОЧНОИ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИЕЙ С ЭЛЕКТРОХИМИЧЕСКИМ ДЕТЕКТИРОВАНИЕМ A.P.Solomonov, E.G.Larsky, E.L.Styskin, Moscow, USSR Evaluation state of aminoergic systems of brain in different patological states of organism have been of great importance for nevrology. We used microbore HPLC with electrochemical detection for determination of 10 substan ces in blood plasma and cerebrospinal fluid (CSF). Columns were 1 x 400 or 2 x 20C mm slurry packed with Nucleosil 5 C18. Mobile phase consisted of 0,05 M phosphate-cytrate buffer, 1 mM EDTA, sodium octyl sulphate and methanol. Concentrations of 2 last comeonents were changed for different groups of substances. Catecholamines were separated from 1 ml plasma by alumina adsorption with additional extraction by 0,1 M HC104 5-Hydroxyindoles were analysed from 0,5 ml plasma or CSF after protein removal by 5 M НСЮ4. Preliminary separation of acidic and neutral

6. Зак. 872 8) catecholamines metabolites were made on clean-up micro columns with ЭО mkm C18 silica. Recovery of standards added to plasma was 51-65 % for catecholamines, 61-72 % for 5-hydroxyindoles and 81-94 % for acidic metabolites. Described method is very simple, economical and sensitive one. By determination the number of substances it was possible to apply results for analy sis of activity aminoergenic systems of brain for diag nostic and control for medical treatment in nevrology and psychiatry.

116 P.4-05 ANALYSIS OF SOME METABOLITES OF THE FUNGUS OUOEMANSIELLA MUCIDA BY GAS CHROMATOGRAPHY АНАЛИЗ НЕКОТОРЫХ МЕТАБОЛИТОВ ГРИБА "OUDEMANSIELLA MUCIDA" МЕТОДОМ ГАЗОВОЙ ХРОМАТОГРАФИИ M.Wurst, M.Jurkova, I.Zemanova, V.Musilek, Prague, CSSR

GC methods for the separation and determination of some groups of metabolites, e.g. mixtures of lower fatty acids (I), higher fatty acids (II), sterols (III), soluble neutral sugars (IV) and aromatic carboxylic acids (V), in extracts of the mycelium, cultivation medium and fruiting bodied were worked out. Mixtures (II) were analyzed as methyl derivatives, mixtures (III - v) as trimethylsilyl derivatives (TMS) The mixtures were separated mostly on silicone statio nary phases (GE SE-52, GE XE-60, OV-17, MPHT) of different polar properties under isotheraic conditions (200°C, I, II; 250°C, III; 280°C, IV; 160°C, V) and using also programmed temperature (160 - 200°C at 2°C/min, II; ISO - 280°C at 3°C/min, IV; 120 - 260°C at 2°C/min, V).

Identity of individual components in the mixtures was verified chromatographically and MS and NMR spectra of some compounds were measured. Compounds in the mixtures were determined using methods of internal normalization and internal standard. Relative standard deviation of the measurement was verified with the mixture of aromatic metabolites (V) and did not exceed 2 %. The elaborated methods were used to determine mixtures of metabolites in the study of the metabolism of lipids, sugars and aromatic compounds produced by the fungus Oudemansiella mucida as related with biosynthesis of the antifungal antibiotic mucidin.

82 117 P.4-09 SOME METHODICAL PRINCIPLES INCREASING THE QUANTITY OF INFORMATION AND THE RELIABILITY OF THE RESULTS С А CHROMATOGRAPHIC ANALYSIS OF SEVERAL BIOLOGICALLY ACTIVE SUBSTANCES РАЗРАБОТКА МЕТОДИЧЕСКИХ ПРИНЦИПОВ ПОВЫШЕНИЯ ИНФОРМАТИВНОСТИ И ДОСТОВЕРНОСТИ РЕЗУЛЬТАТОВ ХРОМАТОГРАФИЧЕСКОГО АНАЛИЗА РЯДА БИОЛОГИЧЕСКИ АКТИВНЫХ СОЕДИНЕНИЙ V.I.Trubnikov, V.N.Khokhlov, S.I.Niculitcheva, Yu.M.Sapozhnikov, L.P.Tikhomirova, I.A.Kotova, O.G.Vintina, V.B.Yudovitch, Moscow, USSR Different methodical ways improving the quality of the instrumental chromatographic analysis of phenolcarboxylic acids, amino acids, low-molecular weight peptides and bio logically active derivatives of cyclopentanperhydrophenan- threne were suggested and experimentally tested. To secure a high reproducibility of the results obtained by TLC with densitometric detection (TLC-D) and HPLC methods solvent systems consisting of azeotropic and isobaric components or mixtures consisting of components with adjacent boiling points are advisable. To eliminate the effect of "false" peaks in GC analyses of amino acids and steroids an injector without septum was used. To increase the quantity of information obtained analy sing phenolcarboxylic acids in the wines and propolis it is advisable to use ultrafiltration, TLC-D and HPLC combi ned, while when analysing the synthetic hormones the TLC-D, HPLC and GC combined are recommended.

118 P 4—12 PARTITION CHROMATOGRAPHY OF PHARMACEUTICAL-PRODUCTS ON FORMAMID-SATURATED SILICA GEL - COLUMNS РАСПРЕДЕЛИТЕЛЬНАЯ ХРОМАТОГРАФИЯ ФАРМАЦЕВТИЧЕСКИХ ПРЕПАРАТОВ НА КОЛОНКАХ С СИЛИКАГЕЛЕМ, НАСЫЩЕННЫМ - ФОРМАМИДОМ H.Potter, M.Hulm, Dresden, GDR HPLC of pharmaceutical-products has received consider able attention for the purty of this compounds. Most modern techniques use some form of high-performan ce liquid chromatography (normal-phase-chromatography and reversed-phase chromatography). This paper describes the use of a formamide-saturated silica gel - column (Lichrosorb Si60 Merck; 5 jam) in the partition chromatography mode with a formamide-saturated quaternary solvent system, optimized for maximum resolution.

83 119 P.4-13 THE CORRELATION BETWEEN THE MOLECULAR STRUCTURE OF CARDIAC GLYCOSIDES, STEROID HORMONES AND CARBOHYDRATES AND THEIR RETENTION IN HPLC СВЯЗЬ СТРУКТУРЫ МОЛЕКУЛ СЕРДЕЧНЫХ ГЛИКОЗИДОВ, СТЕРОИДНЫХ ГОРМОНОВ И УГЛЕВОДОВ С ИХ УДЕРЖИВАНИЕМ В ВЭЖХ V.Ya.Davydov, Moscow, USSR The molecules of cardiac glycosides include steroid and carbohydrate parts. So it is possible to assume that the retention of these compounds is determined by the contributions of glycone and aglycone in accordance with their structure. But the cardiac glycosides molecules as well as other molecules can be thought as consisting of a number of functional groups which bring their own ave rage contribution into retention. To evaluate these cont ributions it is necessary to solve the system of linear equations by the least squares method for a number of car diac glycosides and steroid hormones, the retention volumes of which were determined at standard conditions. Experimental values of the logarithm of retention volumes are compared with calculated values In Vr «£ n^a^,where

a± = In (Уп)А ie the contribution into the retention of i-group and n^ is the number of functional groups in a molecule. Such comparison shows that the values are in a quite good agreement and the method can be used to evaluate the retention of components under study. The correlation of biological activity of cardiac glycosides and the logarithm of retention volumes was expressed by a linear equation, the coefficients of which have been determined. This equation shows that biological activity is related with hydrophilic-hydrophobic proper ties of cardiac glycosides and consequently with the transport of these compounds to receptor, 120 P.4-15 ISOLATION OF CLINICALLY IMPORTANT SUBSTANCES FROM BIOLOGICAL MATERIAL ON SORBENTS AND MICROCOLUMNS OF CZECHOSLOVAK ORIGIN ВЫДЕЛЕНИЕ КЛИНИЧЕСКИ ВАЖНЫХ СОЕДИНЕНИЙ ИЗ БИОЛОГИЧЕСКИХ МАТЕРИАЛОВ НА СОРБЕНТАХ ЧЕХОСЛОВАЦКОГО ПРОИЗВОДСТВА P.Bozek, K.Sebefcova, D.Berek, I.Novak, J.Zemek, Bratislava, <5sSR Prior to their HPLC analysis, the biological materials have to be deproteined either by extraction or by sorp tion on a tailored sorbent followed by the selective elution. Very effective purification of biological samples presents the liquid-solid phase extraction: material to be extracted is sorbed on an appropriate sorbent such as e.g. Kieselguhr, bare or modified silica gel etc. and subsequently eluted with organic solvents. Sorbents are

84 packed into the disposable devices of various construc tions . In the present study, we have tested the preseparation and preconcentration microcolumns SEPARCOL, available from Centre of Chemical Research, Slovak Academy of Sciences, CS-842 38 Bratislava, Czechoslovakia, that were packed with various sorbents and the results vere compared with the data obtained with commercial cartridges. For example, purine derivatives (theophylline, theobro mine and caffeine) from serum were effectively extracted on the packing based on the microcrystalline cellulose. On the other hand, porphyrins were purified and preconcent- rated on SEPABCOL packed with C^g alkylated silica with the same efficiency as with SEP-PAK C^g cartridges (Waters Ass., USA). Selective preseparation of 5-hydroxyindolace- tic acid and serotonin from urine was achieved with Lichroprep C^g and Amberlite CG II, resp., as a packing material. Finally, estriol from urine was sorbed on the porous carbon sorbent with high surface area. The inter fering substances were washed out and, subsequently, estriol was eluted with a mixture of methanol and chloro form. Selectivity of such preseparation was better than in case of C18 alkylated silica gel. 121 P.4-16 FLUOROGENIC LABELLING OF COMPOUNDS WITH 8-BROMOMETHYL- BENZ [dJPYRIDO [l,2-a)PYRIMIDIN-6-ON (BrBPP) ПОЛУЧЕНИЕ ФЛУОРЕСЦЕНТНО МЕЧЕНЫХ СОЕДИНЕНИЙ С ПОМОЩЬЮ 8-БРОММЕТИЛ-БЕНЗ 1<ШИРИД0 Ц,2-а1 ПИРИМИДИН-6-0НА (ВгВРР) R.Wintersteiger, G.Giibitz, A.Hartinger, O.S.Wolfbeis, Graz, Austria Fluorescence derivatization in combination with DC or HPLC is a very sensitive method for the quantitation of biological and pharmaceutical interesting compounds. This paper reports on the possibility of the fluores cence labelling with 8-bromomethyl-benz[d]pyrido[l,2-a]- pyrimidin-6-on. The synthesis of this new reagent results from 2-bromopyridine and 5-methylanthranilic acid and subsequently bromination with N-bromosuccinimide. BrBPP shows high fluorescence intensity and high reactivity. Its suitability as a derivatization reagent was tested with substances of different chemical classes. Screening experiments with about 100 compounds were carried out using 50 Jul dioxane as a reaction medium. Monocarboxylic acids, dicarboxylic acids, barbiturates, amines, sulfonami des, phenoles and some alcohols from highly fluoresceing products. The method was used both in combination with TLC and with HPLC. For TLC development methanol-ethyl acetate (60 + 40 or 75 + 15) as an eluent is well suited. The derivatization can also be performed directly on the thin layer plates by spotting the sample spot with reagent. By dipping the plate in a mixture of polyethylene glycol in

85 chloroform an enhancement of fluorescence intensity and stability may be obtained. The HPLC system consists of a RP-8 column and methanol- water (80 + 20) as an eluent. The fluorescence detection was carried out at 365 ran excitation and 435 nm emission. In all cases the calibration graphs are linear over a range of more than two orders of magnitude. The value for the relative standard deviation of the method was less than 3 %. Detection limits are in the picogram range. The applicability of the method is demonstrated by the separa tion and determination of numerous compounds. 122 P.4-19 OPTIMIZATION OF SEPARATION OF BASIC DRUGS ON RP-HPLC ОПТИМИЗАЦИЯ РАЗДЕЛЕНИЯ ЛЕКАРСТВ ОСНОВНОЙ ПРИРОДЫ С ПОМОЩЬЮ 0БРАЩЕНН0-ФА30В0Й ВЭЖХ B.Gober, M.Wolff, D.Kersten, Berlin, GDR As a reason for the irregular retention behaviour of basic drugs in reversed-phase liquid chromatography the presence of residual unreacted silanol groups at the sur face of alkylsilica bonded phases is discussed. A retention of basic drugs without interference by silanophilic effects - so that regular retention beha viour is attained - is an important supposition for inves tigation and selection of relevant parameters for optimi zing chromatographic separations. The silanophilic inte ractions of a basic drug (i.e. localanesthetics, analge tics, proteins and other drugs) can almost completely be suppressed by a compound with stronger silanophilic inte ractions; thus a regular retention behaviour due to sila nol masking can be effected independent of the pH value by use of mobile phases containing alkyltrimethylammonium compounds. Mathematical methods of optimization of such chromatographic separations are discussed. 123 P.4-20 HPLC DETERMINATION OF QUINIDINE AND /3S/-3-HYDROXYQUINIDINE щ SERUM AND URINE AFTER ADMINISTRATION OF RETARD PREPARATIONS ОПРЕДЕЛЕНИЕ ХИНИДИНА И /3s/-3-OKCHXHHHJMHA В СЫВОРОТКЕ И МОЧЕ С ПОМОЩЬЮ ВЭЖХ J.Klima, J.Grafnetterova, J.Sedivy, Prague, CSSR Quinidiite is widely used in the treatment of cardiac arrhytmlas. Monitoring of its levels is important as the therapeutic and toxic effects are related to serum quinidine concentration. Biological half-life of quinidine is short and therefore preparations with prolonged absorption have been introduced to permit longer interdose intervals. Two retard peroral preparations: Chinidin - retard (Spofa, CSSR) and Oftochinidin" retard (Boehringer, FR6) were ad ministered i.i a single dose of lOOO tag (4 table tee) to 6 healthy volunteers in a crossover study. The concentra tions of quinidine and /3S/-3-hydroxyquinidine in serum 86 and urine were determined by HPLC method using fluores cence detection ( A EX " 340 nm; ЛЕМ " *18 nm> • After protein precipitation (1 ml serum + 3 ml methanol), an aliquot of clear supernatant was injected into column (Separon C18 10>um; 250 x 4,6 mm) at 35°C. The mobile phase was the mixture Qf methanol: 0,85 t H,POj in 0.01 M

KH2P04 (30:70, v/v) and the flow rate was adjusted to 2 ml/min. Concentration valuee were estimated from peak areas compared with simultaneously determined standard curves for quinidine and /3S/-3-hydroxyquinidine. The mean serum levels of quinidine and its metabolite were higher (p<0.05) after administration of Optochini- dinR in the time interval 1-6 h. This fact is in correla tion to lower elimination of these drugs in urine collec tion period 0-3 h. There were no statistically significant differences in serum levels and excreted ar>ounts of quini dine and its metabolite during the following time inter vals after administration of both preparations. The phar macokinetic analysis of serum quinidine and /3S/-3-hydroxyquinidine concentrations were performed on a digital com puter ADT 4316 using one-compartment model with first- order absorption. There was no significant difference between the two preparations in the absolute bioavailability calculated from the area under the serum concentration - time curve (AUC). The serum concentrations after Chinidin retard and OptochinidineR reached the therapeutical range (1.0-3.5 mg/1). In this respect both preparations seem to have almost an equal therapeutical value in clinical practice.

124 P.4-22 AMINO-ACID ANALYSIS BY HIGH PRESSURE LIQUID CHROMATOGRAPHY АМИНОКИСЛОТНЫЙ АНАЛИЗ С ПОМОЩЬЮ ВЭЖХ G.Fiedler, E.Radda, Vienna, Austria Abstract not arrived on time for printing. 125 P.4-23 ROUTINE N-TERMINAL ANALYSIS OF PEPTIDES ON PICOHOLE LEVEL WITH FLUOROMETRIC DETECTION OF DNS-AMINO ACIDS СЕРИЙНЫЙ N-КОНЦЕВОЙ АНАЛИЗ ПЕПТИДОВ НА ПИКОМОЛЬНОМ УРОВНЕ С ФЛУОРИМЕТРИЧЕСКИМ ДЕТЕКТИРОВАНИЕМ ДНС-АМИНОКИСЛОТ N.V.Levina, H.H.Muradov, I.V.Nazimov, Moscow, USSR A technique was developed for the N-terminal analysis of peptides (10 picomoles) as 1-dimethylaainonaphthalene- -5-sulfonyl-(DNS)amino acids, involving three rapid proce dures: peptide dansylation (15 min), hydrolysis of DNS- peptides (15 min), separation of DNS-amino acids by RP HPLC (26 min). The conditions of dansylation (5-lO-fold DNS-C1 excess, 6 jal of reaction mixture volume) precluded the formation

87 of large amounts of side products which complicated the subsequent chromatographic separation of DNS-amino acids. Under the conditions proposed (5,7 M HC1 and trifluoroacetic acid 2 2:1, v/v, 190°) hydrolysis of DNS-peptides took IS min. Optimization of the eluent buffer, its molarity and pH allowed the complete RF HPLC separation of the mixture of 22 mono- and di-DNS-amino acids. Due to the low content of side products in the sample as well as to mild condi tions of separation of DNS-amino acids, the efficiency of the chromatographic column remained the same after about 3000 injections. Use of fluorometric detector allows reliable detection of 10 picomoles of DNS-amino acids. These techniques (dansylation, hydrolysis, separation) were employed for quantitative determination of the N-ter minal amino acids of poorly hydrolysed dipeptides (Val-Val, Ile-Val, Leu-lie), Hydrolysis kinetics for these sequences and similar ones was studied that allowed us to reduce the total time of the N-terminal amino acid analysis of membra ne protein fragments from 18 h (conventional time) to l.Sh, and, at the same time, and to increase the yields of DNS-amino acids by 1.5-?.2.

126 P.4-24 DETERMINATION OF TREONINE, TRIPTOPHAN, LYSINE AND GLUTAMIC ACID BY CHROMATOGRAPHIC METHODS ОПРЕДЕЛЕНИЕ ТРЕОНИНА, ТРИПТОФАНА, ЛИЗИНА И ГЛУТАМИНОВОИ КИСЛОТЫ ХРОМАТОГРАФИЧЕСКИМИ МЕТОДАМИ T.I.Struchalina, O.P.Kutkina, A.M.Bogher, Frunze, USSR Extraction of amino acids from natural raw materials reguires effective test procedures. Chromatographic techni ques are widely used for this purpose. When modifying analytical sample preparation procedure, the composition of intermediates in the microbiological synthesis of treonine, triptophan and glutamic acid has been studied using the AAA-881 analyzer (Czechoslovakia) and recommendations have been worked out as to production of valuable feed additives from process wastes. Lyzine, glycine, isoleucine, leucine and other amino acids have been found present among the associated amino acids. At the same time we have studied resolution of amino acids from model mixtures by electrophoresis and thin- layer chromatography techniques. By varying the pH and composition of the buffer solution sharp resolution of amino acids from the model mixture of lysine, glycine and treonine was achieved' in the course of 15 to 20 minutes using the EFA-I apparatus with the electrophoresis cham ber operated under the voltage of 800 V and current of 20 to 25 mA. A solution of mixed phosphoric, boric and acetic acids of pH 2,2-2,4 was used as the buffer. Adjust ment of pH was carried out with an alkali solution. A mixture of 8 amino acids (valine, lysine, glycine. 88 serine, treonine, isoleucine, glutamic acid and triptophan) with the buffer solution pH 2,2 was separated in 4 groups or. the OE-201 apparatus (Hungary) . In this case triptophan remained closer to the starting line than all the other amino acids cind could be determined quantitatively with the electrophoresis chamber operated under 70O V, 12 mA and running time 90 minutes. When current was brought down to 5-7 mA and running time reduced to 60 minutes, separa tion of lysine and triptophan was observed, while all the other amino acids were developed in a single spot. In the acetate buffer of pH 3,8 separation of glutamic acid from neutral amino acids was achieved. Besides, we have studied a number of systems for separa ting amino acids using thin-layer chromatography techni ques. The best results in separating mixed amino acids were obtained for the system isopropyl alcohol:ethyl ace tate :ammonia:water 20:20:1,5:25. Double distilation allowed to separate leucine and isoleucine in this system. The results of the study have been presented as recommended procedures for laboratory schedules.

127 P.4-25 HPLC SCREENING METHOD OF ECDYSTEROIDS ••КРИНИНГ ЭКДИСТЕРОИДОВ МЕТОДОМ южх M.Bathori, K.Szendrei, Szeged, Hungary Steroids, possesing moulting hormone activity are struc turally related to the compounds of 14»C-hydroxy-5^-cholest- -7-ene-6-one. These hormones» interfere with the growth processes of insects. Recently, the most interesting insect hormones, the acdysone (2/J,3j3,14«4,22R,25-penta- hydroxy-5j8-cholest--7-en-6-one) and crustecdysone (2$, 3jJ, 14o£,20,22R,25-hexahydroxy-5j8-cholest-7-c-n-6-one) as well as their precursors and metabolites are the target of intensive research. The possibility of using insect moulting steroids for genetic control of insects or perhaps as chemotherapeutic agents for parasites has increased the interest in the study of these hormones including the procedures how they can be detected, isolated and produced. Although these compounds were firstly isolated from insects (Borabyx mori) and crustaceans (Jasus lalandei), considerable amounts of these hormones have been found in the axtracts of several plants (Amaranthaceae, Polypo- diaceae, Taxaceae, Aizoaceae, Labiatae, etc.) as well. Crude extracts of plants belonging to six plant fami lies were screened in order to detect their ecdysteroid content. The analyses were done by high-performance liquid chromatography (Liquochrom 2010, Labor MIM, Budapest, Hungary) and the compounds separated were followed by UV detection at 254 nm. Octadecyl silica and acetonitril-phosphate buffer at pH 2.1 (9:11) were used as stationary and mobile phases, respectively.

89 128 P.4-26 HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC .DETERMINATION OF EPINEPHRINE, NOREPINEPHRINE, ISOPRENALINE AND CORRESPONDING SULFONIC ACIDS IN INJECTION SOLUTIONS ОПРЕДЕЛЕНИЕ ЭПИНЕФРИНА, НОРПИНЕФРИНА, ИЗОПРЕНАЛИНА И СООТВЕТСТВУЮЩИХ СУЛЬФОНОВЫХ КИСЛОТ В ИНЪЕКЦИОННЫХ ПРЕПАРАТАХ МЕТОДОМ ВЭЖХ М.DoleZalovcf, M.Novik, Prague, CSSR Colorimetric, spectrophotometry and fluorometric determination of catecholamines in pharmaceutical preparr- tions lack specificity and an extensive sample preparatit is often required. Gas chromatographic methods are tedioi. and time consuming. High-performance liquid chromatography doesn't suffer from these disadvantages. All published HPLC methods to determine various catecholamines are simple, rapid and specific. Catecholamines are separated from their degrade tion products, impurities and antioxidants. Catecholamine sulfonic acids, the degradation products formed by an interaction of catecholamines and bisulfite used as an antioxidant, give peaks at or very closed to the solvent peak with all described HPLC procedures. So no information about the content of these degradation products can be obtained. In our work HPLC in reverse phase mode is used for simultaneous determination of catecholamine and the corresponding sulfonic acid in injection solutions of epinephrine, norepinephrine and isoprenaline. Both UV or electrochemical detector can be utilized. The presence of oxidation degradation products or likely impurities doesn't interfere with the assay. 129 P.4-33 A NEW DERIVATIZATION AGENT FOR THE DIAGNOSIS OF ENZYMATIC DEFECTS OF STEROID BIOSYNTHESIS НОШИ АГЕНТ ДЛЯ ДЕРИВАТИЗАЦИИ ПРИ ДИАГНОСТИКЕ ЭНЗИМАТИЧЕСКИХ ДЕФЕКТОВ БИОСИНТЕЗА СТЕРОИДОВ Z.Juvancz, K.Ujszciszi, A.H.Csarnyi, К.Payer, а.вЛуот, D.Knausz, Budapest, Hungary The congenital adrenal hyperplasia is an inborn error of steroid metabolism caused by various enzymatic deficit cies in the biosynthesis of Cortisol. The characteristic symptom of the disease is the abnormility of the externa!» genital. The profile chromatograms of urinary steroids ar used to identify exactly the given enzymatic defects. The urine contains various steroid compounds in higher quantity, depending on enzymatic shortage as compared to normal cases. A new silylation agent/ without enolizatio. was used. An aliquot of the urjne samples (0-24 h) were enzymati- cally hydrolyzed with ^-glucuronidase aryl sulphatase, to liberate the steroids. The hadrolyzed samples were puri-

90 s

fled on a short column, which is similar to Sep-Pak C,B. The silylation was carried out with the N-substitutfd ellyl carbamate - a new silylation agent. Another poster deals with the chemical nature of the silylating agent (See: O.Xnausz, K-subetituted silyl carbamates as silyla- ': .ing agent for gas chromatographic analysis). Gas chromato- i raphic analysis were done with Perkin Elmer F 22. Split- i ess injection was applied. The GC-MS investigations were g arried out by VGHM 12 PIA type mass spectrometer, coupled " о PYE 104 gas chromatograph. | The new derivatization agent, N-substituted silyl car- i,amate, is very suitable for analysis of steroids. The )*ilylation was carried out without enolization of keto j.jroups. The one step derivatization is very useful due to I ime and material saving in routine work, and it produced I ess side products.

! 30 P.4-34 I • HPLC OF NUCLEOTIDES. PROGRESS IN APPLICATION, 1 METHODS DEVELOPMENT AND PROBLEMS ! ВЭЖХ НУКЛЕОТИДОВ. УСПЕХИ В РАЗВИТИИ И ПРИМЕНЕНИИ И НЕРЕШЕННЫЕ ПРОБЛЕМЫ i A.N.Wulfson, S.A.Yakimov, Moscow, USSR The mechanism of chromatographic separation of nucleoti- I за involves ionic, hydrophilic and hydrophobic interac- I ions. Therefore ion-exchange and reverse-phase HPLC are I pplied as well as normal-phase adsorption chromatography I >f fully protected nucleotides. The wide application of [these methods in biochemistry, molecular biology and 'jenetlc engineering is resulted from a broad variety of (nigh performance packing materials, which allow to separa te the complex mixtures of nucleotides according to their size and composition. The latest developments are the separation of the mixtures from 30 and even 50 oligomer- homologues, large fragments of DNA, RNA and plasmides. The study of retention (K1), composition, pH and eluent concentration (C) reveals the universal dependence log K' = a + b log C, that simplifies the optimisation of HPLC methods. The ound correlation of selectivity with K' allows to study rid choose the sorbents on the basis of objective parame- •:*rs. The methods of purposeful control of selectivity «able to increase resolution and informability of chroma- •graphy and they happen to be especially necessary for jvelopment of two-dimensional HPLC. , The developed method for prediction of optimal gradient • lution program with a given retention and resolution .Hows to design rapidly the optimal procedure for analy- | is or isolation of the components of interest from the jomplex mixtures The main problems in this field are: (1) the mixed mode mechanisms of nucleotides interactions with the sorbent hat leads to complexities in selection of a mobile phase :omposition and (2) the complex dependence of retention from heterocyclic composition of oligonucleotides. Both problems slow down development of informability of chrom ,- tography, which is necessary for identification of the components of separated mixtures, 131 P.4-35 PHVSICO-CHEMICAL FEATURES OF CHROMATOGRAPHY OF MACROMOLECULS ФИЗИКО-ХИМИЧЕСКИЕ ОСОБЕННОСТИ ХРОМАТОГРАФИИ МАКРОМОЛЕКУЛ Yu.A.Eltekov, Moscow, USSR Retention of macromolecules on column material In quasi-equilibrium conditions of chromatography is contrc- led mainly by sizes and chemical structure of macromole cules and/or their associates and also by the composite of mobile phase, pores distribution and surface chemist!. of column material. The combination of different parame ters of chromatography system allows to change the direc tion of separation process to the distribution of macromolecules on their sizes or to the distribution of macr - molecules on their adsorption energies. The decrease o' adsorption activity of column materials and/or the in crease of polarity of eluent leads to the transition fr. adsorption chromatography to the sieve one. The generalized calibration of column for liquid sic • chromatography . (LSiC) as K-function (K - distribution coefficient of polymer in column) on R - ratio coil size of flexible macromolecular chains to the average diameter of opening into pores of column material has been sugges ted. Dependences К on R have been considered for chromato graphy systems containing the column materials (silica gels, silochroms and macroporous glasses) with of diffe rent pore diameters from 5 to 200 nm and surface chemistry of different polymers (polystyrene, polyoxyethylene, poly ethylene, polyethyleneglycole, dextranes and proteines). The effect of the composition of mobile phase, of the temperature and internal diffusion of macromolecules on the parameters of LSiC of polymers was discussed. 132 P.4-36 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF HYDROXYETHYL STARCH ВЫСОКОЭФФЕКТИВНАЯ ЖИДКОСТНАЯ ХРОМАТОГРАФИЯ ОКСИЭТИЛИРОВАННОГО КРАХМАЛА M.A.Chlenov, O.A.Yurin, T.N.Telkova, V.A.Dombrovsky, Moscow, USSR Amylopectin starch, modified by hydroxyethyl groups - hydroxyethyl starch (HES) is widely used as plasma expan der, cryprotector and as hemodynamic component of oxygen- carrier blood substitutes. It has been shown in this work that HPLC can be applied for investigation of HES structu re, biological properties and molecular weight parameters. 2-0-Hydroxyethyl-D-glucose and D-glucose have been 92 shown to be the main products of total acid hydrolysis of

HES. The separation was carried out on Lichrosorb NH2 column using MeCNiH^O - 85:15 as a mobile phase. The peak area rationing of alkyl glucoses to free glucose allows to determine the degree of HES substitution. The molecular weight parameters of HES samples have been determined by high performance size exclusion chroma tography (HPSEC) on SynChropak GPC 10O, 50O and 1000 columns connected in series as well as on Spherogel TSK 3000 and 5СЮ0 PW columns, using aqueous sodium azide as mobile phase. The column set was calibrated with dextrans and polyethyleneglycols with known molecular weight para meters. The Л~ and M„ - values of HES samples have been calculated automatically by SP-41O0 minicomputer using universal calibration approach. The data obtained exhibi ted a good correlation with the values found by other methods. It has been also shown that HPSEC can be applied for determination of changes of HES molecular weight parame ters in the course of its circulation in blood stream. 133 P.4-37 STEP-BY-STEP ANALYSIS OF POLYCONDENSATIONAL OLIGOMERS AND POLYMERS BY LC-METHOD ПОСТАДИЙНЫЙ АНАЛИЗ МЕТОДОМ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ ОЛИГОМЕРНЫХ И ПОЛИМЕРНЫХ ПРОДУКТОВ ПОЛИКОНДЕНСАЦИОННЫХ ПРОЦЕССОВ V.V.GuryanOva, A.V.Pavlov, Moscow, USSR Exclusion and adsorption chromatography permits to determine the detailed composition of polycondensation products at all stages of synthesis of following polymers: polycarbonate, polysulfone, polytbutylen terephtalate), aliphatiche polyamide. The theoretical conception of oligomer separation in "critical regime" was used. It gives the optimal conditions for separation under investigation mixtures in gradient regime, the adsorption interaction's of end-functional groups and macromolecular chains being taken into account. It permits to receive the simultaneously information on MWD and functional type distribution for oligomer products. Experimental chromatograms of oligomers fits well the cal culated ones. For rigorous evaluation of MWD the precise GPC method is proposed based on addition the internal standard into a probe. If a narrow-distribution polystyrene standard is not available the package of program for calculation of calibration parameters is developed. It is based on appli cation of the well characterized polymer sample with wide MWD. In many cases the application of LC allows to ascertain the interrelation of MWD and various polymer properties, particularly, optical properties. With the use of the double-detection method - UV in the range of 360-380 nm and the refractometer - we found the effect of the high

93 and low molecular-weight fractions of a polymer on the change of optical characteristics of a sample due to destruction and crosslinking processes. 134 P.4-38 ADSORPTION EFFECTS IN LIQUID CHROMATOGRAPHY OF POLYMERS АДСОРБЦИОННЫЕ ЭФФЕКТЫ В ЖИДКОСТНОЙ ХРОМАТОГРАФИИ ПОЛИМЕРОВ A.M.Skvortsov, A.A.Gorbunov, Leningrad, USSR It has been found experimentally, that except the GPC case, there is an extensive range of conditions, in mixed solvents mostly, when forces of attraction act between chain units and the surface of porous adsorbent. There

exsists the so-called "critical energy of adsorption" -£e, which separates the molecular-sieve and adsorption chroma tography. Using mgdel of a flexible Gaussian chain in a slit-like pore we have constructed an equilibrium theory of chroma tography at an arbitrary energy of polymer-adsorbent inte raction and at any values of the ratio of chain radius R to pore width D. The polymer-adsorbent interaction is given by the parameter H, connected with the adsorption energy £ by the ratio _:l H ~ b (£-£c) , where b - is the size of a chain segment. Thus, H charac terises the deviation of the system from the critical sta te (the origin of adsorption). From the theory it follows that there are five diffe rent regimes of chromatography, covering the whole range of R, D and H parameters. For each regime analytical ex pressions have been obtained for the distributional coeffi cient K, as well as for the monomer density profile inside the pore, and also for the fraction of chain units adsor bed on the pore walls.

135 P.4-40 MATHEMATICAL MODEi,. :NG AND OPTIMIZATION OF GRADIENT LIQUID CHROMATOGRAPHY OF BIOLOGICALLY ACTIVE SUBSTANCES МАТЕМАТИЧЕСКОЕ МОДЕЛИРОВАНИЕ И ОПТИМИЗАЦИЯ ГРАДИЕНТНОЙ ХРОМАТОГРАФИИ БИОЛОГИЧЕСКИ АКТИВНЫХ ВЕЩЕСТВ V.A.Pasechnik, S.А.Roshchin, E.A.Polyakova, Leningrad, USSR Analytic and numeric methods of mathematical modelling and optimization of gradient elution liquid chromatogtaphy separations are developed. They are based on the physical conservation laws, theory of convective diffusion, kine tics of adsorption and principles of formulating the optimal separation criteria. Analytic expressions are derived which determine the position and the width of zone for simple gradient programs. New approach to optimiza tion of retention time, wiith of zone and separation

94 criteria for pairs and groups of zones is proposed. The application of numeric methods and computer techni ques gives possibility to consider more general forms of gradient programs without using the moments of distribu tions. The functional dependence of the chromatographic curve on hydrodynamlc, transport and adsorption parameters of chromatographic columns for different gradient programs is investigated. Practical applications of proposed principles of opti mization to the solving the problem of optimal separation of solutions of biologically active substances are dis cussed.

136 P.4-41 STUDY OF ADSORPTION EQUILIBRIUM IN CHROMATOGRAPHIC COLUMN BY FRONTAL CHROMATOGRAPHY ИЗУЧЕНИЕ ДИНАМИЧЕСКОГО РАВНОВЕСИЯ В ХРОМАТОГРАФИЧЕСКОЙ КОЛОННЕ МЕТОДОМ ФРОНТАЛЬНОЙ ХРОМАТОГРАФИИ - Yu.V.Kazakevitch, Yu.A.Eltekov, Moscow, USSR The automatic home-made chromatographic system was described. This system allowed to study adsorption of non- electrolytes from solutions by frontal and elution chroma tography. Frontal method permits, to determine in one sing le experiment only one point on adsorption isotherm which corresponds to the equilibrium concentration of front. The void volume of chromatographic column was calcula ted by frontal curve of pure adsorptiv, because in this case the value of Gibss excess adsorption is equal to zero. The chromatographic systems benzene-n.heptane-silica gels were chooser) for the evaluations of the possibility of application of whole frontal method to the measuring of adsorption isotherm. The adsorption of benzene on silica gels with the various pore diameters in the wide range of concentrations was determined. The dependence of adsorp tion of benzene on flow rates and on the pore diameters of adsorbent was found. For the silica gel with the average pore diameter about 10 nm the coincidence of adsorption isotherms measured by static and dynamic methods was found only for flow rate below 15 cm/s. For the samples of silica gel with the various pore diameters the equilibrium values of linear flow velocity was found above which the sorption dynamics became nonequilibrium. This chromatographic system was used for the investiga tion of adsorption isotherm by step-by-step method. The coincidence of the two methods was shown for the convex adsorption isoterms, and also the possibility of their joint application for the investigation of the dynamics of the adsorption front movement along the column.

The isotherm of adsorption of CH3CN from H20 on rever sed-phase adsorbent was measured.

9S 137 P.4-42 SIMULTANEOUS SEPARATION OF NITROGENOUS BASES, NUCLEOSIDES AND NUCLEOTIDES BY REVERSED-PHASE HPLC USING ION PAIRING ON RADIALLY COMPRESSED COLUMNS ОДНОВРЕМЕННОЕ РАЗДЕЛЕНИЕ РИБОНУКЯЕОТИДОВ, НУКПЕОЗИДОВ И АЗОТИСТЫХ ОСНОВАНИИ МЕТОДОМ ОБРАЩЕННО-ФАЗОВОЙ ИОН-ПАРНОИ ВЭЖХ НА КОЛОНКАХ С РАДИАЛЬНЫМ УПЛОТНЕНИЕМ A.M.Pimenov, Yu.V.Tikhonov, I.S.Meisner, R.T.Toguzov, Moscow, USSR Investigation of the metabolism of free necleotides and their derivatives in normal organs and tissues and during different diseasee determines the necessity of a quantita tive analysis of the metabolic pool. The use of ionic and hydrophobic interaction mechanisms in a single HPLC system enables a simultaneous.separation of charged (nucleotides) and neutral (nucleosides and their bases) compounds. A radially compressed NovaPak C18 column and an ion pairing reagent (tetrabutylammonium phosphate, TBA) in a phpsphate buffer/acetonitrile mobile phase were employed for separation of purine and pyrimidlne ribonuc leotides, nucleosides, and their bases. The effect of TBA and acetonitrile in the mobile phase on the retention of purine and pyrimidine nucleotides was studied. The optimum conditions of gradient elution of some biologically important nucleotides, nucleosides and nitrogenous bases were selected and the possibility of a precise control of the retention time of the compounds imder study through variation of pH and the ionic strength of the mobile phase and also of the time of equilibration of the column with the initial buffer was studied.

The retention mechanisms on a C18 column are discussed together with the prospects of the use of ion pairing reagents (of the polymethylsuJ.phonic acids type) for a simultaneous separation of DMA and RNA components with respect to their basicity degree and hydrophobity.

The gradient elution method in a NovaPak Сд8 - TBA - acetonitrile - containing buffer chromatographic system was employed for the analysis of the nucleotide pool of acid-soluble fractions of animal and human tissues in experimental and clinical pathology.

138 P.4-43 APPLICATIONS OF CHROMATOGRAPHIC METHODS FOR THE CONTROL OF THERMAL STABILITY OF POLYORETHANES ПРИМЕНЕНИЕ ХРОМАТОГРАФИЧЕСКИХ МЕТОДОВ ДЛЯ КОНТРОЛЯ ТЕРМИЧЕСКОЙ СТАБИЛЬНОСТИ ПОЛИУРЕТАНОВ J.Simon, Veszprem, Hungary F.Barla, A.Kelemen-Haller, F.Farkas, Gy6"r, Hungary The thermal stability of polyurethanes (PUR) is their very important characteristic from the point of view of their practical application. The most widely used control method is the thermogravi-

96 metry. It give» information about the weight-changing of the PUR during heating in different gas-environments. The pyrolysis G.C. is also applied for that purpose at high temperature, generally from 300°C up to 1000°C, with few seconds of handling time. This sample-handling repre sents only very extreme heat-loading conditions. The reactor G.C. can be used for the simulation of the everyday heat-loading of the materials, because the examinations can be carried out from room-temperature up to 250°C, in time period of several minutes up to several hours in different gas environments. We summarize the findings of our research, carried out by the combination of the above listed methods.

139 P.4-45 INVESTIGATION BY CHROMATOGRAPHY OF THE HIGH MOLECULAR RESIN COMPOUNDS OF THE ORGANIC MATTER FROM RECENT AND ANCIENT SEDIMENTS IN THE HOESIAN PLATFORM (NORTH BULGARIA) ХРОКАТОГРАФИЧЕСКОЕ ИССЛЕДОВАНИЕ ВЫСОКОМОЛЕКУЛЯРНЫХ СМОЛЯНЫХ КОМПОНЕНТОВ ОРГАНИЧЕСКИХ ОСТАТКОВ В НЕДАВНИХ И ДРЕВНИХ МОРСКИХ ОТЛОЖЕНИЯХ ЧЕРНОГО МОРЯ /СЕВЕРНАЯ БОЛГАРИЯ/ R.S.Koleva, Sofia, Bulgaria The aim of the investigation was (in connection with crude oil and natural gas perspectives) to clarify the possibilities of the high molecular compounds to produce hydrocarbons. Samples obtained from recent sediments in the Black Sea and from Mezozoic rocks in deep wells were extracted with chloroform. The extracts free from asphaltenes were sepa rated into fractions by column chromatography. The frac tions of benzene resins were further chromatographed on A^Oj/silica gel in a glass column with hexane, hexane- benzene, benzene-methanol, diethyl ether. The total ben zene resins and their fractions were examined by elemental analysis, IRS, GC, GC-MS, TLC. As a result of the complex investigation the presence of the following compounds is established in the benzene resins: normal alkanes, alkylbenzenes, aliphatic and aromatic esters, sterols, aromatic ketones, multinuclear compounds with condensed benzene nucleus. As the samples were with equal diagenetic oxidation, but with different level of maturity of the marine plank- tonic organic matter the investigation proved that under suitable pressure and thermal conditions the hydrocarbons might migrate from the rocks.

7. Зак. 872 97 140 P.4-44 SEPARATION OF LIPOPHILIC FRACTIONS BY HPLC ПРИМЕНЕНИЕ ВЭЖХ ДЛЯ РАЗДЕЛЕНИЯ ЛИПОФИЛЬНЫХ ФРАКЦИЙ B.P.Lapin, N.A.Pisareva, T.E.Rubtsova, M.L.Zhakevich Moscow, USSR An improved scheme was developed to investigate lipo philic fractions in different hydrobionts using a combina tion of modern instrumental methods. As part of this technique it was shown that it is possible- to separate the total lipids fraction on a single column with silica. By utilizing the flow rate gradient instead of mobile phase gradient, and by using of n-hexane:isopropanol:ace tate buffer (500:500:37,5), we made an elution of all compounds including the most polar phospholipids. Waxes, esters of sterines, mono-, di- and triglycerides, free fatty acids, carotenoids, fat-soluble vitamins, hydrocar bons, lecithin and others were detected. As the final sta ge of analysis, we use the injection of pure isopropanol through the sample loop of the injector unit. This step helps us to elute the most polar components from the column. Results show that it is possible to utilize certain photometric detectors, which are relatively simple and easily available for preparative type separations. Because there are unsaturated compounds in total lipids fraction, such work can be done using fixed wavelength 254 nm. Acetone has been shown to be acceptable mobile phase constituent for short-wave UV detection at 213 nm. 141 P.5-01 MICROBORE COLUMN LIQUID CHROMATOGRAPH MODEL 310 МИКРОКОЛОНОЧНЫЙ ХРОМАТОГРАФ МОДЕЛЬ ЛО K.Les'niak, Warsaw, Poland Posters informing about principles of operation, construction of main blocks and main features of the instrument. Examples of application are also presented. SPECIFICATIONS Pump and programmer - syringe pump with stepping-motor - pump volume: 32 cm'. - maximum working pressure: 30 MPa - flow rate for filling and empting: 2 cm /min - adjustable flow: 0-200 jul/min - pressure stop-fJcw regulation within the 0-30 MPa range - power requirements: 90 VA, weight: 25 kg Detector UV-254 - optical path lenght: 5 mm, cell volume: 1 pi - absorbance ranges: 0.01. 0.05, 0.25, 1.25 AUFS - baseline noise: 4 x 10~5 AU - recorder output: lOOO mV 98 - integrator outputs 1000 mV/AUFS - power requirements: 8 VA, weight: 4 kg Columns with various sorbents - column with the dimensions 1 x 250 mm Constant volume injection valve: 0.4 jul. 142 P.5-03 SOME APPLICATIONS ON ENVIRONMENTAL AND BIOCHEMICAL ANALYSIS OF NEW ТУРЕ OF AMPEROMETRIC DETECTOR IN HPLC ПРИМЕНЕНИЕ НОВОГО ТИПА АМПЕРОМЕТРИЧЕСКОГО ДЕТЕКТОРА ДЛЯ ВЭЖХ В АНАЛИЗЕ ОКРУЖАЮЩЕЙ СРЕДЫ И В БИОХИМИИ P.Kovif, Brno, C*SSR A new type of amperometric detector in HPLC was develo ped. The cell of the detector excels by very small dimen sion (cell volume is less than 30 nl), so that it can be used in low dispersion liquid chromatography system with microbore columns. An important aspect of the detector is two-, resp, three-electrode configuration with glassy- carbon working electrode. This detector connected with 150 x 1,4 mm glass columns was applied for selective and sensitive LC analysis, for instance phenols, chlorophenols, catecholamines etc. in water and biochemical materials (human serum blood). Typical minimum detectable amount of this LC-EC system is less than 1 pmol in direct injection on column. 143 P.5-04 LASER FLOOROMETRIC DETECTOR FOR HPLC ЛАЗЕРНЫЙ ФЛУОРИМЕТРИЧЕСКИЙ ДЕТЕКТОР ДЛЯ МИКРОКОЛОНОЧНОЙ ХРОМАТОГРАФИИ V.A.Mostovnikov, A.F.Lobazov, S.V.Nechaev, Minsk, USSR B.G.Belenkii, E.E.Kever, E.M.Korolyova, Leningrad, USSR The laser fluorometric detector is designed to analyze DNS-derivatives of aminoacids together with a high-effi ciency liquid chromatograph with a peak volume no greater than l/il. As the excitation source, a helium-cadmium laser is used which oscillates at a wave-length of 325 nm. The detector has the following spacifications: Detection limit at s/n =2 ,, for DNS-Ala 2-10 mole Detected volume 0.1 to 0.4 yul Post column volume no more than 0.5/ll Base-line deviation no more than 2 percent Standard base line deviation from the limiting value of the recorder scale for the maximum sensitivity range no «more than 3 percent Linearity coefficient , (in a dynamic range of 103) 1 + 0.01 Lag O.T s Measurement accuracy 3 percent In chromatographing DNS-aminoacids in a 0.5 x 170 mm column, the detection limit of substance being analyzed at 99 the chromatographic peak is 2 lO"15 mole. Such high sen sitivity has been obtained for the first time. It should be noted that the small detector volume makes it possible to use the high-performance microcolumns to full advantage.

144 P.5-07 HIGH-PERFORMANCE DUAL-WAVELENGTH MICROCHROMATOGRAPHY OF POLYPEPTIDES OP THE NEURON STRUCTURES ВЫСОКОЭФФЕКТИВНАЯ ДВУХВОЛНОВАЯ МИКРОХРОМАТОГРАФИЯ ПОЛИПЕПТИДОВ НЕЙРОННЫХ СТРУКТУР N.I.Mikichur, M.B.Shtark, Novosibirsk, USSR The use of dual-wavelength detection permitted applica tion of HPLC for the study of proteins and- peptides of separate brain structures and individual neurons in diffe rent model systems. Registration of optical density is carried out in quartz cell - a capillary of O.3 mm i.d., which is the extension of a metal column of approximately 1 /il volume. A reversed phase of aminochrome and nucleosil Co are used as lorbente. The sample volume is approximately 0.03 jul. This method makes it possible to reveal the existence of a functionally conditioned biochemical heterogeneity within single brain structure (areas: CA, and fascia den- tata Hippocampus) and even the heterogenity of command neurons H.pomatia, responsible for certain physiological functions. The results obtained help to clear up the molecular mechanism of neuronal plasticity which lies in the basis of processes of adaptation, education, memory etc.

145 P.5-15 MICRO-COLUMN LIQUID CHROMATOGRAPHY WITH MULTI- WAVELENGTH PHOTOMETRIC DETECTION. II.COMPUTER TREATMENT OF TIME-SHARE MULTI-WAVELENGTH DETECTION DATA МИКРОКОЛОНОЧНАЯ ЖИДКОСТНАЯ ХРОМАТОГРАФИЯ С МНОГОВОЛНОВОЙ ФОТОМЕТРИЧЕСКОЙ ДЕТЕКЦИЕЙ П. КОМПЬЮТЕРНАЯ ОБРАБОТКА ДАННЫХ МН0Г0В0ЛН080Я ДЕТЕКЦИИ В РЕЖИМЕ РАЗДЕЛЕНИЯ ВРЕМЕНИ G.I.Baram, M.P.Perelroyzen, A.P.Zenkov, Novosibirsk, USSR Our previous communication (J.Chromatogr. 264 (1983) 69-90) has described the micro-column liquid chromato - raph OB-4. The detector of this instrument is a precise double-beam microspectrophotometer with a special device for rapid change of wavelengths according to cyclic programs, i.e. a programmable spectrophotometry detector. Computer treatment of data obtained with such detectors has some peculiarities because discrete values of absorbances are obtained, at the selected wavelengths at time intervals (time-share multi-wavelength detection), and 100 these discrete values must be interpolated to obtain con tinuous chromatograms, and also the running values of adsorbance ratio. Our computer system consists of the mini-computer SM-4 having an eight-colour monitor with a resolution 256 x 256, and an eight-colour plotter. The programs are written in Fortran and in Assembler. The random access memory stores up to б 000 absorbance data, each consisting of five deci mal digits. The system works in the dialogue mode and allows: - on-line presentation of running chromatograms on the colour TV screen of the monitor with different colours denoting absorbances at different wavelengths; - control, and, if necessary, manual correction of data; - control of the time schedule of the instrument; - interpolation between discrete absorbance values and calculation of absorbance ratio from interpolated data at selected time intervals; - separate subtraction of baselines at different wave lengths ; - digital filtration of noise .(smoothening)> - detection of peaks, calculation of retention times at each of the wavelengths, and of integrals; - display of the results on monitor screen at the selected scale; - collection of libraries of crude and treated chromato grams, 146 S.6-2 SUPERCRITICAL FLUID CHROMATOGRAPHY FOR GROUP SEPARATION OF HYDROCARBONS СВЕРХКРИТИЧЕСКАЯ ЖИДКОСТНАЯ ХРОМАТОГРАФИЯ ДЛЯ РАЗДЕЛЕНИЯ УГЛЕВОДОРОДОВ W.Ecknig, H.-J.Polster, Berlin-Adlershof, GDR Supercritical fluid chromatography (SFC) is success fully applied for separations of high boiling crude oil and coal liquification fractions using high efficient micro bore and capillary columns. However, besides the resolution of individuals the SFC Includes a natural physico-chemical potential for group separations caused by the decreasing of the separation factor Л. with increa sing column pressure. Using pressure programming, methods have been elabora ted for a fast iso-n-paraffin group separation of an indu strial hydrocarbon fraction containing Cj0- C^Q paraffins on molecular sieve 5A, for group separation of paraffins, olefins and aromatics in the chain length range Cg - Cie on modified X-type molecular sieve and for separation of hydrocarbon species on Lichrosorb RP 18. The efficiency parameters of micro bore columns and classically packed columns are compared. Beside smaller theoretical plate height and shorter time of analysis the peak symmetry is improved, which is important for the quantitative interpretation of the chromatograms.

101 Equipments for supercritical fluid chromatography inclu ding a flame ionization detector in series with a dead volume free flow restrietor are presented. An absolutely

pulse-free C02 flow, in the range up to 1 ml/min, is achieved by a thermostated pressure wessel. The stainless steel or glass micro bore columns (100- 300 mm x 1 mm I.D.) are filled with microcrystalline molecular sieve (particle size 2-3 jtim) or LiChrosorb RP 18 (particle size 5 лип). The columns are packed by the slurry technique. 147 P.2-60 SELF-ASSOCIATION INDUCED OPTICAL RESOLUTION BY LIQUID CHROMATOGRAPHY ОПТИЧЕСКОЕ РАЗРЕШЕНИЕ, ИНДУЦИРОВАННОЕ САМОАССОЦИАЦИЕЙ, В ЖИДКОСТНОЙ ХРОМАТОГРАФИИ S.Hara, Tokyo, Japan When chiral solutes take part in solute-solute interac tions in achiral solvents, these self-associations can result in chiral differentiation of D and L solutes in enantiomerically enriched mixtures since, by external comparison, each individual enantiomer is in different average environment due to its predominant association with major enantiomer. We found that enantiomeric mixtures of N-acetylvaline tert-butyl ester in carbon tetrachloride solution exhibit distinct H nmr signals for the D and L enantiomers, i.e., self-induced chemical shift nonequivalence. Nmr study demonstrated that observed sense of nonequivalence is induced through the formation of diastereomeric dimers, where associative interactions are bidentate NH—0«C (ester) hydrogen bonds. Though the main contributor for this phenomenon is intrinsic magnetic nonequivalence between the sensor nuclei of instantaneous diastereomeric dimers formed under conditions of fast-associate exchange, we confirmed the contribution of association stereo-selec tivity: that is, additional stability imparted to the homochiral dimer. This contribution premoted the magnitude of chemical shift nonequivalence with lower sample tempe ratures. Thus, one must assume that the stability differen ce between the diastereomeric dimers of the solute enantio mers is perceived through the chromatographic technique using the chiral mobile phase solvents on a silica gel column. When the solute enantiomers are associated with one single enantiomer of these as the chiral additive, it seems reasonable to expect that the diastereomeric dimers are formed in self-association equilibria while they passing through the column, and the enantiomers are sepa rated. These behaviors will be observed applying radio- labelled solute enantiomers and a detection of radioacti vity in the column eluent. We report self-induced chiral differentiation of 1*C-labelled racemic solute with a silica gel liquid chromatography using the chiral mobile phase solvent containing the unlabelled L enantiomer.

102 This separation should allow us to determine the mechanism by which stereo-selectivity is attained in the column. A.Dobashi, S.Hara, J.Am, Chem. Soc, in press. A.Dobashi/ S.Hara, J.A». Chen. Soc., submitted. 148 S.14-5 DETERMINATION OF CHEMICAL COMPOSITION PARAMETERS OF STATISTICAL COPOLYMERS BY MULTI-DETECTOR GPC ОПРЕДЕЛЕНИЕ ХИМИЧЕСКОГО СОСТАВА СТАТИЧЕСКИХ СОПОЛИМЕРОВ В МУЛЬТИДЕТЕКТОРНОЙ ГПХ R.Tymczynski, zabrze, Poland A multiple detector • system consisting of an infrared flow-through detector and an automatic viscometric detector has been used for characterizing by gel permea tion chromatography the compositional and molecular mass heterogeneities in statistical copolymers of styrene and n-butyl methacrylate synthesized to various degrees of conversion. A computer program establishing the chemical composi tion and molecular mass distribution parameters of the copolymers investigated has been developed. The proposed approach makes it possible to present the copolymer chemi cal inhomogenelty changes as a function of molecular mass. Notwithstanding the relatively small differences bet ween the reactivity ratios of the comonomers, the nultiple- detector GPC method has been found sufficiently sensitive to show an increase of the chemical inhomogeneity of the copolymers studied on increasing the degree of conversion, ,as predicted by general copolymerization theory. 149 P.2-61 EFFECT OF MOLECULAR STRUCTURE OF SOME NATURAL COUMARIN DERIVATIVES ON RETENTION IN HPLC AND HPTLC REVERSED-PBASE SYSTEMS ВЛИЯНИЕ МОЛЕКУЛЯРНОЙ СТРУКТУРЫ НЕКОТОРЫХ ПРОИЗВОДНЫХ КУМАРИНОВ НА УДЕРЖИВАНИЕ В 0БРАЩЕНН0-ФА30В0Й ВЭЖХ И В ВЫСОКОЭФФЕКТИВНОЙ ТСХ M.Bieganiwska, Lublin, Poland The quantitative relationships between capacity factor and solvent composition in reversed-phase systems for the rational optimization of both isocratic and gradient elution are given by the simple semiempirical equation which is a modified form of the equation derived by Soczewinski and Wachtmeister: L°9 *w,org 5lo'ki " b t org' where *P is the volume fraction of the polar modifier (O.Ol ж vol.%) and b is a constant empirically derived

from the system used (b - log к„ - log k0rg)s У and org denote components of the eluent-water and organic modifier. Coumarins, furocoumarins and pyranocoumarins

103 ternary solvent mixtures composed of water and organic modifiers, as methanol, acetonitrile, dioxane, propanol, The chromatographic results are presented as plots of Rj, or log k' vs percentage concentration of organic modi fier in the mobile phase. The plots are linear for almost the whole concenration range investigated, and spread fanwise owing to increased selectivity at higher water contents in the eluent, especially for less polar solutes as asthol and imperatorin, the compounds possesing the greatest hydrophobic properties due to large isopentenyl radical. The selectivity can be directly estimated from the plots (Alog к' =log kA - log k?). The sequence of solutes on RP-18 HPTLC plates and tor HPLC using Lichro- sorb RP-18 was found to be identical, however, for KP-2 plates there were some changes in the sequence. For iden tical adsorbents the correlation of HPLC and HPTLC reten tion data of investigated compounds is very good, but the use of the HPTLC-RP-2 data for correlation gave in terior results: a smaller correlation coefficient was obtained.

150 P.4-55 QUANTITATIVE DETERMINATION OF PSYCHOTROPIC DRUGS WITH HIGH PERFORMANCE LIQUID CHROMATOGRAPHY КОЛИЧЕСТВЕННОЕ ОПРЕДЕЛЕНИЕ ПСИХОТРОПНЫХ ЛЕКАРСТВ с помощью вета M.Przyborowska, Lublin, Poland Various external causes lead sometimes to disturbances in the normal functioning of the nervous system. The diseases are cured with the psychotropic drugs. Methods of quantitative determination of some derivatives of phenothiazine and 4-furobutyrophenone in blood were elabo rated. 1. 5 ml blood sample was taken from patients treated with chlorpromazine and extracted according to the elabo rated method; the extract was evaporated to dryness and dissolved in a known amount of heptane (100 - 200 ,ul). The sample was introduced into an alumina column (d^= 10 pm) and eluted with a saturated solution of acetonitrile in heptane. Nanogram quantities of chlorpromazine were determined from an analytical curve. 2. Quantitative determination of haloperdol was carried out using a column packed with octadecyl silica (LiChrosorb RP 18, B.Merck, Darmstadt, FRG). As eluent mixture of methanol and aqueous phosphate buffer, pH 8,2, was used. Analytical curves were determined for haloperidol and for fluphenazine as an internal standard. After extraction from blood samples, nanogram quantities of haloperidol were determined from peak heights and the analytical curves.

104 GAS CHROMATOGRAPHY

151 S.l-03 PHYSICO-CHEMICAL AND ANALYTICAL ASPECTS OF ADSORPTION PHENOMENA IN GAS-LIQUID CHROMATOGRAPHY ФИЗИКО-ХИМИЧЕСКИЕ АСПЕКТЫ АДСОРБЦИОННЫХ ЯВЛЕНИИ В ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ V.G.Berezkin, Moscow, USSR The advance of gas-liquid chromatography (GLC) is con nected closely with the use of new ideas about the roie of adsorption in chromatographic processes. The simplest model of sorbent in GLC contains throe phases:. 1) a sta tionary liquid phase that absorbs (dissolves) the substan ces being separated; 2) an interface of gas-stationary liquid phase that adsorbs the separated substances, and 3) an interface of stationary liquid phase - solid support that also adsorbs the separated substances. The development of an additive equilibrium retention theory that takes into account both adsorption and absorp tion of the compounds being separated reguires considera tion of the contribution of retention on individual bulk and surface phases of a real sorbent and experimental verification of the eguations obtained, including a check of this equation by the independent methods. Concepts of adsorption arid absorption mechanism of retention of the separated compounds in GLC have undergone revolutionary transformations in retention mechanism of GLC as a result of studies of adsorption phenomena by many scientists from several countries. Some of the most important results connected with the adsorption phenomena in GLC are listed below: 1. Development of gas chromatographic methods of stu dying the thermodynamics of interaction between the vola tile dissolved substance and non-volatile solvent. 2. Gas chromatographic studying of adsorption of vola tile compounds on the interphase of stationary liquid phase with support and gas phase. 3. New more universal values of retention and methods of identification of chromatographic zones in GLC. A, New classification of chromatographic methods taking into account the role of solid support. 152 S.4-5 THE PROBLEMS IN ANALYSIS OF TRACE ORGANIC TOXICANTS IN ENVIRONMENTAL OBJECTS ПРОБЛЕМЫ АНАЛИЗА МИКРОПРИМЕСЕЙ ОРГАНИЧЕСКИХ ТОКСИКАНТОВ В ОБЪЕКТАХ ОКРУЖАЮЩЕЙ СРЕДЫ P.E.Tulupov, ObninF' , USSR One of the peculiarities in analysis of trace organic toxicants in environmental objects lies in the fact that the information obtainable from the analysis can be only «05 used as reference data to evaluate the environmental qua lity (air, water, soil) and to predict the change in the quality, Therefore along with the problems due to choice of concentration conditions, sample preparation for the analysis, selection of chromatographic conditions, identi fication and quantitative analysis it is essential to evaluate the contributions from toxicants transport inside and between the adjacent environments (air-water-soil) and also the transformation into other compounds, which may appear to be of more ecological hazard in comparison with reference pollutants. The paper considers the concenration features for different organic toxicants from air, water and soil and their preparation for analysis as well as the findings of chromatographic analysis. Several compounds serve to illustrate the role of trasport and transformation pro cesses in assessment of environmental quality.

153 S.4-3 GAS CHROMATOGRAPHIC DETERMINATION OF NITROGEN- CONTAINING ANTIEPILEPTIC DRUGS AND OPIUM ALKALOIDS USING THERMOAEROSOL DETECTOR ГА30ХР0МАТ0ГРАФИЧЕСК0Е ОПРЕДЕЛЕНИЕ АЗОТСОДЕРЖАЩИХ ПРТ^ВОЭПИЛЕПТИЧЕСКИХ ЛЕКАРСТВЕННЫХ СРЕДСТВ И OlbliriHX АЛКАЛОИДОВ С ПРИМЕНЕНИЕМ ТЕРМОАЭРОЗОЛЬНОГО ДЕТЕКТОРА B.A.Rudenko, N.V.Lenchic, D.N.Dzhabarov, Moscow, USSR Gas chromatographic determination of antiepileptic drugs (phenobarbital, hexamidin, diphenalhydantoin and opium alkaloids (codein, morphin, papaverin, narcotin) was carried out using selective to nitrogen-containing compounds thei.~.waerosol detector. The limit of detection was Ю-10- 10"11 g which is circa factor 1000 less than results, obtained with the use of flame ionization detec tor. On determination of 10~9- 10-1°g of the above men tioned compounds the relative standard deviation was 0,15-O,2 (n = 7). The results achieved allowed to dimi nish the biosubstrat (blood plasma) volume to 0.1 - 0,2 ml that is of importance in pediatry. The considerable simplification of analysis technique and the shortening of time and labour expenditure was achieved due to GC-determination without derivatization with use of relatively short glass packed columns (0,3 m in length) and fused silica open tubular columns (10 m length) coated with OV-17 in isothermic conditions at 260°C or with temperature programming. Inerton N AH DMCS with 5 * of SE-30 was used in packed columns. The analysis time was 3 ir.in with packed and 10 min with open tubular columns.

104 154 S.6-1 GAS CHROMATOGRAPHY IN RESEARCH OF KOSHOS ГАЗОВАЯ ХРОМАТОГРАФИЯ В КОСМИЧЕСКИХ ИССЛЕДОВАНИЯХ L.M.Muchin, V.N.Khokhlov, Moscow, USSR Abstract not arrived on time for printing 155 S.6-5 DETERMINATION OF ADMIXTURES BY CHROMADISTILIATION ХРОМАДИСТИЛЛЯЦИОННОЕ ОПРЕДЕЛЕНИГ ПРИМЕСЕЙ S.M.Yanovskit, Moscow, USSR Chromadistillation (CD) of admixtures provides good op- portunutuies for the introduction of large samples into the chromatographic column. Analysis of the mechanism of process which take place in the entrance of the column (including "solvent effect" in capillary columns) shows that all the methodical ver sions of CD are present: the thermal, restrictive, iso thermal elution, etc. On the basis of CD a number of methods was elaborated for the analysis of admixtures in water and some solvents. By direct introduction of 2 cirr of a liquid sample into the preliminary column, packed with an inert mater**!, it is possible to analyse about 0,005 mg/1 of n-alkanes Сб-С« in water. Extraction from water by a volatile sol vent (hexane) permits to analyse there petroleum products

• up to C21 alkanes. Compression of the admixtures on the boundary between two major components of the water-ethanolic mixtures, permits to separate without deterioration of the efficien cy of separation and to transfer to the chromatographic column (packed or capillary) light (Cj-C-) or «юге heavy (to C,,) alcohols. The method was applied for analysis of pollutants in water with concentrations below the permissible limit (0,1 mg/1) as well as the admixtures in wines and products of fermentation. Application of circulation CD permits to introduce samples up to Ю ml. 156 S.7-2 THE SEPARATION AND IDENTIFICATION OF POLYCYCLIC HYDROCARBONS BY GAS ADSORPTION CHROMATOGRAPHY РАЗДЕЛЕНИЕ И ИДЕНТИФИКАЦИЯ ПОЛИЦИКЛИЧЕСКИХ УГЛЕВОДОРОДОВ МЕТОЛОМ ГА30-АДС0РБЦИ0НН0Й ХРОМАТОГРАФИИ E.V.Kalashnikova, V.I.Nazarova, R.S.Petrova, K.D.Shcherbakova, Moscow, DSSR Among polycyclic hydrocarbons there are many structural isomers *ith similar physical properties which are diffi cult to separate by gas liquid chromatography. At the same time high sensitivity of adsorption on graphitized thermal carbon black (GTCB) to the geometrical structure of adsor- 107 bate molecules allows this adsorbent to be successfully applied in gas adsorption chromatography for the separa tion of stereoisomers. But usual packed columns are not efficient enough if the mixtures to be chromatographed consist of a large number of isomers with near boiling points, such as compounds with three fused rings. There fore capillary columns packed with CTCB have been used to separate these isomers. The investigated families of isomers include the deri vatives of bicyclo(2.2.1)heptane and the hydrocarbons with molecules consisting of three fused cyclohexane rings or one cyclopentane and two cyclohexane rings (perhydrophe- nanthrene, perhydroanthracene, perhydrofluorene, perhydro- phenalene and cyclopentanodecalir,) and of four rings (steranes). The sequence of elution of the isomers from' a column with GTCB has been ascertained on the basis of their structure. It has been shown that the identification of chromatographic peaks can be made with the aid of mole cular statistical calculation of retention volumes, pro vided that the geometry of the isomers is known. On the other hand, if the identification is already accomplished by other methods, then the comparison of experimental and calculated results can help to estimate the true geometry of the molecule.

157 S.7-3 GC ANALYSIS OF WATER MISCIBLE SOLVENTS VIA DIRECT AQUEOUS INJECTIONS АНАЛИЗ СМЕШИВАЮЩИХСЯ С ВОДОК РАСТВОРИТЕЛЕЙ МЕТОДОМ ГАЗОВОЙ ХРОМАТОГРАФИИ ПУТЕМ ПРЯМОГО ВВОДА ВОДНОГО РАСТВОРА K.Lakszner, L.Szepesy, L.Pcdmaniczky, Budapest, Hungary The major pollutants in pharmaceutical and distillery waste waters are the miscibls, low molecular weight vola tile solvents such as alcohols and ketones. For the deter mination of the above water pollutants mainly static or dynamic headspace methods are being used. The very thick film (5-10 ,ul) recently developed immo bilized nonpolar capillary columns offer new possibilities in the analysis of aqueous samples. Due to immobilization, very large water samples can be injected without damaging the stationary phase film and thick films permit satisfac tory separations also at higher temperatures. This paper presents the results of an investigation aimed at the quantitative determination and the detection limits of such solvents with direct aqueous injection techniques. Direct injection techniques have been compared with head space analysis and other techniques as regards the limit of detection and ease of use for routine analysis of water pollutants

toe 158 S.7-1 CHROMATOGRAPHIC STUDY OF DITHIOCARBAMATES ХРОМАТОГРАФИЧЕСКОЕ ИССЛЕДОВАНИЕ ДИТИОКАРБАМАТОВ J.Garaj, Bratislava, USSR Dithiocarbamates (DTC) have been commonly used not only in analytical chemistry but also in industry and in agri culture as an important group of agricultural fungicides that are used on seeds and crops throughout the growing season. With their application a problem of elaboration of methods of identification and determination of their resi dues in environment has emerged. Besides.DTC residues, mainly their degradation products have been placed into the class of hygienically hazardous substances. They may produce goiterogenlc, oncogenic and teratogenic effects after being applied to laboratory animals. In the analysis of DTC residues asd their degradation products in the environment, crops and food chromatogra phic methods (GC, HPLC, TLC) predominate. The methods are reliable in ppm level using either a precleaning step and preconcentration step (extraction), or several methods require deriva'tization step prior to analysis. Derivatiza- tion procedure» significantly increase the sensitivity. Chromatographic methods have been also used for the study of chemical transformation of complexes of transi tion metals with dithiocarbamates as ligands due to the exchange of ligands and central atoms. The results were compared with those obtained by other physical methods.

159 S.12-2 IMPROVEMENT OF GC IONIZATION DETECTORS СОВЕРШЕНСТВОВАНИЕ ИОНИЗАЦИОННЫХ ДЕТЕКТОРОВ ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ J.Gregoire, Suresnes, France By means of some results from the important research work we have been doing In order to develop the DI 700 new series of GC apparatuses, we show the way to improve a flame ionization detector (FID) and an electron capture detector (ECD), FID: we justify the importance of the efficiency of oxlge- nation, and we introduce the involved improvements as to sensitivity, linearity, value and constancy of response factors, especially In the case of faintly ionisated compounds at trace levels. ECD: we introduce the method allowing the characteristic curve of an electronically linearisated ECU to be studied (frequency versus, reference intensity). We give the physical meaning of the slope and of each particular point of this curve. He explain the Involve J genesis of the main Improve ment (patent -pending): no change in sensitivity versus modification of emission ability of the N1 63 source. This modification is caused by the source pollution. 109 160 S.12-3 DETERMINATION OF INORGANIC GASES USING NOBLE GAS DETECTOPS AND SPECIAL MODIFIED ELECTRON-CAPTURE DETECTORS (ECD) ОПРЕДЕЛЕНИЕ НЕОРГАНИЧЕСКИХ ГАЗОВ С ПРИМЕНЕНИЕМ ДЕТЕКТОРОВ С БЛАГОРОДНЫМИ ГАЗАМИ И МОДИФИЦИРОВАННОГО ДЕТЕКТОРА ЭЛЕКТРОННОГО ЗАХВАТА Р.Рорр, G.Oppermann, Leipzig, GDR In many fields e.g. air pollution measurements or puri ty control of gases, the determination of small concentra tions of inorganic compounds is necessary, but the detec tion is complicated by the fact that the sensitivity of the thermal conductivity 'detectors is not high enough to ensure detection limits below 1 ppm. For such problems noble gas detectors (in first line helium detectors) with detection limits within the ppb-range are usable and for some substances the use of special modified electron-cap ture detectors is favourable. The advantages and disadvan tages of the helium detector and the analytical possibili ties of ECD modifications like electron attachment spect roscopy are discussed. Experimental results of 02, N2, H2,

CO, C02, N20 trace determinations in pure gases with a helium detector developed at the Central Institute for Isotope and Radiation Research of the Academy of Sciences of the GDR and results of 02, N20 or S02 measurements with ECD modifications are given. 161 S.12-4 MULTIPURPOSE COLD INJECTOR FOB HIGH RESOLUTION GAS CHROMATOGRAPHY МНОГОЦЕЛЕВОЙ ХОЛОДНЫЙ ИНЖЕКТОР ДЛЯ ГАЗОВОЙ ХР0МАТ0ГРА4ИИ ВЫСОКОГО РАЗРЕШЕНИЯ C.A.Saravalle, F.Kunari, S.Trestianu, A.Sironi, Milano, Italy The cold injector presented here represents an original approach towards a multipurpose sampling system designed to overcome the injection discrimination problems over a virtually unlimited range of matrices analyzed by high resolution gas chromatography. Unlike other systems based on cold split/splitlese hardware, the injector proposed is designed on a modular concept to make it ideally complementary to the already well established cold on-column injector with secondary cooling introduced and patented by Carlo Erba Strumentazio- ne in 1978. The hardware is in fact very easily fittable to the bottom of the cold on-column injector and its design makes it ideal for cold split/ splitless injections. Basically the system contains a short specially desig ned and treated, precolumn, a splitting line and also provides the possibility for em optional auxiliary carrier gas line for solvent back flush.

110 This multipurpose cold injector is based on a 1980 patent by Carlo Erba Strumentazione which quotes as follows: "the temperature of the precolumn representing the injection zone is controlled such that the sample remains in the liquid state until the injection is comp leted and then a controlled heating and vaporisation of the sample is achieved in order to transfer it totally (splitleBS mode) or partially (split mode) inside the capillary". The injection zone is independently temperature cont rolled from that of the column oven: it is cooled before the sample introduction by the built-in line of the on-column injector secondary cooling and it is quickly electrically heated to accomplish the total sample transfer into the column. Besides manual injections, the system is fully compa tible with the recer.tly developed autosampler for cold on-column injections. In its standard configuration it allows sample sizes ranging from 0,2 to 10 /ll to be auto matically injected even under remote computer control. Typical examples illustrating performance of the system are reported. 162 S.12-5 SOME ASPECTS OF THEORY AND PROSPECTS OF DEVELOPMENT OF IONIZATION GAS CHROMATOGRAPHIC DETECTION AND IDENTIFICATION OF SUBSTANCES НЕКОТОРЫЕ ВОПРОСЫ ТЕОРИИ И ПЕРСПЕКТИВЫ РАЗВИТИЯ ИОНИЗАЦИОННОГО ГА30ХРОМАТ0ГРАФИЧЕСКОГ0 ДЕТЕКТИРОВАНИЯ И ИДЕНТИФИКАЦИЯ ВЕЩЕСТВ V.A.Rotin, B.G.Gelman, Moscow, USSR The problems of the theory of detection on the basis of Penning effect in helium and neon are considered in the report. It is shown that the abnormal weakening of Penning effect caused by substances with relatively high potentials of ionization: hydrogen, argon, nitrogen and 'sulphur dioxide are observed in neon when operating in saturation current mode. It is determined that this phenomenon is observed in high purity inert gases after their contamination with some substances: organic com pounds, moisture etc. It is indicated that this phenomenon is directly connected with the processes of the inert gas excimer molecules formation and their extinguishment when colliding with the molecules under detection. The mentioned regularities are used for the elaboration of the methods or the quantitive interpretation of chromatograms and identification of substances during chromatographic analysis of the Venus atmosphere. The problems of the identification of substances on the basis of the regularities of flarae-ionization detection are considered. The possibility to use the ionization efficiency dependence of the content and structure of organic molecules for identification of the classes of compounds and individual substances is shown.

Ill 163 P.1-04 THE ALGORITHM OF GC/COMPUTER IDENTIFICATION OF SULFUR CONTAINING COMPOUNDS IN MIXTURES ON THE BASE OF GC-DATA АЛГОРИТМ ГХ/ЭВМ ИДЕНТИФИКАЦИИ СЕРОСОДЕРЖАЩИХ СОЕДИНЕНИЙ В СЛОЖНЫХ СМЕСЯХ D.N.Grigoryeva, T.A.Misharina, R.V.Golovnya, Moscow, USSR The identification of components in complex mixtures on the base of sorption-structure correlations may be carried on by computer. In the present paper the algorithm of GC/ computer identification of sulfur containing compounds in the complex mixtures is given. The algorithm consists of two parts. The uniform equa tions for calculating of retention indices of the 28 homo logous seria of mercaptanes, sulfides, JL, (t)-dl (alkylthio) alkanes, esters of thioacids obtained on stationary phases Apiezon M, OV-17, Triton X-305 at 130°C are used for iden tification of volatile compounds of natural objects. The algorithm includes an information retrieval system for determination of indices of substances not belonging to the homologous eeria obtained on the same column. The criteria of reliable GC/ computer identificetior of sulfur containing compounds were elaborated. The reliabi lity of GC/computer identification increases If the GC/ data obtained for separated fractions were used. The relia bility of the algorithm was tested on a special mixture of 23 components which are difficult to separate: mercapta nes, sulfides, disulfides, thloacetates, thiophenes, dithianes, dithiolanes and furanes. 164 P.1-5 A THEORY OF CHROMATOGRAPHIC DETERMINATION OF ADMIXTURES ТЕОРИЯ ХРОМАТОГРАФИЧЕСКОГО ОПРЕДЕЛЕНИЯ ПРИМЕСЕЙ V.I.Kalmanovsky, Moscow, USSR Two situations occurring in the course of admixtures determination have been considered: (1) Peaks of interest are amplitude-comparable with the neighbouring peaks on chromatogram; (2) Peaks of admixtures elute "on the slope" of interfering component peak. Mathematical models have been offered. The first case is defined as superposition of two individual peaks each being described by the difference of two probability inte grals. The second case is represented by superposition of a similar Individual admixture peak and exponential slope of interfering component peak. The model parameters are: relative content of the component to be determined, volume of analysed sample, as well as separation characteristics which include resolution R and dispersion ratio for the first model and ratio of time constants for the second model. The validity of the models is confirmed by the ex perimental results. The notion of detection limit constricted by insuffi cient separation has been introduced. The possibility is 112 discussed to define it as concentration at which the infle xion point of output curve vanishes, or as concentration at which the derivative at the inflexion point is equal to zero. The relationships between detection limit, degree of broadening, quality of separation and model parameters have been presented for both cases. The sufficient separa tion criterion has been established which provides the realization of the detection limit for the detection system employed. The results are supported by experiments using test mix ture within concentration range Ю"' to 10~- % (Uy mass) .

165 P.1-06 SORPTION REGULARITIES OF AMINO COMPOUNDS IN STATIONARY PHASES CONTAINING COMPLEXING AGENT ИССЛЕДОВАНИЕ СОРЙШИ АМИНОСОЕДИНЕНЛИ В УСЛОВИЯХ К0МПЛЕКС00БРА30ВАНИЯ В НЕПОДВИЖНОЙ ФАЗЕ A.A.Anderson, P.A.Mekss, M.V.Shymanska, Riga, USSR A method of simultaneous evaluation of the sorbate complex formation constants K* and coefficients of parti tion in the bulk of stationary liquid phase KL and of adsorption at phase boundaries KGL and Kg has been propo sed. Sorption characteristics of aliphatic, aromatic ami nes and nitrogen-containing heterocycles in Apiezon M containing additions of substances being able to form the weak complexes with sorbate (octadecanol, methyl stearate) have been evaluated. In the completing process with octadecanol and methyl stearate amines act as electron donors (correlation of K* with values of ionization potentials or energies of HOMO of amines). Nevertheless the influence of electron-accep tor activity of sorbate molecules on K* is observed in some cases (dependence of K* values on H atom number in amino group or on energies of LVMO), Quantum-chemical calculations confirm the possibility of formation of the donor-acceptor bonds of the both types. Partition coefficients of amines in the bulk of sta tionary liquid phase are correlated with molecular mass and the Palm's sterical constants. Interfacial sorption of amines in the systems under study occurs to be essentially suppressed due to the superficial activity of the polar component of stationary phase and to the modification of supports surface by the latter molecules.

8. 3IK. 872 113 166 P.1-07 GAS CHROMATOGRAPHY OF AROMATIC COMPOUNDS! INFLUENCE OF MOLECULAR ORBITAL PARAMETERS ON RETENTION ГАЗОВАЯ ХРОМАТОГРАФИЯ ОРГАНИЧЕС-"Т< СОЕДИНЕНИЙ: ВЛИЯНИЕ ПАРАМЕТРОВ МОЛЕКУЛЯРНЫХ Л>БИТАЛЕЙ НА УДЕРЖИВАНИЕ A.Garcla-Raso, F.Saura-Calixto, J.Garcia-Raso, Palma de Mallorca, Spain M.A.Raso, Madrid, Spain The gas chromatographic behaviour of aromatic compounds was studied by the authors previously, considering para meters such as Van der Waals volume and connectivity index /1/. On the other hand, the usefulness of molecular orbital parameters to the study of chromatographic reten tion-chemical structure relations were previously reported /2,3/. In the present communication the influence of molecular orbitals on gas chromatographic retention of alkylbenzenes and monoeubstltued benzenes is considered.

Values of total energy, Et, binding energy, Eb, energy of highest occupied molecular orbital, EHOMO, energy of lowest unoccupied molecular orbital, Едодо' electronic energy, Ee, charge over the aromatic ring, Qc6, and interaction energy, E, defined by Klopman's equation /4/ were calculated by the complete neglet of differential overlap (CND0)/2. Total energy or binding energy are the principal factors determining the GLC retention of alkylbenzenes, as can be observed in the following equation:

I - 244.33 + 68.64 Eb (I - Kovats retention indices on squalane at 86°C) N (number of compounds used) « 31; r (correlation coeffi cient) - 0.980) s (standard deviation) » 24.ОЗ.

Similar results were obtained for I versus Et. In order to yield higher correlation and lower stan dard deviation, other molecular parameters such as Ee, EH0M0 and ELUM0' besides E. or &., were considered. Hiis equation is not applicable to monosubstitued benzenes y-X (X - F, CI, NH2, N02, CN, CHO, COOEt, CH2C1,

CH2-COOCH3, CH2CN,CH2-CH2OH, С(«СН2)-СНз, СН2-ОСНз, CH20H). In this case is necessary to consider electrostatic parameters:

I - 563.72 + 0.60 E_ - 2062.74 ELUMO - 2156.88 Qc6 N - 14 r - 0.903 s - 57.79 This expression can represent .or explain the general chromatographic behaviour of these compounds, but without prediction or analytical purposes. That is to say, chromatographic retention of alkylbenze nes is directly related to Eb, while the values of charge over the aromatic ring is the most important factor for f -X compounds. 1. F.Saura-Calixto, A.Garsia-Raso, Intern. J, Environ. Anal. Chem., 17,279, 1984. 2. A.Garcia-Raso, F.Saura-Calixto, M.A.Raso, J. Chromatogr,,3£2, 107, 1984. 114 3. L.Buydene, D.L.Massart, P.Geerlings, Anal. Chem.,55_, 738, 1983. 4. G.Klopman, J. Am. Cham. Soc., 20., 223, 1968.

167 P.1-12 RELATIONSHIPS BETWEEN HEATS OF FORMATION AND GAS CHROMATOGRAPHIC RETENTION FOR ORGANIC COMPOUNDS ВЗАИМОСВЯЗЬ МЕЖДУ ТЕПЛОТОЙ ОБРАЗОВАНИЯ И ГАЗОХРОМАТОГРАФИЧЕСКИМ УДЕРЖИВАНИЕМ ОРГАНИЧЕСКИХ СОЕДИНЕНИИ F.Saura-Calixto, A.Garcia-Raso, J.Garcia-Raso, Palma de Mallorca, Spain The binding energy, Еь, is a determining factor in chro matographic retention. Ед, represents the difference bet ween the total energy ot the compound and the sum of the energy of all constituents atoms; for it retention indices, I, must be related to the heat of formation of the solute, &Hf, /1/, In a previous paper the authors studied these correlations for aliphatic hydrocarbons and alkylbenzenes /2/. On the other hand, it is well known that molecular topo logy determines molecular and thermodynamic properties. Recently, information theory provides molecular values having greater discriminatory power for isomers than the known topological indices /3,4/. The purpose of this communication is to study I-AHf relationships for alkylnaphthalenes and oxygen-containing organic compounds, considering information theory in order to improve correlations. Experimental values of I were plotted versus ДН* corre sponding to the gas state, expressed in К cal/mol /5/. Straight lines with high correlation were obtained for homologous series in all cases. To improve correlations and to obtain more general equations including larger number of isomers the mean information index of distances, 1^, is introduced. Some equations obtained were: Alcohols: I - 54O.01 + 9.91 ДН* + 28.45 1° + 158.47 n N « 13 r « 0.999 s = 14.86 " Esters (linear and branched methyl, ethyl, propyl and bu- tvl esters)*

I • 454.37 + 3.08 &Hf + 197.91 ij* + 69.04 n N = 19 r " 0.984 s = 41.77 Aldehydes and Ketones: „

I = -220.02 - 5.44 AHf + 190.98 I~ + 18.60 n N - 11 r = 0.997 s - 12774 where n = number of carbon atoms of the compounds; N * number of compounds employed; r * correlation coeffi cient and s « standard deviation.

115 All compounds with дН, available in literature were employed. Alkylbenzenes and alkylnaphthalenes are also considered. I values correspond to polar and non-polar stationary phases.

uHf values are important in basic research and techno logy, but few data i>re known for organic compounds. Expres sions of this type could be used for empirical predictions. 1. F.Saura-Calixto, A.Garcia-Raso, M.A.Raso, J.Chromatogr. Scl.. 22. 22, 1984. 2. F.Saura-Calixto, A.Garcia-Raso, J.Garcia-Raso, J.Chromatogr. (in press)i 3. D.Bonchev, "Information Theoretic Indices of Characte rization of Chemical Structures", Research Studies Press, John Wiley 4 Sons, 1983. 4. N.Trinajstic, "Chemical Graph Theory" C.R.S. Press,1983. 5. D.R.Stull, E.F.Weetrum Jr., G.C.Sinke, "The Chemical Thermodynamics of Organic Compounds", John Wiley 4 Sons, 1969.

168 P.1-08 RELATIONSHIPS BETWEEN RETENTION CHARACTERISTICS AND MOLECULAR WEIGHT OF THE STATIONARY PHASES IN GAS-LIQUID CHROMATOGRAPHY ЗАВИСИМОСТЬ ХАРАКТЕРИСТИК УДЕРЖИВАНИЯ ОТ МОЛЕКУЛЯРНОГО ВЕСА НЕПОДВИЖНЫХ ФАЗ В ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ A.N.Korol, G.V.Fllonenko, Kiev, USSR Relative retention data and relative molar heats of solution of paraffins, alcohols and ketones were determi ned on the hydrocarbon (for paraffin solutes) and polyethy lene glycol (for polar solutes) stationary phases of diffe rent molecular weight. Two types of the relationships were obtained: i.e. the linear relationship for the non-polar systems (between log r and 1/M) and for the polar systems (between r and 1/M). These relationships were confirmed by the retention data taken from McReynolds handbook. These relationships may be applied in order to calculate retention data when using the stationary phases with diffe rent molecular weight within the homologeous series. Taking into account' that polyethylene glycol stationary phases are manufactured by different companies and that the products have different molecular weight distribution the relationship between relative retention and 1/M aids to differentiation of origin of the polyethylene glycol stationary phase. The reasons of the different relation ships between relative retention and 1/M (reciprocal molecular weight) are discussed. Taking into account these relationships some rules are formulated in order to obtain the interlaboratory reproduced retention data for the polyethylene glycol and paraffinic stationary phases.

116 169 P.1-09 CHARACTERIZATION OF LIQUID PHASES IN GAS-LIQUID CHROMATOGRAPHY BY THE RETENTION OF NEIGHBOURING n-ALKANES ХАРАКТЕРИСТИКА ЖИДКИХ ФАЗ В ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ ПО УДЕРЖИВАНИЮ СОСЕДНИХ Н-АЛКАНОВ R.I.Sidorov, MOSCOW, USSR The ability of molecules of stationary liquid phases (SLP) in gas-liquid chromatography (GLC) to paticipate in intermolecular interaction between each other is considered as the evidence of their chromatographic polarity. Relative

retention of neighbouring n-alkanes t' z+i/t*R , = ti^ 2 (t'Cn2 is the retention index of methylene group) is used to determine this polarity. Chromatographic polarity, #CH2 *s estimated from the equation ^«t'cHj'sq " ^'cH^ph 9Гсн„ " loo ^'cH^sq where subscripts "sq" and "ph" correspond to squalane and polar phase respectively. It is shown that бГсНз te a

f1 - effective content or polar-'groeps in SLP molecule, A.. • their interaction energy. MCH2 valuee for several organic SLP are presented in the paper. The obtained polarity values correlate with Гд1values. The advantages of the method are: clearness of physical sense, low labour consumption, sufficient accuracy, actual independence of analysis temperature and compliance with the notion of liquids polarity. Dependence on GLC adsorption effects and on SLP mole cular mass may be referred to as shortcomings of the method. In some cases W сНэ value» ваУ be used for a priori estimation of retention values for compounds with an error, not exceeding Ю Kovatch index units.

170 P.L-IO STRUCTURE-RETENTION CORRELATIONS OF ISOMERIC DECAHYDRO- AND PERHYDROPYRENES IN GLC СВЯЗЬ СТРУКТУРЫ И УДЕРЖИВАНИЯ В ГХХ ИЗОМЕРНЫХ ДЕКАГИДРО- И ПЕРГИДРОПИРЕНОВ W.Engewald, R.Herzschuh, G.Mann, Leipzig, GDR There exist two constitutional isomers I and II of decahydropyrene with 3 and 2 different configurations, respectively: /—i /—-i

b cP. a 117 With respect to some unresolved problems on the struc ture of the configurational isomers we studied this prob lem by means of selective hydrogenation of pyrene and hexahydropyrenes and investigation of the reaction pro ducts using GC retention, GC-MS and 13C-NMR, The retention indices were measured on a glass capillary column coated with methylsilicone OV-1, It was established that the retention sequence of all 5 stereoisomeric decahydropyrenes which includes a range of about 100 i.u. can be explained by considering both the degree of substitution of the benzene ring and the stereo- structure of the saturated rings: All isomers derived from the constitution I which corresponds a trialkylated ben zene have 1- - sr retention values than the isomers of the constitution II (tetraalkylated benzene). In the case of the isomers with the constitution I the retention depends on the configuration of the bridged carbon atoms 8, 12 and 16. For instance, the stabilomere in which all of these carbon atoms possess equatorial C-C - connections, are eluted at first. In the case of the isomers with the constitution II the retention depends on the arrangement of the carbon atoms 5 and 8, The isomer with the 1.4-trans arrangement of the C-C - bond are eluted before the isomer with the 1.4-cis arrangement. The hydrogenation of decahydropyrenes under kinetic or thermodynamic controlled conditions yields different mixtures of the stereoisomers perhydropyrer.es, the reten tion of which has aleo been measured and correlated to their stereochemistry,

171 P.1-11 GAS CHROMATOGRAPHIC RETENTION AND SOLUTION THERMODYNAMICS OF n-ALKANES IN POLYDIMETHYLSILOXANE OV-lOl ГАЗОХРОМАТОГРАФИЧЕСКОЕ УДЕРЖИВАНИЕ И ТЕРМОДИНАМИКА РАСТВОРЕНИЯ Н-АЛКЕНОВ В ПОЛИМЕТИЛСИЛОКСАНЕ OV-lOl S.A.Rang, T.V.Strenze, K.R.Kuningae, Tallinn, USSR The specific retention volumes, retention indices and retention index increments of 77 Cg- Сц n-alkenes were determined on OV-101 and correlated with the molecular structure of isomers. Equations are presented which des cribe the dependence of retention parameters on column temperature, on the carbon number of isomers in a homolo gous series and on the physico-chemical properties of sorbates. The thermodynamic characteristics of solution (partial molar free enesgy, enthalpy and entropy) were calculated and relationships between these data and the chemical structure of n-alkenes investigated. A number of common regularities in the gas chromato graphic behaviour of n-alkenes on OV-lOl and squalane have been found, e.g., the elution of isomers is in agree ment with the shift of the double bond from the center to the second carbon atom of the chain in a given series. 118 For pairs of cis- and trans-configuration the elution order depends on column temperature, the position of the double bond end the chain length of the molecule. Diffe c rent from squalane, C10" li 1-alkenes are eluted first from the OV-101 column followed by other isomers. The increase in retention by 5-11 index units when changing squalen» to OV-101 is related to the increase of the intermolecular interaction of n-alkenes with OV-101. The retention indices measured and their correlations were used in the analysis of high purity chemical reagents, products of organic synthesis and various technological products. Retention indices were measured on glass capillary column (50 m x 0.25 mm), at 12 different temperatures within the range of 30-140°C, the linear rate of the carrier gas (helirm) was 16,7 cm/s, the split ratio ca 1:200. Specific retention volumes were measured on glass column (2.5 x 3 mm) with 8 % OV-101 on Chromaton N Super, the carrier gas(helium) flow rate was 35, ml/min. The authors are indebted to Dr. A.E.Korol for delive ring the column packing with OV-101. 172 P.1-13 RELATIONSHIP BETWEEN THE GAS CHROMATOGRAPHIC RETENTION INDICES AND THE PHYSICO-CHEMICAL PROPERTIES OF AROMATIC HYDROCARBONS AND THEIR NITROGEN DERIVATIVES ВЗАИМОСВЯЗЬ МЕЖДУ ХРОМАТОГРАФИЧЕСКИМИ ИНДЕКСАМИ УДЕРЖИВАНИЯ И ФИЗИКО-ХИМИЧЕСКИМИ СВОЙСТВАМИ АРОМАТИЧЕСКИХ УГЛЕВОДОРОДОВ И ИХ А30ТПРОИЗВ0ДНЫХ V.M.Nablvach, V.A.Geraslmenko, Yu.S.Berlizov, V.P.Dmitrikov, Dnepropetrovsk, USSR The systematic study of the relations between the reten tion indices and the molecular structure allows to evalua te both the physico-chemical properties and the structural peculiarities of an analysed substances from the gas chromatographic data as well as the intermolecular interac tions in gas-liquid chromatography. These investigations were carried out on the packed and the capillary columns with the stationary phases of va rious chromatographic polarity at lOO -180°C temperature range. The Xovats retention indices both more than 80 homologues of benzene, naphthalene, Ыphenyl and 5C alkyl derivatives of pyridine and quinoline were measured and their structural increments were calculated. It was shown that retention index values and the elu tion sequence of isomeric hydrocarbons are defined by the steric factors, which depend on the length and the mutual position of the alkyl groups. The retention index incre ment depends on the stationary phase polarity and the screening degree of the nitrogen atom by the methyl group in the pyridine and quinoline molecule. The selective separation of these isomeric substances is achieved by the formation of the hydrogen bonds between the hydroxyl- containing phases and the nitrogen of the heterocycles.

119 The correlation equations of the retention index and various physico-chemical parameters (boiling point, mole cular refraction, ionisation potential, logarithm of the dissociation constant and oth.) of the tested substances were found. The obtained data were used for an investiga tion of both the coal-tar nitrogen bases and the isola ted distillates. 173 P.1-16 THE STUDY OF REACTION OF TRANSMETALLIZATION WITH BIS(PENTAMETHYLDISILANIL)CARBODIIMIDE ИССЛЕДОВАНИЕ РЕАКЦИЙ ПЕРЕМЕТАЛЛИРОВАНИЯ С УЧАСТИЕМ БИС/ПЕНТАМЕТИЛДИСИЛАНИЛ/-КАРБОДИИМИДА A.C.Gordetzov, O.P.Chereshnya, S.E.Scobeleva, E.A.Mishina, Gorky, USSR By liquid chromatography and UP.-spectroscopy the reac tion of bis(pentamethyldisilanil)carbodiimide with diffe rent elementorganic substances or' IV В group was inves tigated. It was shown, that the high efficiency of separation and the possibility to realise the reaction at mild conditions permits to investigate the complex mixtures of products of reaction of carbodiimide with thrimethyl- silyl-, thriethylsilyl-, thrigermil-, thributylstanile-, halogenides, oxides, isocyanates and isothiocyanates. The use of high performance liquid chromatography and UR-spectroscopy gave the information about a direction of reactions of transmetallization. 174 P.1-18 PYROLYSIS GAS CHROMATOGRAPHIC ANALYSIS OF AEROSOL PARTICLES ПРИМЕНЕНИЕ ПИРОЛИТИЧЕСКОИ ГАЗОВОЙ ХРОМАТОГРАФИИ ДЛЯ АНАЛИЗА АЭРОЗОЛЕЙ E.E.Sotnikov, Moscow, USSR Pyrolysis gas chromatography was used for rapid recog nition of specific chemical composition and identifica tion of kind of aerosol particles. The pyrolyser was an electrically heated quartz tube packed partially with filtering medium. Investigated air passed through filter bed and aerosol particles precipitated on filter. Then nitrogen or helium passed through tube, gas flow was stopped and pyrolysis was carried out at 70Ooc in statical conditions. After completion of pyrolysis the volatile components of the degradation products directed to the chromatographic column. Different filtering medium was studied: powder quartz, porcelain, molecular sieves(particle size 0.25-0.5 mm). The best filtering medium for precipitation of solid aero sol is powder quartz and for £. recipitation of liquid

aerosol is the powder molecular sieves. The% effect!vness of the precipitation of cotton, wool and meal aerosols (particle size 5-20 .Aim, air flow-rate 3-5 1/min and 120 quartz powder bed thickness 10 mm) was equal 95 %. The selective combined detector (SCD) was used for group identification of the pyrolysis products. SCD is the combination of flame-ionization (FID), flame-photometric (FPMD) and negative superficial ionization (NSID) detec tors. The relation of the peak heights recorded from three registration channels provided information about the class of the substances: NSID is a selective detector for nitro gen and halogen compounds and FPMD for sulfur, phosphorus and nitrogen compounds. This information facilitates recognition of a kind of aerosol particles. The examples of determination of a kind of a dust in air and dustness are discribed. 175 P.1-19 STUDY OF THERMAL DEGRADATION OF POLYMERS BY CORRELATION CHROMATOGRAPHY ИССЛЕДОВАНИЕ ТЕРМОДЕСТРУКЦИИ ПОЛИМЕРОВ МЕТОДСМ КОРРЕЛЯЦИОННОЙ ХРОМАТОГРАФИИ E.Kullik, M.Kaljurand, Tallinn, USSR Correlation chromatography may be used to determine the low-temperature thermal degradation kinetics of ther mally stable polymers. While other techniques (sample concentration or single injection) fail, correlation chromatography can provide valuable information about the thermal behaviour of polymeric materials at temperatures 100-5CO°C where the degradation product evolving rate and detector signal level is low. Correlation chromatography is applied for investigation of the thermal degradation kinetics of polyimide film at temperatures below 450°C. Due to its detector noise sup pression property correlation chromatography appears to be a reasonable alternative to single injection or sample concentration. 176 P.l-20 ANALYSIS OF COMPOSITE MATERIALS USING PYROLYSIiS - GAS CHROMATOGRAPHY ПРИМЕНЕНИЕ ПИРОЛИТИЧЕСКОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ ДЛЯ АНАЛИЗА КОМПОЗИЦИОННЫХ МАТЕРИАЛОВ K.V.Alexeeva, L.S.Solomatina, Moscow, USSR The analysis of composite synthetic polymer materials by pyrolysis-gas chromatography is described. "Biochrom- 26" chromatograph with a filament pyrolyzer and a flame ionization detector was used in the investigations. Depending on the analytical task, both flash pyrolysis of composite material samples and stepwise heating followed by chromatographic separation of released pro ducts were employed. The technique used in the experiments makes it possible to simultaneously determinate separately synthetic poly mers in any proportions and high-boiling ingredients (antioxidants, plasticizers, etc.) in composite materials. 121 The detected individual characteristic pyrolysis pro ducts enable the determination of the polymer type in the presence of another high-molecular-weight compounds and mineral fillers. The established quantitative ratios of characteristic pyrolysis products are indicative for individual grades of a polymer in samples of a complex composition. The possibility of the determination of minor quantities of polymers and non-polymer additives and poly mer impurities in composite materials on a ~ 1 % detection level was demonstrated. Examples of a simultaneous determination of synthetic polymers and ingredients in composite polymer materials are presented.

177 P.l-21 APPLICATION OF STAGE HEATING FOR GAS CHROMATOGRAPHIC ANALYSIS OF RESIDUAL SOLVENTS AND IDENTIFICATION OF ELEMENT COMPOSITION OF POLYMER ПРИМЕНЕНИЕ СТУПЕНЧАТОГО НАГРЕВА ДЛЯ ГА30ХР0МАТ0ГРАФИЧЕСК0Г0 АНАЛИЗА ОСТАТОЧНЫХ РАСТВОРИТЕЛЕЙ И ИДЕНТИФИКАЦИИ ЭЛЕМЕНТНОГО СОСТАВА ПОЛИМЕРОВ E.E.Sotnikov, G.K.Torosyan, T.G.Lanskova, Moscow, USSR Application of gas chromatography for the analysis of residual solvents, monomers and identification of element composition of polymer using stage heating in static reac tors and selective combined detectors (SCD) has been considered. At heating above glass transition temperature of a poly mer sample under static conditions the residual solvents are evaporated and monomers stable at these temperatures are evolved. pyrolysis was carried out at 650-8OO C. Using flame ionization, suface ionization and two flame photometric detectors with different light filters we exactly deter mined specific N, CI, S, P compounds in complex mixtures containing residual solvents, monomers and polymers pyro lysis products. Polysulfones were identified by specific sulfur-contai ning products, polyamines and polyimines - by nitrogen- containing products, polytetrafluoroethylene - by perfluo- rocarbon, polyvinylchloride -by chlorinated hydrocarbon. Separation of pyrolysis products and residual solvents was carried out on glaits and PTFE columns packed Polysorb 1 or Chromosorb Ю2. Identification of other products evolved at heating and pyrolysis was carried out by chromato-mass-spectrometry. Advantages and limitations of stage heating for polymer analysis were shown. The examples of determination of element composition of polymer and residual solvents analysis in polyester polymers are given.

122 178 P.1-22 IDENTIFICATION OF THE DECOMPOSITION PRODUCTS FROM SOME COPOLYMER RESINS ВУ PYROLYSIS GAS CHROMATOGRAPHY ИДЕНТИФИКАЦИЯ ПРОДУКТОВ РАЗЛОЖЕНИЯ НЕКОТОРЫХ СОПОЛИМЕРНЫХ СМОЛ МЕТОДОМ ПИРОЛИТИЧЕСКОИ ГАЗОВОЙ ХРОМАТОГРАФИИ A.GrObler, L.Fazakas, Budapest, Hungary Flash and step-wise pyrolysis of copolymers and polymer blends and'gas chromatographic identification of their decomposition products were carried out on stationary phases of different polarity. Styrene/butyl aerylate, divinyl benzene/acrylic acid, diallyl phthalate/vinyl acetate copolymers, homopolymer blends and some combinations thereof were involved. Suggestions are given for some structural features of the pyrolyzed resins on the basis of the identified decomposi tion products.

17Э P.1-23 К COMPARISON OF THE PRODUCTION OF POLYCYCLIC AROMATIC HYDROCARBONS FROM THE COMBUSTION OF POLYMERIC MATERIALS СРАВНЕНИЕ СОСТАВА ПОЛИЦИКЛИЧЕСКИХ АРОМАТИЧЕСКИХ УГЛЕВОДОРОДОВ, ОБРАЗУЮЩИХСЯ ПРИ СГОРАНИИ ПОЛИМЕРНЫХ МАТЕРИАЛОВ M.A.Elomaa, S.Salo, J.Lindberg, Helsinki, Finland Some common polymeric materials were combusted in labo ratory scale in order to gain an insight into their incli nation to produce soot and polynuclear aromatic hydrocar bons (PAH). Corabuuted materials were polypropylene, poly styrene, cellulose, and wood. Polymer samples of about SO mg were subjected to a sudden temperature of 973 К in an air stream. Soot was collected in glass fiber filters. PAHs were extracted from the soot, and the extracts were examined after separation on an alumina column by capil lary gas chromatography /1/. A group of PAHs was identified by means of mass spectro metry and by a retention index comparison /2/. In the conditions that were used, different polymeric materials proved to have much similarities but still differ clearly from each other with regard to their PAH emission. 1. M.A.Elomaa, S.H.Salo, J.J.Lindberg. The Effect of Mixing Polymers on Their Combustion, Die Angewandte Makromolekulare Chemie.,1985, in press. 2. M.L.Lee, D.L.Vassilaros, C.M.White, M.Novotny, Anal. Chem,51, 758,(1979).

123 180 P.1-24 THE APPLICATION OF GAS CHROMATOGRAPHY TO LOW - TEMPERATURE OXIDATION OF COAL SUBSTANCE STUDY ПРИМЕНЕНИЕ ГАЗОВОЙ ХРОМАТОГРАФИИ ДЛЯ ИССЛЕДОВАНИЯ ПР0ДУКТ03 НИЗКОТЕМПЕРАТУРНОГО ОКИСЛЕНИЯ УГЛЕЙ K.C^ip, В. Tar aba, Ostrava, CsSR The application of gas chromatography as a dynamic pul se method to study the interaction of different rank black coals with aerial oxygen is described. Values of the oxidation rate k within the temperature interval (20-85°C) on the basis of the chromatographic data were calculated, as well as global activation energies E ,- from graphical curves of In k ~1/T (Arrehenius equation) were obtained. The different linear intervals in the slo pe of In k ~ 1/T curve, were found for all coals studied. The changes in the slopes have been explained as changes in the oxidation mechanism at the given temperatures. The apparent activation energies E- calculated for the observed oxidation stages have Been compared with publi shed data where EL* was determined by other experimental techniques. The main advantage of the method described in this paper is the possibility to distinguish physically sorbed oxygen from chemically sorbed one on natural black coals, the suitability to values k and E. determination, the sufficient sensibility and'last but not least the possi bility to observe accurate isothermal conditions during measurements. 181 P.1-25 THERMAL DEGRADATION OF GENERATOR INSULATING MATERIALS BY PYROLYSIS GAS CHROMATOGRAPHY ТЕРМИЧЕСКАЯ ДЕГРАДАЦИЯ ИЗОЛЯЦИОННЫХ МАТЕРИАЛОВ ДЛЯ ГЕНЕРАТОРОВ С ПОМОЩЬЮ ПИРОЛИТИЧЕСКОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ K.Torkos, J.Borossay, G.Nagy, Budapest, Hungary The operational safety of turbine generators heavily depends in addition to mechanical characteristics, on the properties of the applied insulating materials. During the past decade an important research activity has been in progress an aim at understanding the thermal degrada tion of insulating polymers /1/ and at developing fault diagnostic systems based on the detection of degradation products /2,3/. Thermal degradation in the temperature interval 450 - 700°C of some insulating materials of epoxy resin and polyester base was investigated by us with the method of pyrolysis gas chromatography. The identification of ali phatic, aromatic hydrocarbons and phenol products origi nating during the pyrolysis was carried out by means of packed columns. The resulting CO and CO, were determined by methanization on Ni catalyzer (detector - FID) /4/. We determined also the C.-Cj hydrocarbons. The degradation 124 products unambiguosly characterize the chemical structure of the polymer. The results can be applied in the quality control of the polymers to be used, or in the control of the techno

logical processes of the fabrication. The C,-C3 hydrocar bons, being permanent gases, make it possible Co immedia tely singnal any failure occuring in the cooling gases of working generators. 1. A.N.Freedman, J.Chromatogr.,157, 85-96 (1987). 2. G.Metzger, R.Fournie; R.G.E.,8"5"7 271-276(1976). 3. W.V.Ligon, J.L.Webb, Anal. cKim 5_3, 1655-1658 (1981). 4. K.Torkos, J.Borossay, A.Szekely, J.Chromatogr., 286, 317-321 (1984). 182 P.1-26 INCREASING THE RELIABILITY OF IDENTIFICATION OF POLYMERS BY PYROLYSIS GAS CHROMATOGRAPHY УВЕЛИЧЕНИЕ НАДЕЖНОСТИ ИДЕНТИФИКАЦИИ ПОЛИМЕРОВ С ПОМОЩЬЮ ПИРОЛИТИЧЕСКОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ G.Stoev, I.Mladenov, Sofia, Bulgaria The interlaboratory reproducibility of gas chromato graphic data in the identification of polymers by pyrolysis gas chromatography is unsatisfactory. This is due to the lack of control on the properties of the columns and to the inaccurate reproduction of the chromatographic conditions, In the suggested method, the retention charac teristics of the capillary columns used are controlled through factor g , while the working conditions are controlled by internal standards in the programs. The probability for the coincidence of polymers of different types at the identification with five characteristic pyrolysis products is about 1 10~. 183 P.1-27 SPECIFIC INTERACTIONS OF ALKENES WITH CHEMICALLY BONDED PHOSPHINE COPPER COMPLEXES IN GAS CHROMATOGRAPHY СПЕЦИФИЧЕСКОЕ ВЗАИМОДЕЙСТВИЕ АЛКЕНОВ С ХИМИЧЕСКИ СВЯЗАННЫМИ ФОСФИНМЕДНЫМИ КОМПЛЕКСАМИ В ГАЗОВОЙ ХРОМАТОГРАФИИ W.Szczepaniak, Poznan, Poland Transition metal cations are able to interact specifi cally with unsaturated compounds and compounds containing hel .Toatoms /1,2/, As a continuation of our investigations of specific interactions between unsaturated hydrocarbons and phosphine-transition metal complexes two packings containing GuCl, and CuBr, were prepared. The salts were bonded to the silica surface (Porasil C) through 1-tri- ethoxy silyl-2(diphenylphosphine methyl phenyl)ethane /3/. The packing containing bonded phosphine without metal salt was used as a reference.

Retention data (к* and IR) for ~50 hydrocarbons

125 (linear, branched and cyclic) as well as'for —30 chloro- derivatives of alkan?s and alkenes allowed to find how the specific interactions of ОГ-type depend on the struc tures of the adsorbat* (electron-donor site) and of the electron-acceptor centre i.e. bonded metal complex. The values of the specific interactions increased for the

sequence: -PPh2 < -PPh2 CuClj < -РРП2 CuBr2. Differences in the stability constants of the J"-complexes formed during chromatographic process make possible to separate hydrocarbons with very slight differences in structure e.g.' cis- and trans-isomers. The values of specific inte ractions depend on a nature of an anion coordinated with ttis transition metal cation. This can be explained by both electronic and steric effects of the anion. Stronger interactions were observed in case of the packing contai ning CuBr2. 1. W.Szczepaniak, J.Nawrocki, W.Wasiak, Chromatographia, 12 (1979) 484. 2. W.Szczepaniak, J.Nawrocki, W.Wasiak, Chromatographia, 12 (1979) 559. 3. wTwasiak. W.Urbaniak, W.Szczepaniak, Polish patent pending P-246727. 4. W.Wasiak, W.Szczepaniak, Chromatographia. 18 (1984) 205. 184 P.2-28 OIL SHOWS ANALYSER (O.S.A.) s GEOCHEMICAL TOOL FOR PETROLEUM RESEARCH АНАЛИЗАТОР НЕФТИ - ПРИБОР ДЛЯ ГЕОХИМИЧЕСКОГО ИССЛЕДОВАНИЯ НЕФТИ J.Gregoire, Suresnes, France Fundamental research made by French Institute of Petro leum and Belgium Labofina on kerogens led to the develop ment of a method and of the corresponding patented equip ments available from Delsi instruments. We describe the O.S.A. apparatus which is derived from the Rock-Eval device (R.E.). Both apparatuses are based on the temperature pyrolysis of small rock samples in an inert atmosphere. They are used as well as on the well site or in the laboratory. In addition to measurement of the R.E. parameters (except S3) used to characterise the source rock quality and the organic matter maturation. O.S.A. gives the total organic carbon content and the light hydrocarbon gases content of the cuttings. Geochemical logging by the O.S.A. permits in about 20 to 30 minutes per sample: - quantitative analysis of oil and gas potential of source rocks • - evaluation of source rock quality (type of organic matter) - determination of source bad thickness - determination of organic matter maturation (imature zone, oil zone, gas zone) - determination of depth reguired for oil generation

126 - Identification of cumulated and migrated hydrocarbons (oil and/or gas shows) - distinction between true and false gas and oil shows (which where not possible by the very analysis of gase ous hydrocarbon shows carried by the drilling mud) - stratigraphic correlations - screening of representative samples for more complete geochemical study.

185 P.2-29 THE CHROMATOGRAPHIC INVESTIGATION OF LUBRICATING OILS AND TECHNOLOGICAL LIQUIDS ХРОМАТОГРАФИЧЕСКОЕ ИССЛЕДОВАНИЕ СМАЗОЧНЫХ МАСЕЛ И ТЕХНОЛОГИЧЕСКИХ ЖИДКОСТЕЙ D.E.Diskina, Novokuibyshevsk, USSR Some chromatographic procedures and their combinations with other physico-chemical methods are used for the investigation of composition, properties and work parame ters of industrial oils, their technological liquids and additives. The methods of investigation of evaporativity and ther mal oxidation stability of lubricating oils based on the combination of gas chromatography and thermal analysis have been proposed and the corresponding devices have been constructed. The chromatographic method of evaluation of lubricating efficiency of additives is worked out. The method of investigation of the composition of the waste heat carrier containing the non-evaporable substan ces includes the combination of thermal analysis, liquid chromatography and programmed temperature gas chromatogra phy with quantitative interpretation of chromatograms obtained by double internal standard. 186 P.2-30 GAS CHROMATOGRAPHIC DETERMINATION OF COMPOSITION OF UNFRACTIONAL NATURAL HYDROCARBON MIXTURES ГАЗОХРОМАТОГРАФИЧЕСКОЕ ОПРЕДЕЛЕНИЕ СОСТАВА НЕФРАКЦИОНИРОВАННЫХ ПРИРОДНЫХ СМЕСЕЙ УГЛЕВОДОРОДОВ M.L.Sazonov, M.Kh.Lunskii, L.I.Zhiltsova, I.L.Paizanskaya, N.I.Chuvilyaeva, Moscow, USSR This report deals with the problems of determination the light hydrocarbon content (Cj-Cg) of crude oils, con densates and organic matter in' rocks and formation waters by multidimensional gas chromatography. Using the back flush system which is common for all these techniques,.the necessary information have been obtained without preliminary treatment of natural samples. Some analytical characteristics such as sensitivity, relia bility, efficiency of resolution, decreasing sample size and assay duration have been improved. Imformation about crude oils and condensates fractional composition can be received for 10-15 mln. The light 127 hydrocarbons up to Cg (196 compounds) in unfractional oils and condensates are defined by using the capillary columns. The analysis of formation water samples is carried out by their direct injection into the gas chromatograph. The organic matter in rocks is investigated by two ways using pyrolysis and organic solvent extracts. For the pyrolysis experiment, the whole rock sample is pre viously powdered and then heated at an appropriate tempe rature. The sample organic extracts are examined without vaporization. The high sensitivity and precision of light hydrocarbons determination is reached by using the sample cold trapping and the direct injection of large liquid volumes of sample. The schemes and typical chromatograms obtained from the natural sample analysis are given. 187 P.2-31 DETERMINATION OF ENANTIOMERIC PURITY OF TERPENOIC HYDROCARBONS USING el-CYCLODEXTRIN COMPLEXES BY GAS-LIQUID CHROMATOGRAPHY ОПРЕДЕЛЕНИЕ ЭНАНТИОМЕРНОЙ ЧИСТОТЫ ТЕРПЕНОВЫХ УГЛЕВОДОРОДОВ С ИСПОЛЬЗОВАНИЕМ ^,-ЦИКЛОДЕКСТРИНОВЬК КОМПЛЕКСОВ МЕТОДОМ ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ D.Sybilska, Warsaw, Poland Terpenee occur in large amounts in most plants; they are of special interest, because of their industrial applications and on account of their chemical and biolo gical properties. Since the most commercially available terpenes derived from natural sources are of variable enantiomeric purity, many attempts at development of methods for exact determination of this purity have been made. We have recently reported /1/ on successful resolution of oL- and jS-pinene into enantiomers, using oL-cyclodextrin (oL-CD) as a chiral-selective agent in gas-liquid chromatography with formamide as a matrix medium. The pre sent communication reports our further attempts at applica tion of the same principle in enantiomeric separation of other terpenoic hydrocarbons. The model compounds tested were: el- and jS-pinene, cis- and trans-pinane, el- and ol-phellandrene, A3-carene and limonene. The aim of this work was to develop a new, possibly general method for the determination of enantiomeric purity of terpenoic hydrocarbons. It was found under optimized conditions that complete separations into enantiomers can be achieved for all investigated compounds with the exception of limonene. Such system modified by el-CD results in a rise of enantioselectivity, while at the same time the loss of effi ciency of the original column is comparatively small. The reliability and accuracy of the method developed are F~ correlated with specific rotation measurements. The scope and limitations of the method are discussed from the 128 1 ;

standpoint of the structure of the investigated compounds and of their ol-CD inclusion complexes. 1. T.Koecielski, D.Sybilska, J.Jurczak, J. Chroraatogr., 280, 131 (1983). 188 P.2-32 CHROMATOGRAPHIC FRACTIONATION OF NEUTRAL ASPHALTENES IN SEDIMENTARY ROCKS , ХРОМАТОГРАФИЧЕСКОЕ ФРАКЦИОНИРОВАНИЕ НЕЙТРАЛЬНЫХ АСФАЛЬТЕНОВ В ОСАДОЧНЫХ ПОРОДАХ G.P.Stoyanova, Sofia, Bulgaria Consecutive fractionation of chloroforre extracted disse minated organic matter (DOM) in sedimentary rocks was performed to let more comprehensive insight in its chemi cal composition. Three samples from Central North Bulgaria with mesosoic age were studied. Sapropelic, humic and humic like facial genetic type of DOM, low degree of deagenetic oxidation (q <50 %) and low rank of mesokatagenesis were established. The asphaltenic compounds were precipitated by petro leum ether and fractionated in neutral aephaltene (ethanol insoluble part) and asphaltenic acids (ethanol soluble ?art). The object of the investigation were neutral asphalte- les. They were fractionated by column chromatography (A1,0,, II degree of activity, Brocktnann) ; petroleum ether, senzene,acetone,ethanol-benzene (1:1, v/v), methanol and Uethyl ether were employed as eluents. Narrow fractions «ere characterized by elemental analyses and IR spectra. К separation according to the functionality was considered. 189 P.2-33 APPLICATION OF CHROMATOGRAPHY TO INVESTIGATION OF ALKYLBENZENES FROM BULGARIAN OIL i ПРИМЕНЕНИЕ ХРОМАТОГРАФИИ ДЛЯ ИССЛЕДОВАНИЯ АЛКИЛБЕН30Л0В В БОЛГАРСКОЙ НЕФТИ G.Xovachev, Sofia, Bulgaria Interest in the origin of aromatic hydrocarbons was enchanced by the discovery and utilization of petroleum. Aromatic hydrocarbons are constituents of oils and studies of their origin are viewed as a part of studies of the irigin of petroleum. Investigation of their structures 'ill be useful to clear the concepts of the chemistry of rarbon evolution, metabolic processes, distribution of iydrocarbons in nature and some possible precursors of oil. In relation to the study of organic compounds, samples :f bulgarian oil have been investigated. Using column and •in-layer chromatography a mono-aromatic fraction was olated. Capillary gas chromatography on Dexil-4O0 was •ed to separate a homologous series of alkylbenzenes from j^O to Сзо- Computerized combination gas chromatography -

•ак. 872 t» mass spectrometry was applied for qualitative evaluation of the above mentioned alkylbenzenes. On the basis of chemical analysis and the library sear ching process, we are able to mention the similarity bet ween the investigated alkylbenzenes from the oil and natural carotens. This carotene were probably one of the starting materials for oil formation.

190 P.2-34 GAS CHROMATOGRAPHIC DETERMINATION OF ORGANIC IMPURITIES IN THE SOLVENTS FOR LIQUID CHROMATOGRAPHY ГА30ХР0МАТ0ГРАФИЧЕСК0Е ОПРЕДЕЛЕНИЕ ОРГАНИЧЕСКИХ ПРИМЕСЕЙ В РАСТВОРИТЕЛЯХ ДЛЯ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ V.Yu.Zelvenskii, A.S.Lavrenova, S.I.Samolyuk, L.V.Borodai, Moscow, USSR The gas chromatographic techniques have been developed for next highly purified solvents for liquid chromatogra phy: acetonitrile, methanol, ethanol, methylene chloride, pyridine, formic and acetic acids, dimethylformamide, dimethyleulfoxide, and tetrahydrofuran. The limites of organic impurities determination are 5>10"*- 5»10~5%. There are results for compositions of UV-absorbing conta mination and other organic impurities. The approaches to selection of optimal conditions for the main component and limiting impurities resolution are discussed. The summary table including resolution conditions and quanta - tlve results is represented.

191 P.2-35 GC ANALYSIS OF WASTES FROM THE PRODUCTION OF CARBON TETRACHLORIDE AND TETRACHLORETUYLENE ГА30ХР0МАТ0ГРАФИЧЕСКИЙ АНАЛИЗ СТОЧНЫХ ВОД ПРОИЗВОДСТВА ЧЕТЫРЕХХЛОРИСТОГО УГЛЕРОДА И ТЕТРАХЛОРЭТИЛЕНА I.Beranek, M.Uhlir, Vsti nad Labem, SsSR In addition to the expected compounds (such as hexa- chlorethane, hexachloro-1,3 butadiene and hexachloroben- zene) a number of other compounds in significant concen trations have been found by analysing the distillation residue from the high-temperature chlorolysis of propylene. The precision of the analytical method employed is such that it can be used in monitoring and control of the technological regime of chlorolysis on the basis of rela tive amounts of the main components in the waste. The required identification of additional compounds in the residue from chlorolysis is certainly justified from the point of view of environmental protection.

130 192 P.2-36 TRACE ANALYSIS OF POLYCHLORINATED BIPHENYLS IN ENVIRONMENT АНАЛИЗ СЛЕДОВ ПОЛИХЛОРБИФЕНОЛОВ В ОКРУЖАЮЩЕЙ СРЕДЕ T.Buzinkaiova", J.Krupiik, Bratislava, 6SSR The polychlorinated biphenyls (PCBs) are industrial compounds narked in Czechoslovakia under the trade name Delor ЮЗ, Ю5 and 106. They contain a complex mixture of closely related chlorinated biphenyls varying in chlo rine content. PCBs are chemically and biologically resis tant environmental contaminants and are therefore incor porated into various biological life cycles. Gas chromatographic separation of components present in Delor containing samples were carried out on various glass capillary columns. Qualitative analysis was based on GC-MS combination. The influence of the change of splitting ratio on separation efficiency was studied. To get rid of compounds bleeding from the septum, what is absolutely necessary in trace analysis, a septum bleed cap was installed. Modifi cation of injector has no influence on the number of theo retical plates at the given splitting ratio, but the peaks from septum bleeding completely dissapeared. Various methods of PCBs extraction from geological waters and butter were tested. The results of quantita tive analysis were calculated with a new method utilizing results obtained with FID and BCD detectors enabling the determination of PCBs after biodegradation, too.

193 P.2-37 THE DETERMINATION OF SOME ORGANIC TRACES IN WATER WITH THE SORPTION ENRICHMENT ОПРЕДЕЛЕНИЕ НЕКОТОРЫХ ОРГАНИЧЕСКИХ ПРИМЕСЕЙ В ВОДЕ МЕТОДАМИ ХРОМАТОГРАФИИ С ПРЕДВАРИТЕЛЬНЫМ СОРБШЮННЫМ КОНЦЕНТРИРОВАНИЕМ Ifu.S.Filippov, N.F.Tolikina, N.N.Senin, G.A.Smolyaninov, S.A.Volkov, Moscow, USSR For the determination of butyl acetate in urban sewage a samples were passed through an adsorbent packed in the trapping column. After accumulation of trace the trap ping column was washed by small volume of acetone and washing liquid was analized on the gas chromatograph. The stationary phases DC 550 and Apieson L. The limit of determination O.Ol mg/1, relative reproducibility was not exceed 15 %. The possibility of employing a number of adsorbents (Polysorb 1, Chготовоrb 102 and modificated aluminium silicate) for the enrichment of butyl acetate from water has been investigated. Aluminium silicate is the most effective sorbent (90 * extraction). The influence of va rious parameters on the effectivity of the enrichment has

131 been Investigated. This method was used on stations of biological purification. The determination of substituted phenylureas (fenuron, doeanex, monuron, kotoran, linuron, diuron) in natural water was performed by high-performance liquid chromato graphy on a column packed with Partisyl ODS, using an acetonitril-water mixture (10s3) as mobile phase. To in crease the sensitivity, the water sample was passed through a column with 1 g of Chromosorb-102, and the sub stances adsorbed were eluted with 3 ml of acetone. After the enrichment (enrichment extent 300, yeld 74 ») detec tion limit was lO"4 - 10~5 mg/1, relative error was 10 %. Porapak Q and Chromosorb-104 do not desorb components investigated very effectively (yeld < 30 %).

194 P. 2.-38 THE ANALYSIS OF MERCAPTAN TRACES IN AIR BY GAS-LIQUID CHROMATOGRAPHY АНАЛИЗ МИКРОПРИМЕСЕЙ МЕРКАПТАНОВ В ВОЗДУХЕ МЕТОДОМ ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ P.E.Tulupov, L.B.Alekseeva, V.M.Goncharyonok, Obninsk, USSR Mereaptans are referred to weakly stable compounds, however, a pronounced objectionable order and a high toxi city call for controlling air pollution by then. When controlling air pollution by me reaptans the problems arise not only from concentration of their traces but from sample conservation between the stages of concentra tion and analysis. This report discusses the kinetics of mercaptan sorp tion from air using liquid, thin-film and solid sorbents, the kinetics of their transformation in a sorbed state, sample preparation for analysis and analysis by gas-liquid chromatography. Derived are the kinetic equations to compute mercap tan concentrations at stages of sorption and conservation as concentrated samples prior to analysis. 195 P.2-39 SIMULTANEOUS DETERMINATION OF TOTAL OXYGEN DEMAND AND TOTAL ORGANIC CARBON IN HATERS ОДНОВРЕМЕННОЕ ОПРЕДЕЛЕНИЕ ОБЩЕГО РАСХОДА КИСЛОРОДА И ОБЩЕГО ОРГАНИЧЕСКОГО УГЛЕРОДА В СТОЧНЫХ ВОДАХ I.Kamenikova', V.Rezl, Brno, £sSR Basic characteristics of the degree of loading of waters with wastes include the determination of biochemi cal, chemical, and total oxygen demand (TOD) and of total organic carbon (TOC). Particularly the last two methods (TOD, TOC) are rapid, amenable to automation and prospec tive from the viewpoint of analytical control of waste waters and their purification. Since they can provide in principle differing results, both determinations are performed separately, TOD determination usually coulomet- 132

I ricly. Reaction frontal GC is suitable for simultaneous determination both of TOD and of TOC. After removal of inorganic carbon, a sample of analyzed water, 10-100 ul, is injected into a stream of helium containing oxygen and then it is transported into a reaction zone filled with

granulated Sn02 at 950-1000°C. The present impurities are oxidized at oxygen expense . The reaction products are freed from water and nitrogen oxides, the remaining mixtu re of helium, oxygen, carbon dioxide, eventually of nitro gen, is collected in a dilution chamber and homogenized under the constant state conditions. It is thsr. injected into a chromatographic column packed with Porapak Q, oxygen and nitrogen are then separated from carbon dioxide by frontal technique and detected by a katharoroeter measu ring cell. Then the mixture passes through a layer of ascarite, and copper at 500°C, and through anhydron in order to remove oxygen and COj. A reference cell then detects nitrogen concentration. The loss of oxygen then corresponds to TOD, C02 content to TOC. The detection sensitivity is 1 and 10 ppm for TOC and TOD, respectively, the analysis time is about 6 min.

196 P.2-40 TRACE ANALYSIS OF SOME CHLORINATED HYDROCARBONS IN SEA WATER BY GAS-LIQUID CHROMATOGRAPHY АНАЛИЗ СЛЕДОВ НЕКОТОРЫХ ХЛОРИРОВАННЫХ УГЛЕВОДОРОДОВ В МОРСКОЙ ВОДЕ МЕТОДОМ ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ M.Mohnke, P.Franz, Leipzig, GDR K.-H.Rohde, L.BrUgmann, Rostock-Wamemunde, GDR In September/October 1980, May/June 1981, and in June/ July 1983 the distribution of some pesticides (hexachloro- benzene, the isomers of hexachlorocyclohexane, DDT and its metabolites DDE and DDD) and of polychlorinated biphenyls were investigated at more than 30 stations in the Baltic Sea (Baltic Monitoring Programme), North Sea, and North Atlantic. These expeditions were carried out with the GDR research vessel "A. v. Humboldt". The devices for deep water (1 500 meter) and surface sampling (0,3 millimeter), as well as the sample extrac tion method,the procedure for concentration of pesticides, and sample clean up and their composition are described in the poster. Samples were analysed with QF 1 packed column and SE 54 glass capillary column with on column injection technique using Hewlett-Packard 58Э0А with 63Ni - ECD. The results of both methods were in good agreement. The results of investigations have been surr.tiarized as mean values, concentration ranges and standard deviations.

133 197 P.2-41 CHROMATOGRAPHIC ANALYSIS OP PARAFFINIC HYDROCARBONS FROM WATER OF WESTERN AQUATORIOM OF THE BLACK SEA ХРОМАТОГРАФИЧЕСКИЙ АНАЛИЗ ПАРАФИНОВЫХ УГЛЕВОДОРОДОВ ИЗ ВОДЫ ЗАПАДНОЙ АКВАТОРИИ ЧЕРНОГО МОРЯ F.I.Demyanov, V.S.Petrosyan, Moscow, USSR A number of samples for analysis of oil pollution have been prepared by extraction of water of Black Sea western aquatorium between state sea frontiers and 32 E in spring of 1981. Capillary gas chromatography (GC Varian 3700, column 25 m, i.d, 0.25 mm, SE-30, column oven temperature pro grammed from SO to 320°C) was used fer identification and quantitative analysis of pollutant n-alkanes. Molecular weight distribution curves for n-alkanes (containing from 14 to 44 carbon atoms) have minima for octa- and nonadecanes, and their relative contents in the sea water are rather low. Hydrocarbons, polluting upper layer of the Black Sea, can be devided into two groups,

basing on the form of such curves. С,д-С,7 n-alkanes re c ar present, the first, while C2o~ 38 e the main part of the second group. Maxima of the molecular weight distribution curves shift upwards or downwards on the hydrocarbon molecular weight scale, depending on sampling location. The pollution degree decreases with'the distance from the shore, but there is a sharp pollution increase in the vicinity of the Bosphorus strait - Danube delta shipway. The obtained data indicate possible sources of hydro carbon pollution of the Black Sea. 198 P.2-42 GAS CHROMATOGRAPHIC INVESTIGATIONS OF ORGANIC COMPONENTS OF PAPER MILL EFFLUENTS ИССЛЕДОВАНИЕ ОРГАНИЧЕСКИХ КОМПОНЕНТОВ СТОЧНЫХ ВОД ПРЕДПРИЯТИИ, ПЕРЕРАБАТЫВАЮЩИХ МАКУЛАТУРУ, МЕТОДОМ ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ O.I.Kalchenko, V.O.Volovenko, V.P.Svitelski . Kiev, USSR The organic components of paper mill effluents were analysed before and. after treating. The effect of efflu ents treating method on its quantitative and qualitative variations had been investigated. In paper-mill effluents organic compounds, selected from the class of saturated and unsaturated fat acids, hydroxyacids, esters, mononuclear phenols and monocarbo- hydrates have been found. Their amounts vary from 0.01 to 30-40 mg/.l. It was determined that removal of organic acids in course of physico-chemical cleaning increases with the growth of their molecular weights. The removal of acids decreases with the growth of molecular weight in course of biological cleaning.

«34 It is clear that biological cleaning favour the expan sion of fat acids composition in effluents. Gas chromatographic investigations were carried out on immovable phases Reoplex-400, XE-60, Apieson L and Polysorb-1. 199 P.2-43 PATTERN RECOGNITION FOR INTERPRETATION OF COMPLEX CHROMATOGRAMMS IN ENVIRONMENTAL ANALYSIS УЗНАВАНИЕ ПРОФИЛЯ 17РИ ИНТЕРПРЕТАЦИИ СЛОЖНЫХ ХРОМАТОГРАММ В АНАЛИЗАХ ОКРУЖАЮЩЕЙ СРЕДЫ E.Stottmeiater, M.Nagel, D.Feller, P.Hendel, H.Hermenau, Bab Elster, GDR The coupling of gas chromatography and computer enables the application of modern statistical data analysis for interpretation of complex chromatographic data from envi ronmental samples. The use of modern analytical techniques and the automation of measuring procedures lead to an increase of the data stream and enlarge the detectable spectrum of substances in biotic and abiotic environmental structures. On examples from the field of water research, using high resolution capillary gas chromatography, is illustra ted the gain of information by application of multivariate methods of data analysis. Beside methods like "image, ana lysis" - that means searching the chromatogram for the presence of specific peaks - and the identification of complex mixtures statistical pattern recognition methods are used. The chromatogram-library as well as some timesaving algorithms for library search are demonstrated, useful for an application of small microcomputers. By me^ns of some tools from the FUZZY-theory the interpretation of complex chromatograms becomes more objective. Automation of the interpretation and classification processes have two potential benefits, consisting in increased producti vity and in replacing the intuition and judgement of indi vidual analysis with well-defined decision rules imple mented by computer.

D.Feller, M.Nagel, E.Stottmeister, R.Koch: Pattern recog nition Verfahren in der Umweltanalytik and Okotoxikologie: Methodische Grundlagen. Acta hydrochim. et hydrobiol. (in press). D.E.Knuth: The Art of Computer Programming. Vol. Ills Sorting and Searching. Reading, Mass., 1973. B.R.Kowalski (ed.): Cheroometrie, Dordrecht 1984, Varmuza,K.: Pattern Recognition in Chemistry, Berlin,1980,

135 200 P.4-11 DETERMINATION OF ANTIEPILEPTIC DRUGS BY GAS-LIQUID CHROMATOGRAPHY ОПРЕДЕЛЕНИЕ АНТИЭПИЛЕПТИЧЕСКИХ СРЕДСТВ МЕТОДОМ ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ J.Volmut, Bratislava, CSSR Gas-liquid chromatography is today the most used techni que for the monitoring of antiepileptic drugs. However, the preseparation of drugs from biological material in many labs seems to be complicated, tedious and time con suming. In many cases it is necessary to prepare deriva tives . Our work was focused on simplifying procedure without classical extraction and derivatization. A modern concept of column extraction based on the normal or reverse phase liquid chromatography has been used for isolation of drugs from biological material. We have tried several Czechoslo vak sorbents and have found out, that they are suitable for this type of extraction. The advantage of this proce dure is the quick isolation of drugs, a clean extract and diminishing amount of sample and of organic solvents. Nowadays we can avoid derivatiuation of antiepileptic drugs before the gas-liquid chromatographic determination only by using commercial filling materials for packed columns or by utilization of fused silica capillary columns. In our paper we compare a few different columns from the viewpoint of their utilization in the therapeutic drugs monitoring. In conclusion we suggest the quick and simple method for the simultaneous determination of four antiepileptic drugs - phenobarbital, carbamazepine, primidone and phenytoin.

201 P.4-06 GAS CHROMATOGRAPHIC DETERMINATION OF THE MICROQUANTITIES OF ETHYLENE OXIDE AMD METHYL BROMIDE IN THE POLYMER MATERIALS ГАЗОХРОМАТОГРАФИЧЕСКОЕ ОПРЕДЕЛЕНИЕ МИКРОКОЛИЧЕСТВ ОКИСИ ЭТИЛЕНА И БРОМИСТОГО МЕТИЛА В ПОЛИМЕРНЫХ ИЗДЕЛИЯХ В МЕДИЦИНЕ N.I.Baranova, T.V.Lichtman, B.A.Rudenko, Moscow, USSR In last decades the products of various purposes made of the polymer materials have been widely used in medicine, for diagnosis, for prosthetic'appliance of the various organs and parts of the body and also for the complex apparatus with the details of the polymer materials. For example, such apparatus as artificial kidney, artificial heart, the apparatus for the artificial lung ventilation and others. Such medical apparatus are sterilized by gases. Ethylene oxide and its mixture with methyl bromide are used for the purpose. These substances are highly poiso nous , they have general toxic effect on the organism and

136 also they can cause the remote consequences (mutagenic and cancerogenic changes) and others. Therefore for the safe application of the sterilized articles the thorough degazation and the control the residual microquantities of the indicated substances are required. Gas-liquid chromatography is the most effective method for this purpose,' because it is sufficiently sensitive, easy, reliable and provides also simultaneous determina tion of the both components. For the determination of the sterilizing gas microquantities in the polymer materials head space analysis has been used. For this purpose the special adapter to the chromatograph LCHM-8MD has been constructed. It is a germetic vessel (volume 50 ml) jointed by two valves with the gas line and the columns. The vessel is thermostated at 90°C for the acceleration of the achievement of the equilibrium distribution of the substance between the polymer and the gas phase. The compressed air was used as the carrier gas in these that simplify its exploitation. In this experiment the absolute calibration detector method was applied using the ethylene oxide and methyl bromide solutions in acetone prepared by weight method. The developed method is simple, useful sensitive and effords fast results what enable its introduction into production and medical establishments. 202 P.4-07 THE QUANTITATIVE ANALYSIS OF SOME O-GLYCOSIDES OF PHENOLS, MONO- AND SESQUITERPENE ALCOHOLS BY GAS CHROMATOGRAPHY КОЛИЧЕСТВЕННЫЙ ГАЗОХРОМАТОГРАФИЧЕСКИЙ АНАЛИЗ НЕКОТОРЫХ 0-ГЛИКОЗИДОВ ФЕНОЛОВ, М0Н0- И СЕСКВИТЕРПЕНОВЫХ СПИРТОВ A.A.Bezzubov, I.A.Egorov, Moscow, USSR V.N.Velev, V.I.Litchev, Sofia, Bulgaria O-glycosides in the form of trimethylsilyl ethers have been analyzed by gas liquid chromatography using 3% methyl and phenylmethylsilicon liquid phase on Chromaton N super (0,125-0.16 mm). Temperature was programmed from 180° to 280°C, 4° per minute. 4-nitrophenyl-j8-D-glucopyranoside and 1-naphthyl-jS-D-glucopyranoside were used as internal standards. The glucosides were synthesized from tetraacetylbrome- glucose, -galactose and -arabinose. Phenol, hydroquinon and resorcin were used as a phenolic components, linalool, geraniol, citronellol were used as a monoterpene alcohols, farnesol, nerolidol as a sesquiterpenes alcohols. The results obtained show that gas liquid chromatogra phy can be applied for quantitative analysis of O-glyco sides with relative error 2,5-4 %. Synthesized O-glycosides have been utilized for the identification and quantitative determination of natural glycosides, isolated from plant material.

137 203 P.4-08 A FAST PROCEDURE FOR THE ISOLATION AND THE GAS CHROMATOGRAPHIC ESTIMATION OF THYROID HORMONAL COMPOUNDS IN HUMAN SERUM БЫСТРАЯ МЕТОДИКА ВЫДЕЛЕНИЯ И ГА30ХР0МАТ0ГРАФИЧЕСК0Г0 ОПРЕДЕЛЕНИЯ ТИРОИДНЫХ ГОРМОНОВ В СЫВОРОТКЕ КРОВИ ЧЕЛОВЕКА P.Husek, V.Felt, Prague, CSSR

Low levels of the main deiodination metabolites (T3, rT,) and also of the deamination and decarboxylation ana logs (triac, tetrac) of thyroxine ('£,) in human serum were hardly to assess before the very sensitive RIAs in the 70s were introduced. Next to this, there was a las ting effort to use the instrumental chromatographic tech niques (GC and later HPLC) for the integral analysis of the whole hormonal spectrum. Using the selective detec tion (GC-ECD) amounts down to femtomoles could be analy zed successfully /1,2/, however, triac and tetrac were not covered. The HPLC is not applicable to the low biolo gical levels. Based on the condensation procedure aiming to derivatization and GC estimation of the protein amino acids /3/ we extended its applicability toward GC analysis of the mentioned hormonal compounds /4/. A complete spectrum of the derivatized compounds of interest can be obtained after few minutes of chemical treatment using a short fused silica capillary column with the OV-1 bonded phase within a • -ge of 180-220 С. The derivatives are injected via the splitless (in heptane) or the "on-column"((in decane) technique and amounts down to femtomoles can be assessed with the electron capture detector. Next to this, a rapid and effective procedure for the isolation of the thyroidal compounds from the human serum was elaborated. It is based on combination of anion-exchange column chromatography with the gel chromatography as the subsequent step. By this means the triac and tetrac metabolites also are preserved and amenable to be assessed. A suitable internal standard for the applica tion range was foand /5/. 1. J.A.Corkill, R.W.Giese, Anal. Chem., 53 (1981) 1667. 2. J.A.Corkill, M.Joppich, A.Nazareth, R.W.Giese, J. Chromatogr., 240 (1982) 415. 3. P.Huiek, J.Chromatogr., 234 (1982) 381. 4. P.Husek, V.Felt, J.Chromatogr., 288 (1984) 215. 5. Р.НиЗек, J.Chromatogr., 288 (1984) 200.

138 204 P.4-10 MEASUREMENT OF LIPOPHILICITY ВУ GLC ИЗМЕРЕНИЕ ЛИПОФИЛЬНОСТИ С ПОМОЩЬЮ ГА30ЖИДК0СТН0И ХРОМАТОГРАФИИ E'.Janos, T.Ceerhiti, Budapest, Hungary Quantitative structure - Activity relationships studies are widely used to design bioactive molecules. These approaches often use lipophilic!ty indices to characterize the investigated molecules. The lipophilicity of a mole cule can be measured by several ways. In our present study we try to measure the lipophilicity by GLC. We used 38 ring substituted aniline derivatives. Xovats' retention indices of aniline derivatives were measured on five gas chromatographic columns of different polarities (3 % Car- bowax 20M, 3 % OV-1, 3 % OV-17, 3 % OV-275 and 3 * Epon 1001 on Supercoport, 80-100 mesh, 180 cm, 2 ram) on three different temperature. Lipophilicity of compounds was determined by reversed-phase thin-layer chromatography (on DC Alufolien Kieselgel 60 F_54' Merck, impregnated with 5 % paraffin oil in n-hexani, developed in methanol- water with different organic phase ration in the mobile phase). Log P values were taken from Hansch-Leo compila tion. Relationships between the different parameters were studied with multivariate mathematical methods. By this way we can establish the applicability of GLC as a technique for the measurement of lipophilicity.

205 P.4-17 ELABORATION OF OPTIMUM METHOD FOR QUANTITATIVE, DETERMINATION OF DISOPYRAMIDE AND ITS DECOMPOSITION PRODUCTS ВЫБОР ОПТИМАЛЬНОГО МЕТОДА КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ ДИЗОПИРАМИДА И ЕГО ПРОДУКТОВ МЕТАБОЛИЗМА E.Vincze-Pil Berezvai, Budapest, Hungary Measurements of Disopyramide have usually, been made by gas chromatography till now in our laboratory. The relati ve standard deviation of the method was high, the deter mination was not decomposition-specific. HPLC methods can be found in literature recently. Only two of them are good for the separation of Disopyramide and its major metabolite, but neither of them is specific for the degradation products of Disopyramide. In pharmaceutical products such degradation occurs during the stability tests as the effect of temperature and moisture. He present a new HPLC method, which has been elaborated and optimized by using an artificially degraded Disopyra mide sample. The effect of the pH of the eluent and the ratio of aqueous - nonaqueous phase were studied, the parameters were optimized.

139 The optimal chromatographic parameters ares solvent: acetonitrile - Mcllvalne buffer {pH - 3) - water= * 105 : 65 : 130, flow rates 1 ml/min, columni 25 cm x 4,6 mm I.D, packed with LiChrosorb RP-8 (particle size 7 м), detection: variable wavelength UV detector set at 262 nm.

206 P.4-18 CHROMATOGRAPHIC STUDY OF THE METABOLISM OF THE POTENTIAL CANCEBOSTATIC DRUG, 5- [2-{N,N-DIMETHYLAMHJO)- ETHOXYj-7-OXO-7H-BENZO(c)FLUORENE ("BENFLURON") ХР0МАТОГРАФИЧЕСК0Е ИЗУЧЕНИЕ МЕТАБОЛИЗМА ПОТЕНЦИАЛЬНОГО ПРОТИВОРАКОВОГО ПРЕПАРАТА - 5-^2-(Н,н-ДИМЕТИЛАМИН0)- ЭТ0КСИ)-7-0КС0-7Н-БЕНЗ(с) ФЛУбРЕНА /"БЕНФЛУРОН'7 I.M.Hale, M.Nobilis, A.Sroler, E.Kvasnickova, Hradec Krilovrf, CSSR For the separation of Benfluron and its metabolites by TLC on silica gel ("Sllufol" Kavalier) the following I conditions were chosen: sluent chloroform, methanol, triethylamine 70:10:5 (v/v/v), bed prewashed with a mixture of cyclohexane and triethylamine 1:1 (v/v), detection mainly by colour and fluorescence of the compounds. For HPLC on octadecylsilyl silica gel, glass columns of Sepa- ron SIX С 18,5jum (150 mm x 3,2 mm) were used with nonylamine buffer, acetonitrile, 2-propanol 2:2:1 (v/v/v) as eluent and 294 and 340 nm for absorptiometry detection and quantitation. The buffer contained 2 ml nonylamine in 1 litre water made to pH 7.6 with orthophosphoric acid. On silica gel columns (Separon SIX, 5 дин) we used chloro form, methanol, triethylamine 73:7:5 (v/v/v) and chloro form with triethylamine 80:5 (v/v). Benfluron (supplied by the Institute or Pharmacy and Biochemistry in Prague) was transformed into a number of oxidation or reduction products on incubation with the postmitochondrial or microsomal' fraction of liver homoge- nate of various mammals in the presence of oxygen and NADH or NADPH. 7-Dihydrobenfluron (enantiomeric lorm not established), M-oxide of Benfluron and a phenolic hydroxylated Benfluron (OH position not known) were identified. With high probability further products correspond to K-demethylated Benfluron, N-demethyiated 7-dihydrobenfluron and to N-oxide of dihydrobenfluron. In urine of mammalians dosed with Benfluron, we found some of these substances and, in addition, several unidentified substan ces exhibiting yellow fluorescence. Liver homogenates of the mouse, rat, hamster, rabbit, rhesus monkey (Macaca mulatta) and man were compared and species differences established, out of which the absence of the phenolic product and the relatively high formation of the demethylated dihydrobenfluron on incubation with liver homogenates of man and monkey were especially interesting.

140 207 P.4-21 CHROMATOGRAPHY STUDY OF PSYCHOTROPIC DRUGS ХРОМАТОГРАФИЧЕСКИЙ ДНАЛИЭ ПСИХОТРОПНЫХ ПРЕПАРАТОВ Z.A.Zurabashvili, Tbilisi, USSR In modern psychiatric clinics neuroleptics, antidepres sants, tranquilizers, stimulants and other psychotropic drugs are simultaneously used. The individualizing and optimising of simultaneous study of their pharmacokinetics is the actual problem. It Is important to find the most simple and effective analysis for Its using in routine clinical practice. Simple and express procedures of simul taneous extraction of various classes psychotropic compounds from biological fluids (whole blood, serum, urine, saliva) have been suggested. This procedure makes possible to obtain each of psychotropic drugs and their metabolites from a small volume of sample. The gas chromatography analysis were carried out by Soviet Tsvet-1000 chromatograph with available materials. We have used different stationary liquid phases solid supporters and carrier gases. The retention volumes were measured on columns prepared with different supporters. The optimal modification value for different stationary liquid phases have been found. The inner standard was controlled by following indices: retention time and the separation criterion. The advanta ges of the photo-ionization detector for psychotropic drugs analysis in biological fluids include simplicity of operation, universal drug response and stability. The results were compared with those of flame-ionization detector.

208 P.4-30 A NEW DERIVATIVE OF 6-OXO-PGFlj. FOR GAS CHROMATOGRAPHY WITH ELECTRON CAPTURE DETECTION НОВЫЕ ПРОИЗВОДНЫЕ б-ОКСО-ПГФ-ы, ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ С ЭЛЕКТРОННОЗАХВАТНЫМ ДЕТЕКТОРОМ E.Tomori, M.Vargek, E.Tormasi, G.Cseh, Budapest, ' Hungary Two sensitive and selective quantitative methods are described and compared for the determination of the endogeneous prostacyclin content of the aorta of rats treated with GYKI-11 674. Thoracal aorta was excised and pooled from 3 rats one hour after treatment with Ю mg/kg of GYKI-11 679. For the transformation of prostacyclin into 6-oxo-PGFij the samples were kept in a 0,05 M Tris (HC1) buffer, pH8,0 at 37°C for 10 min. Prostaglandins were extracted with organic solvents. Samples for gas chromato graphic analysis required thin-layer chromatographic sepa ration prior to conversion to the methyl ester -0-methyloxime-tri-trleethylsilyl-ether derivatives. For gas chromatographic-electron-capture detection the samples were transformed into 0-methyloxime-tri-trifluoroacetyl-hexa- fluoroisopropyl derivatives. Recently we have found this 141

i new derivative Is useful for measuring of prostaglandins. The GC-ECD properties, stability and mass spectrometric- mass chromatographic characteristics of this new deriva tive in relation to the determination of 6-oxo-PGFj, will be discussed. For the quantitative analysis nor

PGFlpt as internal standard was used. Using these methods the endogeneous prostacyclin content of the aorta of GYKI-11 679-treated rats will be detailed. GYKI-11 679, a new hydrazone derivative of antihyper tensive action has been synthesized In our Institute The possible involvement of the synthesizing system of the walls of blood vessels in the antihypertensive action of this new hydrazone derivative has been discussed /1/. 1. G.Cseh, I.K.Szabo, E.Toroori: Biochemical Society Transactions, 1981, vol.9, p. 223

209 P.4-31 NEW ASPECTS IN CHEMICAL DERIVATIZATION OF CARBOXYLIC ACIDS FOR THEIR GAS CHROMATOGRAPHIC ANALYSIS НОВЫЕ АСПЕКТЫ ПОЛУЧЕНИЯ ПРОИЗВОДНЫХ КАРБОНОВЫХ КИСЛОТ ДЛЯ АНАЛИЗА С ПОМОЩЬЮ ГАЗОВОЙ ХРОМАТОГРАФИИ I.Molnar-Perl, M.Pintir-Szakace, Budapest, Hungary While esterification under strictly water-free condi tions is a classical and general demand according to the literature, many practical tasks require the analysis of aqueous solutions The esterification of different homoloque series of carboxyic acids in homogeneous ternary systems of water, butanol and one of the mineral acid is described. It has been established that the normal and/or isobu- tyl esterification of C,-C, fatty acids in aqueous solu tions, in the presence of sulfuri0 c or hydrochloric acid can be used for their quantitative determination in the concentration range of [H,0j/ [n-BuOHj « 0.01-5,3. In the concentration range of O.01-0,8 the water present does not interfere. At [H,0j/[n-BuOHj > 0.8 anhydrous sodium sulfate is used for Binding the water, in the amount of [Na SO.]/[H 0]^0.2. The optimal conditions for the es- terificatiof2 i 2 С,-С dicarboxylic, of aliphatic hydroxy and of aromatic dl1-6 and polycarboxylic acids are given concerning the water content, the quality and the amount of the mineral acids used as catalyst and the amount of anhydrous sodium sulfate for the proposed esterification mixture.

142 210 P.4-32 N-SUBSTITUTED TRIMETHYXSILYLCARBAMATES AS SILYLATING AGENTS FOR GAS CHROMATOGRAPHIC ANALYSIS N-ЗАМЕЩЕННЫЕ ТРИМЕТИЛСИЛИЛКАРБОМАТЫ В КАЧЕСТВЕ СИЛИЛИРУЩИХ АГЕНТОВ ДЛЯ ГАЗОХРОМАТОГРАФИЧЕСКОГО АНАЛИЗА D.Knausz, A.Meszticzky, J.Galacz, B.Csikva'ri, F.Szederke'nyi, Budapest, Hungary Several N-substituted trimethylsilylcarbamates have been synthesized by a new method /1/. These compounds react easily with acidic hydrogen of various organic sub stances according to the reaction:

X 2 RVNCCOJOSiMej + ZH « ZSiMe3 + R R NK + COj, where Z « different organic groups. The use of silylcarbamates in GC determination of the following type of compounds has been studied: volatile alcohols, fatty acids, different alkaloids and steroids, prostaglandines, substituted phenols, bile acids, nucleo sides, 'From these experiments we concluded that advantages of silylation with N-substituted trimathylsilylcarbamates are the following! - Silylation can generally be carried out under mild con ditions. A low excess of the agent is needed. - In most cases no solvent is required because the silyl carbamates are excelent solvents of the compounds to be sllylated. - Water content of the samples doesn't disturb the analy sis. In some cases determination of compounds even in aqueous solution is possible. - As substituted amines are formed during the siiylation as byproducts the following features can be encountered: I) the amine evolved acts as a catalyst (autocatalysis) for the silylation (e.g. in the case of hindered OH groups of alcohols /2/, prostaglandines, steroids); II) no basic compound is needed in the GC determination of addition salts of alcaloids (e.g. scopolamine.HC1); III) the volatile amine elutes in early peaks of the GC, enabling the determination of compounds with low molecular weight (e.g. volatile alcohols, fatty acids). 1. D.Knausz, A.Meszticzky, L.Szaka'cs, B.Csakvari, K.Ujszaszy: J.Organomet. Chem. ,256 (1983) 11. 2. D.Knausz, A.Meszticzky, L.SzaXa'cs, B.Csakvari; J. Organomet. Chem.,266 (1984) 207.

143 211 P.5-05 THE EMPLOYMENT OF MICROCOMPOUND CHROMATOGRAPHS IN INDUSTRIAL AUTOMATIC CONTROL SYSTEM AND THE OPERATION OF HELIUM PRODUCTION ПРИМЕНЕНИЕ МИКРОПРИМЕСНЫХ ХРОМАТОГРАФОВ В ПРОМЫШЛЕННЫХ АВТОМАТИЗИРОВАННЫХ СИСТЕМАХ КОНТРОЛЯ И УПРАВЛЕНИЯ ГЕЛИЕВЫМ ПРОИЗВОДСТВОМ O.M.Bondaryuk, A.V.Lander, L.I. Freidgeira, V.N.Khokhlov, Moscow, USSR The realization of modern technological process for obtaining pure gas demands the employment of microcompound chromatographs. The system solves the problems of microcompound control in the technological process, the opera tion of the manufacturing regime, the optimization of technological documents and registration certificates for commodity output. The data processing is conducted in real-time with the help of the central computer. The efficiency of these systems is demostrated on the example of helium production. The control and operation system of this production contains more than 100 chromatograph canals. Microcompound industrial chromatographs were specially developed and used in the system. The low limit of concentration.meaeured by the conductivity «tas decreased for 102 - 10 times with the help of the fron tal-adsorption and thermodynamic accumulation. The report deals with the problems of metrological, algorithmical and software ensuring of the system. The methodical and designer's features of microcompound chromatographs are mentioned. Special attention is paid to the problems of information flow organization and system calibration.

212 P.5-06 AMPEROMBTRIC FLOW-THROUGH SOLID ELECTRODE DETECTOR. DESIGN, PERFORMANCE AND APPLICATION IN HPLC FOR THE MICROCONCENTRATION ANALYSIS OF ANTIOXIDANTES IN MONOMERS АМПЕРОМЕТРИЧЕСКИЙ ПРОТОЧНЫЙ ДЕТЕКТОР С ТВЕРДЫМ ЭЛЕКТРОДОМ. КОНСТРУКЦИЯ, ХАРАКТЕРИСТИКИ И ПРИМЕНЕНИЕ В ВЭЖХ ПРИ АНАЛИЗЕ МИКРОКОНЦЕНТРАЦИИ АНТИОКСИДАНТОВ В МОНОМЕРАХ A.Ya.Lazaris, L.N.Beloded, A.I.Kalinin, Gorky, USSR At present electrochemical detection in HPLC attracts ever increasing attention of analysts due to its high selectivity, sensitivity and practically unlimited number of possible routes to perform«analysis. Despite a wast number of publication the design of EC detector becomes more sophisticated. Polished glassy carbon is employed as working electrode. Wall-jet detection operates as three electrode scheme. The cell is a slit of a complex shape with an auxiliary electrode being superimposed over the working one. Maxi mum sensitivity of measuring circuits is 0,4 nA/cm,

144 minimum detectable quantity in oxidation mode is 5 10" mg (hydroquinone). Linear dynamic range is more than 104, repeatability is about 1,5-3 %. Detector sensitivity and yield of conversion versus eluent velocity has been studied. Electrode performance has been tested based on polarography in flowing stream with the eluent containing a constant solute concenration. The detector is used for antioxidants analysis in acry lic monomers. Electrochemical antioxidants behaviour and hydrodynamic polarograms have been studied in constant current and differential pulse modes. Analytical proce dure for hydroquinone, p-methoxyphenol and phenothiasine estimation for 5-10"5- l'lO-4 * wt concentration has been proposed.

213 P.5-08 THE NEW IONIZATION METHOD FOR DETECTION 0Г GASES AND VAPOURS НОВЫЙ ИОНИЗАЦИОННЫЙ МЕТОД ДЕТЕКТИРОВАНИЯ ГАЗОВ И ПЛРОВ E.B.Shmidel, L.I.Kalabina, V.N.Khokhlov, L.N.Kolomiets, Moscow, USSR

The sensitivity of electron capture detector for rany problems in gas control of a threshold level is insuffi cient. The more sensitive detection method without disadvan tages limiting the extreme performances of electron cap ture method has been developed. Some aspects of sensitivity mechanism of new method have been considered. The physical model of ionization detection was introduced, which is based on the changes in mobility of the change carriers upon getting into the sensitive volume of the analyzed sunstance. The functional correlation was obtained between the net current signal, concentration of the analyzed substan ce, its electron affinity at a given operation mode and the detector design.

The detector design realizing the new method has been developed. The experiments showed that the data obtained correlated with theoretical conclusions. The obtained value of threshold sensitivity for lindane in crude nit rogen is better than WO"1* g/s. The extreme performances of this method are limited now by instability of gas and electric connections of detec tor. The new ionization method is perspective for gas chromatographic determination of trace amounts of pesti cides in atmosphere.

10.3»к. 872 US 214 P.5-09 EFFECT OF HATER BACKGROUND ON THE RESPONSE OF GAS CHROMATOGRAPHIC DETECTORS ВЛИЯНИЕ ФОНА ВОДЫ НА ОТКЛИК ГА30ХР0МАТ0ГРАФИЧЕСКИХ ДЕТЕКТОРОВ М.Dressier, Brno, CSSR At the direct analysis of water and aqueous solutions by gas chromatography, co-elution of water high compounds under investigation can occur. It is therefore desirable for the purposes of quantitative analysis that the extent of detector response changes due to water background should be known. The effect was followed with the alkali flame ionization, photoionization and flame photometric detectors. The intensity of the influence differs for various detector types, the decrease in the response caused by water background can amount up to tens of per cents. Character of the response, however, makes it possible to use these detectors in steam eolid chromato graphy , 215 P.5-10 APPLICATION OF HELIUM-DETECTOR FOR CONTROL OF ULTRA HIGH PURE HYDROGEN ' ПРИМЕНЕНИЕ ГЕЛИЕВОГО ДЕТЕКТОРА ДЛЯ АНАЛИЗА УЛЬТРАЧИСТОГО ВОДОРОДА H.Merten, M.Schirutschke, Dresden, GDR P.Popp, G.Oppermann, Leipzig, GDR The purity hydrogen is a essential condition for the production of integrated circuits. A simple device with a helium-detector allows to mea sure the following .impurities in hydrogen: oxygen, nitro gen, methane and carbon dioxide. Concentrations of 0,5 ppm„ are detectable. A parallel-columns-technique is used for chromatographic separation. Column 1 contains molecu lar sieve 5A. Column 2 contains a composition of Porapak Q and T. All parameters (length of the columns, temperatu re, flow) are preselected so, that all components enter the detector without overlapping. The authors discuss different ways of operation and give conditions for foolproof behaviour of the helium- detector in the continuous run.

216 P.5-11 THE USE OF A NOVEL DESIGN OF SATELLITE GAS CHROMATOGRAPH FOR MULTIDIMENSIONAL CHROMATOGRAPHY ПРИМЕНЕНИЕ НОВОЙ КОНСТРУКЦИИ САТТЕЛИТНОГО ГАЗОВОГО ХРОМАТОГРАФА ДЛЯ МНОГОМЕРНОЙ ХРОМАТОГРАФИИ D.F.K.Swann, Cambridge, Great Britain Abstract not arrived on time for printing.

146 , 217 P.5-12 THE DESIGN AND PERFORMANCE CHARACTERISTICS OF A NEW LC SYSTEM FOR MICROBORE AND FAST LC КОНСТРУКЦИЯ И ХАРАКТЕРИСТИКИ НОВОЙ СИСТЕМЫ ДЛЯ СКОРОСТНОЙ И МИКРОКОЛОНОЧНОЯ ХРОМАТОГРАФИИ J.Dolphin, Cambridge, Great Britain Abstract not arrived on time for printing. 218 P.5-13 IMPROVEMENT OF INFORMATION RELIABILITY AND FAULTS DIAGNOSTICS IN INDUSTP.IAL CHROMATOGRAPHS ПОВЫШЕНИЕ ДОСТОВЕРНОСТИ ИНФОРМАЦИИ И ДИАГНОСТИКА НЕИСПРАВНОСТЕЙ ПРОМЫШЛЕННЫХ ХРОМАТОГРАФОВ L.A.Rusinov, V.V.Kurkina, Leningrad, USSR The industrial application of chromatographs, especial ly as effective sensing transducers of composition direct ly in the automatic control systems for technological processes, gives priority to the problem of providing the high functional reliability of equipment. Among different methods of improvement of analytical information reliability the most available and quite effec tive are the algorithmic methods, we may distinguish two groups of these methods. The first group includes different statistical methods of reliability control of processing information and is based on traditional methods of logging with revealing of different abnormal features by means of some criteria. The testing in this case is based on the computation of corresponding statistics from sliding samples of M values of monitoring parameter and comparison of these statistics with some threshold limits. The research of the criteria was done by Monte-Carlo method and recomen- dations depended on the conditions of realization were given, considering its application. The methods of the second group are based on the esti mates of signal parameters (or results of processing), which are robust to the possible presence of rough errors, spikes, drift, pulse noise and other factors. Here it is possible to use some transformations of initial data (for example the processing in spectral domain with respect to the signal model and correlation function of noise), or to use other robust methods of estimation. Some simple in realization methods are considered; the results of this algorithms investigation are given. The cause-consequence information graph for typical in dustrial chromatograph was drawn, on the base of its ana lysis the set of the most informative features is found, which provide control of information reliability with high degree of faults identification probability. The diagnostics test designed for realization in micro computers was synthesized and provided the possibility of faults automatic detection in chromatograph apparatus.

U7 It is shownf that for effective operation of control and diagnostic system it is necessary to provide 2-level state control of chromatograph's detector signal and inter mediate and final results of processing, together with the main regime parameters of the analyzer. 219 P.5-16 NEW MULTIDIMENSIONAL GAS CHROMATOGRAPHIC SYSTEMS IN INDUSTRIAL ANALYTICAL CHROMATOGRAPHY НОШЕ МНОГОМЕРНЫЕ ГА30ХР0МАТ0ГРАФИЧЕСКИЕ СИСТЕМЫ В ПРОМЫШЛЕННОЙ АНАЛИТИЧЕСКОЙ ХРОМАТОГРАФИИ V.N.Lipavsky, Moscow, USSR Industrial analytical chromatography is traditionally developed as isotermic multidimentional chromatography. The devlopment of multidimensional gas chromatography in connection with the problems of industrial analytical chromatography is based in this paper on system approach. New difficulties appeared are caused by the application of control systems of data processing in the real time Scale and utilization of big volume of information for active control. It was found that one of the most optimal methods of their solution is development of multidimen sional chromatography for routine analysis. Multidimensional chromatographic systems with columns and switching devices as the main units are described and classified. Multidimensional systems, providing realization and improvement of main chromatographic characteristics are examined. Among them are - separation criteria, system efficiency, reducing of analysis time and measure devia tions, increasing in system safety and sorbent life, chan ges of separation efficiency and the application of vari ous gas mobile phases during the same analysis, indepen dent utilization of thermic factors in different parts of chromatographic system, summing up (concentration) of components, exception of columns overload. Practical examples are represented. 220 P.5-18 CHROMATOGRAPHS AS UNIVERSAL MEASUREMENT SYSTEMS OF PHYSICO-CHEMICAL PARAMETERS OF NATURAL GASES ИСПОЛЬЗОВАНИЕ ХРОМАТОГРАФОВ В КАЧЕСТВЕ УНИВЕРСАЛЬНЫХ ИЗМЕРИТЕЛЕЙ ФИЗИКО-ХИЛИЧЕСКИХ ПАРАМЕТРОВ ГАЗОВ A.M.Ovsepyan, V.P.Zalinyan, A.V.Khitarov, E.S.Ter-Stepanyan, Yerevan, USSR The possibility of construction of universal measure ment systems for determination of physico-chemical para meters of gases on base of the chromatographs realising in automatic regime the analysis of all components of gase system, was shown. On the example of natural gas it was shown, that the realised precision of determination of such parameters as density and specific heat of combustion, calculated on 148 additive property of gases with using of information about the composition of gas, is the best one among the known methods. The precision of determination of different com ponents is normal for gas chromatography and that fact is the most important property of these measurement systems. The developed system includes two columns and two derectors thermoconductivity and the heat of combustion) in combination with integrator and calculator. The system solves one of the most complex problem of gas industry - a problem of automatic determination and the transmission on the high level of system of automatic regulation for operative and high precision information about principal physico-chemical parameters of natural gas. The measurement system has got a good metrological cha racteristics according to the theoretical and experimental data for chromatographs with microprocessor. For measurement system it was presented structure and functional scheme including the chromatographs with auto- control system, fixation of time "on" and "off" of current, control of results of analysis,etc.

221 P.5-17 THE AUTOMATIC MEASUREMENT SYSTEM FOR QUALITY CONTROL OF NATURAL GAS BY GAS CHROMATOGRAPHS АВТОМАТИЧЕСКАЯ ИЗМЕРИТЕЛЬНАЯ СИСТЕМА КОНТРОЛЯ КАЧЕСТВА ПРИРОДНОГО ГАЗА НА БАЗЕ ХРОМАТОГРАФОВ V.P.Zalinyan, A.M.Ovsepyan, Z.A.Arutunyan, E.S.Stepanyan, A.V.Khitarov, Yerevan, USSR New microprocessor technique was a base for development of partial or full automatic multifunctional system of quality control of natural gas using the gas chromato graphs with microprocessors. The original scheme of automatization of analysis by means of chromatographs and microprocessors was a base for development of practically full automatic system of natural gas quality control when.operator is only to press a button.' The other operations including sampling, chan ging of columns and detectors, registrators are automatic. Chromatograph "Zvet 1OO0" enables to analyse on four columns with katharometer and flame-photometric detectors. The regime of analysis is set up by microprocessor. The developed system permits to determine real concent rations of main natural gas components: N,, CO,, H20,

H2S, CH,SH, C-H-SH, from CH. to CgH,- and to give 5ut data on density specific heat of combustion number Webbs viscosity, number of adiabats on printer or display. In communication structure functional scheme, scheme of automatization algorithms of data collection and working up of emerging data, faulty diagnostics, physico-chemical parameters, regulation of the whole system, methods of chromatographic analysis etc, are presented.

i 222 P.5-19 AUTOMATED INFORMATIVE MEASURING SYSTEM "COMPLETE CHROMATOGRAPHIC LABORATORY KHL-1" АВТОМАТИЗИРОВАННАЯ ИНФОРМАЦИОННО-ИЗМЕРИТЕЛЬНАЯ СИСТЕМА "КОМПЛЕКТНАЯ ХРОМАТОГРАФИЧЕСКАЯ ЛАБОРАТОРИЯ КХП-1" P.A.Moroz, V.N.Khokhlov, Moscow, USSR The system consists of five gas chromatographs and microcomputer. Each chromatograph with electronic devices connecting it to computer forms an information channel. The data system provides on-line information processing independently for each -channel at sampling rate up to 20 Hz. The system provides the calculation of sample com position using four different techniques simultaneously. So called automated calibration mode is possible. There is calibration table formed in this mode, which contains calibration coefficients for every mixture component being calculated by the least squares procedure. The sys tem also provides verification mode, which allows to obtain average concentration, it's standard deviation, and resulting error calculated for every verification mixture component. These statistical characteristics are calculated for pre set number of verification analysis. The resulting error ie calculated taking into acount the error of verification mixture preparation. Using test mixtures there following results were obtained for thermal conductivity and flame ionization detectors: standard deviation 0,5 to 1,8 per cent and resulting error 2,0 to 5,5 per cent from current concentration value of test mixture components. KHL-1 system allows user wide abi lity to control the information processing, and executes effective automated processing algorithm. Processing parameters and user directives required enter the system using the dialogue mode.

223 P.5-39 GAS CHROMATOGRAPHIC STUDY OF COMPLEXING OF ALIPHATIC NUCLEOPHILIC SOLUTES ON STATIONARY PHASES CONTAINING CHELATES OF PRASEODYMIUM ГА30ХР0МАТ0ГРАФИЧЁСК0Е ИССЛЕДОВАНИЕ К0МПЛЕКС00БРА30ВАНИЯ АЛИФАТИЧЕСКИХ НУКЛЕОФИЛОВ НА СТАЦИОНАРНЫХ ФАЗАХ, СОДЕРЖАЩИХ ХЕЛАТЫ МЕТАЛЛОВ W.J.Kowalski, Katowice, Poland Coordinative interactions between the chelates of pra seodymium: tris(2,2,6,6-tetramethyl-3,5-heptanedionato)Pr and tris(6,6,7,7,8,8,8-heptafluoro-2,2-dimethy1-3,5-octan- edionato)Pr dissolved in a polydimethylsiloxane stationa ry liquid phase and series of solutes involving homolo gous members of aliphatic alcohols, methyl ketones, methyl ethers and carboxylic acid methyl esters were studied within a temperature range 9C—140°C by means of the isothermal gas chromatography. The thermodynamic equilibrium constants for complexing ISO /1/ were determined from the retention indices of solutes by a regressional correlation procedure /2/. One found a selective Increase of retention parameter values for the given series of solute compounds when chromatographed on the complexing stationary phases. The fluorinated chelate of praseodymiun enhanced more selectively the formation of complex than the methylated one. The interaction intensity of fluorine substituents in the reference stationary liquid phase was considerably lower.Main complexation scheme was discussed. 1. J.R.Conder, C.L.Young, "Physico-chemical measurement by GC", John Wiley and Sons Ltd., New York, (1979), p.222. 2. R.V.Golovnya, D.N.Grigoryeva, J.High Res. Chromatogr., Chromatogr. Commun., £,(1983), 269. 224 P.5-40 APPLICATION OF SEC TO FOLLOW THE POLYCONDENSATIOH COURSE OF UNSATURATED POLYESTERS ПРИМЕНЕНИЕ ГПХ ДЛЯ КОНТРОЛЯ ПОЛИКОНДЕНСАЦИИ НЕНАСЫЩЕННЫХ ПОЛИЭФИРОВ J.Veith, G.Wigand, W.Leukefeld, Schkopau, GDR This paper presents the possible application of effi cient SEC for controlling the manufacturing process of unsaturated polyesters based on phthalic and maleic anhyd ride as well as propylene and butylene. glycol. Waters ALC/GPC244 gel .chromatograph (USA) used is provided with a set of 4 ji - styragel columns (1000-500-500-500 A), an UV-detector (#* 254 run), and a RI detector. Tetrahydro- furan was used as an eluting agent. With the chromatograms separate peaks were obtained for phthalic/maleic anhydride, maleiс acid, and phthalic acid. A whole series of lower condensation steps can be distinguished as peaks or shoulders. It has been shown that in addition to determining the molecular weight distribution and the average molecular weights by use of an efficient SEC, the time for attaining the optimum molecular weight distribution as an important economic factor as well as the content of monomers and low oligomers as a quality factor are easily and readily accessible. 225 P.5-41 POSSIBILITIES OF GAS CHROMATOGRAPHIC INVESTIGATION OF ASSOCIATION PHENOMENA BETWEEN HYDROCARBONS AND ALUMINIUM CHLORIDE IN SOLUTION ВОЗМОЖНОСТИ ГА30ХР0МАТ0ГРАФИЧЕСК0Г0 ИССЛЕДОВАНИЯ АССОЦИАЦИИ УГЛЕВОДОРОДОВ С ХЛОРИНОМ АЛЮМИНИЯ В РАСТВОРЕ H.Pscheidl, E.Oberdorfer, E.MSller, D.Haberland, Greifwald, GDR The association of some alkanes, alkenes, and aromatic compounds on aluminium chloride dissolved in liquid polytetrafluoroethylene an inert and stable phase is investi- 151 gated by meane of a gas chromatographic elution method w.' thin the temperature range of 50-80°C. In all cases it i.j poesible, to measure association phenomena depending on the amount of dissolved aluminium chloride and hydro carbons, respectively, and their molecular structure. The resulting decrease in the vapour pressure of hydro carbons being present as solutes in a liquid phase contai ning a complexing substance effects an increase in reten tion behaviour and is measurable on the other hand in a head space device. The association constants calculated from experimental results of both methods are comparable.

226 P.5-42 GAS CHROMATOGRAPHIC INVESTIGATION OF SILICIC ACIDS SILYLATED IN AQUEOUS SOLUTION ГА30ХР0МАТ0ГРАФИЧЕСК0Е ИССЛЕДОВАНИЕ КРЕМНЕВЫХ КИСЛОТ ПУТЕМ СИЛИЛИРОВАНИЯ В ВОДНОМ РАСТВОРЕ G.Bouguet, D.Knausz, J.Galacz, K.Ujszaszy, Budapest, Hungary The fact that trimethylsilylated derivatives of mono- and disilicic acids as well as some silicic acid oligomers can be determined by gas chromatographic method at. suitab le conditions /1,2/ offers a new possibility for the in vestigation of the composition of silicic acid solutions if the silylation does not cause transformation among the molecules of silicic acids. This can be expected of sily lation is a fast reaction, all of the si Hanoi groups are silylated, and the reaction does not change the concent ration and the pH of the solution. According to our experiences, the N,N-dimethylsilylcar- bamate meets these requirements /3,4/. Its effect on silicic acids was investigated in the aqueous solutions of SMOC-H,). and OSi2(OC,H5)6. The reaction between the solution consisting silicic acids and the silylcarbamate, was very fast under the applied conditions. The silylated products, /(CH,),SiO/4Si and /(CH,)3Si/gSi20 could be well measured by gas chromatographic method from a mixture con taining silylcarbamate in excess. We have found that from diluted acidic solutions the whole quantity of the orthosilicic acid transforms into /(CH,),SiO/.Si. Under the same conditions the disilicic acid partly decomposes into monomers although polymeriza tion also occurs. These reactions are probably spontaneous ones and not the result of the effect of silylation. At least this can be concluded from the result of our experi ments. 1. G.Garzo, G.Alexander, Chromatographia 4_ (1971) 554. 2. G.Garzo, D.Hoebbel, J.Chromatogr., 119 (1976) 173. 3. D.Knausz,• A.Meszticzky, L.Szakacs, 3.Csakvari, K.Ujszaszy, J.Organomet. Chem., 256 \1983) 11 • 4. D.Knausz, A.Meszticzky, L.Szakacs, B.2sakvari, J. Organomet. Chem., 268 (1984) 207.

1S2

/ 227 P.5-43 CHROMATOGRAPHIC STUDY OF INTERMOLECULAR HYDROGEN BONDS IN A SERIES OF ORTHO-TOLUIDIDES OF DIALKYLAMINOACETIC ACIDS ХРОМАТОГРАФИЧЕСКОЕ ИЗУЧЕНИЕ МЕЖМОЛЕКУЛЯРНЫХ ЮДОРОДНЫХ СВЯЗЕЙ В РЯДУ ОРТО-ТОЛУИДИНОВ ДИАЛКИЛАМИНОУКСУСНЫХ КИСЛОТ V.V.Larikov, V.S.Shklyaev, V.I.Pantsurkin, V.I.Ryazanov, Perm, USSR The investigation of H-bond nature is important for the discovery of the mechanism of a pharmacologic effect, the structure and reactivity of organic compounds. The capacity to form intermolecular H-bonds with metha nol, acetonitrile, acetone, diethyl ether and dioxan of orthotoluidides of dialkylaminoacetic acids has been inves tigated in the present paper. Values AH, As and AG with a relative error 5Ъ have been determined. The negative values of these parameters show the formation of H-complexes. It has been stated that the intermolecular bond formation goes along two mechanisms which is caused by the influence of the end radicals. It was shown that the big ger AG value, the smaller was the reactivity of the compounds. The absolute values of the complex formation constants (Kr) of the irtermolecular hydrogen bonds have been calculated by the "two liquid phase" method. The sta bility of H-complexes of the investigated compound series is not high which is demonstrated by the linear dependence

228 P.5-44 A CHROMATOGRAPHIC METHOD FOR EVALUATING PHYSICO-CHEMICAL AND THERMODYNAMIC CHARACTERISTICS OF PLASTIC COLLOID ХРОМАТОГРАФИЧЕСКИИ МЕТОД ОПРЕДЕЛЕНИЯ ФИЗИКО-ХИМИЧЕСКИХ И ТЕРМОДИНАМИЧЕСКИХ СВОЙСТВ ПЛАСТИЧНЫХ КОЛЛОИДНЫХ СИСТЕМ S.A.Leont'eva, E.M.Nikonorov, A.N.Timofeeva, Moscow, USSR Thermodynamic sorption parameters determined by the reverse phase chromatography are suggested for evaluating the thermodynamic state of plastic colloid systems. The stable thermodynamic state satisfies the minimum Gibbs energy, which is the sum of the bulk phase energies AG, and AG,, and the surface energy S' S. We suggest that uG, and AG, should be evaluated using the sorption energies of the system components, i.e. the disperse phase and dispersion medium, while the free sur face energy can be determined from the retention volume and the phase interaction energy of the system components Th£°¥Kermodynamic stability criterion

Na3P04 И ЕГО ИСПОЛЬЗОВАНИЕ ДЛЯ ИДЕНТИФИКАЦИИ I.L.Zhuravleva, B.M.Polanuer, R.V.Golovnya, Moscow, USSR Donor-acceptor interaction of organic bases with inorga nic salts present in sorbent is used for selective GC se paration of them. Alkylpyridines are capable to donor-ac ceptor interaction with Na,PO. but not with KF. Method for determination of temperature limit of a]kylpyridine comp lexes (T,) based on a comparison of thermodynamic data of sorption on two columns with non-polar stationary phase Apiezon (hydrogenated) and equimolar amounts of KF and

Na,P04 is suggested. The isokinetic relationship between 0( ДН), , and G" ( US), , f°r eleven alkylpyridines with a correlation coefficient 0,999 was observed. It shows that the type of complexes with Na.PO. is the same for all compounds. The temperature limit of complexing was calculated from an equation

<Э ( ДН), , = T,- &(iS), 2. Interval of T, values for eleven alkylpyridines is 122- 132°C. x These data can be used for group identification of alkylpyridines in a mixture with other compounds which don't interact with Na,PO.. The method includes determina tion of retention indices on a column with Na,PO. at two temperatures: 100 and 150 С Compounds which nave sharp decreasing of index values (~25 i.u.) are alkylpyridines.

230 P.5-46 GAS CHROMATOGRAPHIC SEPARATION OF VOLATILE COMPOUNDS AS A TOOL FOR REALIZATION OF ABSOLUTE METHOD OF ATOMIC FLUORESCENCE ANALYSIS ГА30ХР0МАТ0ГРАФИЧЕСК0Е РАЗДЕЛЕНИЕ ЛЕТУЧИХ СОЕДИНЕНИИ КАК СПОСОБ РЕАЛИЗАЦИИ АБСОЛЮТНОГО МЕТОДА АТОМНО-ФЛУОРЕСЦЕНТНОГО АНАЛИЗА V.I.Rigin, Krasnoyarsk, USSR The impossibility of manufacturing of the enormous number of standard-reference materials for calibrating of analytical instruments in order to analyze continuously increased number -of analyzed objects urgently reguires development of the absolute methods of analysis, which allows the calibration of instruments without using of 154 standard reference materials, especially for routine environmental analysis. The best and versatile technique of separation in gas chromatographic preconcentration and separation of the determined elements after their conversion into volatile compounds. The indispensable preliminary condition for efficient separation consists in total decomposition of sample and full conversion of determined elements in con ventional unique forms, for instance, simple volatile compounds or volatile metal chelates. The sample was decomposed in autoclave by means of XeF.. This compound undergoes decay at elevated temperatu re to xenon and free fluorine, which converts all the components of sample into fluorides of corresponding ele ments. The volatile fluorides of refractory metals were distilled off autoclave into gas chromatographic column, filled with granulated cerium trifluoride. The deposited in column metal fluorides were separated by means of heating the column, in succession of increasing of the retention coefficients. The blowing out column gases were fed in atomizer of multielement atomic fluorescence spectrometer. The residue in autoclave was treated by HC1 with addi tion of boric acid. The obtained solution was extracted using chelating agents which yields volatile metal chela tes, e.g., di-(trifluoroethyl)-dithiocarbamate. This extract was chromatographed on suitable column using multielement atomic fluorescence spectrometer as detector. The total decomposition of sample and efficient gas chromatographic separation of determined elements provide a basis for increasing of sensitivity of analysis. The offering technique has been used for multielement analy ses of soils, coals, fly-ashes, some environmental objects, ores on contents o.f a gxeat many metals, with sensitivity of determination to Ю "% and relative standard deviation not higher than 0,10. 231 P.5-47 MODIFIED METHOD FOR THE GAS CHROMATOGRAPHIC DETERMINATION OF CYCLOHEXAHONE AND CYCLOHEXANOL IN CYCLOHEXANE OXIDATION PRODUCT МОДИФИЦИРОВАННЫЙ МЕТОД ОПРЕДЕЛЕНИЯ ЦИКЛОГЕКСАНОНА И ЦИКПОГЕКСАНОЛА В ПРОДУКТАХ ОКИСЛЕНИЯ ЦИЮ10ГЕКСАНА МЕТОДОМ ГЖХ W.Czerwiriski, .Warsaw, Poland The method consists in gas chromatographic separations of cyclohexanone and cyclohexanol in conditions elimina ting decomposition of cyclohexyl hydroperoxide being an intermediate in the process. The contents of cyclohexanone are in the range 1-2%, those of cyclohexanol 2-3% and those of cyclohexyl hydroperoxide up to 1% in cyclohexane as the main constituent. Prior to gas chromatography acidic components of the samples (such as oxalic acid and others) are removed by 155 neutralization with ethanolic potassium hydroxide solu tion. Separations are carried out at the temperature of 60°C using a PTFE column packed with polyethylene glycol on textured glass beads. After recording each of the ohro- matograms the column should be conditioned at 110°C before the next injection. Addition of the constituent to be determined is used for calibration. By working wi*-h a precise electronic integrator for peak area measurements a relative standard deviation of about 1,5% can be achieved.

232 P.5-48 DETERMINATION OF VOLATILES MICROIMPURITIES CONSTANTLY FORMING IN POLY'-IERIZING SYSTEMS BY HEADSPACE GAS CHROMATOGRAPHY ГА30ХР0МАТ0ГРАФИЧЕСК0Е ОПРЕДЕЛЕНИЕ МИКРОКОЛИЧЕСТВ НЕПРЕРЫВНО ОБРАЗУЮЩИХСЯ ЛЕТУЧИХ ПРОДУКТОВ РЕАКЦИИ В ПОЛИМЕРИЗАЦИОННЫХ СИСТЕМАХ МЕТОДОМ ПАР0ФАЗН0Г0 АНАЛИЗА R.N.Mokeeva , Yu.A.Tsarfin, Vladimir, USSR H.Ernst, Schwarzheide, GDR Method of continuous gas extraction was suggested to determine the level of volatiles constantly forming in liquid polymerizable systems. Gas extraction of volatile reaction products was car ried out by passing the flow of argon through the polymer!- zate at a rate of 15-60 ml/min till equilibrium between quantities of products formed in the system and extracted from was achieved. From 10.000 to 40.000 g of the polymerizate sample was placed in a 20 ml glass vessel conditioned at 50-130C. Vapour phase was injected by means of a gas distribu tion tap with a 1-ml loop. The entire injection unit was maintained at 120°C. The equilibrium gas phase was monitored by Tsvet-5 chromatograph with flame ionization detector and 3 x 0,3 m glass column packed with 20% _|S-methoxy( .Д-cyanate-ethoxy) diethyl ether on chromosorb W (60-60 mesh) at 55 С and argon rate of 38 ml/min. Equations were suggested to determine the level of vo latiles yielded, specific rates of formation and distribu tion coefficients for the substances analyzed in the polymerizate-argon system. The method applicable for investigation of propylene oxide alkaline polymerizate was used to determine the level of acetone microimpurities formed during the propy lene oxide polyol synthesis. Mean-square errors are: 1.8% for determination of acetone content (at the level of 0.8-10 % - in the poly merizate) , 2.0% for distribution coefficient of acetone in the propylene oxide alkaline polymerizate-argone sys tem, and 1.6% for specific rate of acetone formation.

156

/ 233 P.5-49 DETERMINATION OF SATURATED VAPOUR PRESSURE OF FRAGRANCES USING GAS CHROMATOGRAPHY APPARATUS ОПРЕДЕЛЕНИЕ ДАВЛЕНИЯ НАСЫЩЕННОГО ПАРА ДУШИСТЫХ ВЕЩЕСТВ С ИСПОЛЬЗОВАНИЕМ ГАЗОХРОМАТОГРАФИЧЕСКОИ АППАРАТУРЫ Т.Д.Rudolf1, M.M.Shchedrina, L.O.Mindlin, K.K.Prilepskaya Moscow, USSR Saturated vapour pressure is a basic physical parameter, showing evaporation velocity of the substances. The inethod of determination of saturated vapour pressure has been developed using gas chromatography apparatus with tempera ture-programming. Concentration of saturated vapour in the flow was determined after cooling of vapour-gas mixture, that has been obtained in the injector. The comparative method using standard hydrocarbon having vapour pressure similar to that of the examined substance has been applied. The substance and the standard have been injected in two injectors of the same gac chromatograph simultaneously. The signals from two independent FID corresponding to the saturated vapour pressure of the substance and the stan dard at given temperature have been recorded by the dual pen recorder. So the series of the saturated vapour pres sure values have been obtained in one experiment. The saturated vapour pressure values' for-the studied substan ces at 20°C were found by extrapolation. The saturated vapour pressure values of different fragrances (over 150) related to various classes of organic compounds have been measured (p2° - 1,05-5,6 •Ю~ь mm Hg; 140-7,47»10~* Pa). 234. P.5-50 STUDY OF DILUTE SOLUTIONS OF IMPURITIES IN TITANIUM TETRACHLORIDE BY A GAS CHROMATOGRAPHIC HEAD SPACE ANALYSIS ИССЛЕДОВАНИЕ РАЗБАВЛЕННЫХ РАСТВОРОВ ПРИМЕСЕЙ В ТЕТРАХЛОРИДЕ ТИТАНА \?ОМАТОРАСПРЕДЕЛИТЕЛЬНЫМ МЕТОДОМ V.M.Pryakhina, A.I.Boiko, Zaporozhye, USSR Determination of microquantities of impurities in high- purity chlorides is a serious problem in analytical che mistry. In this paper partition ratios IK*) of impurities in titanium tetrachloride-based dilute solutions were determined in the liquid-gas system. Obtained Ki values enable one to find the conditions of carrying out the head space analysis of impurities in high-purity titanium tetrachloride. The values of maximum activity coefficients (Af**) for the compounds under inves tigation were determined as well. It is essential to point out the change of Ki with the 1 temperature in the range from 293 to 343 K. This permits temperature to be considered as a factor of optimizing the head space analysis of impurities. Changes in enthal py (AH) and entropy (AS) of impurities dissolution in titanium tetrachloride were calculated from the slopes of lg Ki vs. 1/T. 157

i Obtained constants have a decisive value for carrying out the head space analysis with a required accuracy and a sufficient sensitivity. 235 P.5-51 KINETIC INVESTIGATION OF THE CATALYTIC DECOMPOSITION OF ORGANIC PEROXIDES BY PULSE CHROMATOGRAPHY ИЗУЧЕНИЕ КИНЕТИКИ КАТАЛИТИЧЕСКОГО РАЗЛОЖЕНИЯ ОРГАНИЧЕСКИХ ПЕРОКСИДОВ ИМПУЛЬСНЫМ ХРОМАТОГРАФИЧЕСКИМ МЕТОДОМ A.F.Shushunova, Gorky, USSR The kinetics of the catalytic conversion of cumyl, tert-butyl and 1,1-diphenylethyl hydroperoxides under the chromatographic column conditions has been studied by means of modified pulse chromatography. In the course of experiments one part of the column was used as a microreactor, the other - as a separating column. The catalysts were inserted in the microreactor sorbent by different methods. The hydroperoxide solution was directly added into the microreactor by a microsyringe, the carrier gas flow being turned off. The sample was kept inside the microreactor for a definite time (from 0.5 to 30 min). Then the carrier gas flow was turned on again, ana the sample was transported into the separating part of the column, where the separation of the hydroperoxides and their decomposition products occurred. The catalytic cur ves for the consumption of the hydroperoxides as well as for the accumulation of their main decomposition products have been obtained using the results of the experiments. .The reaction rate constants and half-life periods of the hydroperoxides have been calculated.

236 P.£-52 THE NEW CHROMATOGRAPHIC METHOD OF EQUILIBRIUM STUDY OF HETEROGENEOUS REACTIONS IN SYSTEMS CONDENSED PHASE-GAS PHASE НОВЫЙ ХРОМАТОГРАФИЧЕСКИЙ МЕТОД ИССЛЕДОВАНИЯ РАВНОВЕСИЯ ГЕТЕРОГЕННЫХ РЕАКЦИЙ В СИСТЕМАХ КОНДЕНСИРОВАННАЯ ФАЗА- ГАЗОВАЯ ФАЗА L.G.Berezkina, S.I.Borisova, S.V.Melnikova, Moscow, USSR It is developed a chromatographic dynamic method of equilibrium study of heterogeneous reactions with volatile products evolved. The method includes the introduction .of a reactor with the sample into the gas chromatograph cir cuit before the measuring cell oi the detector, the satu ration of carrier gas flow with volatile components and the registration of the outlet equilibrium curves. The constant value of a detector signal and its independence on the conditions of contact of carrier gas and condensed phase - is a criterion of attaining the equilibrium. The method has its advantages, when studying multiple- staged processes, characterised with the formation of con- 158 densed and gas phase of alternative composition. In this case, wa employ flow division after the reactor, combina tion of detectors i conversion or selective absorption of hardly determining components. The dispersion of liquid phase into inert support was used to stabilize and increase contact surface area. The additional advantages: combination of saturation of the flow with Impulse inlet of the volatile component into reactor zone, the usage of isotermic contact temperature for a certain period of time, programmed heating and cool ing; the temperature gradient is absent in the chromatograph between the reactor zone and concentration measuring zone as a result, flow method limitations, connected with data distortion due to thermal diffusion. The method is applied for complex study of equilibrium processes of thermal ammonium phosphates dissociation with volatile ammonia and water, when process of polycondensation of phosphate in the melt is taking place. The depen dences of temperature on vapour pressure of the components and Дн for all phases of the process are determined. The peculiarities and advantages of the method are shown as well upon investigating equilibrium in system

Cr,(SO.)3 - Cr,03 - SO, - SO- - 0,, for which the applica tion of convential methods involves some difficulties. 237 P.5-53 THE GENERATION OF "PIANO" HYDROCARBONS, REQUIRED AS CC STANDARDS BY THE ADVANCED METHYLENE INSERTION REACTION ПОЛУЧЕНИЕ НАБОРА УГЛЕВОДОРОДОВ, НЕОБХОДИМЫ В КАЧЕСТВЕ СТАНДАРТОВ ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ С ПОМОЩЬЮ РЕАКЦИИ ВВЕДЕНИЯ МЕТИЛЕНОВЫХ ГРУПП T.Toth, E.M.Nagyvaradi, Budapest, Hungary A number of standards of different hydrocarbon types (paraffins /P/, i-paraffins /I/, aromatics /A/, naphthenes /N/, defines /0/ - let's call it "PIANO" system - cove ring the range of Се-С.,) are needed for reliable naphtha analysis for retention Index measurements and index-struc ture correlations, for tuning of multicolumn hydrocarbon analysers etc. At the same time such type of compounds are available commercially only in a relatively limited number and for rather high price. Dvoretzky et al. /1/ published an intelligent procedure for producing such compounds using diazomethane in the so called "methylene insertion treaction" (MIR). There are, however, limitations and disadvantages in the .generation of olefines and aromatics because of the cyclopropanisa- tion or ring enlargenlng side reactions. In the present paper this MIR procedure was modified eliminating the above disadvantages and enhancing its productivity: - the disturbing side products can be discovered and/or chemically filtered out; - the productivity can be dramatically increased by using

159 more than one precursors. For example, from a mixture of Cc-C,, n-alkanes treated together, 23 monomethylated paraffin» are produced in one reaction step. All of them can be separated and identified on a methyliilicone column of TZ - 50-80. - the other possibility to produce higher number of isomers is to conduct the reaction to a higher conver sion, thus not only the monomethyl but also the dimethyl derivatives can be produced. From n-heptane all the pre dictable 15 i-nonanes were produced and separated in such a way (even the two diastereomeric pairs.). Up to now 120 "PIANO" hydrocarbons mostly non-aromatics has been generated and the generation of further 100 ones (mostly aromatics) is in progress. 1. I.Dvoretzky, D.B.Richardson, L.R.Durrett: Anal. Chem. 35 (1963) 545-549.

238 P.5-54 THE STUDY OF COMPLEXES BY THE GAS CHROMATOGRAPHY ИССЛЕДОВАНИЕ КОМПЛЕКСОВ МЕТОДОМ ГАЗОВОЙ ХРОМАТОГРАФИИ S.L.Davydova, L.M.Rapoport, Moscow, USSR The complexes of transition metals of type L-M are acti ve participants in catalysis of organic and petrochemical reactions. In such reactions the coordination of organic substrate to metal which is already coordinated with 11- gand L takes place. The gas chromatographic method gives us the possibility of direct determination of substrate affinity to coordination centre of complex under study. For complexes of transition metals of type L-M, where L » polydiethylazelainate, polysalicylaldiminate and others, the physical sorbtion (by Roginekii-Yanowskli method) of number of organic compounds, having the diffe rent affinity to the complex, was determined (the field of low saturation). The affinity degree changes in the following sequence: pyridine 288, anyline 45, acetone 37, dioxane 30, benzene 20, ether 19, ethanol 11, methanol 7, pentane 0,97 mg/g of complex. The data obtained indicate the significant contribution of the forces, determined by coordinative unsaturation of metal atoms to the total energy of molecular interactions. For complexes of transition metals of type b,M, where L = macrocyclic compounds of crown-type, the complete adsorption isotherms (in intervals of nitrogen relative concentrations from 0.1 to 0.8) were measured. It has been shown that in capillar condensation region at 77 К the hysteresis can be observed only for complexes with signi ficant distances between the ligand molecules (hysteresis zone, adsorption 2.5-7.5%, the beginning of adsorption P/P = 0,48 - 0,35).

160 239 P.5-55 QUANTITATIVE HEAD-SPACE ANALYSIS КОЛИЧЕСТВЕННЫЙ АНАЛИЗ ПАРОВ J.Gregoire, Suresnes, France Knowing the concentration of a volatile compound "I" in a sample "S" equilibrium or static head-space analysis (S.H.A.) allows to calculate the partition coefficient "K" of this volatile compound between condensed and gaseous phases. With a simple liquid mixture "M" S.H.A. allows as well to determine the activity coefficient of the volatile mixture. S.F.A. has been used in this way. to show large non linear variations of activity coefficient versus the matrix composition of binary mixtures. On the contrary, it is inferred that activity coeffi cient of the volatile "I" in the sample "S" is equal to activity coefficient of the same "I" in the calibration mixture "H". when S.H.A. is used to estimate the concentra tion of a volatile compound, in a sample to be analyzed. AB a general rule, this assumption is not often valid. Therefore, B. Kolb proposed two stepwise static methods in order to avoid the corresponding quantitative error which resulted from the so-called matrix effects! discontinuous gas extraction (D.G.E.) and multiple head-space extraction (M.H.E.). Applied to the same samples, these static methods can give very different results when compared to the results coming from a dynamic head-space method. We describe the corresponding so-called D.C.I, {desorption - concentration- introduction) stepwise method. We show and explain the discrepancy with the help of various volatile compounds analysis (ethylene oxide, formaldehyde and styrene). It is because not only does the D.C.I, method take into account the solubility and the vapour pressure of a volatile core- pound to be analysed, but over-all its diffusion (which can limit its approach to the area of the sample where the desorption takes place!, that this dynamic method is more valid than the static methods, to eliminate matrix affects and the corresponding quantitative errors.

240 P.5-56 QUANTITATIVE HEAD-SPACE ANALYSIS UNDER NON-EQUILIBRIUM CONDITIONS КОЛИЧЕСТВЕННЫЙ ПАРОФАЗНЫЙ АНАЛИЗ В НЕРАВНОВЕСНЫХ УСЛОВИЯХ

A.G.Vitenberg, N.I.Kalacheva, Leningrad, USSR

Abstract not arrived on time for printing.

11.3ак.872 161 241 P.5-57 GAS CHROMATOGRAPHIC BEHAVIOUR OF FRAGRANCES IN APIESON L AND 1,2, 3-TRIS ( /*-CYANOETHOXY)PROPANE. THE ACTIVITY COEFFICIENTS DETERMINATION ГАЗОХР0МАТ0ГРАФИЧЕСКОЕ ПОВЕДЕНИЕ ДУШИСТЫХ ВЕЩЕСТВ НА АПИЕЗОНЕ L И 1,2,3-ТРИС/БЕТА-ЦИАНЭТОКСИ/ПРОПАНЕ. ОПРЕДЕЛЕНИЕ КОЭФФИЦИЕНТОВ АКТИВНОСТИ T.A.Rudolfi, M.M.Shchedrina, M.A.Dubrova, L.O.Mindlin, Moscow, USSR The behaviour of various sorbates in Apieson L and 1,2,3-tris ( j5-cyanoethoxy)propane have been studied. The character of the sorbates behaviour in the stationary pha se was evaluated using the activity coefficient ( ^&)» which was calculated from the well-known aquation

r= 1.704 10? U"> V° P M Alaboratedby us procedure employing gas chromatography apparatus was used for determination of saturated vapour pressure (P). The specific retention volume (V ) was cal culated from the retention time of the exam'.ned"substances. The activity coefficients have been determined for the series of fragrances (more than 50) belonging to various classes of organic compounds! alkyl substituted cyclohexa- nols and their esters, macrocyclic lactones and oxalacto- nes, macrocyclic diesters, terpenic alcohols and aldehydes. For some substances (more than 50, e.g. aliphatic alco hols and aldehydes, cinnamic aldehyde and cinnamic alco hols derivatives, some heterocyclic compounds) the relati ve activity coefficients У / У have been determined. Basing on the results obtained the dependence of ^"oo on the molecular mass and on the structure of the compounds has been considered.

242 P.5-58 COMPLETE HYDROGENOLYSIS OF ORGANIC COMPOUNDS IN A CHROMATOGRAPHIC REACTOR. A NEW SIMPLE VARIANT OF THE DETERMINATION OF TOTAL ORGANIC CARBON CONTENT IN WATER AND AQUEOUS SOLUTIONS ИСЧЕР.ЫВАЮИИИ ГИДР0ГЕН0ЛИЗ ОРГАНИЧЕСКИХ СОЕДИНЕНИИ В ХРОМАТОГРАФИЧЕСКОМ РЕАКТОРЕ. НОВЫЙ ПРОСТОЙ ВАРИАНТ ОПРЕДЕЛЕНИЯ ОБЩЕГО УГЛЕРОДА В ВОДЕ И ВОДНЫХ РАСТВОРАХ B.V.Stolyarov, E.E.Galev, Leningrad, USSR

Abstract not arrived on time for printing.

162 243 P.5-59 PURITY CONTROL OF CAPROLACTAM BY HEAD-SPACE GAS LIQUID CHROMATOGRAPHY КОНТРОЛЬ ЧИСТОТЫ КАПРОЛАКТАМА ХРОМЛТОГРАФИЕ0 ГАЗОВОЙ ФАЗЫ R.N.Kikolov, p.I.Pishev, A.D.Stefanova, Sofia, Bulgaria Head space sampling technique combined with GLC super ficially bonded (Carbowax 20M) packings has been proposed for purity control of caprolactam production. To determine the content of impurities in the original sample from its vapour phase chromatogram, the conventional procedure of internal normalization has been modified by introducing appropriate conversion factors instead of the commonly used peak-area correction coefficients. A suitable proce dure of determining the conversion factors has been deve loped. The application of GLC head space technique for industrial purity controj of caprolactam has been illustra ted by samples obtained from a Bulgarian chemical plant.

244 Р.5-60 GAS CHROMATOGRAPHIC ANALYSIS OF HIGH PURITY VOLATILE CHLORIDES, HYDRIDES AND PERMANENT GASES ГАЗОХРОМАТОГРАФИЧЕСКИЙ АНАЛИЗ ВЫСОКОЧИСТЫХ ЛЕТУЧИХ ХЛОРИДОВ, ГИДРИДОВ И ПОСТОЯННЫХ ГАЗОВ V.A.Krylov, A.E.Ezheleva, S.G.Krasotskii, G.V.Sokolova, S.D.Alexandrov, Gorky, USSR High reactivity of some inorganic matter as well as similarity in properties of impurities and of base sub stance lead to a number of specific problems while develo ping analytical techniques. It has been found that the substances investigated con tain the impurities of hydrocarbons, hydrocarbon chloro-derivatives, organoelemental compounds, chlorides, hydri des and permanent gases. Identification has been carried out by the method of additives and in some cases the other methods of analysis have been used such as mass-spectrometry and IR-spectroscopy. In this work the most sensitive detectors were applied: electron capture, flame-ionization, flame-photo metric, thermoionic, photoionization and helium detectors. The sources of systematic errors in the analysis of volatile inorganic matter have been considered. It has been shown that the content of moisture impurities in carrier gas as well as that of metal impurities in solid supports and adsorbents may result in the marked under estimation of the analysis results. Another source of errors may be the difference in composition of a liquid and of an equilibrium vapour. The estimation method of possible systematic error 'while liquid phase sampling is suggested. It is shown that application of capillary columns ma

I kss it possible to reduce the absolute limit of detection by at least an order of magnitude; relative limit of detec tion is also reduced when preliminary packed columns are used. Reaction gas chromatography markedly reduces the limit of detection of organic matter. It has. been found that application of carrier gas with residual moisture-of not more than 10 %, chemically resis tant stationary phases and above-mentioned detectors makes it possible to analyse high purity volatile inorganic matter with the limit of detection of lo"5 - 10~°%. The accuracy of determination has been confirmed by analysis of standard mixtures of known content. The values of rela tive standard deviation were in the range of 0,05- 0,3.

245 P.5-61 METROLOGICAL ESTIMATION OF GAS CHROMATOGRAPHIC METHOD USED FOR THE DETERMINATION OF IMPURITIES AND MAIN COMPONENT CHLORINATED HYDROCARBONS Cj- Cj МЕТРОЛОГИЧЕСКАЯ ОЦЕНКА ГА30ХР0МАТ0ГРАФИЧЕСК0Г0 МЕТОДА ОПРЕДЕЛЕНИЯ СОДЕРЖАНИЯ ПРИМЕСЕЙ И ОСНОВНОГО КОМПОНЕНТА

В ЛЕТУЧИХ ХЛОРУГЛЕВОДОРОДАХ Cj- C2 I.P.Ogloblina, V.V.Sergievsky, M.M.Ivanovo, M.T.Djaparidze, Moscow, USSR The determination of the main substance concentration in volatile and gaseous products by chemical analysis methods causes basic problems. The possibilities of gas chromatographic method have been estimated. It was assumed that main component concentration (C - lOO - £c., where C. is concentration of impurity i. The contributions of random and systematic errors in different analysis stages have been estimated using vari ous methods of sampling of gaseous and liquid samples. The precision of various methods for gaseous calibration mix ture preparation was compared. Ampoule-gravimetric, diffu sion and dynamic methods were used for concentration range 0.0001-0.5%. The error structure has been studied. It whas been found that a minimum relative error when determining one impurity component is not less than 5%. Ca libration when sampling with a syringe results in an error within 7-15%, using a sampling valve - 3-5%. For liquid sample calibration error is not less than 7%. The error in the total impurity concentration determi nation is a function of the number of impurities, their concentration and it ranges from 40 to 130%. The concentration of the main substance in methyl chlo ride and vinyl chloride within a range of 99.8-99.0% has been verified using this method. The error of determina tion was 2-3%.

164 246 P.5-62 ISOTOPO-CHROMATO-SPECTRAL ANALYSIS OF INORGANIC GASES ИЗОТОПНО-ХРОМАТО-СПЕКТРАЛЬНЫИ АНАЛИЗ НЕОРГАНИЧЕСКИХ ГАЗОВ V.M.Nemets, A.A.Solov'ev, v.N.Funtov, Leningrad, USSR The analytical process includes the dosed introduction of rear isotope of the component to be determined in sample of the analysed gas, the chromatographic isolation of the isotope mixture, the spectroscopic determination of isotope composition of it and the calculation of con tents of the component to be determined. During the deter mination of low contents of impurities the cryogen-adsorption concentration of isotopes of component to be deter mined is used. The method allows to determine both the molecular composition of the gas sample and its element composition. In the last occasion the isotope balancing of the compo nent to be determined is used taking into consideration all molecular forms which the component contains. For this purpose either catalytic action of the hot surface of platinum or the action of high-frequent non-electrone discharge are used. Isotope isolation is carried out by means of the elu - tion chromatography. The helium or argon are used as car rier gas. The source of the light with high-freguent non electrone discharge is used as chromatographic detector at the pressure of 6-8 10* Pa. It also serves for spectro scopic determination of isotope composition. In order to simplify the coupling of the light sources with chromato graphic column the separation in the column is carried out at the low pressure. The calculation of isotope concentration is carried out according to the formula obtained from solution of material balance equation, which suitable for procedure of isotope dilution. The method is used for the determination of such ele ments as H, N, 0, c, S and molecular forms - H,, N,, CO,,

02, S02, NO. 247 P.5-63 THE HIGH SPEED ANALYSIS OF THE GAS MEDIUM COMPOSITION ЭКСПРЕССНОЕ ОПРЕДЕЛЕНИЕ ГАЗОВЫХ СРЕД T.I.Lomovtseva, I.N.Mylynskaya, G.V.Pogosbekyan, N.A.Romanov, Z.I.Filatova, V.S.Yusfin, Moscow, USSR The possibility to obtain a quasi-continuous analyti cal information concerning the components of the gas me dium with a quickly changing composition is shown using the chromatographic analysis of the converter gas atmos phere. A mode of analysis is offered to receive a rapid in formation about the quantity of hydrogen, oxygen, carbon oxide and carbon dioxide using three parallel selective

165 analytical channels. The packod capillary columns with a diameter from 0.6 to 2.0 n» were filled with the sorbents of a narrow fractional composition. The minimizing of the levels of peak dispersion and the time-constant of detec tion was obtained by the use of the thermal conductivity microdetector and thermoehemical microdetector with cell volume of 30 and 50 jul respectively. The peak width at half height was about 1 second. Only 6,-10 seconds are necessary for complete separation and analysis of the mentioned gases. 248 P.5-64 AROMATIC FRACTION OF GEOTHERHAL HYDROCARBONS АРОМАТИЧЕСКАЯ ФРАКЦИЯ ГЕОТЕРМАЛЬНЫХ УГЛЕВОДОРОДОВ N.V.Porshnev, V.B.Bondarev, Moscow, USSR Views on the genesis of hydrocarbons in geothermal fluid are most contradictory. The problem is usually re stricted to the consideration of C.-C. hydrocarbons. This paper offers the results of tne composition of a hydrocarbon fraction at T. ,,» 68 to 220 С in gas and water condensate samples of the East-Pauxhetskii geothermal Held (Kamchatka, Sept. 1983). To do that the method of preliminary sorption into the polymeric sorbent Tenax GC. The method was chosen since in the commonly used methods part of low-volatile hydrocarbons may be lost over sampling - to detecting stages. According to gas chromatographic measurements, the net concentration of aromatic compounds (accounted to gas composition) in vapour-gas spray is higher than 6x10 mass % and is

comparable with the paraffin fraction C,-C6. In the wa ter condensate the abundances of benzene; toluene; o-, p-, m-xylenes and ethylbenzene; naphthalene are on the average 28, 44, 64, and 375 percent, respectively compa red to those in the gaseous phase. The analysis revealed that in the process of geothermal fluid condensation low-volatile hydrocarbons (such as naphthalene) are subject to considerable redistribution in gaseous and liquid phases. On the other hand, the com position of hydrocarbons in vapour-gas mixtures studied shows that the aromatic fraction forms at a large depth, possibly being a result of P, T-conditions at depth. The use of the method of preliminary concenration in situ of volatiles can improve the analytical potential of geothermal and even volcanic processes in the geochemical studied. 249 P.5-65 THE PURITY PARAMETER, A NEW CONCEPT FOR ACCURATY PURITY VERIFICATION IN HPLC ПАРАМЕТРЫ ЧИСТОТЫ, НОШЕ КОНЦЕПЦИИ ТОЧНОЙ ПРОВЕРКИ ЧИСТОТЫ В ВЭЖХ P.A.Kunz, T.Sheehan, Zug, Switzerland Abstract not arrived on time for printing.

166 250 P.1-64 CHROMATOGRAPHIC ANALYSIS OF THE CATALYTIC NON-TOTAL COMBUSTION PRODUCTS DERIVED PROM THE INDUSTRIALLY EMITTED ORGANIC COMPOUNDS ХРОМАТОГРА«ИЧЕСКИЙ АНАЛИЗ ПРОДУКТОВ НЕПОЛНОГО КАТАЛИТИЧЕСКОГО ПРЕВРАЩЕНИЯ ПРОМЫШЛЕННЫХ ВЫБРОСОВ ОРГАНИЧЕСКИХ СОЕДИНЕНИИ J.Rzepa, M.Sajewicz, Katowice, Poland The separation identification and quantitative determi nation were established by means of gas chromatography of the catalytic non-total combustion products derived from the industrially emitted organic compounds such as ben zene, toluene, ethylbenzene, xylenes, propylbenzene, butylbenzene and phenols (i.e. phenol, cresols, xylenole). The influence of the oxygen amounts in the combusted gaseous mixture, of the combusted substance concentration and of the catalyst type and working parameters was inves tigated upon the quantitative and qualitative characteris tics of the non-total combustion products.

1*7 CAPILLARY CHROMATOGRAPHY

251 S.l-4 COMPUTER OPTIMIZATION OF EXPERIMENTAL CONDITIONS IN CAPILLARY GAS CHROMATOGRAPHY ОПТИМИЗАЦИЯ ЭКСПЕРИМЕНТАЛЬНЫХ УСЛОВИИ В КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАЗИИ С ПОМОЩЬЮ ЭВМ J.Krupcik, JiMocak, D.Repka, S.Gergely, J.Garaj, Bratislava, SSSR One of the goals of chromatographic analysis is the separation of sample components in the shortest time. Se paration of complex mixtures in capillary gas chromatogra phy strongly depends upon the following working parameters: 1. Temperature of separation 2. Stationary liquid selectivity 3. Separation power of chromatographic column. In addition to these main parameters the separation is influenced also by the nature of carrier gas, polarity of the capillary column walls, temperatures of injection-port and detector, construction of a chromatograph (dead volu mes of connections and interfaces) and by few other fac tors. Influence of all additional factors on the separa tion process can be minimized by their standardization at the analysis. In computer optimization several optimization procedu res of the main factors determination has been published. Morgan and Deming/1/ have shown how to optimize temperature carrier gas flow-rate in analysis of model hydrocarbons mixture using sequentail simplex method and carrying out calculation by a computer. Purnell at al. /2/ used a win dow diagram approach to optimize the speed of analysis, temperature and the composition of binary mixtures of sta tionary liquids. He have published /3/ a procedure for computer optimization of temperature and stationary phase selectivity in capillary gas chromatographic separation to 40 polychlorinated biphenyls. Recently we have found a procedure for computer optimization of temperature gradi ent in separation of complex hydrocarbons mixture using linear temperature programmed capillary gas chromatography with Squalane and OV-101 stationary phases. The aim of the paper i. t evaluate the use of up-to- date published computer o^wj-imzation procedures in capil lary gas chromatographic separation of complex mixtures. The possibilities and limitations of the use of various optimization criteria in computer optimization procedures will be discussed in detail. 1. S.L.Morgan, S.N.Deming, J.Chromatogr., 112 (1975) 267. 2. M.Y.B.Othman, J.H.Purnell, P.Wainwright, P.S.Williams, J. Chromatogr., 289 (1984) 1. 3. J.Krupcik, J.Mocak, A.Simova, J.Garaj, G.Guiochon, J. Chromatogr.,238 (1984) 1.

168 252 S.13-1 A NOVEL APPLICATION OF COUPLED, "TANDEM" CAPILLARY GC COLUMNS TOR THE TUNING OF COLUMN POLARITY AND FOR THE DETERMINATION OF RI VALUES IN MULTICOMPONENT MIXTURES. A NEW APPROACH IN NAPHTHA ANALYSIS НОВОЕ ПРИМЕНЕНИЕ СПАРЕННЫХ КАПИЛЛЯРНЫХ ГА30ХР0МАТ0ГРАФИЧЕСКИХ КОЛОНОК ТИПА "ТАНДЕМ" ДЛЯ ТОЧНОГО ВЫБОРА ПОЛЯРНОСТИ И ДЛЯ ОПРЕДЕЛЕНИЯ ИНДЕКСОВ УДЕРЖИВАНИЯ В МНОГОКОМПОНЕНТНЫХ СМЕСЯХ. НОВЫЙ ПОДХОД К АНАЛИЗУ НЕФТИ T.Toth, Budapest, Hungary J.A.Rijks, J.H.M. van Cruchten, Eindhoven, The Netherlands A method Is described for reproducible continuous tuning of polarity of a two-column coupling in series ("tandem" system). It permits a predictable positioning of peaks as well as the determination of I, д I, dl/dT values for both of the two pure constituent columns. The system allows a detailed and highly reliable individual and type analysis of hydrocarbon mixtures. Our "type recognition" correlations in the & I vs. dl/dT plots (published previ ously) greatly help to achieve this. The columns of the tandem system can be coupled in series either directly or via a switching device, and they can be mounted either in a single oven or in independently controllable double-oven instrument (commercially avail able) . A simple mathematical model - based on the addltivity of the relevant k-TM products - was established. It was tested thoroughly, at the highest possible precision level by means of a number of cerfully selected model hydrocar bons. The full mathematical model and the practical details as well as the application, possibilities and limitations of different tandem realisation will be presented in comp lex hydrocarbon analyses such as pyrolysis naphtha and fluid catalytically cracked naphtha. 253 S.13-2 USE OF HIDE BORE CAPILLARY COLUMNS IN GAS CHROMATOGRAPHS DESIGNED FOR PACKED COLUMNS ИСПОЛЬЗОВАНИЕ ШИРОКИХ КАПИЛЛЯРНЫХ КОЛОНОК В ГАЗОВЫХ ХРОМАТОГРАФАХ, ПРЕДНАЗНАЧЕННЫХ ДЛЯ НАПОЛНЕННЫХ КОЛОНОК W.R.Supina, Pennsylvania, USA A.Holfensberger, Gland, Switzerland Wide bore (0.75 mm ID), wall coated open tubular capil lary columns have the large sample capacity of packed columns and the great efficiency of 0.20-0,25 mm ID capil lary columns. Hide bore capillary columns can be used in packed column gas chromatographs, with sample injectors and carrier gas flow regulators designed for packed columns. Specially designed but simple hardware kits en-

169 able an analyst to quickly interconvert the injector and detector connections of a packed column gas chromatograph for installing a 0.75 mm ID column or a packed column. A 0.75 mm ID column can also be used in instruments desig ned for capillary columns. In a capillary column system, 0.75 mm ID columns are compatible with split, splitless, and direct injection techniques. Typical analyses obtained from 0,75 mm ID capillary columns in packed column sys tems will be discussed, and the conversion kits will be described.

254 S.13-3 FUSED QUARTS CAPILLARY COLUMNS: PREPARATION AND ANALYTICAL POSSIBILITIES КВАРЦЕВЫЕ КАПИЛЛЯРНЫЕ КОЛОНКИ: ИХ ПРИГОТОВЛЕНИЕ И АНАЛИТИЧЕСКИЕ ВОЗМОЖНОСТИ V.A.Krylov, V.A.Lazarev, S.C.Krasotskii, Gorky, USSR K.l.Sakodynskli, Moscow, USSR Fused quarts and silica capillary columns are a new trend in capillary chromatography due to the fact that this material has a high inert and mechanical durability. Properties of capillary columns are determined by high quality of raw material. The treatment of the natural quarts dummy perform for drawing the capillary columns was considered. Film stability of stationary liquid phase is the most important property of capillary columns. The instability of liquid films with thickness of more than a micron has been explained by the action of surface tension forces in liquid-gas interface. In case of thin films the influence of surface forces of fused quarts becomes more important. The dependence of film thickness upon the rate of coating by stationary liquid phase using the dynamic method was in vestigated in capillaries with a diameter of 250 urn. The upper boundary of the advancing rate of nonpolar statio nary liquid phase is found which makes it possible to obtain stable films with a thickness of 0.1-0.2 urn. The analytical possibility of fused quarts capillary columns was considered. It was shown, that utilization of concentration detector (BCD) is more preferable than of flow one (FID). Application of capillary columns makes'it possible to reduce the absolute limit of detection by at least an order of magnitude, the relative limit of detec tion is also reduced when preliminary packed columns are used. Chloralcans gas chromatography analysis of a number of high purity water volatile chlorides and of organic solvents was performed with the limit of detection of 10-6- 10"a%. The values of relative standard deviation were in the range of 0.05-0.5.

170 255 S.13-4 REPRODUCIBILITY OF RETENTION DATA IN CAPILLARY GC ВОСПРОИЗВОДИМОСТЬ ДАННЫХ УДЕРЖИВАНИЯ В КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ E.Matisova, Bratislava, CSSR The main advantage of using capillary columns in chro matography is high efficiency. The criteria i'or judging the performance of capillary columns have remained the same over years and they include efficiency, selectivity, inertness and temperature operating range. For identifica tion purposes main stress is given to the reproducibility of retention data. In capillary GC with conventional liquid stationary phases retention data depend on film thickness. In such separations a mixed retention mechanism occurs /1,2/ that is dissolution in liquid phase and adsorption in gas- liquid and liquid-solid interface which depends on proper ties of stationary liquid, analysed solute and surface quality (glass, fused silica).'The adsorption properties of the capillary inner wall surface influence both quali tative (retention) and quantitative analysis when a non- polar stationary liquid is used. The molecule» of polar stationary liquid phase can undergo orientation on polar inner wall surface leading to a decrease of overall pola rity. This effect strongly influences retention mainly on columns with an active inner wall surface and thin films of stationary phase. The reproducibility of retention data of newly applied immobilized stationary phases has been also studied. Liquid crystals are stationary phases of medium pola rity, so mixed retention mechanism is expected. The effect of orientation of liquid crystalline stationary phase molecules is greater than with conventional stationary phases. It was verified /3/ that capacity ratios, Kovats indices and selectivity are significantly dependent on the film thickness of liquid crystalline stationary phase and surface quality. Melting and clearing points of liquid crystal are also dependent on these two factors. 1. J.Krupcik, E.Matisova, J.Garaj, L.Sojak, V.G.Berezkin, Chromatographia, 16 (1982) 166. 2. E.Matisova, J.Krupcik, J.Garaj, Chromatographia.16 (1982) 169. 3. E.Matisova, D.Hudec, J.Garaj, G.Kraus, M.Schierhorn, A.Isenberg, Chromatographia (in press). 256 S.13-5 QUANTITATIVE CAPILLARY GAS-LIQUID CHROMATOGRAPHY OF TRIGLYCERIDES КАПИЛЛЯРНАЯ ГАЗОЖИДКОСТНАЯ ХРОМАТОГРАФИЯ ТРИГЛИЦЕРИДОВ Р.MareS, Prague, £sSR Despite of the last development of liquid chromatogra phy and liquid chrornatography-mass spectrometry, gas- liquid chromatography is still the most sensitive method 171 for the analysis of triglycerides. Short glass and fused silica capillaries have been recently used for the sepa ration of these compounds. Unfortunately, there are some restrictions for the application of capillaries in this type of analysis. Host important one is the low volatility of triglycerides, which affect the introduction of the sample into the column and the quantitative recovery of the separated compounds. We have studied in our laboratory the factors affecting the quantitative analysis of triglycerides up to 63 car bon atoms in the molecule using fused silica column with chemically bonded stationary phase. Dependences of the results on carrier gas flow rate, construction of the injector, injection technique, amount of the stationary phase and column quality were observed. The results of our studies give us the possibility to optimize conditions for the quantitative course of the whole analytical procedure. Under the conditions of cold on-column injection we achieved the. limit of quantitative analysis for the component with 63 carbon atoms about 2-3 ng. Serial reproducibility of ttnalyses for 5 ng samp les of triglycerides was better than 2% except of triara- chidin, for which the c.v. value was 2,4%. These results were achieved on 5 m column with chemically bonded OV-1.

257 S.13-6 A CONCEPT IN MULTIDIMENSIONAL CAPILLARY CHROMATOGRAPHY ТЕОРИЯ МНОГОМЕРНОЙ КАПИЛЛЯРНОЙ ХРОМАТОГРАФИИ E.F.Dawes, P.A.Dawes, Ringwood, Australia Though the technique of multidimensional chromatogra phy has been available for many years, it has not found the widespread use that would be expected for such a powerful technique. Part of this is due to the concern by many chromatographers for the difficulty of the technique. The new system described has been designed with ease of set up and operation in mind. Though based on the Dean's system of pressure balancing it has several modifications that eliminate the critiqal aspects of pressure balancing. The low volume and all fused silica and glass system permits dual capillary column operation either with or without cold trapping on the second column. Packed column to capillary column operation with total or partial trans fer will be shown. The functions that can be carried out are: - heartcutting - backflushing - variable splitting of a sample between different columns and detectors - foreflushing - cryogenic focussing. Some examples of the power of the techniques available will be presented.

172 258 P.5-20 AUTOMATION IN HIGH RESOLUTION CAPILLARY GAS CHROMATOGRAPHY АВТОМАТИЗАЦИЯ ВЫСОКОЭФФЕКТИВНОЙ КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ P.A.Kunz, C.A.Ray, R.X.Simon, Zug, Switzerland Abstract not arrived oa time for printing.

259 P.5-21 UNIVERSAL DEVICE FOR SAMPLING AND PRELIMINARY CONCENTRATING OF SAMPLES УНИВЕРСАЛЬНОЕ УСТРОЙСТВО ДЛЯ ОТБОРА ПРОБ И ПРЕДВАРИТЕЛЬНОГО КОНЦЕНТРИРОВАНИЯ ОБРАЗЦОВ V.A.Minaev, V.V,Pomazanov,Yu.A.Kolchin, V.V.Kobac, A.M.Alymov, Moscow, USSR Device for sampling and preliminary concentrating of samples is used for chromatographic control of contamina tion of environmental objects with microadditlvee of che mical and biological origin. The main principles and mechanism of sampling and concent rating of samples and the results of development and test of device for preliminary concenrating ("UPK-2") are ob served. The characteristics of this device are on the le vel of the best serial modern devices. The technical data of the device are the following: factor of concentration, better than 1000 interval of temperature, °C 5-350 power requirements, H 250 deviation of temperature, С +2 emerge time on the.operation conditions, min ? dlmentions 300x250x150 This device may be used for the concentrating of ex tracts, sampled from environment objects, analysis of cont amination of water air with pesticides, petroleum products and products of microbiological synthesis and other cont aminants .

260 P.5-22 SIMPLE AND FAST DYNAMIC METHOD FOR THE PREPARATION OF HIGHLY EFFICIENT CAPILLARY COLUMNS AND THEIR PRACTICAL APPLICATION ПРОСТОЙ И БЫСТРЫЙ ДИНАМИЧЕСКИЙ МЕТОД ПОЛУЧЕНИЯ ВЫСОКОЭФФЕКТИВНЫХ КАПИЛЛЯРНЫХ КОЛОН.ОК И ИХ ПРАКТИЧЕСКОЕ ПРИМЕНЕНИЕ A.A.Korolev, T.P.Popova, V.E.Sbiryaeva, V.G.Berezkin, M.v.Taodikov, Moscow, USSR Nowadays the dynamic coating method is widely used for the preparation of capillary columns. But the usually used processes have the following disadvantages very long film formation from the SLP solution and long removal of 173 the solvent from the film {tens of hours) and also their strong dependence on experimental parameters. In order to eliminate these drawbacks a fast "superdy- namic" method of coating capillary columns with SLP was applied. In this method the steps of film-formation and solvent removal are combined. The method was used for coating capillary columns 10-40 m in length and 0,15-0.32 mm in inner diameter with various SLP: OV-101, SE-30, SE-52, SKTFT-50, PMS-lOO, PEG-1500, PEG-20M etc. The coa ting procedure is less dependent on experimental parame ters and external conditions, and takes about 15-90 minu tes depending on the column length and type of the sol vent used. The columns prepared by this method have an efficiency of 2000-5000 theoretical plates per metre depending on the column diameter and SLP type and the value of the ca pacity factor is in the range of 2-6, Practical examples of gas chromatographic separation on the columns obtained by the abovementioned method are given.

261 P.5-23 HIGH RESOLUTION CIRCULATION GAS CHROMATOGRAPHY TECHNIQUE USING GLASS CAPILLARY COLUMNS МЕТОД ЦИРКУЛЯЦИОННОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ С ВЫСОКОЙ РАЗРЕШАЮЩЕЙ СПОСОБНОСТЬЮ НА СТЕКЛЯННЫХ КАПИЛЛЯРНЫХ КОЛОНКАХ V.P.Chizhkov, N.V.Sterkhov, Moscow, USSR The ideas on the effects of chromatographic bands im pulse hydrodynamic expansion and contraction in CGC sys tems incorporating gas flow switching device have been developed. The factor of impulse zones expansion for chromatographed compounds has been introduced. Its value depends on capacity factor K*. The relation has been found between the newly-developed factor (b ) and that proposed earlier (b ): Ь = (Ь К' + 1)/(K* + 1).

The dynamics of n-alkanes (C6-C,) peak width change in capillary CGC with КЧ2 has been studied. Theoretical cal culations coincide perfectly with experimental data. The CGC impulse technique using glass capillary columns, characterized by linear increase in efficiency with the increase in number of cycles, has been developed. The possibility has been demonstrated to perform sharp sepa ration of isomers and isotopic compounds with the effi ciency up to n = 2OOOO0O0 theoretical plates. Varieties of circulation techniques with capillary columns back- flush and multidimensional capillary chromatography are discussed.

174 262 P.5-24 GLASS CAPILLARY GAS CHROMATOGRAPHIC EVALUATION OF THS MICROEXTRACTION OF PETROLEUM HYDROCARBONS FROM WATER ОЦЕНКА МЕТОДОВ МИКРОЭКСТРАКЩШ УГЛЕВОДОРОДОВ НЕФТИ ИЗ ВОДЫ С ПОМОЩЬЮ КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ J.Ilavsky', J.Hrivilak, J.Marton, Bratislava, CSSR Analysis of trace amounts of hydrocarbons in water usually requires recovery into a small volume of solvent before determination by gas chromatography. This may be done by solvent extraction or by sorption on macroreticu- lar resin or other materials be fee solvent extraction. In most methods the solvent phase requires further concen tration which may cause losses of compounds and accumula tion of solvent impurities render the procedures impracti cable. Thus microextraction (extraction with small volume of solvent) is becoming more popular, since there is no need for a concentration step and the problems of impuri ties in the solvents and losses of compounds are reduced. In this study the recoveries of petroleuii hydrocarbons in water at the microgram per liter concentrations using the microextraction witb n-pentane at 5 С were investiga ted. Preliminary studies showed that for the most of hyd rocarbons consistent and reliable recoveries were obtained which may be used for their routine analyses in water samples.

263 P.5-25 DEACTIVATION OF SOFT GLASS CAPILLARY COLUMN SURFACES FOR GAS CHROMATOGRAPHIC COATINGS WITH POLAR LIQUID PHASES ДЕЗАКТИВАЦИЯ ПОВЕРХНОСТИ СТЕКЛЯННЫХ КАПИЛЛЯРНЫХ КОЛОНОК ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ ПЕРЕД ПОКРЫТИЕМ ПОЛЯРНЫМИ ЖИДКИМИ ФАЗАМИ J.Hrivnak, Bratislava, CSSR Silanol groups present on soft glass capillary column surfaces were esterified in the presence of polyethylene glycol 400 to form stable, inert, organophilic linkages which were then easily wetted by a variety of polar liquid phases. Such gas chromatographic capillary columns exhibi ted well deactivated surfaces, good thermal stability and high column efficiencies. Application of the method is discussed and the present state of the technique is illustrated by a number of sepa rations, e.g. free fatty acids, phenols etc.

«75 264 P.5-26 PREPARATION OF HIGH TEMPERATURE GLASS CAPILLARY COLUMNS WITH LIQUID CRYSTALLINE STATIONARY PHASES ПОЛУЧЕНИЕ ВЫСОКОТЕМПЕРАТУРНЫХ СТЕКЛЯННЫХ КАПИЛЛЯРНЫХ КОЛОНОК С ЖИДКОКРИСТАЛЛИЧЕСКОЙ СТАЦИОНАРНОЙ ФАЗОЙ Z.Suprynowicz, Lublin, Poland Z.Witkiewicz, Warsaw, Poland Glass capillary columns are very powerful tool in ana lysis of muiticomponent samples of isomeric structure. The allowed temperature of their use is the limitation in some applications. If the necessary temperature of analy sis is %. 573°K the melting point of red-like liquid crys tal must be high enough to protect good thermal stabili ty. Such big molecules of liquid crystals are found to show low solubility which makes difficult the preparation of glass capillary columns by conventional coating from solution. More convenient mode of preparation is to syn thesize liquid crystals directly on capillary wall /1,2/. Three liquid crystals of different chemical nature (end group polarity) were synthesized in soda 1' ; glass columns. The different modes of glass wall pret. ^ .tment were used. /3/. The columns obtained were tested by means of resolution and selectivity towards PAH test solute mixture. 1. F.Janssen, T.Kalidin, J.Chromatoqr., 235 (1982) 323. 2. Z.Suprynowicz, M.Mardarowicz, W.Buda, Z.Witkiewicz (in preparation). 3. Z.Suprynowicz, M.Mardarowicz, W.Buda, Z.Witkiewicz, Chromatographia (in press). 265 P.5-27 SEPARATION OF HOMOLOGOUS SERIES OF HALOGENOPROPIONATEP

OF C,- Clf) ALIPHATIC ALCOHOLS AND HALOGENOPROPYL ESTE. . OF ct- СГ„ ALIPHATIC ACIDS OF THE SAME MOLECULAR WEIGHT ON АЯ SP=400 AND SE-ЗО GLASS CAPILLARY COLUMN

РАЗДЕЛЕНИЕ РЯДА ГОМОЛОГОВ ГАЛОГЕНПРОПИОНАТОВ C-,-Cln АЛИФАТИЧЕСКИХ СПИРТОВ И ГАЛОГЕНПРОПИЛОВЫХ ЭФИРОВ

О,- С,ПАЛИФАТКЧЕСКИХ КИСЛОТ ТОЙ ЖЕ МОЛЕКУЛЯРНОЙ МАССЫ НЛ СТЕКЛЯННЫХ КАПИЛЛЯРНЫХ КОЛОНКАХ С SP-400 И SE-30 K.Koma'rek, J.Churacek, J.Kri'z, Pardubice, CSSR K.Tesafxk, J.Viska, Brno, £SSR The possibility of the GC separation aiid identification of the lower and higher halogenated esters of the same mo lecular weight and summary formula but differing as mole cular "mirror image", i.e. having the same carbon atom number and positions of the halogen atoms in the acid chains of halogenopropionates as in the alcohol chains of halogenopropyl esters was studied. Generally, retention of the corresponding pairs of esters of two homologous series was evaluated., such as of 2-chloropropionates of n-alkanols and 2-chIorcpropyl esters of n-alkanoic acids;

176 2-chloropropionates of i*o-alkanols and 2-chloropropyl es ters of iso-alkanoic.acids; 2,3-dichloropropionates of n- alkanols and 2,3-dichloropropyl esters of n-alkanoic acids; 2,3-dichloropropionatas of iso-alkanols and 2,3-dichloro propyl esters of iso-alkanoic. acids. Kovats retention indices were used to interpret the results. GC separations were carried out on Fractovap model 2150 gas chromatograph (Carlo Erba, Milan, Italy) with an SP-4O0 and SE-30 capillary columns. The column temperatu re was maintained at 80 С for the separation of the lower C,-C, n-alkyl and С,-Сс isoalkyl halogenopropionates and

halogenopropylesters 3f n-C,-Ce monocarboxylic acids and

at 200°C for those of the higher C,-C10 n-alkyl halogeno propionates and halogenopropyl esters or n-Cg-C,- monocarboxylic acids. •

266 P.5-28 PREPARATION, CHARACTERIZATION AND COMPARISON OF GLASS AND FUSED SILICA CAPILLARY COLUMNS WITH IMMOBILIZED STATIONARY PHASES USING GAMMA RADIATION ПОЛУЧЕНИЕ, ХАРАКТЕРИСТИКА И СРАВНЕНИЕ СТЕКЛЯННЫХ И КВАРЦЕВЫХ КАПИЛЛЯРНЫХ КОЛОНОК С ИММОБИЛИЗОВАННОЙ СТАЦИОНАРНОЙ ФАЗОЙ С ПРИМЕНЕНИЕМ ГАММА-РАДИАЦИИ G.Anders, H.G.Struppe, K.Anders, J.Reinhard, Leipzig, GDR After acid leaching or hydrothermal treatment the glass capillary columns (alkali and borosilicate glasses) and the fused silica capillary columns were deactivated by silylation or by polysiloxane deactivation (PSD) and coa ted with different stationary phases (OV-1, PS-255, SE-54 and others). For coating the static vacuum method was pre ferred. The capillary columns were subjected to cobalt-60 gamma radiation. The influence of irradiation was investi gated. The chromatographic performance of these open tubular columns was found to be dependent on the deactivation method, on the stationary phase and thickness of its film. We will discuss such properties of the column as sepa ration power, inertness and high temperature stability (up to 300°C). 267 P.5-29 CAPILLARY GAS CHROMATOGRAPHY AND GAS CHROMATOGRAPHY-

MASS SPECTROMETRY OF С.-С1й ALKYL ESTERS OF ACRYLIC AND METHACRYLIC ACIDS КАПИЛЛЯРНАЯ ГАЗОВАЯ ХРОМАТОГРАФИЯ И ГАЗОВАЯ ХРОМАТОГРАФИЯ-МАСС-СПЕКТРОМЕТРИЯ С,-С,о АЛКИЛЭФИРОВ АКРИЛОВОЙ И МЕТАКРИЛОВОИ КИСЛОТ х 10 P.Matous'ek, А.Нота, O.Dufka, Pardubice, CSSR Acrylic and methacrylic monomers have become major industrial chemicals. A great expansion in their produc tion scale is still occuring. On the other hand broad 12.3w.872 177 variety of applications is associated with health hazard and environmental effects. Gas chromatography has been found to be very useful and most freguent tool used in acrylic and methacrylic monomers analysis. Gas chromatographic behaviour of Cj-C.g alkyl esters of acrylic and methacrylic acids has Been studied on glass capillary columns with non-polar (OV-101, SE-54) and polar (SP-IOOO) stationary phases under isothermal and temperatu re programmed conditions. The possibilities of separation of all 38 esters are discussed in terms of both reten tion indices at different column temperatures and relative retention times at programmed temperatures. The electron impact (EIMS) and chemical ionization (CIMS) mass spectral characteristic features of acrylates and methacrylates studied under various experimental con ditions are presented. 268 P.5-30 IDENTIFICATION AND DETERMINATION OF TRACE LEVELS OF PHTALATE ESTERS IN STERILIZED DISTILLED HATER ВУ CAPILLARY GAS CHROMATOGRAPHY ИДЕНТИФИКАЦИЯ И ОПРЕДЕЛЕНИЕ СЛЕДОВЫХ КОЛИЧЕСТВ ФТАЛАТОВ В СТЕРИЛЬНОЙ ВОДЕ С ПОМОЩЬЮ КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ M.s'ars'unova, J.Hrivftak, A.Hichalkova', J.RemeX, Bratislava, CSSR Concern has arisen over the toxical effects of phtalate esters widely used as plasticizers. Phtalate esters can be leached into intravenous solutions and contaminate human blood and other tissues. Considering this problem, an ana lytical procedure including the mlcroextraction of trace quantities of phtalates from sterilized distilled water, their identification and determination by capillary gas chromatography was used. The obtained results have confirmed that except of the chemicals and distilled water used in the manufacturing process of .intravenous solutions, also black rubber stop pers of the bottles and plastic tubings of the intravenous solution administration sets can be the source of contami nation by phtalate esters. In this case however the mean detected concentration level of the leached phtalates was only 24 yug/l of distil led water. 269 P.5-31 CAPILLARY GAS CHROMATOGRAPHY OF BRANCHED ALKANES КАПИЛЛЯРНАЯ ГАЗОВАЯ ХР0МАТ0ГРА4ЯЯ РАЗВЕТВЛЕННЫХ АЛКАНОВ J.Hrouzek, P.Cellar, I.Ostrovsky, L.Sojak, Bratislava, 2sSR The branched alkanes are important components of petr oleum and their origination and chemical changes are Often studied, too. The retention data of these hydrocarbons are 178 known in the range up to С.-, but there are only a few data for higher branched alkanes. The comparison of reten tion data from the different sources, for this area (up to

C.0) shows significant differences among them. Our goal was tRe measurement of retention data for branched alkanes up to C,_ on squalane, the derivation of structure-reten tion correlations as a mean of the higher branched alka- nts identification, and the study of isomeric alkanes se paration on liquid crystals as a stationary phases. Model mixtures of 126 branched alkanes up to С,- were preparated by using methylene insertion reaction. The homomorphous factor functions H = f/C/ asympthoti- cally decrease with the elongation of the carbon chain for each homologous serie of branched alkanes. There are some deviations for the first members of homologous series, from the regularities of the function H = f/C/, which are directed to lower H values. For these alkanes is characte ristic the presence of tertiary or quaternary carbon atoms on the ends of molecules. Using the extrapolation of H • f/C/ functions it is possible to predict retention indices for the higher branched alkanes (over C,0), with accuracy about 1 i.u.. The function dl/dT » f/C/ for homo logous series has the same character as the function H - f/C/ dl/dT values rise with the number and the magni tude of the substituting alkyls and with their shifting to the carbon chain. It is possible to solve the separation problems of iso meric alkanes with the similar characteristics, as well as the diastereoisomers by using separation systems connec ting the high efficiency of capillary columns with the high structure selectivity of liquid crystals as a sta tionary phases.

270 P.5-32 REPRODUCIBILITY OF RETENTION DATA AND INFLUENCE OF CHEMICAL STRUCTURE ON SELECTIVITY OF MESOGENIC STATIONARY PHASES IN CAPILLARY GAS CHROMATOGRAPHY ВОСПРОИЗВОДИМОСТЬ УДЕРЖИВАНИЯ И ВЛИЯНИЕ ХИМИЧЕСКОЙ СТРУКТУРЫ НА СЕЛЕКТИВНОСТЬ МЕЗОГЕННЫХ СТАЦИОНАРНЫХ ФАЗ В КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ A.Isenberg, G.Kraus, H.Zaschke, Halle-Wittenberg, GDR

W Weissflog,- Leipzig, GDR v L.Sojak, i.Ostrovsky, Bratislava, CSSR Mesogenic compounds are applicated as stationary phases in gas chromatography both to solve separation problems and to Investigate physico-chemical properties of liquid crystals. In order to solve separation problems it is im portant to know the relations between polarity and selec tivity, which are influenced by the chemical structure of the stationary mesophase.

179 Measuring retention data of lath-like is well as of so- called swallowtail liquid crystalline compounds by syste matically variation of terminal and lateral substituents the influence of this structural elements on selectivity and polarity was investigated. The polarities oi investi gated mesogenic stationary phases are compared with those of conventional ones and are strongly dependent on the nature of their terminal substituents. Lateral substitu ents in mesogenic molecules effect a change in selectivi ty by varying the distance between the liquid crystal molecules. Investigations on dependence of reproducibility of retention data of substrates on the conditions of prepa ring of glass capillaries as well as on type of used mesogenic compound have been carried out.

271 P.5-33 CAPILLARY GAS CHROMATOGRAPHY OP ALKYNES КАПИЛЛЯРНАЯ ГАЗОВАЯ ХРОМАТОГРАФИЯ АЛКИНОВ P.Parkas, L.Sojak, Bratislava, SsSR The retention behaviour of 92 linear and branched alkynes up to C.. was studied on squalen» and 4-n-pentyl- acetophenone-to-4-n-pentylbenzoyloxime) liquid crystal as stationary phases in glass capillary columns. From retention indices measured for alkynes on squalane and PBO, structure-retention correlations, important from the viewpoint of characterization, were derived. In crements due to the methylene group (ICH?)• homemorphous factors (H) and temperature increments ox the retention indices (dl/dT) show characteristic dependences on triple bond position, on character of branching and on number of carbon atoms in the molecule.

The observed deviations from the regularities of the dependences of both ICH, - f are dis cussed from the viewpoint of fine structural effects as the propyl effect, the hyperconjugation and the alterna tion of retention. On this basis precise prediction of elution behaviour of the higher homologues is possible. Retention behaviour of the branched alkynes was predicted from analogy of those of branched alkenee and was confir med by measuring of retention indices obtained during methylene insertion reaction of alkynes. The use of PBO liquid crystal as stationary phase in capillary gas chromatography enables separation of some difficult separable pairs of isomers on squalane. The results were applicated by analyses of alkynes which occur in products of n-alkanes dehydrogenation and pyrolysis of gasoline.

It» 272 P.5-34 CAPILLARY GAS CHROMATOGRAPHY RETENTION DATA OF IMPURITIES IN TECHNICAL GRADE ACRYLATE AND METHACRYLATE MONOMERS ПАРАМЕТРЫ УДЕРЖИВАНИЯ ПРИ КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ ПРИМЕСЕЙ В ТЕХНИЧЕСКОМ АКРИЛАТЕ И МЕТАКРИЛАТЕ P.Matousek, V.Moravkova, O.Dufka, Pardubice, CSSR Because of the importance of acrylate and methacrylate monomers in manufacturing of variety synthetic resins, the identity of impurities in these starting materials has become a subject of increased interest. Among other analy tical techniques the capillary column gas chromatography has been used for this purpose. Relative retention times and retention indices of both GCMS identified impurities and unknowns in technical grade methyl-, ethyl-, butyl- and 2-ethylhexylacrylate and methyl-, butyl and 2-hydroxyethylmethacrylate monomers we were determined on glass capillary columns with SE-54, Ucon LB-550-X and SP-100 stationary phases under program med column temperature conditions. Tabulations of retention data obtained together with their precision are presented and factors which affect their reproducibility and usefulness in relation to the sample size and relative retention of the main component are mentioned. 2 P.5-35 STUDY OF RETENTION INDICES OF ALKYLCHINOLINES ON CAPILLARY COLUMNS ИССЛЕДОВАНИЕ ИНДЕКСОВ УДЕРЖИВАНИЯ АЛКИЛХИНОЛИНОВ НА КАПИЛЛЯРНЫХ КОЛОНКАХ Yu.S.Berlizov, P.Buryan, J.Macak, Prague, 6sSR V.P.Dmitrikov, V.M.Nabivac, Dnepropetrovsk, USSR Nitrogen heterocyclic compounds are widely used in pro duction of herbicides, medicines, vitamins etc. As a sour ce of nitrogen bases the tars are employed, which are very complicated mixtures and which are obtained by thermal processing of lignite and bituminous coal. A perspective analytical method for these mixtures is capillary gas chromatography, especially on quartz capil lary columns. The known data on the retention indices of alkylchinolines deals with a relatively limited number of sorbates and stationary phases. In this paper, measurements of retention indices of more than 25 alkylchinollnes are made on capillary glass and quartz columns packed with OV-101 and Ucon-LB S50X, and their thermal and structural increments are calculated. These data make possible to evaluate the character of molecular interactions in the studied systems. The regression equations obtained prove the linear

1(1 dependence of retention indices of methyl- and dimethyl chinolines on their boiling points (the correlation coeffi cient is higher than 0,995). It has been established that the found increments of the CH,-group for various positions allow to calculate the retention indices'of corresponding dimethyl- and trimethyl- chinolines. The determined dependences can be utilised for qualita tive identification of alkylchinolines without a standard in complicated industrial mixtures. 274 P.5-36 GAS CHROMATOGRAPHIC ANALYSIS OF NATURAL STEROIDS AND SYNTHETIC DRUGS WITH CAPILLARY COLUMNS ГА30ХР0МАТ0ГРАФИЧЕСКИЙ АНАЛИЗ ПРИРОДНЫХ И СИНТЕТИЧЕСКИХ ЛЕКАРСТВЕННЫХ СТЕРОИДОВ НА КАПИЛЛЯРНЫХ КОЛОНКАХ V.P.Uralets, V.A.Semenova, V.A.Krokhin, M.A.Yakushin, V.A.Semenov, Moscow, USSR In the studies of steroid drug action on human body it is important to follow the steroid profile changes in bio logical fluids. Capillary gas chromatographic (GO steroid profiling of methoxyamine-trimethylsilyl derivatives allow to detect these changes caused by the use of steroid drugs. This is illustrated by the examples of some anabolic ste roids and synthetic corticosteroids being doping compounds vorbidden in sports use. Analysis is performed with the on-line data system permitting a direst calculation of retention indices. Simultaneous selective and universal detection was used with N-P, EC and PID. Narrow bore (100 urn) fused silica capillary columns showing high inertness were used for separation of under i- vatized urine steroids with the traces of anabolic metabo lites. The latter are reliably, detected by selected ion monitoring (SIM) with mass selective detector (MSD). Anabolic steroid stanozolol and its urine metabolites are difficult substances for normal urine steroid profi ling. Their detection is performed by the use of special derivatisation procedure and selective N-P detection or SIM with MSD. 275 P.5-37 THE METHOD OF THE POLAR LIQUID PHASE FORMATION ON THE INNER SURFACE OF THE GLASS CAPILLARY COLUMN МЕТОД ФОРМИРОВАНИЯ ПОЛЯРНОЙ ЖИДКОЙ ФАЗЫ НА ВНУТРЕННЕЙ ПОВЕРХНОСТИ СТЕКЛЯННОЙ КАПИЛЛЯРНОЙ КОЛОНКИ N.M.Pavlenko, B.A.Vinogradov, Yalta, USSR With the aim of obtaining the Carbowax 20M glass capil lary columns a high degreeof resolution and reproducibili ty (resolution - 300.000 m ) the method of the liquid phase coating on the inner surface of the column was pro posed. The method consists in the covering of the mixture of 0,1% NaCl and 0,25% Carbowax 20M in anhydrous methanol by the high pressure static method on-the inner surface of

182 a column. During this process a thin reproducible layer of a stationary phase is formed. The disadvantage of such columns is the impossibility to use liquid mediums for analysis. The time needed for the column preparation is three hours. To improve the separation of the high polar compounds the adding to the solution of 0.01% KF is recommended.

276 " P.5-38 DEVELOPMENT AND APPLICATION OF QUARTZ CAPILLARY COLUMNS FOR THE ANALYSIS OF MULTICOMPONENT MIXTURES РАЗРАБОТКА И ПРИМЕНЕНИЕ КВАРЦЕВЫХ КАПИЛЛЯРНЫХ КОЛОНОК ДЛЯ АНАЛКЗА МНОГОКОМПОНЕНТНЫХ СМЕСЕЙ Yu.A.Kolchin, V.V.Pomazanov/ V.A.Minaev, Moscow, USSR Application of quartz capillary columns for the analy sis of multicomponent mixtures of biological and chemical objects is the important step for the further development of techniques and practice of capillary chromatography. The problem of reproducible development of quartz ca pillary columns is observed, the techniques of prelimina ry treatment of inner surface of quartz capillary and application of immovable liquid phase into e" 0,15-0,3 quartz capillary is developed. This procedure is adapt able to streamlined production, ease of handling and allows to develop quartz capillary columns with different polarity phases, such as SE-30, OV-17, PEG-20M, CKT, CKT T-50X. Ready for use columns were tested with stan dard mixtures and their effectivity is 4000-6000 theore tical plates for meter. The usage perspectives of quartz capillary columns for separation of multicomponent mixtu res of fatty acids and their derivatives, carbohydrates, aminoacids, easy volatile components, pesticides and al cohols are shown.

183 THIN-LAYER CHROMATOGRAPHY AMD ELECTROPHORESIS

277 PL ELECTROPHORETIC SEPARATION AND ANALYSIS OF LIVING CELLS FROM SOLID TISSUES SY SEVERAL METHODS. HUMAN EMBRYONIC KIDNEY CELL CULTURES AS A MODEL ЭЛЕКТРОФОРЕГИЧЕСКОЕ ОТДЕЛЕНИЕ ОТ ТВЕРДЫХ ТКАНЕЙ И АНАЛИЗ ЖИВЫХ КЛЕТОК С ПОМОЩЬЮ НЕСКОЛЬКИХ МЕТОДОВ. КУЛЬТУРА ЭМБРИОНАЛЬНЫХ ТКАНЕЙ ПЕЧЕНИ В КАЧЕСТВЕ МОДЕЛИ P.Todd, L.D.Plank, M.E.Kunze, Pennsylvania, USA M.L.Lewis, D.Morrison, Houston, USA G.H.Barlow, Chicago, USA J.W.Lanham, C.Cleveland, St.Louis, USA Preparative electrophoresis of living cells has been considered for some time as a potential tool for purifying, from heterogeneous mixtures, subpopulations of cells accor ding to function. Such purification depends upon the reten tion of electrophoretic heterogeneity and the retention of function. Human embryonic kidney cells that had been in monolayer culture for 1-5 generations were resuspended by treatment with trypsin or EDTA and suspended in a variety of electro phoresis buffers ranging in ionic strength from 0.003 to 0.15 g-ions/liter. Analytical electrophoresis using a Zeiss "Cytopherometer" or "Pen Kern 3000" automated light- scattering electrophoretic analyzer indicated that electro phoretic heterogeneity was retained under the full range of conditions tested. Preparative electrophoresis in a density gradient, in continuous flow and in micror;ravity indicated that electrophoretic heterogeneity coincided with functional heterogeneity; for example, some electro- phoretically isolated subpopulatione produced increased levels of urokinase while others produced increased levels of tissue plasminogen activator. Likewise, diffe rent electrophoretic subpopulations were found to have differing sizes, morphologies, and optical properties as assessed by phase microscopy and flow cytometry. Work supported by the U.S.National Aeronautics and Space Administration. 278 S.15-1 GENERAL THEORY OF ISOELECTRIC FOCUSING IN BUFFER pH GRADIENT AND SCOPE OF THEIR APPLICATION ОБЩАЯ ТЕОРИЯ И30ЭЛЕКТРИЧЕСК0Г0 ФОКУСИРОВАНИЯ В БУФЕРНЫХ ГРАДИЕНТАХ рН И СФЕРА ИХ ПРИМЕНЕНИЯ G.V.Troitsky, Simferopol, USSR It has been shown that isoelectric focusing in artifi cial buffer pH-gradients, has some preference over ampholine methods. For example more homogeneous media in which electrophoresis takes place, shows lower probability of artefacts etc. 184 Practical use of the buffer pH-gradients have got some difficulty in creating of the main conditions lifetime of pH-gradients should be much less time of protein focusing. At present they succesaed in three types of systems: 1) Borate-polyols methods (BP-pH-gradients), 2) Steady-state rheoelectrolysis, 3) Immobilins. pH-gradients created by dielectric permiantablity gra dients appear promising. BP-pH-gradients stability car be drawn from next diffe rential equation: dC /dt «СЧИ.Е , where*c - concentration of polyhydroxyl compounds (polyols) , o(. and U. degree of ionisation and mobility of comp lex ion, t- time. Solution of this equation, for simple case of boric acids only, with glycerol or xylite, gives the lifetime of the gradients for 240 h. One can use BP-pH-gradient with borate (pH region'11-6;. In this case pH-gradient lifetime is shorter (20-100 h). Ionisation of polyols d. may be given in next equation:

pH d - {(1 - g) Cg + 10" 7 Cp

Q10"POH £or alkaline region), where C_ and g - concentration and degree of hydrolysis of buffer salts. From this equation in is drawn that for decrease of polyols ionisations which isn't bound with pH of medium should use bases and small concentration of borate. Buffer capacity of the gradients will be depen dent on buffer volume in electrode vessels. In contrast with ampholine, BP-method can be applied in salt medium, for fractionation of low molecular weight compounds: amino acids, peptides and ampholins as\well. Preference of the BP-method as preparative is no doubt. As preparative the method in primary importance due to cheaper reactives as glycerol and boric acid. The BP-pH- gradient was used as preparative method in condition of gravitation in "Salyute-7". This experiment showed the more resolving power in gravitation. 279 S.15-2 THE USE OF PLANAR TECHNIQUES IN PRELIMINARY EXPERIMENTS ON THE CHOICE OF OPTIMAL CHROMATOGRAPHIC SYSTEMS FOR THE ANALYTICAL AND PREPARATIVE SEPARATIONS ИСПОЛЬЗОВАНИЕ ПЛАНАРНОЙ ХР0МАТ0ГРА4ИИ ДЛЯ ВЫБОРА ОПТИМАЛЬНОЙ ХРОМАТОГРАЗЙЧЕСКОЯ СИСТЕМЫ ДЛЯ АНАЛИТИЧЕСКИХ И ПРЕПАРАТИВНЫХ РАЗДЕЛЕНИЙ Е.Soczewinski, Lublin, Poland Paper chromatography and thin-layer chromatography are very useful in the determination of optimal liquid-liquid and liquid-solid systems for the analytical or preparati ve separations. The advantages of planar techniques are their rapidity, low consumption of solvents and materials, possibility to compare the sample with reference compounds

I8S in a single experiment. The main condition of successful application in pilot experiments is the close analogy between chromatographic processes in column and planar ch romatography. Several examples of applications of planar techniques in pilot experiments are described: 1. Preliminary TLC experiments in HPLC analysis (correla

tion between log к'мртс «nd К,1ТтС% values). 2. Stepwise gradient efutlon in TEC TK preliminary optimi zation of continuous gradient HPLC. 3. Determination of extraction coefficients of alkaloids by "moist buffered paper* chromatography for batch counter-current extraction (preparative separations). 4. Zonal TLC as a pilot technique for column preparative separations. The conditions of analogy between planar techniques and HPLC are discussed,

280 S.15-3 STUDIES ON PROPERTIES AND STRUCTURE OF SATIETIN AND ITS FRAGMENTS ВУ MEANS OF LIQUID CHROMATOGRAPHY AND ELECTROPHORESIS ИССЛЕДОВАНИЕ СВОЙСТВ И СТРУКТУРЫ САТИЕЛИНА И ЕГО ФРАГМЕНТОВ МЕТОДОМ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ И ЭЛЕКТРОФОРЕЗА J.Nagy, L.Varady, H.Xalasz, J.Knoll, Budapest, Hungary Satietin, an endogenous substance, which selectively inhibits the food-Intake of the rats deprived for 96 hours /1/. It was proved to be carbohydrate-rich glycoprotein isolated from the plasma of humans by a series of separa tion techniques /1,2/. Isolation and preliminary characterisation of satietin were described previously /3/. This paper's intent is to report the more distinct properties and structural characteristics of the native substance and the fragments obtained by chemical and enzymatical treatments subsequent to isolation by gel chroma tography. The fragments of this endogenous food-intake inhibitor were characterized by means of gel electrophoresis in the absence and presence of sodium dodecyl sulfate, isoelect ric focusing and high performance liquid chromatography. The specific biological activity and the sequence of fragments obtained by chemical and enzymatical cleavages of satietin will be depicted in details. 1. J.Knoll., Physiol. Behav., 23 (1979) 497. 2. J.Nagy, I.Mazsaroff, L.Varady, J.Knoll., in B.Kalasz and L.S.Ettre (Eds.) "Chromatography, the State of the Art", Akademial Kiado, Budapest, 1985, p. 337. 3. J.Nagy, H.Kalasz, J.Knoll,, In Frigerio, A. (Ed.) "Chromatography and Mass Spectrometry in Biomedical Sciences", No.2", Elsevier, Amsterdam, 1983, p.421.

186 281 S.15-4 FORCED-FLOW THIN-LAYER CHROMATOGRAPHY ТОНКОСЛОЙНАЯ ХРОМАТОГРАФИЯ С ПРИНУДИТЕЛЬНЫМ ПОТОКОМ ПОДВИЖНОЙ ФАЗЫ H.Kalasz, J.Nagy, Budapest, Hungary Forced-flow thin-layer chromatography (FF-TLC) is novel variation of the planar separation aethods when the sta tionary phase is totally closed and the aoblle phase is supplied by forced flow of a pump. The separation power of FF-TLC can be optimized accor ding to the flow velocity of the mobile phase. The optimum of flow rate also depends on the particle size of the stationary phase bed. 0.10-0.15; O.12-0.17 and 0.20-0.28 ran/sec have been found as the optima of speed of eluent using Fixion 50X8, TLC silica and HPTLC silica, respecti vely. Examples dealing with the research in chemistry and biochemistry of substances of natural origin and their fragments have proved the wide application field of FF-TLC. 282 P. 3-30 SOME REMARKS ON THE USE OF THIN-LAYER CHROMATOGRAPHY IN QSAR STUDIES НЕКОТОРЫЕ ВОПРОСЫ ПРИМЕНЕНИЯ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ • В ИЗУЧЕНИИ ВЗАИМОСВЯЗИ СТРУКТУРА-БИОЛОГИЧЕСКАЯ ФУНКЦИЯ J.Gasparic, Hradec Kralove', CSSR Thin-layer chromatography on layers impregnated with organic stationary liquide is often used by medicinal chemists in QSAR (Quantitative structure - biological acti vity relationships) studies. Many, authors use it quite mechanically and present calculations based on Rm values without giving any details on conditions under which these values have been obtained. Parameters of experimental conditions are summarized in this contribution which should be taken into conside ration in such investigations! 1) the type of chambers used for development) 2 ) the type of support of the organic stationary phase (silica gel, cellulose etc.); 3) the type of the stationary liquid (liquid paraffin, 1-octanol); 4 ) the degree of impregnation; 5 S the mode of impregnation* 6 ) the composition of the mobile phase; 7 ) properties of the solute (dissociation, polarity etc.). All these factors can affect the results on layers im pregnated with organic stationary phases not only in chan ging the absolute Km values of all solutes in one direc tion, but they can cause also changes in the sequence of the solutes on chromatograms. Thus, it can be concluded that a partition system in this type of thin-layer chromatography involves the fol lowing interactions: solute-support, solute-stationary 187 phase, solute-mobile phase, support-mobile phase, support- stationary phase, mobile phase-stationary phase, provided all experiments are carried out in the same type of the developing chamber. Othervise the vapour phase and its interactions with other parameters should be taken into consideration, too. 283 P.3-31 RARE EARTH METAL SEPARATION BV PLAIN SILICA GEL TLC РАЗДЕЛЕНИЕ РЕДКОЗЕМЕЛЬНЫХ ЭЛЕМЕНТОВ С ПОМОЩЬЮ ТСХ НА СИЛИКАГЕЛЕ E.Fugaerth, Budapest, Hungary Despite of the increasing in number of very promising findings chromatography as a tool for analysing mixtures of metal-ions have not became too popular yet, perhaps, because of every published method has its own range of applicability and at the same time a severe limitations being usually not compatible when any farther metal-ion is present. There are, however, separation problems when similari ties between metal-ion components are so high that for reliable analysis chromatography is a must. That is the case for the rare earth elements. Altough their first suc cessful chromatographic separation dates back to the early 60s* interest on the topic is ceaseless. This led to have now three kind of very effective separation based on a rather different principle. These are partition by usage of dialkylphosphates, ion-exchange (mainly as EDTA comp lexes) and partition on sxlanized silica gel. The third method - developed by H.Specker et al. - is really attrac tive as it requires readily available materials only, no special treatment or high care for successful results compared with the other two methods. This work was iniciated by that of Specker and can be divided into two parts: the first is dealing with thorough investigation of the chromatographic pattern of the Lanta- nida elements by systematic changes of many variables cen tered on Specker's results, while the second part shows how n more commercial sorbent - plain silica gel - instead of Rt>-2 material is amenable to solve the same task. Stu dies will be supported by arranged series of photocopies allowing a true inspection/comparision.

284 P. 3-32 ADVANTAGES OF OVERPKESSURED THIN LAYER CHROMATOGRAPHY ON THE ANALYTICAL AND PREPARATIVE SCALE SEPARATION OF SESQUITERPENE LACTONES ПРЕИМУЩЕСТВА ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ ПОД ДАВЛЕНИЕМ ДЛЯ АНАЛИТИЧЕСКОГО И ПРЕПАРАТИВНОГО РАЗДЕЛЕНИЯ ЛАКТОНОВ'СЕСКВИТЕРПЕНА X.Kery, Gy.Turiak, G.Verzar-Petri, Budapest, Hungary Separation and quantitative determination u£ sesquiter pene lactones in complex plant mixtures is of a difficult 188 and timeconsuming task. The problem of the efficient separation is due to the very similar behaviour of the different sesquiterpene lactones as well as to the pre sence of other constituents in the plant extracts. This paper investigates some separation performance data obtained under different conditions with a newly de veloped technique in the area of thin layer chromatogra phy, overpressured thin layer chromatography (OPLC) both on the analytical and preparative scale. Extracts, prepared from different Asteraceae plants, rich in different groups of sesquiterpene lactones (germacranolides, guaianolides, eudesmanolides, etc.) have been used as models. The compounds, separated were identified by comparison with authentic samples and /or spectroscopic techniques. By the aid of CHROMPRES S-10 apparat (LABOR Instrument Works, Budapest, Esztergom, Hungary) we successfully sepa rate the model compounds using the solvent systems: Car bon tetrachloride-Acetonitrile 4:1,5; Benzene - n-Hexane - Acetonitrile 3:2:2,1; Benzene - n-Hexane- Ethylacetate 3:2:2,1; n-Hexane-Ethylacetate 7:3: Carbon tetrachloride- Acetonitrile-Chloroform 4,5:1,5:0,5; etc. Chromatography was carried out on precoated plates (HPTLC, Silica gel 60 F254 tlerck' Darmstadt) with impregnated edges. The spots were visualized at UV light and using selective reagents. Determination of the lactones was carried out by SHIMADZU type densitometer. Advantages of OPLC both on analytical and preparative scale are discussed, and the benefit of the pronounced shorter migration times in 0P.X is emphasized and demonst rated both on sesquiterpene lactones with small differen ces in selectivity (tatridines, etc.) and complex lactone mixtures with a large number of compounds have to be sepa rated.

285 P.3-33 TLC QUANTITATION OF THE LIPIDS OF RATS LIVER HOMOGENATES AFTF.R EXPOSURE BY THICHLOROETHYLENE КОЛИЧЕСТВЕННОЕ ОПРЕДЕЛЕНИЕ ЛИПИДОВ В ГОМОГЕНАТАХ ПЕЧЕНИ КРЫС ПОСЛЕ ВОЗДЕЙСТВИЯ ТРИХЛОРЭТИЛЕНА МЕТОДОМ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ J.Gartzke, H.Hohn, D.Burek, Berlin, GDR Lipid extracts of liver homogenates were spotted on different ammoniumsulfate impregnated silica gel thin- layers (Merck, Art.Nr. 5548 and 555 3) together with estra diol as internal standard. After a two stage one dimen sional development in benzene:dioxane = 94:6 (v/v) and carbontetrachloride , the lipid classer were quantitated by densitometry, factorization for their different carbon number and calculation with the internal standard. The method is compared with direct enzymatic methods. The method, results and application are discussed.

189 286 P.3-34 THIN LAYER CHROMATOGRAPHY DETERMINATION OF DIURNAL RHYTHM OF EXCRETED PORPHYRINS IN PORPHYRIA CUTANEA TARDA ОПРЕДЕЛЕНИЕ С ПОМОЩЬЮ ТОНКОСЛОЙНОЙ ХРОМЛТОГРАФИИ ДНЕВНОГО РИТМА ЭКСКРЕЦИИ ПОРФИРИНОВ ПРИ PORPHYRIA CUTANEA TARDA P.Martasek, M.Jirsa, V.Kordac, P.Kotal, Prague, CSSR

Porphyrias are enzymopathies of haem synthesis pathway. In the seventieth years thin layer chromatography (TLC) of porphyrin methylesters^played an important role both in the recognition of haem synthesis and, above all, in the diagnostics of individual porphyrias. Basic chromatogra phic characteristics of individual porphyrias are presen ted. In porphyria cutanea tarda (the most frequent porphy ria in Europe) five main porphyrins are excreted by urine; these in the form of methylesters are in TLC divided accor ding to the number of carboxyl groups (4-8) , the lowest

Rf being that of uroporphyrin (8-COOH porphyrin).

In this study we tried to reveal the daily rhythm of individual porphyrins in 10 patients with porphyria cuta nea tarda (9 men, 1 woman, range 40-65 years, mean 53.4, SD 9.2) . The patients were hospitalized having the same daily and diet regime. In 4-hour intervals samples of urine were collected by spontaneous miction. Samples of 100 ml of urine were acidified by acetic acid and urine porphyrins were adsorbed on talc (10 g to 100 ml of urine). No porphyrins were found in the filtrate. TLC of methyles ters was carried out according to Doss /1/ and the fluorometric scans were obtained using Vitatron TLD 100 densito meter. The total porphyrins were determined as described earlier /2/.

The highest quantity of total porphyrins was excreted from midnight to noon. Uroporphyrin and 7-C00H porphyrin were proved to have the same daily rhythm and follow the same maxima as the total porphyrins. Contrary to that, the highest quantity of coproporphyrin (4-COOH porphyrin) and 5-C00H porphyrin was excreted from 12 to 8 p.m., when the lowest quantity of uroporphyrin and 7-С00И porphyrin was excreted. No statistically significant differences were found in the excretion of 6-C00H porphyrin. 1. M.Doss (197]) : Z. klin. Chem,,8, 197-207. 2. P.Martasek, M.Jirsa, V.Kordac (1982); J. clin.Chem. clin. Biochem.,20, 113-117.

190 287 P.3-35 APPLICATION OF HORIZONTAL SANDWICH CHAMBER FOR THIN-LAYER CHROMATOGRAPHY OF COMPLEX MIXTURES ПРИМЕНЕНИЕ ГОРИЗОНТАЛЬНОЙ КАМЕРЫ ТИПА СЭНДВИЧ ДЛЯ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ СЛОЖНЫХ СМЕСЕЙ G.Matysik, E.Soczewiriski, Lublin, Poland Complex mixtures of organic substances, e.g., plant extracts, can only partly be separated by thin-layer chro matography. The separation efficiency (expressed by the number of spots separated on a given distance of develop ment) can be markedly increased by the use of special modifications of the development process. A modification of Brenner-Niederwieser horizontal sand wich chamber with a glass distributer of the eluent (or sample) is described. The construction of the chamber permits the use of special techniques which increase the separation efficincy: 1. Stepwise and continuous gradient elution 2. Continuous development 3. Band application of the sample with the glass distri buter (micropreparative separations). The advantages of the modifications of the TLC techni que are illustrated in the analysis of plant extracts. 288 P.3-36 SIMULTANEOUS ANALYSIS OF NEUTRAL AND PHOSPHOLIPID CLASSES IN BIOLOGICAL SPECIMENS BY MONODIMENSIONAL THIN-LAYER CHROMATOGRAPHY USING A NOVEL DEVELOPING SYSTEM AND "IN SITU" SPECTROFLUOROMETRY ОДНОВРЕМЕННЫЙ АНАЛИЗ НЕЙТРАЛЬНЫХ И ФОСФОЛИПИДОВ В БИОЛОГИЧЕСКИХ ОБРАЗЦАХ МЕТОДОМ ОДНОМЕРНОЙ ТСХ С ПРИМЕНЕНИЕМ НОВОЙ СИСТЕМЫ РАСТВОРИТЕЛЕЙ И СПЕКТРОФЛУ0Р0МЕ7РИИ "IN SITU" R.Segura, X.Navarro, J.Prat, S.Martin-Ayuso, Barcelona, Spain Lipids are essentail constituents of all cells, the phospholipids have a particular biological significance as for they are the major components of cellular membranes and play a special role in the structure and metabolism of lipoprotein complexes. Up to now, it has been rather difficult and cumbersome to separate and quantify the different classes or species of phospholipids and few attempts have been made in order to achieve the simulta neous separation of all major lipid classes (non-ionic, or neutral, plus phospholipids). After numerous assays, we have found a chromatographic system that allows the simultaneous separation of more than 15 different lipid classes by monodimensional thin- layer chromatography on silica gel plates. With this sys tem we have analyzed the lipid constituents from a great variety of biological samples, ranging from plasma lipo proteins to blood cells (erythrocytes, leukocytes, plate lets) , from cereal grains (wheat, barley,...) to milk, dried fruits, eggs, etc... 191 The different lipids are extracted from the biological specimens by 2-propanol and an aliquot of the alcoholic sample was applied at the origin of the chromatographic bed. The different phospholipid classes are separated by means of a developing system containing water, ethanol and dichloroethylene; with this system the neutral lipids move with, or very closed to, the solvent front which is allowed to migrate up to the middle of the plate. After drying the solvent, the plate is introduce in a second tank containing 1,2-dichloroethane as the only component of the mobile phase; with this second development the frac tionation of the different neutral lipids is obtained. The total time involved in these procedures is two hours. The different lipid components are rendered visible by means of an original procedure (SEGURA et al., To be pub lished) in which the compounds are transformed into fluo rescent derivatives through thermal treatment of the chromatoplates, after they have been impregnated with an aque ous solution of aluminium dihydrogen phosphate. The fluo rescent derivatives, thus obtained, are easily detected, in rather low quantities, by examining the chromatoplates under ultraviolet radiation and are evaluated by means of a spectrofluorometer equipped with an attachment that allows the scanning of thin-layer chromatoplates,

289 P.3-37 NEW HYBRID METHOD: COMBINATION OF THIN-LAYER CHROMATOGRAPHY WITH X-RAY FLUORESCENT MICROANALYSIS НОВЫЙ ГИБРИДНЫЙ МЕТОД: СОЧЕТАНИЕ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ С РЕНТГЕНОФЛУОРЕШЕНТНЫМ МИКРОАНАЛИЗОМ M.P.Volynets, V.G.Berezkin, N.P.Ilyin, Moscow, USSR A new way of identification and determination of the content of elements in zones of plane chromatograms based on the combination of two analytical methods: separation (thin-layer chromatography, TLC) and determination (x-ray fluorescent microanalysis, RFM) is suggested. The main area of application of the new TLC-RFM method is the determination of nano- and micro amounts of inorganic and_ elemento-organic compounds in small zones (less than 1 mm ). Unlike the acknowledged direct ways of determination of separated substances on TL-plates (ex. densitometrie, luminescent, fluorimetric, radioactivational) the TLC-RFM method does not demand additional processing on the chromatogram before determination. After the chromatogra phic separation on plate is over, scanning of the sorbent layer by the collimated beam of the primary x-ray radia tion is carried out with the help of an x-ray fluorescent microanalyzer, and the characteristic radiation of the determined element is registered. The method is characterized by a high selectivity. The absolute limit of detection makes Ю -10"10 g. Locality of scanning is 0.1-1.0 mm. The method is non-destructive, the chromatogram remains intact. Simultaneous registration of several elements in each 192 zone is possible. Combination of TLC and RFM makes wider the possibilities of each method: it enlarges the group of analyzed substances and the number of determined ele ments; it gives the opportunity to estimate the quantita tive correlation of element* with different valency and other forms of preliminarly separated elements; it helps to establish the completeness of chromatographic separation and to study distribution of components within the limits of each zone, etc. The method is universal, J.t can be applied for the study of plane chromatograms on any bed. It can be used for the analysis of various natural and industrial micro- objects, in physico-chemical studies of the sorption pro cess, etc. The possibilities of the method are illustrated on the example of determination of microamounts of nickel and cobalt after chromatographic separation and determination of various ion forms of molybdenum which coexist in hydro chloric acid solutions.

290 P.3-38 PHYSICO-CHEMICAL ASPECTS OF INORCANIC ION SEPARATION CONDITIONS OPTIMIZATION BY THIN-LAYER CHROMATOGRAPHY METHOD ON ALUMINIUM OXIDE ФИЗИКО-ХИМИЧЕСКИЕ АСПЕКТЫ ОПТИМИЗАЦИИ УСЛОВИИ РАЗДЕЛЕНИЯ НЕОРГАНИЧЕСКИХ ИОНОВ МЕТОДОМ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ НА ОКСИДЕ АЛЮМИНИЯ D.S.Gaibakyan, Yerevan, USSP. J.K.Rozylo, Lublin, Poland In the literature there are few works concerning the problem of the optimization of the separation conditions of inorganic ions on different sorbent-carriers, while for organic compounds they are sufficiently discussed. The perfection of this method and its development are strictly connected with the output of some theoretical approaches to predict the conditions of the separation of ions, which can enlarge the limits of its application and arise its effectiveness. Such approaches can be sol ved using calculation methods on the ground o-f a strict physico-mathematical model of sorption processes. In the report must be discussed some possibilities of a new approach to a particular case of.the separation of rare elements ions from their mixtures using different solvent systems as mobile phases It has been shown the relative complexity of the ion separation mechanism on aluminium oxide layer because simultanously many processes occur such as adsorption ion exchange, hydrolysis of multivalent ions and complexformation in the presence of different ligands in the solutions of mineral and organic acids. Some parameters of these processes have been taken into account to predict and to optimize the conditions for the separation of inorganic ions from their mixtures.

13.3ак.872 193 291 P.3-62 CONTINUOUS THIN LAYER CHROMATOGRAPHY (CTLC) WITH FLUORIMETRIC FIBER-OPTIC DETECTION IN POLYAROMATIC COMPOUNDS DETERMINATION ПРОТОЧНАЯ ТОНКОСЛОЙНАЯ ХРОМАТОГРАФИЯ /ПТСХ/ ПОЛИАРОМАТИЧЕСКИХ СОЕДИНЕНИЙ С ФЛУОРИМЕТРИЧЕСКИМ ДЕТЕКТИРОВАНИЕМ НА ОСНОВЕ КВАРЦЕВОЙ ВОЛОКОННОЙ ОПТИКИ A.S.Kanev, G.S.Katychin, A.M.Vorontsov, Leningrad, USSR The results of CTLC nanogram level determination of polyaromatic compounds are presented, CTLC is a combination of column liquid chromatography (CLC) and thin layer chromatography (TLC). In CTLC a samp le is injected on the layer with steady eluent flow. The fluorimetric quantitative detection of individual compound zones was realized when they were moving by use of quartz fiber optic excitation system at various wavelengths. The comparison of CTLC, CLC and TLC is discussed in

terms of HETP and resolution factors; for CTLC the Rf value is of no use. 292 P.3-39 THIN LAYER CHROMATOGRAPHY OF SOME FOOD PRODUCTS ТОНКОСЛОЙНАЯ ХРОМАТОГРАФИЯ НЕКОТОРЫХ ПИЩЕВЫХ ПРОДУКТОВ I.I.Gavrilyuk, Kishinev, USSR The quantity and quality content of sugars and organic acids in food products: juices (tangerine, lemon, apple, water-melon),natural wine and cognac has been analysed by thin layer chromatography. It has been shown that the main components of food products under study are glucose, fructose, arabiiiose being present in some less amount. The presence of insignificant quantities of galacturonic acid in natural wine and water-melon juice and of mannose in lemon and tangerine juices should be considered as their speoific peculiarity. Besides, raffinose has been discove red in water-melon juice, and in tangerine juice galactose has been discovered. The set of organic acids is widely represented in juices and natural wine. Lemon and water-melon juices are charac terized by a large amount of citric acid as compared with the malic one discovered in minor amounts. In apple juice malic and citric acids are practically in equal quantiti- tes. When studying natural wines it has been established that the content of succinic acid is twice as much as tartaric and malic ones, and citric and chlorogenic acids have been found in trace amounts. The application of thin layer chromatography for the study of quality and quantity composition of sugars and organic acids in food products gives the possibility to increase the control efficiency of the industrial food production quality.

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1. 293 P.3-40 CLATHRATION OF П-ALKANES ON THIN-LAYER CHROMATOGRAPHY PLATES КЛАТРАЩШ л-АЛКАНОВ НА ПЛАСТИНКАХ ДЛЯ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ G.Kovachev, T.Mlncheva, M.Dacheva, Sofia, Bulgaria The separation of n-alkanes from other branch hydrocar bons using urea adducts or the so called "clathration process" Is well known and commonly used In practice. It is very useful to apply this method to the organic geo chemistry/ where separation of pure n-alkanes is necessa ry as well as their identification and calculation of the odd/even ratio - CPI (carbon preference index). The last approach is very often used in organic geochemistry. But the method is limited quantitatively and the limit is about 10-20 mg. To go over the problem we used special TLC plates treated with urea. A 0,5% solution of n-alkanee in chloroform was put on a Merck TLC plate covered with, silica gel. Then the plate was treated with 7% solution of urea in methanol for 20 minutes. The dryed plate was developed in hexane/dlethyl ether (1:1). Identification was made using Rodamin GG or charring a standard of n-alkanee on the outside end of the plate. The spot was scraped, dryed and put on a small glass column with silica gel and eluted with diethylether. Using this method we are able to run and separate samples of less than 10 mg. Gas liquid chromatography was used to show the results obtained from this kind of clathration of n-alkanes.

294 P.3-41 HPTLC PRE-COATED PLATES CN F 254s - A MEDIUM POLAR MODIFICATION FOR HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY ПЛАСТИНЫ ДЛЯ ТСХ МАРКИ CN F 254s - НОШИ МАТЕРИАЛ ДЛЯ ВЫСОКОЭФФЕКТИВНОЙ ТСХ W.Jost, H.E.Hauck, W.Fischer, Darmstadt, FRG In thin-layer chromatography chemically modified silica gel stationary phases have achieved an increasing importan ce and a growing number of applications in the last years. In this field especially the hydrophobic modified, reversed phase pre-coated plates have established themsel ves. But besides these non-polar stationary phases also the polar modified, hydrophilic pre-coated plate with amino modification has opened interesting possibilities of separation in recent past. A further completion of the modified HPTLC pre-coated plates represents the newly developed, medium polar HPTLC pre-coated plate CN F 254s. In this case silica gel 60 was used as base material for a propyl cyano modification. The chromatographic properties of the HPTLC pre-coated plates CN F 254s are characterized by means of velocity

195 coefficients, retention data of suitable substances and separations, which demonstrate the range of their applica bility. The velocity coefficients К of four different solvent systems (methanol/water, sthanol/water, acetonitrile/water, acetone/water) are determined. In this case the ratio between each organic compound and water is varied from pure organic solvent to pure water. To classify the polarity of the HPTLC pre-coated plates CK in relation to HPTLC pre-coated plates silica gel 60, NH , RP-2, RP-8 and RP-18 the K -values of some steroids in 2two different solvent systemsf are compared with each other. According to i-he extent of the polarity of the HPTLC pre-coated plate CN compounds with medium polarity can be separated with success on these plates. First results in this field were obtained with the following substance classes: alkaloids, drugs, FTH amino acids, dansyl amino acids, steroids and gallic acid esters.

295 P.3-42 STRUCTURE-RETENTIOH-CORRELATION OP DIHYDROPYRAZOLO- TRIAZIN-ON DERIVATIVES IN THIN LAYER AND GAS CHROMATOGRAPHY КОРРЕЛЯЦИЯ СТРУКТУРЫ И УДЕРЖИВАНИЯ ПРОИЗВОДНЫХ ДИГИДРОПИРАЗОЛО-ТРИАЗИНОНА M.Bidld-Igldy, F.Szederkenyi, E.Tihanyi, Budapest, Hungary Structure-retention correlation of the title compounds was studied by TLC and GC methods. The investigated com pounds contained alkyl and cycloalkyl •ubstituents on the N atoms and on one of the С atoms of the triazin ring respectively. The separability and retention order of isomer derivatives was investigated and interpreted. The change of retention values was studied as a function of the position and the carbon atom number of substituting groups respectively. TLC experiments were carried out both on silica gel and on C, reversed phase. GC studies were performed on differently8 substituted polysiloxane phases. The structure retention correlation were compared in the case of silica gel and reversed phase TLC. Linear correlation was found between the carbon atom number of the substituent and the R„ value of the compound, when the substituting alky] and cycloalkyl-groups was placed on the N-atom farer from the keto group or on the C-atom, respectively. The retention properties of these compounds in reversed phase TLC and apolar/moderately polar statio nary phase GC were compared. The retention order was generally the same using the two methods but in the GC the volatilization effect of N-methylation resulted in some alteration.

196 296 P.3-43 CROSSED IEF-IMMUNOELECTROPHORESIS CHARACTERIZATION OF CELLULOLYTIC ENZYMES FROM GLIOCLADIUM SP ХАРАКТЕРИСТКА ЦЕЛЛЮЛАЗ ИЗ GLIOCLADIUM SP МЕТОДОМ ПЕРЕКРЕСТНОГО ИММУННО-ЭЛЕКТРОФОРЕЗА M.Szakace-Dobozi, A.Halasz, Budapest, Hungary A crude cellulase preparation from Gliocladium sp. was immunochemically characterized and the components with cellulolytic activity were identified. More than twenty different antigenic components could be detected by crossed IEF-immunoelectrophoresis, The cellulase complex was also analysed for cellulolytic activity after prepa rative isoelectric focusing using Ampholine pH 3-10. The separated 28 components were characterized by fused rocket Immunoelectrophoresis with polyspecific antisera. The main protein components in the enzyme complex were found to be endoglucanases with pi values between 3 and 5. Sera produced against a single endoglucanase reacted with the other isoenzymes of the same activity indicating that they are antigenically related proteins differing only slightly in pi values. Five exoglucanases with pi valu->* between 2.4 and 3.4 were also characterized. Two protein components were identified/ cellobiase I and II. 297 P.3-44 MATHEMATICAL METHODS IN THE DESCRIPTION OF PROCESSES AND ARTIFACTS OF ELECTROPHORESIS МАТЕМАТИЧЕСКИЕ МЕТОДЫ В ОПИСАНИИ ПРОЦЕССОВ И АРТЕФАКТОВ ЭЛЕКТРОФОРЕЗА V.G.Babski, Kiev, USSR M.Yu.Zhukov, V.I.Yudovich, Rostov-Don, USSR Electrophoresis is a special group of electromigration phenomena appearing during the motion of charged partic les or molecules in the electrical field in a liquid or structured medium. These phenomena should be described in terms of physics of continua, using the general equa tions of hydroelectrothermodynamics of multicomponent reacting mixtures! The assumption of local chemical equi librium makes it possible to simplify these equations and to reduce them to mathematical models of subsequent electrophoretical methods: - Isotachophoresis: the dynamics of zones (times of zones reforming, velocities of zones boundaries and time of complete separation) is studied; the suggested algo rithm refers to the theory of quasilinear hyperbolic equa tions . - Isoelectric focusing: the difference between the point of zero electric charge of molecule and the point of its zero electrophoretic mobility is analyzed; for multicomponent mixtures of carrier ampholytes an analytical

J»7 dependence of natural steady-state pH-gradient from the relative composition of mixture is found; the dynamics of artificial stepwise and continuous pH-gradients of borate- polyol type is studied; the effect of different factors on resolving power of isoelectric focusing is studied. - Zonal electrophoresis; three types of zone broadening are studied - diffusion (at small concentrations of a sample), electromigration (at great concentrations of a sample) and convection when stabilizing measures are Insuf ficient to prevent the onset of thermal or concentration convection; boundary of stability electrophoretic process is evaluated. 298 P.3-45 SUPERLOWER BORATE-POLYOL pH GRADIENTS OF HIGH RESOLVING POWER СВЕРХУЗКИЕ БОРАТ-ПОЛИОЛЬНЫЕ рН ГРАДИЕНТЫ ВЫСОКОГО РАЗРЕШЕНИЯ G.Yu.Agitsky, G.V.Troltsky, V.F.Petrenko, T.K.Sharaeva, Simferopol, USSR It is shown, that borate-polyol isofocusing system con sisting of boric acid, weak base and sucrose allow to qet superlow pH gradients in the range of 0.1 from to 1.0 pH units. Adjustment of the pH gradient width is carried out by changing the ratio of boric acid and weak base, using the weak base for the corresponding pH region, the changing general concentration buffer solution and the width of sucrose concentration gradient. Buffer system obtained allow to perform isoelectrofo- cusing in the pH range from 2.0 to 12.0. Stability of the pH gradients in electric field is very high due to low borate-sucrose complexes mobility. These systems were used for the separation albumin acid minor fractions and other proteins in columns and gels. Polyethylen glycol of low molecular weight as complex forming substance allows to obtain pH gradients in the width of 0.01 to 0.1 pH unit. Potential regions of appli cation of such"width pH gradients are under study. 299 P.3-46 CHROMATOGRAPHIC AND ELECTROPHORETIC METHODS OF MODIFICATED ALBUMIN REVEALING ХРОМАТОГРАФИЧЕСКИИ И ЭЛЕКТРОФОРЕТИЧЕСКИИ МЕТОДЫ ВЫЯВЛЕНИЯ МОДИФИЦИРОВАННОГО АЛЬБУШНА S.N.Borisenko, G.A.Kasymova, Simferopol, USSR We have worked out the new methods of chromatographic and electrophoretic analysis of modificated forms of albu min . They are based on the denaturated and chemical modifl oated albumin ability to be separated from native form, after proper treatment in TCA-ethanol system. After such treatment, native part of albumin is washed out from the carrier (chromatographic paper or acetatcellulose films) 198 and modifioated part remains fixed on the carrier. The amount of modifiested albumin after chromatography or electrophoresis can be detected and quantitative analysis can be carried out with colorimetric or densytometric methods. Methods worked out are simple and accesible for serial simultaneous analysis of large amount of samples. The method can be applied to albumin investigations after thermal and chemical modification, when physico- chemical properties of this protein are undergone consider able changes. Under these conditions conformational and other properties changes occur not in all electrophoretic fraction, but only in a number of molecules, which are detected by these methods. Such form of albumin was found to be present in persons who are directly associated with toxic preparatures (for example with superficially active substances) at the chemical enterprises. Therefore its separation and the possibility of quantitative analysis is of great importance. Revealing of modificated forms of albumin with its quantitative analysis may be used as toxic test at unhealthy enterprises and in some other con ditions.

300 P. 3-47 DESIGN AND EXPERIENCE OP ISOELECTRIC FOCUSING IN ТИЕ HORIZONTAL MULTICHAHBER APPARATUS КОНСТРУКЦИЯ И ОПЫТ Й30ЭЛЕКТРИЧЕСК0Г0 ФОКУСИРОВАНИЯ В ГОРИЗОНТАЛЬНОМ МНОГОЯЧЕИСТОМ АППАРАТЕ I.F.I.iryukhin, O.G.Kosik, S.P.Klintsova, V.I.Kuprienko, Simferopol, USSR Isoelectric focusing (IEF) in the horizontal multicham- ber apparatus enables to carry out a separation of mixture of ampholytes, precipitating at the isoelectric point. IEF in this device is effected in the medium devoid of sub stances used for the construction of density gradient in vertical columns. A principle of design is presented and its realization in the specific running devices for analytical and prepa rative purposes. Using a method of specific detection of proteins and their quantitative estimation by radial immunodiffusion technique we have obtained isoelectric spectra (IES) of the number of proteins in biologic fluids without their preliminary isolation. Specific features of human serum immunoglobulin IES are studied in subjects with different blood groups. In the process of immunization of man and animals the dynamics of accumulation of specific antibodies, isolated by preparative IEF in pure form, was followed up. Only by the indicated approach we have succeeded in detec ting the anomalous by some physico-chemical properties, of lg G fractions. Method is successfully used for the separation of pro tein fragments.

1» 301 P. 3-48 APPLICATION OF ELECTROPHORETIC AND ISOELECTRIC FOCUSING METHODS IK THE STUDY OF IMMUNOGLOBULINS ИСПОЛЬЗОВАНИЕ МЕТОДА ЭЛЕКТРОФОРЕЗА И ИЗОЭЛЕКТРИЧЕСКОГО ФОКУСИРОВАНИЯ ДЛЯ ИЗУЧЕНИЯ ИММУНОГЛОБУЛИНОВ O.G.Kosik, S.P.Klintsova, I.F.Kiryukhin, V.P.Fesenko, V.E.Krotenko, Simferopol, USSR For an elucidation of changes in the structure of immu noglobulins we have carried out a study of Fc-fragments of immunoglobulins in the blood serum of healthy subjects and in the state of functional disorders of human organism. Investigation was conducted by the method of two-dimensio nal electrophoresis according to our modification. Immuno globulin G (IgG) fraction was subjected to the papain hydrolysis just in the gel. Fc-fragments were isolated in the process of electrophoresis in the second direction with the use of nonionic detergents. Isoelectric focusing (IEF) in the horizontal multichaT- ber apparatus was used for the analysis of immunoglobulir distribution in the whole blood serum, and for studying the changes of IgG Fc-fragments we have used IEF in the tubes with borate-polyol system. It was shown that IgG isoelectric spectra of human blood serum after thyroidectomy were changed depending on the severity of the process with significant accumulation of "alkaline" IgG fractions. In diabetes, hepatitis, hepa- tocirrhosls, ulcerative disease of stomach and duodenum we have observed the appearance of more acidic components in IgG Fe-fragments. And in this case there is a correla tion depending on the degree of severity of the disease.

302 P.3-49 QUANTITATIVE DETERMINATION OF FRACTIONAL COMPOSITION OF SOME HORMONE PREPARATIONS BY POLYACRYLAMIDE GEL ELECTROPHORESIS AND ISOELECTRIC FOCUSING КОЛИЧЕСТВЕННОЕ ОПРЕДЕЛЕНИЕ ФРАКЦИОННОГО СОСТАВА ГОРМОНАЛЬНЫХ ПРЕПАРАТОВ МЕТОДАМИ ЭЛЕКТРОФОРЕЗА И ИЗОЭЛЕКТРОФОКУСИРОВАНИЯ P.A.Babich, G.I.Fedotova, V.P.Pakhomov, N.V.Makarov, Moscow, USSR By the combination of polyacrylamide gel electrophore sis (PAGE) and isoelectric focusing (IEF), the fractional composition of insulin, glucagon, andecaiin, thymosin and some other protein preparations has been determined. The following standard preparations have been selected as a criteria of purity of these hormones: monocomponent insu lin and proinsulin ("Novo", Denmark), glucagon, thymosin and trypsin ("Serva", FRG). The quantitative estimation of the protein fractional composition after PAGE was car ried out on a "Cellosystem" densitometer ("Sebia", France). IEF was performed on Multiphore ampholine plates (pH 3.5- 9.5 and 2.5-6.0, "LKB", Sweden). The parameters of PAGE

200

1. to be optimized were gel concentration, buffer composition, diameter and length of tubes and sample weights. It has been demonstrated that IEF was more relevant for determina tion of the protein composition than PAGE. 303 P.3-50 COMBINATION OF ELECTROEXTRACTION AND LIQUID CHROMATOGRAPHIC METHODS FOR ANALYSIS OF ALKALOIDS СОЧЕТАНИЕ ЭЛЕКТРОЭКСТРАКЦИОННЫХ И ЖИДКОСТНО-ХРОМАТОГРАФИЧЕСКИХ МЕТОДОВ АНАЛИЗА АЛКАЛОИДОВ P.P.Gladyshev, V.S.Volodin, Alma-Ata, USSR Chromatographic analysis of natural compounds presuppo ses their quantitative isolation from object in question and further division. HPLC secures a rapid division of a alkaloids if they isolate from compounds of another nature. Alkaloids are isolated usually from plants with extraction of organic solvents. To obtain a sum of alkaloids purifed from ballastic substances it uses reextraction with diffe rent solvents. These methods are long and require great expenditure of manual labour. This problem is solved easier with electroextraction method. With this purpose it worked out analytical membrane electroextractor which allows to isolate automatically a sum of alkaloids from plants raw material and theirs industrial semiproducts. The method is based on using of electrophoretical and mo lecule-size effects. It is more reasonable to relize a chromatographic division ionizable organic compounds using water and water-organic eluents. In this case addi tional possibility is appeared to control the division with the help of pH variation. All of this secures successful combination of considering methods. This is showed on example of analysis of opium and glaucium alkaloids. Mechanisms of sorption of alkaloids on ion-exchangers and reversed-phase sorbents and optimization methods of chro matographic division are discussed.

304 P.3-51 TLC DETERMINATION OF 7fi- (4-CARB0XYBUTANAMID0 ) CEPHALOSPORANIC ACID ОПРЕДЕЛЕНИЕ МЕТОДОМ ТСХ 7Й -/4-КАРБ0КСИБУТАНАМИД0/ ЦЕФАЛОСПОРАНОВОЙ КИСЛОТЫ A.Pisafikova', J.Machova', K.PlhacTtova, Prague, CSSR A combined method for analytical determination of 7j8 — (carboxybutanamido)cephalosporanic acid )or glutaryl-7- ACA) is reported. The nethod is based TLC chromatography combined with spectrophotoraetric indication. The 7-aminocephalosporanic acid (7-ACA) is an impor tant intermediate in the preparation of semisynthetic cephalosporins. It is produced by chemical or enzymatic hydrolysis of a side chain of cephalosporin C. The compounds has been chiefly prepared by removing chemically the side chain from the naturally occuring

201 cephalosporin C. The study of enzymatic deacylation of cephalosporin С to 7-ACA revealed the difficulties at the selection of strains with cephalosporin С acylase and low yields of 7-ACA in one-step bioconversion /1,2/. I was been reported, that cephalosporin С was oxidatively deami- nated enzymaticaly /3,4/ with D-amino acid oxidase or chemically /5/ to glutaryl-7-ACA and this was subsequently deacylated to 7-ACA /6/. Cephalosporins and 7-ACA can sensitively be detected in OV spectrum region. Chemical and biological detections can also be applied for their determination. Host spectrophotometric methods г ?е not specific and therefore can not be used for determination of these oephem compounds or, as the case may be, of products of their degradation in mixture. For this reason chromatographic separations are more convinient. A quantitative thin-layer chromatographic methods for substances of cephalosporin type, except for glutaryl-7-ACA, are described by several authors /7-10/. Vandamme /11/ presents various solvent systems for cepha- losporines and 7-ACA. Magnem /12/ uses for chromatographic determination of cephalosporins *he TLC method with spect rophotometry indication. Individual zones are eluted and measured on a spectrophotometer. The agreement with the biological method is satisfactory. We applied this method for the determination of glutaryl-7-ACA in which case the detection with ninhydrin can not be used (7-ACA Is ninhydrin positive). The proper determination was carried out by combining the TLC separation with spectrophotometric detec tion. The absorption maximum for glutaryl-7-ACA was found through measurement of a standard substance on a spectro photometer OV Beckman ACTA and the results were evaluated • on a Hewlet Packard computer. The results of the measurements are presented in tables from where one can see accuracy and reproducibility of the results obtained with the aid of the present method. 1. F.M.Huber, R.R.Chauvette, B.G.Jakson. in "Cephalospo rins and Penicilins" e. E.H.Flynn, Acad.Press N.Y. London,1972, p.27.. 2. H.Goi, T.Niwa, C.Hojiri, M.Miyado, M.Seki, Y.Yamada, Jap. pat. No.53-94093 (1978). 3. P.Mazzeo, A.Romeo., J. Chem. Soc., Perkin I, 2Q. 2532 (1972). 4. O.A.Gilbert, B.H.Arnold, A.Fildes, Brit.pat. No.l, 272,769, 1972. 5. N.Suzuki, T.Sowa, H.Murakami, Jap.pat. Announce, 51- 86,490 (1972). 6. Y.Shibuya, K.Matsumoto, T.Fujii, Agric. Biol. Chem, 45. 1561 (1981). 7. B.Loder et al.: Biochem. J.,22, 409 (1961). 8. H.P.Fechtig et al.: Helv.Chia.Acta,U, 1Ю8 (1Э68). 9. L.P.Harrelli et al.: Chem. Biol.,609 (1972). 10. G.M.Overvliet et al.: Pharm.Weekbl. 109, 21, 489 (1974). 11. E.J.Vandamme et al.sJ.Chromatogr.,71, 141 (1972). 12. T.M.Magnem et al.: Medd.Norsh.Farm.Selsk.39,3,221(1977).

202 305 P.3-52 ANALYTICAL DETERMINATION OF DIASTEK20MERS OF EPHEDRINE BY THIN-LAYER CHROMATOGRAPHY АНАЛИТИЧЕСКОЕ ОПРЕДЕЛЕНИЕ ДИАСТЕРЕОМЕРОВ ЭФЕДРИНА МЕТОДОМ ТСХ U.Machova, A.Pifsar'ikova, M.Vondra'cek, Prague, c;;SR Natural ephedrine, an important sympatomimetic drug, contains four diastereomers among which only two possess ing positive pharmacological activities. These are (L)(-)- (lR,2S)-l-phenyl-2-methylamino-l-propanol/(-)-ephedrine/ and (L)(+)-(lS,2S)-l-phenyl-2-methylamlno-l-propanol /(+)-pseudo-ephedrine/. (-)-Ephedrine is currently produ ced by the fermentation process yielding 1-hydroxy-l-phenyl-propanone /ketol/ followed by the amination hydrogenation of the latter /1/. (+)-Pseudo-ephedrine can be easily obtained from (-)-ephedrine /and vice versa/ by chemical methods /2/. The production of both particular isomers therefore requires fast and reliable analytical determina tion of individual isomers in a mixture containing some technological impurities, e.g., ketol and N-methylbenzyl- amine, byproduct of the amination hydrogenation. Although many of the published chromatographic methods separate ephedrine diastereomers from other drugs /3/, the separation of isomers from each other and their quantitati ve determination in a mixture is the topic of current inte rest. The promissing results werw obtained by chromatogra phic methods including the GLC chromatography /after functiona.lization of both isomers/ /4,5/, the ion-pair high-performance liquid chromatography /3.6/, the thin- layer chromatography /3/ and also by the 13C NMR spectro scopy /3/. We report on the determination of (-)-ephedrine and (+)-pseudo-ephedrine by thin-layer chromatography on silica gel using a ninhydrine reagent for colour develop ment of spots /3/. The densitoaetric evaluation of the spots allowed the determination of minimum 0,3 ug of (- (-)-ephedrine and 1,0 ug of (+>-pseudo-ephedrine in 1 ul of the solution. Results of analyses of the crude techno logical products, for various developing solvent mixtures as well as experimental procedure will be presented in detail. 1. J.Netrval, M.Vodnansky: Kvasny prumysl.28, 131 (1982) 2. S.Ikuma: J.Pharm.Soc.Japan,75, 52 (1955ТГ 3. S.Barkan, J.D.Webwr, E.SmitKT J.Chromatogr., 219, 81 (1981) and references therein. 4. K.Yamasaki, K.Fujita, H.Sakamoto, K.Okada, H.Yoshida, O.Tanaka: Chem.Pharm.Bull., 22, 2898 (1974). 5. K.Kruse, W.Francke, W.A.KonTg: J.Chromatogr.. 170, 423, (1979), 6. K.Sagara, T.Oshima, T.Misaki: Chem.Pharm.Bull.. 31, 2359, (1983).

203 306 P.3-53 EMPLOYMENT OF A FREE-FLOW ELECTROPHORETIC APPARATUS FOR SEPARATION AND CONCENTRATION OF PROTEINS ИСПОЛЬЗОВАНИЕ ЗЛЕКТР0Ф0РЕТИЧЕСК0Г0 АППАРАТА ПРОТОЧНОГО ТИПА ДЛЯ РАЗДЕЛЕНИЯ И КОНЦЕНТРИРОВАНИЯ БЕЛКОВ S.A.Strukun, V.P.Zav'yalov, Moscow Region, USSR A.V.Gavryuehkln, V.N.Brezgunov, Hoscow Region, Obolensk, USSR Separation of proteins was studied in borate-polyol pH gradients, namely In the borate-mannltol pH gradients,with an initially constant ratio of analytical concentrations of polyol and borate ions as well as in the pH gradients, obtained by the formation of the concentration gradients of boric acid in solutions of borax and mannitol. The gra dients indicated were shown to change drastically under the Influence of the electric field. Thus the Isoelectric focusing of the proteins was not achieved, and the separa tion resulted from electrophoresis in the pH gradients. The quality of such separation is higher than that, ob tained by conventional electrophoresis, and approximates that, achieved with isoelectric focusing, but the fractio nation is attained mora rapidly. The results of the sepa ration were dependent on the field strength and residence time, ratio of the flow rate of the sample to that of the buffer, properties of the membranes between the electrodes and separation chamber, chemical composition of the gra dients, in particular, presence of salts, and in case of the borate-mannitol gradients - on the point of sample injection into the pH gradient. The free-flow electrophoretic apparatus was shown to be suitable for rapid concentration of proteins in the stepped pft gradients combined with conductivity gradients. In the gradients, formed by solutions of borax and boric acid one can concentrate alkaline and neutral proteins together with the simultaneous removal of acidic proteins. 307 P.5-02 SELECTIVITY AND SENSITIVITY OF THIN LAYER CHROMATOGRAPHIC DETECTION SYSTEMS СЕЛЕКТИВНОСТЬ И ЧУВСТВИТЕЛЬНОСТЬ СИСТЕМ ДЕТЕКТИРОВАНИЯ ДЛЯ ТОНКОСЛОИЬОИ ХРОМАТОГРАФИИ K.Fodor-Csorba, F.Dutka, Budapest, Hungary The multiresidue methods are of great importance in the analysis of pesticides. Selective and sensitive procedures are needed for the determination of the quality and quan tity of the pesticides which are present in subnanogram or lower level in food extracts. If the detection system is rut selective enough, some compounds may have disturbing effect on the determination of the pesticides studied. The method of the Association of Official Analytical Chemists (AOAC) is an internationally widely used multi-

204 residue procedure for the determination of the residues of organochlorine and organophoaphorus insecticides. We have extended this AOAC method for the determination of residues of thlocarbamate herbicides of plant origin and we have developed a new and sensitive (50 ng/spot) detec tion using Glbbs reagent. The selectivity of the thin layer chromatographic detec tion systems involved in the AOAC method:

AgN03-2-phenoxyethanol 4,4'-nitrobenzylpyridine, and the new detection of the thiocarbamates by Gibbs reagent: 2,6-dichlorobenzoquinone-N-chloroimine 2,6-dibromobenzoquinone-N-chloroimine were studied in this paper. The sensitivity of these methods were studied, further the detection limit of the organophosphorus insecticides was enhanced. It was estsv 1 ished that the Gibbs reagent detected not only the thiocarbamates but the organophospho rus insecticides containing sulfur atoms with the limit of 5 ng/spot.

205 SORBENTS

308 S.5-1 RECENT DEVELOPMENTS IN THE SYNTHESIS AND CHARACTERIZATION OF REVERSED PHASE HPLC SILICA PACKINGS ПОСЛЕДНИЕ ДОСТИЖЕНИЯ В СИНТЕЗЕ И ИЗУЧЕНИИ 0БРАЩЕНН0-ФА30ВЫХ СИЛИКАГЕЛЬНЫХ СОРБЕНТОВ К.К.linger, Mainz, FRG Reversed Phase HPLC (RP HPLC) has become increasingly important in the separation of ionic or ionizable com pounds, particularly biologically active analytes, e.g. amino acids, peptides and proteins. In order to adapt RP HPLC for the separation of this type of compound (with the alms of high resolution, high mass recovery, maintenance of bioactivity as well as acceptable column reproducibi lity and stability) the bonding chemistry was thoroughly re-investigated utilizing modern surface analysis methods as a tool of characterization. Recent studies in our laboratory indicate that macroporous low-surface area silicas free of metal impurities, prepared by low temperature agglomeration processes, are best suited as base material. They provide an improved surface homogeneity, a uniform pore size and sufficient accessibility to higher molecular weight solutes in gra dient RP HPLC. A systematic examination of the modifica tion procedures showed that silanlzation of silica at room temperature, employing reactive silanes and catalysts, was more advantageous with regard to defined surface structure and ligand density than a high temperature modification. A large number of chemical and physical methods were applied in the characterization of native and n-alkyl modified silicas (e.g. 29Si and 13C NMR CP/MAS, thermal analysis, calorimetry, inductively coupled emission spectroscopy, adsorption techniques etc); however, the interplay of the roles of residual polar sites and bonded n-alkyl groups in RP HPLC could not be fully exploited.

309 S.5-3 NEW SORBENTS AND THEIR APPLICATIONS IN ANALYTICAL AND PREPARATIVE HPLC НОВЫЕ СОРБЕНТЫ И ИХ ПРИМЕНЕНИЕ В АНАЛИТИЧЕСКОЙ И ПРЕПАРАТИВНОЙ ВЭЖХ J.Coupek, Prague, CSSR Abstract not arrived on time for printing.

206 ЗЮ S.5-4 THE BONDED IAVER STRUCTURE AND THE SELECTIVITY OF CHEMICALLY MODIFIED SILICA SORBEHTS FOR CHROMATOGRAPHY СТРУКТУРА ПРИВИТОГО СЛОЯ И СЕЛЕКТИВНОСТЬ ХИМИЧЕСКИ МОДИФИЦИРОВАННЫХ КРЕМНЕЗЕМНЫХ СОРБЕНТОВ ДЛЯ ХРОМАТОГРАФИИ S.M.Staroverov, A.A.Serdan, G.V.Lieichkin, Moscow, USSR The latest achievements In contemporary liquid chroma tography to a great extent are due to the creation of silica-based sorbents. Such sorbents proved to be of inte rest also for gas chromatography. The effects of geometrical structure, composition of organosilicon anchour groupping, length of hydrocarbon chain, nature of bonded compound functional groups, and end capping on the selectivity of sorbents for high perfor mance liquid chromatography have been systematically studied. The calculations of critical parameters for sili cas made to achieve the reproducibility of surface proper ties of the «orbente. The critical analysis of two main methods for «orient synthesis, i.e; immobilization and a surface assembling, is also presented. The influence of an eluent composition, end capping and temperature on the mobility of bonded groups and their structure are elucidated by NHR and EST spectroscopy. The cases of the Interaction of bonded functional groups with surface silanol groups {so called arch struc ture") were found. This phenomena was used for obtaining thermostable and low specific phases for gas chromatogra phy. The sorbents are notable for their low resistance to mass transfer and enable to work with carrier gas contain ing water vapours. In the study the sorbents were successfully used to solve a number of HPLC problems: analysis of isomeric alkylphenols, polunuclear aromatic compounds, oligonucleo tides, inorganic ions, enanthiomers of some unsubstituted amino acids. 311 S.15-5 SORBENTS "ARMSORB-Sil" FOR LIQUID AND THIN LAYER CHROMATOGRAPHY СОРБЕНТЫ И НОСИТЕЛИ "АРМСОРБ Сил" ДЛЯ ЖИДКОСТНОЙ И ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ A.A.Beglaryan, S.G.Babayan, J.G.Martirossyan, E.M.Sarkissyan, S.S.Ayrapetyan, Yerevan, USSR The aim of this research was to develop a set of silica gel carriers and sorbents ("ARMSORB-Sil" 6, 10 and 30, where number refer to pore size in nm) possessing narrow pore size distribution, and to obtain the modified forms

Ся, C., and Ot for purposes of liquid chromatography based ОЙ "ARllSORB-Sil" 10.

207 Directional synthesis process has been developed in order to obtain the sorbents of desired porous structure, and a pilot plant has been erected. Sorbent obtaining orocess involves the following stages: - silicasole obtaining from the alkaline-siliceous solu tion; - silicone dioxide particle growth in alkaline aqueous medium; - gel formation (pH 5.6-6.2); - xerogel freezing <-25°c) and defreezing; - xerogel filtration, washing and drying; - silica gel calcination at 450-500 С; - grinding and fractioning. Parameter estimation of "AEHSORB-Sil" sorbents porous structure has been carried out using adsorption and mer cury porosimetry methods. The obtained results evidence about homogenious character of mesoporous structure for the synthetized sorbents, with narrow distribution in pore size. Silica gel modification has been made by a method of single-stage covalent immobilization over the hydroxylated carrier surface. Data analysis concerning the physico-chemical proper ties of sorbenta, and the results of TLC plate HPLC column testing, prove the high industrial significance of the synthetized materials. 312 S.9-1 FIBROUS ION EXCHANGE SORBENTS FOR PREPARATIVE CHROMATOGRAPHY ВОЛОКНИСТЫЕ ИОНООБМЕННЫЕ СОРБЕНТЫ ДЛЯ ПРЕПАРАТИВНОЙ ХРОМАТОГРАФИИ V.S.Soldatov, Minsk, USSR Application of fibrous ion exchange materials opens new possibilities in preparative chromatography. These mate rials consist of monofilaments slightly differing in diameter, which ranges from 3 to 50 mem in different fib res. They can be prepared in various physical forms such as pulp consisting of short fibres for filling columns, non-woven materials, cloths, filaments, conveyer belts. This feature makes it possible to oarry out easily the processes involving a moving sorbent. A small diameter and a uniform fibre thickness promise good chromatographic characteristics of these materials. It is an important feature of these materials that the resistance of columns, filled with fibrous ion exchanger, can be varied in a wide range since, natural elasticity of the fibres permits retaining the integrity of an ion exchanger layer with different filling degree. Geometric dimensions of the layer are independent of swelling. Therefore, there are no problems associated with variation in a volume, when pas sing from one ionic form to the other. The synthesis, physico-chemical, mechanical and sorp tion properties of synthetic ion-exchange fibres of diffe rent types such as strong-base cation exchangers, strong- 206 base and weak-base anion exchangers, have been described. The applicability of the above materials for separation of ions of metals, anions, amino acids has been shown.

313 S.9-3 INORGANIC IOM-EXCHANGE ATERIALS, THEIR SYNTHESIS, PROPERTIES AND APPLIC7AION НЕОРГАНИЧЕСКИЕ ИОНООБМЕННЫЕ МАТЕРИАЛЫ, ИХ СИНТЕЗ, СВОЙСТВА И ПРИМЕНЕНИЕ V.V.Volkhin, M.V.Zilberman, N.Ph.Kalinin, S.A.Onorin, Perm, USSR Many insoluble inorganic compounds of different classes, for example, hydroxides, sulphides, ferro-cyanides, mixed oxides, etc. exhibit ion-exchange properties. An important peculiarity of inorganic ion-exchangers is the supplemen tary ability, if compared with the ion-exchange resins, to vary the selectivity of the exchange. Thus, not only the nature of compound (the type of functional group) but also the surroundings of a sorption centre affect the selectivity of the exchange, this is due to the fact that insoluble inorganic compounds are, in most cases, three- dimensional polymers. Based on this, cationic exchangers, capable of separating light alkali elements from heavy alkali and alkali-earth ones, as well as heavy alkali me tals and thalhium - from other alkali and multiply charged elements, were producted. Another possibility of separation depends on the diffe rence of the position of the element in the solubility series. Application of sulphides, hydroxides, phosphates and ferro-cyanides to solve the problems of separation of d-elements is based on the above difference. In addition to the difference in solubility, one may use the differen ces in the redox potentials of the elements under separa tion. The removal of the element from the solution takes place due to variation in solubility when the valency state is changing. Now the problems of determined synthesis and selection of inorganic ion-exchange materials to solve a number of practical tasks, for example, pressing of hydromineral raw materials, sewage and industrial solution treatment, have been handled. Since stability characteristics of insoluble inorganic compounds rarely meet the requirements to ion-exchangers, methods of pelletizing of inorganic ion-exchange materials with the binding agent have been brought to a commercial level. These methods of pelletizing help to produce gra nules of a regular (spherical or ellipsoidal) shape, their mechanical resistance is close to that of ion-exchange resin. The pilot production was set up to produce inorga nic sorbents.

14. Зак. 872 209 314 S.lO-1 CHEMICALLY MODIFIED POLYMERIC SORBENTS FOR GAS CHROMATOGRAPHY ХИМИЧЕСКИ МОДИФИЦИРОВАННЫЕ ПОЛИМЕРНЫЕ СОРБЕНТЫ ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ K.I.Sakodynskii, L.I.Panina, S.B.Makarova, Moscow, USSR

The ability of porous polymeric sorbents for chemical transformations gives the opportunity to create new types of sorbents, based on already known ones and to alter their properties and spheres of application broadly. The gas chromatographic properties of some chemically modified sorbents on the basis of macroporous copolymers of styrene and divinylbenzene, 2,5-methylvinylpyridlne and divinylben2ene, glycidic methyl acrylate and divinylbenze ne have been studied. The reactions of chloromethylation, amination, nitration, sulphation, phosphorylation are used for synthesis of these sorbents. The specificity and selec tivity of the sorbents are estimated. It is shown, that the connection of the functional groups of different chemi cal nature to one and the same polymeric matrix as a result of polymeranalogous transformations influences the retention volumes of components as well as the order of their elution from the chromatographic column.

The important role of geometric and chemical structure of polymeric matrix to the surface of which the functional groups are being connected is pointed out. The. use of the polymers with less rigid structure, with less content of the cross-linking agent for modification gives the oppor tunity to receive sorbents with higher specificity of the molecular interactions. The method of polymeranalogous reactions permits to add to the original polymeric sorbent a few functional groups of different chemical nature simultaneously and receive polyfunctional sorbents with new set of gas chromatogra phic properties. The gas chromatographic behaviour of sorbents with NO, and SO,H, NH, and S03Na - functional groups is studied. The chemical modification of polymeric sorbents, provi ding the opportunity of connection of the functional groups with different distribution of electronic density leads to the receiving of 2 and 3 type sorbents, according to their ability for specific interactions. Examples of the use of chemically modified sorbents to solve analytical tasks of gas chromatography are given.

210 315 S.10-2 GAS CHROMATOGRAPHY ON POROUS POLYMERS. II.RETENTION- STRUCTURE CORRELATIONS FOR AROMATIC SUBSTANCES CHROMATOGRAPHED ON SORBENTS OF DIFFERENT POLARITY ГАЗОВАЯ ХРОМАТОГРАФИЯ НА ПОРИСТЫХ ПОЛИМЕРАХ. П. ЗАВИСИМОСТЬ УДЕРЖИВАНИЯ ОТ СТРУКТУРЫ АРОМАТИЧЕСКИХ СОЕДИНЕНИИ ПРИ ХРОМАТОГРАФИИ НА СОРБЕНТАХ РАЗЛИЧНОЙ ПОЛЯРНОСТИ A.Patrykiejew, Lublin, Poland Kovats retention indices and thermodynamic functions for several benzene derivatives have been determined, using columns packed with porous polymeric sorbents of different polarity. The sorbents used were copolymers of 1,4-divinylbenzene and 1,4-dimethacryloyloxymethylnaphtha- lene. It has been found that by changing the molar ratio of monomers one can obtain porous polymers of different polarity. The correlations between solute physico-chemical proper ties and the retention data have been determined. In par ticular, we have studied the influence of chemical nature of substituents attached to aromatic ring ав well as their mutual positions on the solute - sorbent interactions. It has been found that the increase in the sorbent polarity leads to higher selectivity towards various types of ben zene derivatives. It has been also shown that solute-sorbent interactions were nonspecific irrespective of the polymer polarity. One of the most interesting results was a clear corre lation between temperature coefficient of retention indi ces and the chemical nature of substituents attached to aromatic ring. This fact may be practical importance as it enables to improve resolution by a proper choice of temperature. 316 S.10-3 RECENT ADVANCES IN INVESTIGATION AND APPLICATION OF MESOGENIC PHASES IN GAS CHROMATOGRAPHY ПОСЛЕДНИЕ ДОСТИЖЕНИЯ В ИЗУЧЕНИИ И ПРИМЕНЕНИИ МЕЗОГЕННЫХ ФАЗ В ГАЗОВОЙ ХРОМАТОГРАФИИ G.Kraus, Halle-Wittenberg, GDR Chromatographic separations are being performed success fully with mesogenic phases and won more importance and application in science and industry. Their selectivity is affected both by structure of the mesophase, which inte racting with substrate makes their molecular geometry felt, and by chemical structure of the liquid crystal, like every conventional phase in gas chromatography causing different interactions with substrates of different pola rity. Especially difficult separation problems arise with mixtures of geometric isomers having closely related structures and slight differences in their vapor pressures. The relationship between selectivity and alternation of

211

I retention of isomeric n-alkenes and the separation of iso meric n-alkylbenzenes on nematic phases was Investigated. The alternation of selectivity goes parallel to alterna tion of clearing points and of degree of order. Lateral substituente in mesogenic molecules effect a change in selectivity. Not completely planar structures of molecules with short terminal groups cause an optimum selectivity. Systems for classification and optimization of liquid cry stalline stationary phases and possibilities for utiliza tion have been developed. The selectivity od mesogenic stationary phases as pha ses with regulative selectivity can be used for rapid separations of isomers. Many of separations carried out on mesophases are or can be important for research and indust ry. Some examples in comparison with conventional stationa ry phases will be given. 317 S.10-6 POLYSILOXANE STATIONARY PHASES IN GAS CHROMATOGRAPHY ПОЛИСИЛОКСАНОВЫЕ СТАЦИОНАРНЫЕ ФАЗЫ В ГАЗОВОЙ ХРОМАТОГРАФИИ H.Rotzsche, H.Clause, K.-D.Muller, H.Steimann, Radebeul/Dresden, GOR Polyorganosiloxanas have continued to be the dominant group of stationary phases used in gas chromatography. In a survey, methyl-, phenyl- fluoroalkyl-, and cyanoalkyl- substituted polysiloxanes will be examined as well as car- borane-, polyester-, and chiral polysiloxanes. Molecular interactions (Rohrschneider-McReynolds-constants, molecu lar retention indices), molecular weights, thermal stabi lity, viscosity and film formation and their influence on selectivity, column efficiency, and life, resp., will be summarized. Since silanol groups, both on the substrate ar.d in the liquid phase, affect the quality of a gas chro matographic column, methods for their determination have been developed, and recent results will be reported. The final part of the paper will deal with the possibilities of cross-linking.and immobilizing silicone stationary phases.

318 S.lO-4 AN INCREASE OP THE EFFICIENCY OF CHROMATOGRAPHIC COLUMNS FILLED WITH ZEOLITES FOR SEPARATION OF CERTAIN ORGANIC COMPOUND MIXTURES ПОВЫШЕНИЕ ЭФФЕКТИВНОСТИ ХРОМАТОГРАФИЧЕСКИХ КОЛОНОК, ЗАПОЛНЕННЫХ ЦЕОЛИТАМИ, ПРЕДНАЗНАЧЕННЫХ ДЛЯ РАЗДЕЛЕНИЯ СМЕСЕЙ НЕКОТОРЫХ ОРГАНИЧЕСКИХ СОЕДИНЕНИЙ T.G.Andronikashvili, V.G.Berezkin, N.A.Nadiradze, L.Ya.Laperashvili, Tbilisi, Moscow, USSR Two effects have been used in combination to increase the separative capacity of the chromatographic column: the use of zeolites in a surface-layer variant and "active" carrier gas. 212 Separation of the model mixtures consisting of satura ted hydrocarbons C--C„, alcohols C.-C-, complex esters and ketones was carried out on zeolites N3Y and derived hydro gen (НУ) and cadmium (CdY) forms. In the process of chro matographic separation on HY the most symmetrical separa tion curves are obtained, and CdY is characterized by the ability to enter into specific relations with adsorbate molecules. Chromatographic columns LKM-72 (FID), 0,5m long, dia meter 0.004m were filled with zeolites in the variant of volumetric sorbents,0.5-1.0xlo" m granulation and in the surface-layer variant, the dusty zeolites covered on the s^id layer chromosorb-W, granulation 0.25-О.ЗОхЮ m. hc-iium, nitrogen, carbon dioxide were used as a carrier gas. It was found that the best separation of the model mixture takes place at the use of the surface-layer fil ling of the columns and the"active"carrier gas of the carbon dioxide. In this case the effectiveness of the column per lm for different compounds was 1000 theoreti cal plates averagely, and the symmetry of the chromatograms was close to unity. It was also shown that the zeolite cation nature in the intermolecular interactions in the system of adsorbate- adsorbent is revealed weaker than at the use of such tra ditional inert carrier gases as helium and nitrogen. Dependence of the time and succession of the elution of certain mixture components upon the zeolite cation nature and the carrier gas was established. Selectivity of the homological rows was defined: n-hydrocarbons, al cohols, esters and ketones to zeolite cations depending upon the nature of the carrier gas. 319 S.10-5 A STUDY OF THE STEREOSPECIFIC PROPERTIES OF CYCLODEXTRINS AS GSC STATIONARY PHASES ИССЛЕДОВАНИЕ СТЕРЕОСПЕЦИФИЧЕСКИХ СВОЙСТВ ЦИКЛОДЕКСТРИНОВ В КАЧЕСТВЕ СТАЦИОНАРНЫХ ФАЗ В ГАХ E.Smolkova-Keuleraansovi, L.Feltl, E.Neumannova', Prague, CSSR The ability of cyclodextrins (CDs) to selective interac tion with a great variety of substances depending on the shapes and sizes of their molecules provides wide possibi lities to use them as selective stationary phases and, on the other hand, to study chromatographically the inclusion properties of these substances. In continuation to previous research, a comparative study was carried out of the stereospecific properties of ot-, yS- and }f-CD, using a set of positional isomers of aromatic hydrocarbons (xylenes, diethyl- and triethylben- zenes, cymenes, ethyl- and propyltoluenes). The dependences of the relative retentions on the boi ling points of the studied substances were examined and the retention mechanism was discussed on this basis. The 213 elution order of variously substituted aromatic hydrocar bons / affected by the steric arrangement of the substances and by the size of the cyclodextrin cavity, can be utili zed for separations of complex mixtures, which is demonst rated by chromatograms.

320 P.1-37 PRINCIPLES OF PREPARATION OF SELECTIVE POLYMER SORBENTS FOR ION EXCHANGE SEPARATION ПРИНЦИПЫ ПОЛУЧЕНИЯ СЕЛЕКТИВНЫХ ПОЛИМЕРНЫХ СОРБЕНТОВ ДЛЯ ИОНООБМЕННОГО РАЗДЕЛЕНИЯ A.A.Efendiev, Baku, USSR General principles of preparation and regulation of sorption properties of coirplex-forming polymer sorbents for ion exchange separation have been observed. A new approach to the problem has been developed. This approach is based on the use of opportunities provided by the poly mer nature of the sorbents and consists in regulation of their sorption characteristics via directed variation of the polymer structure, distribution of functional complexing groups in macromolecular chains and their spatial lo cation in three-dimensional polymer matrix. It has been shown that crosslinked copolymers of a si milar composition, containing two types of functional complexing groups, may exhibit different sorption properties depending on distribution of the groups in macromolecular chains. There has been developed a new method of preparation of complex-forming polymer sorbents selective to certain metal ions. The method is based on the use of memory of polymer composition and consists in conformational prtar rangement of macromolecules of non-crosslinked polymer complexons to a favorable for sorbing metal position, fol lowed by fixation of conformations optimum for the metal uptake by means of intermolecular crosslinking. Prearranged macromolecules may "keep in mind" their background i.e. the conformations advantageous for the uptake of the ions of the particular metal and this results in an essential improvement of the basic sorption characteristics of the sorbents - exchange capacity, selec tivity and rate of the uptake. Investigation of sorption properties of the sorbents prearranged for the uptake of ions of different metals has been carried out. It has been shown that the sorbents exhibit higher selectivity with respect to metal for the ions of which they had been prearranged. Using the method of conformational prearrangement a number of sorbents containing phosphorylic, carboxylic, amine and imine functional complexing groups with a higher selectivity to copper, cobalt and nickel ions have been obtained.

214

i. 321 P.1-31 GENERATION OF ANION-EXCHANGE SITES ON OCTADECYLATED SILICA GEL ГЕНЕРАЦИЯ АНИОНО-ОБМЕННЫХ ГРУПП НА ОКТАДЕЦИЛСИЛИКАГЕЛЕ R.veepalec, H.Brzobohata, M.Vrchlabsky, Brno, CSSR During the study of pH effect on the separation of in organic anions /1/, in acidic mobile phases with pH « 4 retention of, e.g., NOj , N0| , Br" and I", was observed on octadecylated silica gel even then when the mobile pha se did not contain tetrabutyiammonium counter ion. Ion exchange on protonized residual silanols was considered as possible mechanism of this retention. The accuracy of this model was tested on unmodified silica gel and LiChrosorb RP 18. The sites, which give rise to retention of inorganic anions in mobile phases with pH < 4 even in the absence of tefcrabutylammonium counter ion in the chromatographic system, were generated by the preparation of LiChrosorb RP 18 with tetrabutyiam monium solution in dilute phosphoric acid (pH = 2.7), The preparation with the acid alone or with the solution of tetrabutylammonium phosphate in neutral phosphate buffer did not affect LiChrosorb HP 18 in this way. On unmodified silica gel, sorption sites were not generated even by the action of tetrabutyiammonium phosphate solu tion in phosphoric acid. The anion exchange sites thus originate due to the interaction of phosphoric acid and tetrabutylammoniura ion with octadecyl groups, 1. R.Vespalec, J.Neca, M.Vrchlabsky, J.Chromatogr., 286 (1984) 171. 322 P.3-03 OCTADECYL SILICA GEL SORBENTS FOR HPLC WITH HIGH DENSITY OF COVERAGE ОКТАДЕЦИЛСИЛИКАГЕЯЬ ВЫСОКОЙ ПЛОТНОСТИ ПОКРЫТИЯ В КАЧЕСТВЕ СОРБЕНТОВ В ВЭЖХ B.Buszewski, J.Garaj, I.Novak, D.Berek, A.JuraSek, Bratislava, CSSR L.Nondek, Prague, CSSR The conditions for bonding octadecyl silylgroups to silica gel were systematically investigated with the aim to prepare HPLC sorbent with high degree of coverage. Three types of octadecyl dimethyl si lanes were applied, viz. monochloro, monoethoxy and S,N-dimethylamino deriva tives CH, CH, CH, I 3 I 3 I 3 CH,

R - Si - CI, R - Si - OCH,, R - Si - H' (R = C10H,,) I I 3 1 NCH, 18 37 J сн3 сн3 сн3 The above sllanes were bonded to the new type of strictly spherical silica gel SG-7 /1,2/ with mean pore diameter 5*12 nm, particle diameter D„ « 7 + 2 um, pore volume 215 2 ml.g and specific surface area about 420 m .g . The conditions of sililation were optimized as to tempe rature, duration of reaction,reaction medium and concentra tion of silylation agent.High degree coverage (N 4 /lmol- m~') sorbents were prepared with very good repeatibility and the subsequent testing showed their sufficient mechani cal and chepical stability as well as surface homoaeneity. The chromatographic measurements revealed that the above materials can be utilized in preparation of high perfor mance liquid chromatographic columns with excellent sample capacity and interesting retention characteristics. 1. D.Berek, I.Novak - US Pat. 4255286. 2, I.Novak, D.Berek - OS Pat. 4 382070. 323 P.3-04 SEPARCOL SI С 18- DISPOSABLE CARTRIDGE FOR ISOLATION OF DRUGS AND THEIR METABOLITES FROM BIOLOGICAL FLUIDS СМЕННЫЙ ПАТРОН - СЕПАРК0Л SI С 18- ДЛЯ ВЫДЕЛЕНИЯ ЛЕКАРСТВ И ИХ МЕТАБОЛИТОВ ИЗ БИОЛОГИЧЕСКИХ ЖИДКОСТЕЙ L.Soltys, I.Novik, D.Berek, Bratislava, c'sSR Numerous authors have demonstrated the advantages of the treatment of biological samples, prior their chromato graphic studies, by microparticulate silica hydrophobised by chemically bonded С 18 alkyl groups. Such isolation technique was reported also for simple, rapid, convenient, efficient and selective preseparation of the drugs and/or their metabolites from the body fluids /1-6/. The above sorbent is usually packed in the disposable cartridges of different constructions that are marketed by several com panies (Waters Assoc., Baker Chem.Co., Chrompack BV, etc.) In the present work, we have tested new type of the disposable cartridges packed by С 18 silica, that are available from Centre of Chemical Research, Slovak Academy of Sciences, CS-84238 Bratislava, under the trade name SEPARCOL SI С 18. Separcol cartridges are very universal and versatile in use for they allow alternative use of either gravity, or vacuum, or buill- in piston or even a syringe to transport both the sample and eluent. The pharmacokinetics and metabolism of ethimizol, a xanthine-related respiratory analeptic, broadly spreaded in clinical practice of the USSR were studied by HPLC and HPTLC /1-3/. Analysis of biological samples revealed that besides quantitative recovery of ethimizol itself, the isolation of its metabolites by means of SEPARCOL SI С 18 was very efficient, as well. The adsorption-desorption process, is not only highly efficient, but it is usually also more selective than any other isolation technique. It was demonstrated that, in this way, the local anaesthetics of Czechoslovak provinien- ce, penta-, hepta-, and carbisocaine, can be isolated high ly selectively from various biological matrices /4-6/. The substances trapped by the SEPARCOL SI С 18 solid support, including local anaesthetics and simultaneously numerous compounds, were further purified by flushing off the great 216 majority of endogens with acetonitrile which does not elute the drugs studied. Finally, the drugs were eluted by methanol and sebsequently quantitatively analysed by GLC, In this way the GLC analysis of local anaesthetics isolated from biological fluids are much more simple. 1. L.Soltes et al.: Pharmacology, 26 (1983) 198. 2. L.Soltes et al.! J. Chromatogr., 273 (1983) 213. 3. L.Soltes et al.: Xenobiotica 13 (1983) 683. 4. L.Soltes et al.: Meth. Find. Exptl.Clln. Pharmacol., 5 (1983) 461. 5. L.Soltes et al.: J. Chromatogr., 286 (1984) 223. 6. V.Marko et all.: J.Chromatogr. (in press). 324 P.3-05 SYNTHESIS AND INVESTIGATION OF POROUS SILICA MODIFIED WITH PYROCARBOd СИНТЕЗ И ИССЛЕДОВАНИЕ ПОРИСТЫХ КРЕМНЕЗЕМОВ, МОДИФИЦИРОВАННЫХ ПИРОУГЛЕРОДОМ N.K.Bebris, Yu.S.Nikitin, A.A.Pyatygin, N.K.Shonia, Moscow, USSR Liquid, chromatography that makes use of porous silica with a surface modified with a thin pyrocarbon coating is described by Kiselev et al., and Guiochon et al. Leboda made several attempts to use these adsorbents in gas chro matography. However, these prowisinq adsorbents and car riers have not yet to find wide chromatographic application; knowledge of their adsorption and chromatographic proper ties is scant. We have investigated how the conditions of porous silica modification with pyrocarbon, obtained by pyrolytic benzene decomposition, influence the amount and the character of depositing of pyrolysis products, and the resultant change in adsorption and chromatographic properties. Increase of temperature (from 750 to 900°C) and duration of pyrolysis, as well as of benzene concent ration in carrier helium is accompained by the increase in the amount of carbon on silica surface. Observed changes in adsorbent pore structure (decrease of specific surface and volume of pores) is explained not only by pyrocarbon coating of pore surface, but also, in a number of instan ces, by caking of the silica itself. Discussed are the optimal conditions for carbosilica gel and carbosilochrome synthesis. Gas chromatography and adsorption studies have shown that pyrocarbon modification leads to considerable (as compared to silica) increase in the contribution of non specific interaction, and a decrease in the contribution of specific interaction» to retention. Silica modified with pyrocarbon approach acetylene and thermal carbon blacks in retention characteristics, but are less homoge neous geometrically and chemically. We also studied the influence of polyaromatic hydrocar bons (PAHs) on carbosilica gel in the normal and reversed- phase types of liquid chromatography. Described is the

217 great affinity of carbosilica gel to PAHs and its high sensitivity to geometric (steric) structure of separated substances. 325 P.3-06 RETENTION OF SOME ORGANIC COMPOUNDS ON CARBON ADSORBENTS УДЕРЖИВАНИЕ НЕКОТОРЫХ ОРГАНИЧЕСКИХ СОЕДИНЕНИЙ НА УГЛЕРОДКК АДСОРБЕНТАХ N.A.Eltekova, Moscow, USSR НРЬС and adsorption of aromatic oxygen-containing com pounds, such as anisole, benzaldehyde, phenol, aliphatic alcohols and ketones from dilute solutions on carbon adsor bents were studied at 293 K. The dependences of retention volumes of organic compounds on the chemical structure of their molecules, the nature of adsorbents and the polarity of solvents were obtained. The behaviour of organic com pounds molecules in the adsorbent pores considering their interactions with solvent molecules was discussed. The correlation dependences between the Hildebrand's parameter and the change of adsorption molar free energy of organic compounds molecules on carbon adsorbents were found. The comparison of chromatography activity and selec tivity of different chemical structure adsorbents for orga nic compounds of different types was made. The "quasi-equilibrium" conditions of chromatographical process were found on the basis of comparison of values of retention volumes with adsorption equilibrium constants, calculated from static adsorption isotherms of organic compounds at low concentrations and at low surface cove rage. 326 P.3-07 OBTAINING OF NON-POLAR ADSORBENTS BY SURFACE MODIFICATION OF THERMAL CARBON BLACK WITH n-HYDROCARBONS IN HIGH FREQUENCY PLASMA "ПОЛУЧЕНИЕ НЕПОЛЯРНЫХ СОРБЕНТОВ ПОВЕРХНОСТНОЙ МОДШКАЦИЕЙ ТЕРМИЧЕСКОЙ САЖИ УГЛЕВОДОРОДАМИ В ВЫСОКОЧАСТОТНОЙ ПЛАЗМЕ T.B.Gavrilova, I.V.Parshina, Moscow, USSR N.Petsev, St.Ivanov, I.Topalova, Ch.Dimitrov, Sofia, Bulgaria It is well known that non-polar adsorbents are highly sensitive to the geometrical structure of adsorbetes. Graphitized carbon black, formed by pure carbon, is one of the adsorbents used in this field. As its surface con tains no functional groups, molecules with different elec tronic structure are adsorbed mainly due to the disper sion forces. Careful investigation of the graphitized ther mal carbon black (GTCB) shows however that its surface includes some chemical and geometrical nonuiniformity. Furthermore it is highly mechanically fragile.

218 In order to investigate non-polar adsorbents on the basis of GTCB and in order to improve geometrical and chemical uniformity of such surfaces a modification with n-hydrocarbons was carried out. Monolayer capacity was calculated from Van-der-Vaals si ze of the molecules it was found also by means of adsorption from solution and by gas chromatography. It was shown that all the results were in good correla tion. The modification of carbon black samples, being undergone different preliminary treatment, with n-hydrocarbons was carried out in low temperature high-frequency plasma. Plasma induced modification appeared to influence the adsorption and chromatographic properties of the ad sorbent. The possibility to use these adsorbents for GC analysis is also demonstrated. 327 P.3-08 APPLICATION OF CLINOPTILOLITE-CONTAINING TOFFS OF BULGARIA IN GAS CHROMATOGRAPHY ВОЗМОЖНОСТЬ ПРИМЕНЕНИЯ В ГАЗОВОЙ ХРОМАТОГРАФИИ НЕКОТОРЫХ КЛИНОПТИЛОЛИТСОДЕРЖАЩИХ БОЛГАРСКИХ ТУФОВ T.G.Andronikashvili, Tbilisi, USSR D.K.Benev, O.A.Bogdanova, Burgas, Bulgaria Cation-decationated samples from cllnoptilolite-containing tuffs, originating from "Bell Plast" and "Zelezni vrata - Trass", Bulgaria, are obtained by methods of ion- exchange and acid-treatment. All the obtained samples are investigated by X-ray and thermal methods of analysis. It is shown, that • ion-exchan ge, acid treatment under certain conditions and thermal activation of samples to 673-723 K, does not influence substantially the crystallity and the samples structure. Chromatographic properties of initial clinoptilolite- containing tuffs from regions said, as well as modified ; forms using model mixtures: H2,02,CH<,CO,C02; He,H2,02,N2, . СВ., СО, CO- and benzene, toluene, xylene are investigated using chromatographs "Zvet - Ю2' and LHM - 8MD (USSR). It is shown that the conditions of decationing unfluen- ce substantially on effictivity crlterions of the obtained samples, used as adsorbents for the gas chromatography. 328 Р.З-ОЭ PLASMA MODIFICATION OF ADSORBENTS OF DIFFERENT CHARACTER ВУ MOLECULES WITH ELECTRON DONOR PROPERTIES МОДИФИКАЦИЯ ПЛАЗМОЙ РАЗЛИЧНЫХ АДСОРБЕНТОВ МОЛЕКУЛАМИ • С ЭЛЕКТРОНОДОНОРНЫМИ СВОЙСТВАМИ N.Petsev, St.Ivanov, I.Topalova, Chr.Dimitrov, Sofia, Bulgaria T.B.Gavrilova, E.V.Vlasenko, T.M.Roshchina, Moscow, USSR The surface of non-polar (thermal carbon black TG-10) and of polar (silochrom - Sio-) adsorbents has been modified 219 by adsorption of molecules, which are electron donors. The modification was carried out by different ways: (1) the deposition of a monolayer from solution and (2) the deposition of monolayer followed by a treatment in high frequency low temperature plasma. It was shown that the way of modification has consider able influence on the chromatographic properties of the adsorbent-s upport. The surface properties of the modified adsobents were compared with corresponding properties of the original materials. For this purpose the GC retention data were determined for molecules taking part only in dispersion interactions as well as for molecules which interact electrostatically, by hydrogen bonding or by complex with charge transfer. It is shown too, that by depositing a thin layer of pyrene on the adsorbents-supports surface it is possible to obtain adsorbents with new properties. By the authors' opinion on the TG-10 surface there are adsorption layers of flatly orientated polyaromatic mole cules. All adsorbents-supports after modification with pyrene and then with plasma show a decrease of adsorption poten tial. The adsorbents obtained by the way described were used for GC analysis of some complex mixtures. 329 P.3-10 SYNTHESIS, CHARACTERIZATION AND APPLICATION OF CEKACHROM, UNPOLAR AND POLAR POROUS COPOLYMERS, USED AS STATIONARY PHASES IN CAS CHROMATOGRAPHY СИНТЕЗ, ХАРАКТЕРИСТИКА И ПРИМЕНЕНИЕ СЕКАХРОКА, НЕПОЛЯРНЫХ И ПОЛЯРНЫХ СОПОЛИМЕРОВ, ПРИМЕНЯЕМЫХ В КАЧЕСТВЕ СТАЦИОНАРНЫХ ФАЗ В ГАЗОВОЙ ХРОМАТОГРАФИИ H.Hoffmann, G.Scbwachula, M.Henke, K.Haeupke, Bitterfeld, GDR Fine grain CEKACHROM-types with high porous structure, developed in VEB Chemiekombinat Bitterfeld were proved as very efficient stationary phases in gas chromatography. CEKACHROM 1, 2 and 3 are unpolar styrene-ethylstyrene- divinylbenzene-, respectively ethylstyrene-divinylbenzene- copolymers' with different porous structures, adapted to their purposes (specific surface between 100 and 550 m /g). CEKACHROM 4, 5 and 6 are polar copolymers, containing out or ethylstyrene and divinylbenzene, acrylic acid ester, respectively acrylonitrile with graduated polarity and different porosity (specific surface between 20 and 350 mz/g). The porous structures were impressively characterized by porosimetry, transmission- and scanning electron micro scopy. Thermal investigations by derivatography confirmed the stability in current operation under inert atmosphere of the polar copolymers up to 2O0°C and unpolar ones up to 250°C.

220 Using a modern classification-method we got from all CEKACHROM-products two grain size-ranges (0.16 to 0,225 and 0.225 to 0,315 mm). These polymer fractions allow exact and quantitative separations of mixtures containing polar components (Hater, alcohols, aldehydes, ketones, esters, carboxylic acids, aromatic compounds with traces of water, water with traces of chlorinated hydrocarbons, amines, nitriles, nitroparaffines etc.).

330 p.3-11 MACROPOROUS CATION EXCHANGE RESINS IN GAS CHROMATOGRAPHIC ANALYSIS МАКРОПОРИСТЫЕ КАТИОНИГЫ В ГАЗОХРОМАТОГРАФИЧЕСКОМ АНАЛИЗЕ L.I.Panina, G.P.Terekhova, S.B.Makarova, O.N.Vlasovskaya, Moscow, USSR The peculiarities of retention of compounds of diffe rent classes on macroporous sulpho cation exchange resins on the basis of styrene-divinylbenzene copolymers in dif ferent ion forms as well as the possibilities of their use for gaschromatographic separation and analyses of organic compounds are investigated. The gas chromatographic properties of these resins shoug- ly depend upon the nature of ionogenic group, which influ ences retention volumes and Kovats retention indices of compounds as well as the order of elution of components from gas chromatographic column is established. High selectivity of macroporous sulpho resins to water and polar compounds is found out. The possibility of using these sorbents to separate paraffin, aromatic and olefinic hydrocarbons, for the purposes of group analysis of nonpolar, midpolar and highpolar compounds, for separation of compounds with close phisical and chemical properties, for trapping of trace amounts of compounds, for defining the existance of admixtures in water and solvents is shown. Hearly manifested selective properties of macroporous sulpho resins give the opportunity to use the columns of small length- 20-60 cm for separation. With the help of the example of separation of nitrous oxides, paraffin, olefinic and aromatic hydrocarbons it is shown, that the order of elution of substances on such sorbents often differs from the order of elution on cer tain polymer sorbents and stationary liquid phases. This facilitates the identification of admixtures of organic compounds and their determination in water and organic mediums. The use of macroporous cation exchange resins in gas chromatography broadens the chances of gas chromatographic method in analysis of chemical compounds.

221 331 P.3-12 POLYCOMPLEXONATES OF METALS ON THE BASIS OF VINYL DERIVATIVES OF PYRIDINE - SELECTIVE SORBENTS FOP. GAS CHROMATOGRAPHY П0ЛИК0МШ1ЕКС0НАТЫ МЕТАЛЛОВ НА ОСНОВЕ ВИНИЛЬНЫХ ПРОИЗВОДНЫХ ПИРИДИНА - СЕЛЕКТИВНЫЕ СОРБЕНТЫ ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ L.I.Panina, Z.A.Evseeva, V.B.Kargman, N.B.Galitskaya, Moscow, USSR

The gae chromatographic properties of polycomplexona tes of copper, silver and mercury on the basis of copoly mers of methylvinylpyridine and divinylbenzene, methylvinylpyridine, divinylbenzene and glycldlc methyl aerylate of porous structure are studied. The dependence of speci fic retention volumes upon the number of carbon atoms in the nolecule for homologous series of compounds of diffe rent classes on these sorbents, separation of organic substances having similar physico-chemical characteris tics are determined.

The differency in gas chromatographic behaviour of original samples - sorbents based on vinyl derivatives of pyridine- and polycomplexonates of metals on this basis is shown. The specificity of these sorbents is estimated. The high selective properties of studied polycomplexona tes to sulphur-, halogen-, nitrogen-containing compounds, to compounds with nonsaturated connections are found out. The general gas chromatographic feature, characteristic of all mentioned metal polycomplexonates, such as high specificity of molecular interaction and selective reten tion of polar compounds as well as individual features, characteristic of their gas chromatographic behaviour, determined with the nature of the metal, coordinatingly connected with polymer matrix are pointed out. It is shown, that the increase of concentration of metal in polycomplexonates studied leads to the intensifi cation sorbents specific properties.

It is suggested to use the developed sorbents for the definition of admixtures of compounds in organic solvents, air and sewage, for the trapping of trace amounts of com ponents from gases. For example, with the help of poly- complexonate of copper the method of separation SO, and H,S, which are toxic air pollutant , is found. The pecu liarity of this method is the selective retention of H-S (after n-nonane) and rapid elution of SO,, which allows to use this way for investigation of some catalytical processes, tests of exausts and for the trapDlng of H,S traces from air. " z

222 332 P.3-14 NEW SELECTIVE PYRAZOLIC S0R3ENTS FOR THE NOBLE METALS CHROMATOGRAPHY НОШЕ СЕЛЕКТИВНЫЕ ПИРАЗОЛ СОДЕРЖАЩЕ СОРБЕНТЫ ДЛЯ ХРОМАТОГРАФИИ БЛАГОРОДНЫХ МЕТАЛЛОВ S.B.Makarova, A.V.Smirnov, A.L.Bratslavskaya, M.L.Petrova, Moscow, USSR The new type of polymeric high selective pyrazolic sorbents for sorption of the noble metals from the diffe rent solutions have been created. The sorbents are obtained by two ways: a) the copolymerization of N-vinyl-3(5)-methylpyrazol with one of the divinyl monomers; b) the treatment of halogenmethylated copolymers of styrene and divinylbenzene having either macroporous or gel structure by 3(5)-methylpyrazol in the presence of proto- philes. The both types of sorbents are characterized by the high sorption capacities for platium family metals, gold and silver. The sorbents have high selectivity for these metals and high chemical and thermal stability. The optimum pH range of the sorption solutions for the platinum family metals have been found out. The desorption conditions for concentrating some of the metals from 20 to 50 times have been established. It has been shown that the introduction of additional amino-groups of the different kinds considerably increa ses kinetic characteristics of the sorbents. 333 p.3-13 ION-EXCHANGED HEMOSORBENTS FOR PURIFICATION BIOLOGICAL FLUIDS ИОНООБМЕННЫЕ ГЕМОСОРБЕНТЫ ДЛЯ ОЧИСТКИ БИОЛОГИЧЕСКИХ ЖИДКОСТЕЙ ОРГАНИЗМА Yu.A.Leikin, Moscow, USSR Processes of the sorbtional extraction endo- and exo- toxic components (TC) from biological fluids (BF) are widely used in practical medicine for the body detoxifying and homeostasis correction at the disfunction of some organs of a human body. The existence in the solution of the protein molecules, which effectively from the stable complexes with TC, comp licates the problem of effective and selective extraction TC from blood, plasma, liquivora by the methods of ion- exchanged chromatography. So its necessary to get the sor bents with selective and superselective ligand groups and to form their chemical and porous structure; may be use ful as well the universal non-selective sorbents on the basis of activated carbon. Due to high reactivity, the possibility of introduc tion of different activing groups, purposeful formation of the phisical porous structure, the polymer sorbents make it possible to settle successfully the task by using 223 the achievements in the field of ion-exchancjed and affini ty chromatography, The иве of the different ion-exchanged resins and affin •orbents for the TC extraction from blood, plasma, liqui- ora has been investigated. Frontal chromatography has been used to work out the methods of purifying BF on the fol lowing ionic ТС: К , NH., Mg+Z, CaTZ, amins, phenols, lactats, fatty acids, BL, proteases. The factors of physical structure such as grade of cros sing, porosity, the type of a crossagent, chemical struc ture of the ionogenic groups, influence the equilibrium of the process as well its kinetic parameters. Comparable investigations of the alteration in the affinity of some heavy metals for ion-exchanged and complex-forming catio- nits in the gradient of the concentration of albumin and other complexons binding metals, have been carried out. Sorbents, splitting protein-metal complex for effective extraction Hg(II), Pb(II), Cu(II) from the complex-forming solutions and biological fluids have been found.

334 P.3-15 TEPARON AND TEPASORB - NEW SELECTIVE SORBENTS FOR GAS CHROMATOGRAPHY ТЕПАРОН И ТЕПАСОРБ - НОВЫЕ СЕЛЕКТИВНЫЕ СОРБЕНТЫ ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ E.J.Heinsoo, Tallinn, USSR Most polymeric sorbents used In gas chromatography have a hydrophobic surface. Therefore they separate well com pounds of low and medium polarity. Nevertheless, they axe the basic CC materials which are used for analysis of polar inorganic gases (CO, C02, ammonia, water etc.). But the retention times for these gases on Porapac Q, for example, are too short and efficiencies hot very high, therefore too long columns and too low temperatures are needed. There are several polymeric sorbents which are more polar, but their surface is significantly smaller than that of Porapac Q and that is why they cannot be used in the analysis of gases. The new polymeric sorbents of TEPARON and TEPASORB series contain aliphatic amino groups and thus they are polar, hydrophilic and with.alkaline properties. Their surface area is up to 500 m /g. When strongly polar com pounds (water, ammonia etc.) are analyzed, the efficiency of the above sorbents is significantly higher than that of all other commercially available GC sorbents. Teparon and Tepaeorb can be recommended for the analysis of most not-too-highly-boiling polar substances (the upper tempe rature limit of the sorbents is 160-180°C). The peaks of alcohols and amines are almost ideally symmetrical. Due to the large surface and high efficiency of Teparon, very short columns can be used. So, a mixture of C0-C0--NH,- -H-0 can be analyzed on a 2-f£,^Teparon-22 column in less than one minute. * * Because of alkalinity of the surface of the sorbents, 224 amines and other alkaline substances have very short reten tion times, while those of amphoteric (e.g. water) and acidic (e.g. CO,) compounds heavily depend on column temperature. The retention index of CO, on Teparon-22 column i« about 120 at 150°C, but at 50°C it is over 800 already. In practice it enables us to shift the peaks of acids on free places on the chromatogram by varying the column temperature by few degrees only. Several types of Teparon and Tepasorb sorbents are being,produced. They differ in surface area (from 100 to 500 m /g) and polarity. Less polar sorbents hay° efficiency as like as Porapak Q while analysing of nonpolar compounds.

335 P.3-16 CHROMATOGRAPHIC RESOLUTIONS ON MIXED STATIONARY PHASES- THE CHOICE OF THE OPTIMAL CONSTITUTION ХРОМАТОГРАФИЧЕСКОЕ РАЗРЕШЕНИЕ НА СМЕШАННЫХ СТАЦИОНАРНЫХ ФАЗАХ - ВЫБОР ОПТИМАЛЬНОГО СОСТАВА W.Czelakowski, P.Kusz, A.Andrysiak, Kedzierzyn-Kozle, Poland We shall report a method tending towards achieving the optimal constitution of mixed stationary phases in GLC when the retention times are attainable only. The method has been empirically tested for chemical resolutions of chlorinated C, - C, hydrocarbons.

336 P.3-17 GAS CHROMATOGRAPHY ON MONOLAYERS OF PORPHIRINE AND CROWN-ETHERS DERIVATIVES ГАЗОВАЯ ХРОМАТОГРАФИЯ НА МОНОСЛОЯХ ПРОИЗВОДНЫХ ПОРФИРИНА И КРАУН-ЭФИРОВ E.V.Zagorevskaya, N.V.Kovaleva, Moscow, USSR The adsorbents for gas chromatography have been prepa red by adsorption modification of graphitized thermal carbon black (GTCB) and of carbochrom with monolayers of derivatives of porphirine and crown-ethers. The adsorption modification of GTCB - nonporous adsorbent with homoge neous surface - is being intensively studied at present because of the importance in analytical gas chromatography. The carbochroms used in this work (pyrolitically deposed carbon on the surface of GTCB) are much more stable mecha nically than carbon blacks. The adsorption properties of the prepared adsorbents were characterized by gas chromatography of different clas ses of organic substances. Retention volumes per m of adsorbent surface at zero, sample size and molar differen tial heats of adsorption were determined for all adsorbates. For substances whose molecules can interact with

1S.3IK.872 225

I. functional groups of the molecules of the modifying sub stance the contributions of the energy of specific interac tion q _ to the total heat of adsorption were determined. It has been shown that the selectivity of adsorbents prepared by modification of carbochroms with monolayers of the derivatives of porphirine towards different classes of organic compounds depends on the nature of the central cation atom in the molecule of metalporphirine; this per mits preparing adsorbents which are selective to structu ral isomers, oxygen- and halogensubstituted compounds. The modification of the surface of carbon blacks and carbochroms with polycyclic polyether dibenzo-18-crown-6 results in the considerable decrease of the retention volumes and heats of adsorption of all compounds and in the increase of selectivity towards isomeric chlororganic compounds, for instance, chlorobenzenes, hexachlorocyclo- hexene isomers. The advantages of the prepared materials in analytical applications include shorter analysis times, improvement in peak symmetry and separation and high temperature stability.

337 P.3-18 THE HIGH TEMPERATURE STATIONARY PHASE ON THE BASIS OF SILOXANE POLYMERS OF STAIR'S STRUCTURE ВЫСОКОТЕМПЕРАТУРНЫЕ НЕПОДВИЖНЫЕ ФАЗЫ НА ОСНОВЕ ЛЕСТНИЧНЫХ СИЛОКСАНОВЫХ ПОЛИМЕРОВ I.P.Yudina, G.N.Semina, Moscow, USSR During last period we have researched a number of diffe rent hightemperature selective stationary phases for gas chromatography on the basis of various siloxanes polymers. One those stationary phase is a polymer on basis of cyclic siloxane tetramer with phenyl group at silicium, connecting with linear siloxane with methylphenyl and dimethyl-groups, so called "stair's polymer" (trade name "Lestosil"). The main physico-chemical properties of this stationary phase have been investigated. It was shown that phase is selective at high temperature. The maximum working tempe rature is 500 С according data of TGA-analysis and FID. It has been carried out the comparison of the proper- -ies of "Lestosil" and well known siloxane phases as OV with' phenyl groups with determination of constant of McReynolds. "Lestosil" is specific for aromatic compounds, alcohols, amines and free fatty acids. "Lestosil" was used successfully for separation of high temperature polar compounds and isomers of them, amines and fatty acids.

226 338 P.3-19 THERMODYNAMIC ASPECT OF LIQUID CRYSTALS APPLICATION IN CHROMATOGRAPHY ТЕРМОДИНАМИЧЕСКИЙ АСПЕКТ ИСПОЛЬЗОВАНИЯ ЖИДКИХ КРИСТАЛЛОВ В ХРОМАТОГРАФИИ V.A.Smirnov, V.I.Klopov, N.B.Kuznetsova, Ivanovo, USSR K.V.Egorova, M.S.Esin, B.I.Pakhomova, V.G.Goldina, N.F.Belyaev, Kuibyshev, USSR It was shown, that essential differences in thermodyna mic function of dissolution of nonmesomorphic substances in liquid-crystal solvents can be applied in practice for assessment of efficiency of chromatographic separation of different classes of organic compounds. To choose stational phases we take as a principle basic ideas of liquid-crystal state and characteristic feature of intermolecular interactions in binary systems of mesogene-nonmesogene. Influence of the nature of cent ral and terminal groups of nematic and cholesteric liquid crystals molecules on their orientational order degree and thermodynamic parameters of dissolution of n-paraffins, aliphatic and alicyclic alcohols, aromatic acids and their esters, as well as aldehydes, ketones, terpenes and ste roids were established. On the base of lows of thermody namic function change of organic compounds sorption diffe rent mechanism of intermolecular interactions with stational liquid-crystal phases was established. It was shown that essential contribution to the entalpy of dis solution polar nonmesogene is given by process complex of formation. Comparative assessment of the obtained results gives the possibility of theoretical selection of liquid-crys tals stational phases for gas chromatography. 339 P.3-20 THE PROPERTIES OF 4-OCTYL-4•-CYANOAZOBIPHENYL, 4-OCTYL-4'-CYANOAZOXYBIPHENYL AND 4"-CYANOBIPHENYL 4-ALKYLBIPHENYLCARBOXYLATES-4 AS STATIONARY PHASES IN GAS CHROMATOGRAPHY СВОЙСТВА i»-OKTHJI-V -ЦИАНА30БИФЕНИЛ-, 4-ОКТИЛ-4' -ЦИАН- А30КСИБИФЕНИЛ-, И 4"-ЦИАН0БИФЕНИЛ-4-АЛКИЛБИФЕНИЛ- КАРБОКСИЛАТОВ-4 В КАЧЕСТВЕ НЕПОДВИЖНЫХ ФАЗ В ГАЗОВОЙ ХРОМАТОГРАФИИ Z.Witkiewicz ,Warsaw, Poland 4-Octyl-4'-cyanoazobiphenyl (polymesomorphism: S_, S„,

S, and N), 4-octyl-4'-cyanoazoxybiphenyl (SE, S_, S.) and

4*-cyanobiphenyl 4'-alkylbiphenylcarboxylates-4 witn Cr

to C7 alkyls (SA1, N) and C. and Cg alkyls (SA1, N_, SA2' N' have been tested. These compounds were deposited on Chromosorb W HMDS in an amount of 10% and placed in glass columns 2.1 m long and 4 mm in diameter. Diethylben- zene, fluorotoluene and dimethylnaphthalene isomers were used as test substances depending on the mesophase teiripe-

227 rature range of the particular liquid crystals. The reten tion tines were measured for these isomers in vapour form. The results are presented in the form of t_ « f(T) and r, _ " '№ characteristics. From these plots we can con clude that: - The phase transition temperatures determined chromatographically are in agreement with those determined thermo- optically and calorimetrically. - The chromatographic method provides additional informa tion as to the character of phase transitions in the solid and in the raesophase. - The test substances reacted in different ways with the particular kinds of mesophase of the same liquid crystal. A change of the relative retention was observed which resulted even in the change of the order of elution of the isomers. These changes of the order of elution were obser ved in the vicinity of the phase transition points. It was found that for esters the selectivity was highest in the SA1 Pnase or in tne vicinity of the H_ S,, transition. The retention properties measured in the column heating and cooling cycles differed for certain kinds of mesophase. The selectivity was usually higher at a given temperature In the cooling cycle as compared with the heating cycle. The tests performed throw new light on the possibili ties of applying gas chromatography for testing liquid crystals with many phase transition points and provide information about the properties of these liquid crystals as stationary phases.

340 P.3-21 GAS CHROMATOGRAPHY OF KETHYLNAPHTHALENES ON LIQUID CRYSTALLINE STATIONARY PHASE ГАЗОВАЯ ХРОМАТОГРАФИЯ КЕТИЛНАФТАЛИНОВ НА ЖИДКОКРИСТАЛЛИЧЕСКОЙ СТАЦИОНАРНОЙ ФАЗЕ Z.Suprynowicz, A.Patrykiejew, Lublin, Polai.d Liquid crystals are known to be very good stationary phases for separation of isomeric compounds. In particular, the separation of positional isomers can be achieved much easier than with conventional Isotropic liquid phases. This work reports the results of our study on the sepa ration of isomeric dimethylnaphthalenes on capillary column coated with liquid crystalline stationary phase. Besides, we have also studied the retention mechanism and solute - stationary phase interactions. It has been found that separation of diraethylnaphtha- lenes depends strongly on the temperature of analysis. The order of elution of isomers has not been the same as pre dicted by the shape parameter consideration. A simple model of retention of dimethylnaphthalenes by liquid crys talline stationary phase has been proposed. This model predicts a correct order of elution for the studied iso mers. Besides, it has been found that solute- stationary phase interaction is highly specific for a given positions of methyl groups attached to aromatic skeleton. 228

I. 341 P.3-22 THE APPLICATION OF LIQUID CRYSTALS AS SORBENTS IN LIQUID CHROMATOGRAPHY ПРИМЕНЕНИЕ ЖИДКИХ КРИСТАЛЛОВ В КАЧЕСТВЕ СОРБЕНТОВ В ЖИДКОСТНОЙ ХРОМАТОГРАФИИ A.A.Aratskova, Ya.I.Yashin, Moscow, USSR S.P.Vetrova, Gorky, USSR The possibilities for application of liquid crystals as sorbents in liquid chromatography to separate the isomers of aromatic compounds have been studied. Two types of liquid crystals were applied on the Silasorb-600: 4-ethoxybenziliden-4'-butylaniline (EBBA) from the solution in chloroform and cholesterol oleate from alcoholic solution. The EB'-tA has the nematic mesophase in the range 36.5-77.5°C, but cholesterol oleate - in the range 20-33°C. The retention times, as well as the capacity and selectivity coefficients for nitrotoluene isomers, bromnitrobenzenes and dinitrobenzenes have been measured over these temperature ranges. It has been established, that with increasing tempera ture within the phase transition, the retention of aroma tic compounds isomers on the EBBA - modified silica gel substantially differs from that on pure silica gel: this retention remains constant for dinitrobenzene and even increases for bromonitrobenzene and nitrotoluenes. On the cholesterol oleate - modified silica gel, the separation selectivity of nitrotoluene meta/para -isomers within the phase transition is decreased that as a whole results in the better separation of these isomers at the temperature 55°C, than at 26°C. The data obtained showed the possibility to use liquid crystals for separation of the complex isomers with simi lar boiling temperatures in liquid chromatography. 342 P.3-23 MODIFIED ZEOLITES FOR CHROMATOGRAPHIC ANALYSIS OF ATMOSPHERIC POLLUTION МОДИФИЦИРОВАННЫЕ ЦЕОЛИТЫ ДЛЯ ХРОМАТОГРАФИЧЕСКОГО АНАЛИЗА АТМОСФЕРНЫХ ЗАГРЯЗНЕНИЙ O.S.Banakh, R.I.Baransky, I.Ya.Golos, V.I.Rogovik, L.M.Charnetskaya, Lvov, DSSR On the basis of synthetic zeolites of Y, X, L, A types and the Carpathian natural aordenites modified models, containing different amounts of exchanged cations of alkali and alkali-earth metals, as well as silver and copper, have been prepared by means of ion exchange and acidic treatment. Using helium and nitrogen as carrier gases in the one metre columns of "Tsvet" gas chromatographs (models 4-67 and 162) packed with above mentioned zeolites at temperatures of 20-300°C at intervals of 20°, separation conditions of the mixtures, consisting of low boiling and hydrocarbons gases, oxides of carbon, nitrogen

229 and sulphur, have been studied and their retention volumes and retention indices have been calculated. Depending on the nature of cations and ion exchange degree the obtained zeolites may serve either as effective adsorption filters for extraction from the air of a great number of pollutants and for their concentration at room temperature or as adsorbents for separation and chromato graphic analysis of the concentrated substances. Condi tions of quantitative determination of oxides of carbon and nitrogen have been defined with the use of У types zeolites, having the average degree of initial ions re placement by cations of silver. Especially high affinity to retention of CO molecules and C,-C, olefines at room temperature by natural morde- nite as well as zeolites of X and У types, containing 10- 20% of silver cations, have been determined, effective extraction and concentration of these gases present in the air on the level of maximum admissible concentrations having been thus provided. The quantitative desorption of the above mentioned pollutants and their transmission by the carrier gas for a chromatographic analysis are achie ved after heating of the column-concentrator up to 240-280°C.

343 P.3-24 EVALUATION OF DEGREE OF MODIFICATION OF DIATOMACEOUS SUPPORTS FOR GAS-LIQUID CHROMATOGRAPHY ОЦЕНКА СТЕПЕНИ МОДИФИКАЦИИ ДИАТОМИТОВЫХ НОСИТЕЛЕЙ ДЛЯ ГА30ЖИДК0СТН0И ХРОМАТОГРАФИИ G.I.Pekov, N.D.Petsev, Sofia, Bulgaria It is known that diatomaceous type supports for gas- liquid chromatography are weak adsorbents because of silanol groups on their surface. By chemical Modification a great part of surface silanol groups are removed and adsorption capacity of these supports decreases strongly. In present investigation is shown a possibility for relative evaluation of degree of modification of diatoma ceous type supports for GLC by «easuring of dye adsorption. Adsorption of methyl red in benzene solution on Ster- hamol is studied. Work is carried out to get right and reproducible results for degree of modification. The method of evaluation is checked up on commercial untreated anc! chemically modified diatomaceous supports as well as on laboratory modified saaples 'GLC packings trea ted at high temperature in an inert athiaosphere and then soxhlet extracted). The obtained results for adsorption are compared with previous data for retention of test solutes on supports in question. The changes in retention are in accordance with the changes in adsorption capacity

230 344 P.3-25 MODIFICATION OF SILICA SURFACE WITH MIXTURE VS. SUBSEQUENT MODIFICATION WITH HEXAMETHYLCYCLOTRISILAZANE AND HEXAMETHYLDISILAZANE СРАВНЕНИЕ МОДИФИКАЦИИ ПОВЕРХНОСТИ СИЛИКАГЕЛЯ ПОСЛЕДОВАТЕЛЬНО И СМЕСЬЮ ГЕКСАМЕТИЛЦИКЛОТРИСКЛАЗАНА И ГЕКСАМЕТИЛДИСИЛАЗАНА W.Szczepaniak, Poznan, Poland An application of mixtures caused a better deactivation of a silica surface than use of a single silylating rea gents. Now, it is interesting to see whether the same reagents applied subsequently can produce similar deactivation effects. The silica was modified with hexamethylcyclotrisilazane (HMCTZ) and hexamethyldisilazane (HMDS) by a method descri bed earlier. Since the 2:1 mixture of HMCTZ and HDDS gave the best results the same ratio of the reagents was applied in all experiments. The sample* of silica were modified as follows: A - with the 2:1 mixture of HMCTZ and HMDS; В - subsequently with HMCTii and with HMDS', С - subsequently with HMDS and with HMCTZ. Elemental analysis of the modified silica* were carried out and specific retention volumes of a number of hydro carbons were determined. An extent of the silica surface "deactivation" (i.e. a suppresion of the surface ability of specific interactions) was estimated by a comparison of relative retention volumes and specific heats of adsorption &(UH) of unsaturated hydrocarbons. The best deactivation effects were achieved when the mixture of the silylating reagents was used (sample A). The results shown here can possibly be utilized for the deactivation of chromatographic supports or capillary columns.

345 P.3-26 INFLUENCE OF A CARRIER GAS ON THE RETENTION VOLUME IN GAS-SOLID CHROMATOGRAPHY ВЛИЯНИЕ ГАЗА-НОСИТЕЛЯ НА УДЕРЖИВАЕМЫЙ ОБЬЕМ В ГА30-АДС0РБЦИ0НН0Й ХРОМАТОГРАФИИ D.P.Poshkus, Vilnius, USSR The virial equation describing the effect of adsorbate- carrier gas intermolecular interaction both in the gas phase and in the adsorben state on the retention volume V, in gas-solid chromatography is derived. The virial terms which account for the pair-wise interactions between adsorbate and carrier gas molecules in the gas phase and in the adsorbed state on graphitized thermal carbon black (GTCB) - an adsorbent with very homogeneous surface - have 231 been estimated theoretically for some adsorbates (argon, methane, benzene, cyclohexane) and carrier gases (helium, hydrogen, nitrogen, argon). On the basis of the obtained vlrial equation and calculated results, the influence of

various carrier gases on VA for various adsorbates, on GTCD has been considered. It has been shown that in the

temperature range in which the measurements of VA were carried out, the adsorbate - carrier gas interaction increases V, by some tenths of percent, when helium is used as a carrier gas, and this increase is mainly due to the adsorbate - carrier gas Interaction in the gas phase. However, V. is decreased by the adsorbate - carrier gas interaction, when other carrier gases (hydrogen, nitrogen, argon) are used, the effects of the adsorbate - carrier gas interaction In the gas phase and in the adsorbed state being of the same order of magnitude. In the last case the effect of pair-wise interactions between adsorbate and carrier gas molecules on V. may reach several percent when the pressure drop in a chromatographic column is small and the column outlet pressure p • 1 atm. This effect is directly proportional to pQ and is increased by the coeffi cient

4 3 I* - (3/4) [(PA/P0) - 1 / lot) when p0» 1 atm or Pj/Pj,» 1. 346 P.3-27 IONIC ADSORBENTS IN CAPILLARY GAS CHROMATOGRAPHY ИОННЫЕ АДСОРБЕНТЫ В КАПИЛЛЯРНОЙ ГАЗОВОЙ ХРОМАТОГРАФИИ L.D.Belyakova, L.H.Strokina, Moscow, USSR When nonpolar nonspecific adsorbents are used, the sepa ration of isomeric hydrocarbon molecules is basically determined by the geometry of the molecules. Isomeric orga nic compounds with similar geometry but different electron distribution were separated on capillary columns packed with inorganic salts, nonporous ionic adsorbents. On the surface of BaSO. crystals there are double - charged Ba2+ cations, whereas the negative charge is dis tributed over large complex SO. anions. In the adsorp tion of molecules with electric moments on BsSO. a non specific interroolecular interaction (dispersion and elec trostatic induction) is accompanied by a specific intermolecular interaction between the (electrostatically oriented) dipole of the adsorbate and the ionic adsorbent. In order to decrease the specificity of this adsorbent toward molecules possessing quadrupole and/or dipole moments it was modified by solution of alkaline metal hali- des. Capillary columns packed with the modified BaSO. were successfully used to separate all xylene and butene Iso mers , isomers of cis ana trans octene-2, dimethylnaphthalenes, some nitrogen - and oxygen - containing heterocyc- 232 lie compounds. The elutlon order of these compounds on the BaSOj depends on the dipole moments of their molecules. 347 P.3-28 ADSORBENTS FOR RAS CHROMATOGRAPHY АДСОРБЕНТЫ ДЛЯ ГАЗОВОЙ ХРОМАТОГРАФИИ N.E.Bolotina, S.M.Rakhovskaya, S.A.Yegorova, Saratov, USSR Adsorbents most frequently used for gas chromatography (silica gel, alumina gel) readily sorb water and other polar substances from industrial mixtures being analysed, which causes changes and deterioration of their adsorbing properties, requires much time for reaching the equilibri um and reduces the life time of chromatographic columns. Therefore the search and study of new homogeneous non- hygroscopic adsorbents having different surface chemistry appear to be significant problem. In the present work the study of the surface properties of single and binary Ni(II), Co(II), Cd hydroxides and oxides has been carried out by gas chromatography. Their interaction energy characteristics with molecules of diffe rent classes (saturated and aromatic hydrocarbons, oxygen containing substances) were calculated. Specific interac tion energy in the series of the sorbents studied increa ses in accordance with donor-acceptor properties of the cations: Nibutane-butylene fractions, off- coming cracking gases, phenol production products et al.) on Co(II) and Cd hydroxides, CdO and binary Ni(II) - Cd (3«) oxides. The proposed adsorbents are moisture and heat resistant (over 250°C for oxides), they need no preliminary thermal treatment, modification or coverage with liquid phases; they are mechanically resistant providing extended service life of chromatographic columns. 348 P. 3-29 ADSORBENTS FOR OXYGEN-CONTAINING COMPOUNDS CHROMATOGRAPHY АДСОРБЕНТЫ ДЛЯ ХРОМАТОГРАФИИ КИСЛОРОДСОДЕРЖАЩИХ СОЕДИНЕНИИ V.G.Pastushenko, S.S.Krivolapov, Tula, USSR T.B.Gavrilova, Moscow, USSR The study of carboxyl-containing substances adsorption on the surface of graphitized thermal carbon black (GTCB) has resulted in the fact of their adsorbed state high association which presents difficulties in their direct 233 chromatography. The quantative a.ialysis of oxygen-contai ning compounds and, more particularly of carboxylic acids Is mostly preferable for different enterprises of chemical food and medical industry. The adsorbents based on the commercial .black TG-10 and PM-15 were obtained by their modification. The monolayers were applied on the surface of the starting carbon black by means of solution adsorption: a) in the form of chains of aromatic dicarboxylic acids (1,4-benzenedicarboxyiic, 2,6-naphthalenedicarboxylic, 4,4'- diphenylsulfonedicarboxylic); b) in the form of nets of aromatic tri- and tetracar- boxylic acids (1,2,4-benzenetricarboxylic, 1,3,5-benzenetricarboxylic, 1,2,4,5-benzenetetracarboxylic); c) and subsequently liquid phases and the said acids. The adsorption of carboxyl-containing compounds on the modified adsorbents surface was studied. The retention characteristics of different oxygen-containing adsorbents (alcohols., ketones, aldehydes, aromatic and aliphatic acids esters) on their surfaces were obtained. The chromatographic separation of different materials with similar boiling points, geometric isomers and stereo isomers of oxygen-containing compounds was carried out.

734 PREPARATIVE CHROMATOGRAPHY

349 S.2-1 PREPARATIVE LOW PRESSURE LIQUID CHROMATOGRAPHY OF PHYSIOLOGICALLY ACTIVE SUBSTANCES (CRITERIA AND EFFICIENCY) ПРЕПАРАТИВНАЯ ЖИДКОСТНАЯ ХРОМАТОГРАФИЯ ФИЗИОЛОГИЧЕСКИ АКТИВНЫХ ВЕЩЕСТВ ПРИ НИЗКОМ ДАВЛЕНИИ /КРИТЕРИАЛЬНЫЕ ПАРАМЕТРЫ И ЭФФЕКТИВНОСТЬ/ G.V.Samsonov, V.S.Pirogov, A.T.Melenevsky, Leningrad, USSR Preparative high-performance large-scale economic chro matography is based on the use of selective high capacity sorbents and on the theory of successive displacement of substances from the sorption column with the formation of sharp boundaries of substance zones. These methods acquire particular importance in the chromatography of organic physiologically active substances slowly diffusing in sor bents. High sorption selectivity is most soundly based for systems containing polymeric ion exchangers and organic counterions. Thermodynamic analysis of selective interac tion in these systems showed that the entropy factor is of major importance. The criteria for the formation of sharp boundaries of ion zones include thermodynamic, kinetic and geometric parameters. With their aid the conditions for complete displacement with sharp zone boundaries are calculated. The solution of the problsm taking into account longitu dinal mixing leads to new cr. .eria including longitudinal diffusion and other factors. In this case it becomes necessary to carry out the process on granules the diamet- re of which is not very small. The use of original sorbents consisting of microdisperse ion exchanger grains included in a porous structure allows us to solve the problem. It makes possible to accomplish fine isolation of physiologically active substances, such as hormones, vitamins, nucleoside phosphates, etc. on low-pressure columns.

350 P.3-54 A PROMISING METHOD OF RARE PURE GASES PREPARATION FOR CHROMATOGRAPHIC PRACTICE ПЕРСПЕКТИВНЫЙ МЕТОД ПОЛУЧЕНИЯ ЧИСТЫХ БЛАГОРОДНЫХ ГАЗОВ ДЛЯ ХРОМАТОГРАФИЧЕСКОИ ПРАКТИКИ I.V.Bondarenko, V.L.Budovich, E.A.Myagkov, B.P.Okhotnikov, Moscow, USSR A solution of some chromatographic problems requires an application of rare pure gases having less than 10"^% vol. of the total contaminant of inorganic dases (N,, 0-, H,, CO, CO,, etc.). Such a problem is a sensitive analysis oi inorganic gas traces. Detectors employed based on 135 Penning's effect require pure He or Ne as a carrier gas. He, Ne may be purified in an electric discharge, This method relies on cathaphoresis effect - composition varia tion of gaseous mixture in an electric field. This change is a result of contaminantal ions flow and is important, when the ionization energy of contamihantal gas is less than the energy of" metastable level of the carrier gas. The positive ions flow towards the cathode and they are compensated in the closed vessel with an diffusive flow of neutral particles into the volume. If the electrodes are hollow and used as gas exit from the chamber, an oppo site flow of neutral particles will decrease and the contaminantal gas concentration in gas flowing away through the cathode will increase. Accordingly the cont aminant concentration in gas flowing away through anode becomes less than the initial one. For the full contaminant removal with the present method it's necessary to have contaminantal ions concentra tion higher than those of main gas and the degree of ioni zation have to be close to unit. Ion concentration depends on discharge type and current value. Discharge chamber construction and current value depend on initial contaminant concentration and required purity level. The experimental results show the presented method may be used for the full contaminant removal of He and Ne with low initial contaminant concentration. In this case the total ionization and decontamination can be simply obtai ned in the corona discharge regime. 351 P.3-55 OPTIMIZATION OF PREPARATIVE SEPARATION OF HYDROXYBIPHENYL ISOMERS ОПТИМИЗАЦИЯ ПРЕПАРАТИВНОГО РАЗДЕЛЕНИЯ ИЗОМЕРОВ ОКСИБИФЕНИЛА K.Gazda, B.Makuch, Gdansk, Poland The separation of hydroxybiphenyl isomers has been carried out on silica gel. The choice of separation con ditions has been impeded due to slight solubility of the para isomer. Three kinds of the mobile phase have been employed, their elution strenght being selected in such a manner that the selectivity coefficient in each type of the phase varied in a broad range (from ca. 3 to 10). The rate of broadening of descending part of a peak has been investigated as a function of mass overloading and solu bility of the sample in the mobile phase for similar k' values. Since the rate of approaching the start of a chromatogram by ascending part of a peak does not solely depend on the k' value for an analytical sample, the dependence between broadening of this part of the peak and an active component of the mobile phase has been looked for. The optimum column loadability has been estab lished for assumed degree of purity, taking into account column throughput and the volume of fractions.

236 The preparative separations have been carried out in a column with the bed dimensions of 50 x 300 mm, 352 P.3-56 ISOLATION OF DNA BINDING GLYCOPROTEID FROM SEA URCHIN SPERMATOGENESIS CELLS ВУ PREPARATIVE ELECTROPHORESIS ВЫДЕЛЕНИЕ ГЛИКОПРОТЕИДА, СВЯЗЫВАЮЩЕГО ДНК ИЗ КЛЕТОК РАННЕГО СПЕРМАТОГЕНЕЗА МОРСКОГО ЕЖА, С ПОМОЩЬЮ ПРЕПАРАТИВНОГО ЭЛЕКТРОФОРЕЗА V.P.Shirokov, Vladivostok, USSR A purification procedure of DNA binding and it to be acid soluble making glycoproteid is presented. The total preparation containing some DNA binding activity was ob tained by lysis of the sea urchin spermatogonia using the ammonium sulphate fractionation. Further purification was achieved by preparative electrophoresis on a column (40 x 500 mm) using 4B Sepharose equilibrated with buffer con sisting of 0.1 H Tris-HCl, pH 8.9, 4% /-mercaptoethanol, 1% SDS, 0.001 M EDTA. The upper and low electrode compart ments were filled with buffer consisted in 0.01 м Tris- glycyn, pH 8.4, 0.2» SDS. The upper electrode compartment buffer and the column one are separated by a membrane, two channels coming off to remove the gases formed through an electrophoresis procedure, as well as for filling the buffer and loading a protein. By using the membrane it is possible to achieve the sufficient pressure on a column as well as to change a buffer system whose resolution ability in partitioning the preparation being increased. This method has allowed to obtain ten protein frac tions one of which is a DNA binding glycoproteid, the specific activity being 250 times higher. The procedure presented for preparative electrophoresis of DNA binding glycoproteid has advantage over other ex perienced methods and it allowes to obtain a protein purified up to 80%. 353 P.3-57 CHROMATOGRAPHIC SEPARATION OF INFLUENZA VIRUS AND HOST PROTEINS DURING PRODUCTION OF INACTIVATED VACCINE ХРОМАТОГРАФИЧЕСКОЕ РАЗДЕЛЕНИЕ ВИРУСА ГРИППА И БЕЛКОВ ХОЗЯИНА ПРИ ПРОИЗВОДСТВЕ ИНАКТИВИРОВАННЫХ ВАКЦИН N.Yu.Polyanskaya, A.B.Gebrune, E.A.Fridman, Leningrad, USSR One of the basic principles of whole virus influenza vaccines construction is the most complete separation of the virus from the host proteins for instance chick em bryo. Immunochemical methods of virus preparations purity control, developed at Pasteur's Institute (Leningrad) allow to carry out this principle during production of real vaccines. Influenza virus and host antigenes elution profiles have been studied in the processes of gel-permiation and adsorption chromatography on macroporous 237

i. silicates. It is stated that by ireans of gel-permiation chromatography contents of on? of the most allergenic antigenee of chick embryo - о ralbumine - nay be brought in the final product up to 0.03 mkg/ml independently of vi rus of preliminary concentration method. Simultaneously during gel-permiation chromatography there is no separa tion of influenza virus and high-molecular Hpoglycopro- teln host components. Adsorbtion chromatography provides high degree of virus purification of ovalbumine and non- ovalbumine impurities, but this process in accompanied by rather high virus losses (up to 50 p.c). Combination of quite different processing methods allow to get prepa rations, fit for vaccinating against influenza.

354 P.3-58 CHROMATOGRAPHY OF VIRUSES ON POROUS GLASSES AND SILICAS ХРОМАТОГРАФИЯ ВИРУСОВ НА ПОРИСТЫХ СТЕКЛАХ И КРЕМНЕЗЕМАХ • V.H.Kolikov, B.V.Mchedlishvili, Moscow, USSR Porous glasses and silicas (silochroms and silica gels) have a rigid structure, are chemically and.thermo-resistant; their pore size is strictly controlled and comparable with virus particle sizer. This permits using successfully these carriers for virus chromatography. The paper is concerned with virus adsorption on porous glasses and silicas the surface of which is modified (phy sically and chemically) by organic macromolecules with different functional groups. It is shown that the main contribution into the virus adsorption on these adsorbents is made by electrostatic attraction,hydrogen bonds and hyd rophobic interactions. Contribution of each of the above intermolecular interactions into the virus adsorption was assessed. Universal calibration curve of the virus distribution coefficients showing the ratio of their dimensions, d, to the pore diameter of porous glasses, D, was obtained. It is shown that in the conditions close to equilibrium, optimal values of the adsorbent pores for gel-chromatography and adsorptional chromatography should be chosen, respectively, from: D t = 2d, and D t = (6 - 8)d. It is shown that the large-scale cKrematdgraphy of viruses on porous glasses and silicas is successfully carried out for the eluent feed rate up to V = (6 - 8) cm/min and the carrier granules sizes d = 0.05 - 0.30 irm. For the adsorptional chromatography of viruses on such carriers one should use V = (1-2) cm/min. A possibility is shown to use the non-equilibrium gel-chromatographic purification of viruses from admixture proteins for V = (10 - 100) cm/min and D/d ^-10 (d = 0.2 mm) . The research results were used to Bevelop methods of pu rification viruses of admixture proteins using adsorptio nal affinity and gel-chromatography in 50 to 150 cm long columns with the diameter of 5 to 15 cm. Adsorption and affinity chromatographies yield 10 to 100 multiple con centrations of viruses. As a result of gel-chromatography 238 the content of admixture proteins in virus suspensions drops to (Ю-2 - 10" ) mg/ml. In Industry, purification and concentration of viruses is better done by combined methods of adsorption and gel-chromatography on porous glasses and silicas with membrane separation processes. 355 P.3-59 PREPARATIVE ON-LINE OPLC SEPARATION OF NATURALLY OCCURRING COMPOUNDS РАЗДЕЛЕНИЕ ПРИРОДНЫХ СОЕДИНЕНИИ МЕТОДОМ ПРЕПАРАТИВНОЙ ТСХ ПРИ ПОВЫШЕННОМ ДАВЛЕНИИ Sz.Nyiredy, K.Mikita, Budapest, Hungary _ B.Meier, С.A.J.Erdelmeier, O.Sticher, Zurich, Switzerland Preparative chromatography is an important tool for the isolation of naturally occurring compounds as well as for the separation and purification of synthetic products. The preparative on-line OPLC requires 20 x 20 cm pre- coated plates, which edges are obliquely scratched off an impregnated with a suitable polymer suspension in order to prevent solvent leakage during overpressure. To ensure that the mobile phase migrates in a linear front, a chan nel is scratched out from the layer. A second channel placed at a distance of 18 cm from the inlet channel is permitting the collection of the eluent. Using the plates prepared as decribed above, the eluting compounds are drained from the chamber through an outlet valve. This technique is allowing the connection of a flow detector, registration of a chromatogram and collection of separated compounds with a fraction collec tor; so that OPLC can be turned into an on-line prepara tive method /1/. The mobile phase for preparative separations had been optimized using the "PRISMA"-model /2/ in preassays on TLC plates in an OPLC chamber /3/. When the average of the particle size of pre-coated TLC plates and the sample loading is strongly increased compared to the analytical loading, it is recommended to decrease the solvent strength of the mobile phase. The use of this on-line method is demonstrated for rapid isolation (e.g. frangula emodin from Rhamnus frangula) and preparative separation of naturally occurring compounds (e.g. furocoumarins from Heracleum sphondylium). Sample loadings on a single plate were between 50 and 5O0 mg. 1. Sz.Nyiredy, C.A.J.Erdelroeier, O.Sticher: Instrumental Preparative Planar Chromatography in Planar Chromato graphy (Ed. R.E.Kaiser et al) Huthig Verlag, Heidelberg- Basel-New York (1985), in press. 2. Sz.Nyiredy, C.A.J.Erdelmeier, B.Meier, O.Sticher: Planta roed. 3 (1985), in press. 3. Sz.Nyiredy, C.A.J.Erdelmeier, O.Sticher: Advances in Liquid Chromatography (Ed. E.Tyihak), in press.

239 356 P.3-60 OPTIMIZATION OF OPLC-SEPARATION OF POLAR NATURALLY OCCURRING COMPOUNDS BY THE "PRISMA"-DESIGN ОПТИМИЗАЦИЯ РАЗДЕЛЕНИЯ ПОЛЯРНЫХ ПРИРОДНЫХ СОЕДИНЕНИИ ТСХ ПРИ ПОВЫШЕННОМ ДАВЛЕНИИ НА СИСТЕМЕ "ПРИЗМА" K.Dallenbach-Tolke, B.Meier, O.Sticher, Zurich, Switzerland Sz.Nyiredy, Budapest, Hungary The "PRISMA"-design was recently developed in our labo ratory for mobile phase optimization for HPLC /1/, TLC /2/, OPLC /3/, CLC /4/ and SCLC /5/. Until now, it was described for different apolar and semipolar naturally occurring com pounds. Now, we use this model also for the optimization of mobile phases and for the separation of very polar com pounds, e.g. flavonoid glycosides, ginsenoside.s and other substances. The pre-assay for OPLC were carried out with TLC. In general the mobile phase contained three organic solvents. Water and/or acetic acid were used as modifiers, which - besides - increased the solvent strength and eliminated the tailing. The optimum TLC mobile phase was transferred to OPLC without any modification. The OPLC separations were carried out with TLC and HPTLC plates with different flow rates using a Chrompres-10 chamber (Labor MIM, Budapest-Esztergom).For the qualitative analysis, the substances were detected with suitable spray reagents and/or a Shimadzu CS-920 scanner. The developed OPLC-separations allow the analysation of flavonoids from Betula pendula (Ph. Helv. VII), Betula pubescens (Ph. Helv. VII) and Ginsenosides from Panax ginseng (Ph. Helv. VII). 1. Sz.Nyiredy, B.Meier, C.A.J.Erdelmeier, O.Sticher: HRC i CC 8/4 (1985), in press. 2. Sz.Nyiredy, C.A.J.Erdelmeier, B.Meier, O.Sticher: Planta med. 3 (1985), in press. 3. Sz.Nyiredy, C.A.J.Erdelmeier, O.Sticher: Advances in Liquid Chromatography (Ed. E.Tyihak), in press. 4. Sz.Ilyiredy, C.A.J.Erdelmeier, B.Meier, O.Sticher: GIT Supplement Chromatographic (1985) , in press. 5. Sz.Nyiredy, C.A.J.Erdelmeier, O.Sticher: Z.Anal.Chem. in press. 357 P.3-61 APPLICATION OF DROPLET COUNTER-CURRENT CHROMATOGRAPHY (DCCC) FOR THE PREPARATIVE SCALE SEPARATION OF GERMACRANOLIDES AND EUDESMANOLIDES ПРИМЕНЕНИЕ КАПЕЛЬНОЙ ПРОТИВОТОЧНОИ РАСПРЕДЕЛИТЕЛЬНОЙ ХРОМАТОГРАФИИ ДЛЯ ПРЕПАРАТИВНОГО РАЗДЕЛЕНИЯ ГЕРМАКРАНОЛИДОВ И ЭУДЕСМАНОЛИДОВ Gy.Turiak, A.Ke'ry, P.Tetenyi, Budapest, Hungary Droplet counter-current chromatography (DCCC) has pro ven to be a powerful, ideal method for the separation of various classes of naturally occurring compounds. The 24P work presented here is part of study for further adapta tions of this technique on the field of sesquiterpene lactones. Preparative scale separation of crude extracts prepared from different Asteraceae plants, rich in sesquiterpene lactones have been used as models. Comparison of the chromatographic behaviour of the comp lex lactone mixtures on TLC and DCCC has been studied, and different, new type solvent systems were selected in the descending and/or ascending mode of fractionation. (Chlo- roform-Benzene-Methanol-Water 5:5:7:2; Carbon tetrachloride-Acetonitrile-Water 6,5:3,5:2,5; Chloroform-Trichloro ethylene-Acetonitrile-Water-Methanol 2:4:2:2:5; etc.) Samples in the range of 0,4-2,0 g were easily handled, the fractions afforded isolation of pure lactones from the germacranolide (tatridines, parthenolide, 9-ol-hydroxy parthenolide, costunolide, artemorin, taurin) and eudesma- nolide (reynosin, santamarine) groups, which were identi fied by spectroscopic techniques and comparison with authentic samples. The sesquiterpene lactones were separated far more readily than by the conventional repeated column chromato graphy. Therefore DCCC is recommended for preparative scale isolation of the biologically active sesquiterpene lactones. 358 S.4-2 CHROMATOGRAPHIC METHODS OF LEUKOCYTE INTERFERON PURIFICATION ХРОМАТОГРАФИЧЕСКИЕ МЕТОДЫ ОЧИСТКИ ЛЕЙКОЦИТАРНОГО ИНТЕРФЕРОНА V.A.Pasechnik, Leningrad, USSR The interferon system is a essential element of nonspe cific immunity against virus infections, therefore inter ferons are perspective therapeutic agents. Human leukocyte, or «(.-interferon, is the family of 12-16 proteins with similar primary structure. The present work is devoted to developing of methods of obtaining «(.-interferon preparations for clinical trials and laboratory practice. Three types of prepara tions are considered: a) with activity lO* - Ю-* ME/mg of protein for the local administration; b) with activity 10 - 10 ME/mg for the parenteral administration, and c) the homogeneous preparation with activity 2-4-10 ME/mg. The preparation of a) type was obtained by single-stage GPC (gels similar to AcA-54, Acrylex P-60, etc.). The b) type preparations were obtained from the a) preparations either by ion-exchange chromatography on carboxycationits (Solosa K24/40), or by reusing GPC (gels similar to Sepha- cry1-200, TSK HW-50, etc.). The preparations obtained by different methods had the different spectra of impurities. Homogeneous interferon preparations were obtained from the a) preparations either by preparative HPLC (Spherisorb ODS 5 /jm i-propanol linear gradient) or by combination of

1/2 16.3«к. 872 241 immunoaffinity chromatography on monoclonal antibodies with ion-exchange chromatography (CM-cellulose, Solosa К 24/40).The obtained preparations have been characterised according to their biological activity, immunological properties (RIA), by electrophoresis in PAAG. T-*. impu rities, contained in the preparations and effecting their biological activity, have been analysed. The number of the preparations contained the factor D of the complement system and other biologically active compounds.

359 P.1-42 APPLICATION OF ION-EXCHANGE COLUMN CHROMATOGRAPHY TO THE PURIFICATION OF PECTIC PREPARATIONS ПРИМЕНЕНИЕ ИОНООБМЕННОЙ КОЛОНОЧНОЙ ХРОМАТОГРАФИИ ДЛЯ ОЧИСТКИ ПЕКТИНОВЫХ ПРЕПАРАТОВ M.Il.Popova, Ch.G.Kratchanov, Plovdiv, Bulgaria Based on theoretical considerations of the causes of strong absorbtion properties of pectins to polar (espe cially positively charged) mineral and organic impurities, the possibility of their removal by ion-exchange chroma tography was studied. Due to that purpose, the applicabi lity of ion-exchange column chromatography to the purifi cation of various plant pectins was experimentally tested. Industrial samples of apple, sunflower and carrot pectins were used. Diluted hydrochloric acid, ethanol, methanol and acetone were applied as eluent. The influence of different factors (temperature, acid concentration, and duration of elution was considered. It was established that along with the ion-exchange, hyd ro lyses, esterification and degradation processes take place. Their speed depends on the type of the solvent, acid concentration, and temperature. As a result pectins with decreased contents of mineral and nitric substances, and increased gelling strength were obtained. The amino acid composition of some washed pec tins was determined. The pectins purified by ion-exchange chromatography were also characterized by IR- and PMR-spectra and optical activity.

242 CHROMATOGRAPHY OF NATURAL PRODUCTS

360 S.ll-4 DETERMINATION OF ORGANIC ACIDS AND FATTY ACIDS IN FOODS ОПРЕДЕЛЕНИЕ ОРГАНИЧЕСКИХ И ЖИРН11Х КИСЛОТ В ПИЩЕВЫХ ПРОДУКТАХ P.A.Biacs, J.Bodnar, L.Zsoldos, A.Hoschke, F.Hajdu, Budapest, Hungary The food industry needs development of simple and in formative methods for quality control. The application of HPLC methods is very intensive nowadays. Reduction or eli mination of extraction and formation of derivatives is necessary for simplifying the methods of food testing. We have investigated the organic acid and fatty acid content in the red wine, sauer cherry juice, rhubarb, to mato, red pepper, onion, cabagge and savoy. Our aim was the determination of organic acids and fatty acids without forming derivatives. These materials are sensitive to heat and usually there is no chance to carry out CLC analysis without formation of derivatives. Separation of organic acids was carried out using ion- pair chromatography. The conditions were isocratic, but the optimization of parameters was making use of gradient elution. To characterize the complex systems, a computer simulation method has been worked out. 361 P.1-48 TRACE ANALYSIS OF ETHYLENETHIOUREA IN FOODS BY HPLC АНАЛИЗ СЛЕДОВ ЭТИЛЕНТИОУРЕТАНОВ В ПИЩЕВЫХ ПРОДУКТАХ №Т0Д0М ВЭЖХ J.Lehotay, E.Brandsteterova', J.Caraj, Bratislava, USSR Metallic derivates of ethylenebisdithiocarbamic acid (EBD'JC) have been extensively utilized as agricultural fungicides. One of decomposed products of these fungicides is ethylenethiourea (ETU) which has the influence on the growth of the plants and has toxic properties. The objective of this work was to develop a new method for determination of ETU in apples, potatos and tomatos. Following characteristics were deemed desirable for such a method: applicability when very low ETU airounts are present and speed compatible with routine measurement requirements. The methanol extracts were evaporated by vacuum evapo rator. Dry residues were dissolved in chloroform-methanol (1:1). The precipitate was removed on a centrifugal sepa rator. The clear solution was injected in liquid chromatograph. The extracts were analysed on Separcon C-18 column (5 jum) in methanol-water (10:1) as mobile phase. The detec-

243 tion of analysed compounds from extracts was made by UV detector at 254 nm. Part of the controversy associated with EBDTC-ETU resi dues studies is the question of ETU methodology with res pect to stability of ETU during workup of samples. To test the error of analytical procedure which is caused by ETU degradation during extraction the two methods were used: standard addition of ЕТИ in foods and standard addition of ETU in extract. The results have shown the concentra tion of ETU slowly decreases. The extraction yield was 75%.

362 P.1-49 SOME COMPLEXITY MEASURES OF FOOD GC ANALYSIS НЕКОТОРЫЕ ОЦЕНКИ СЛОЖНОСТИ РЕШЕНИЯ ЗАДАЧИ ГАЗОХРОНАТОГРАФИЧЕСКОГО АНАЛИЗ/. ПИЩЕВЫХ ПРОДУКТОВ D.A.Ugolev, A.I.Zaporozhets, Leningrad, USSR Much progress has been made in recent years in food GC and GC-MS analysis. The "users" of these analytical methods are specialists in various fundamental and applied disciplines: biochemistry, medicine, ecology, crirtinalogy, agronomy, food technology, etc. The search of a good ana lytical strategy and design of multi-purpose quick and cheep GC-analytical methods remain one of the most pres sing problems both to "users" and GOanalysis specialists. The ideology of "pattern recognition" (Varmuza, 1980) together with head-space GC-analysis of volatile organic compounds (VOC) (Zlatkis et al., 1982) promote overtly simplistic solutions to some very difficult problems, which "users" are interested in ("medicine diagnostics", "quality control", "taxonometry" etc.). The main trends in our recent work are a design of a soft ware and solu tion of these very important for "users", but incorrect for analytical chemistry specialists problems as a GC- head space VOC-pattern recognition problem and to propose to use and optimistic VOC-profile GC procedure of food, which is equivalent to animals odour description of food as a finite automaton (Ugolev et al., 1983, 1984). It is the purpose of present paper to make a next step in this direction to propose a formal objective complexity mea sure both for the problem solving (head-space GC-analysis) algorithm and for the different systems (VOC-fractions a and foodstuffs as a whole) to be analysed. The VOC-frac tions of various food-stuffs and their changes during shelf-life have been studied for a great variety of conditions. GC and GC-MS capillary analysis were used. The computational complexity'of SC head-space analysis (as a "pattern synthesis" problem algorithm) and the stability and "indefinity" of experimental data have been evaluated for the first time. The boundaries for GC (GC-MS) analysis traditional and "pattern recognition" data interpretation are proposed.

244 363 P.1-50 THE GLASS CAPILLARY COLUMN METHOD OF THE DETERMINATION OF THE PHENOLCARBONIC ACIDS IN WINES ОПРЕДЕЛЕНИЕ ФЕНОЛКАРБОНОВЫХ КИСЛОТ ВИН НА СТЕКЛЯННОЙ КАПИЛЛЯРНОЙ КОЛОНКЕ N.M.Pavlenko, B.A.Vinogradov, Yalta, USSR For the analysis of the phenolcarbonic acids and their precursors in wines 50 m long glass capillary column with SE-30 was used. To improve the stability of the column under the conditions of high temperatures (200 - 300 C) the column was successively treated with the solution of 0,5% HF, 5% HC1, anhydrous methanol and hexan, and the inner surface of the column was sililated with TMCS. Sta tionary phase was formed by the high pressure static method. The column obtained has a high resolution and operates without changes in efficiency during two years. After following isolation and sililation with HMDS the phenolcarbonic acids were.determined as TMS-derivatives. 364 P.1-51 A COMBINATION OF CHROMATOGRAPHIC METHODS FOR THE ANALYSIS OF PHENOLIC COMPOUNDS IN WINES СОЧЕТАНИЕ ХРОМАТОГРАФИЧЕСКИХ МЕТОДОВ ДЛЯ АНАЛИЗА ФЕНОЛЬНЫХ СОЕДИНЕНИИ В ВИНАХ Yu.A.Klyachko, E.I.Benkovitch, V.S.Padalkina, E.S.Trybitsina, Moscow, USSR Phenolic compounds (catechines, flavonols, anthocyanes, leucoanthocyane, phenolcarboxylic acids) both bound and in a free state may be found in the composition of wines and are of great importance. The definition of phenolic compounds is connected with their separation. Different chromatographic methods are used for this purpose, liquid chromatography both plate and column methods being most advantageous. In the present work the combination of high performan ce liquid chromatography with reflective spectrophotomet ry in thin-layer cellulose was used. Optimized thin-layer plates having many advantages such as sensitivity and speed were employed. Chromatography was carried out in two directions: the solvents were the mixture of buthanol with acid and water. Such combinations of methods enabled us to identify four cathechines in the poly-phenol fraction from diffe rent wines epicathechine, cathechine, hallocathechine, epicathechinehallate, three leucoanthocyane, aglicones- leucodelphynidine, leucopethunidine, leucopeonidine, four anthocyanine pigments, which as aglycones contain delphy- nidine. pethunydine, malvidine and peonidine. Quercethine, camppherol, myricethine, isoquercitrine are found in the composition of flavonols. Phenolcarbon acids are represented by hallic, vaniline, caffeic, phe- rulic, p-oxybenzoic and other acids.

245 365 P.1-52 CHROMATOGRAPHIC METHODS FOR STUDY OF GEORGIAN WINES COMPOSITION КОМПЛЕКС ХРОМАТОГРАФИЧЕСКИХ МЕТОДОВ ИЗУЧЕНИЯ СОСТАВА ГРУЗИНСКИХ ВИНОМАТЕРИАЛОВ I.Sh.Shatirishvili, Tbilisi, USSR The wines are so complicated compositions that for full evaluation it is not enough only one version of chromato graphy. A combination of chromatographic methods for study of wines composition was developed , at first for composition of georgian ordinary and chateau wines including Gurgaani, Zinandali, Zolikauri, Saperavi e.a. By head-space chromatographic analysis with three detec tors: flame ionization, electron-capture and flame-photo metric the components were determined influencing on

flavor (acetaldehyde, athylpropionat, alcohols C2-Cg et al.). Selective detectors permit to determine diicetyl the content of that changes not strongly) and dimethylsul- phide (the concentration of that changes in different wines till 10 times). Direct sampling of wine in quartz capillary columns filled by SE-30 and PEG 20M with cutting of involative part gave us an interesting information, but high content of water and ethanol makes direct chromatography quite difficult. Aminoacid composition of georgian wines were determi ned by usual method without deproteinisation. It was shown about 37 aminoacids with high concentration of proline. By ionic chromatography in georgian wines .NH* , К , Na , SO, and high fat acids were determined. Phenolcar- bon acids were determined by thin-layer chromatography on "ARMSORB'-silica gel and preliminary separation of some component by membranes. Biopolymer in wines have influence on composition of liquid fase and high sorptlbility to ward a lot of components. 366 P.1-61 ELUTION CHROMATOGRAPHY FOR THE OBJECTS OF FOOD AMD MICROBIOLOGICAL TECHNOLOGY ХРОМАТОГРАЗИЧЕСКИЕ МЕТОДЫ РАЗДЕЛЕНИЯ ОБЪЕКТОВ ПИЩЕВОЙ И МИКРОБИОЛОГИЧЕСКОЙ ТЕХНОЛОГИИ Yu.A.Sultanovich, G.B.Kolesnic, S.L.Gramolin, Moscow, USSR The mixture which of components with wide intervals of boiling temperature are typical for the objects of food and microbiological industry. The results of variants of elution chromatography to increase the speed of Information obtained and also its volume are given. The theory of gas chromatography with programming of mobile phase flow rate is worked out and number of gas 246 schemes of chromatographs for realising this variant of chromatography are improved. The optimum field of another variant intensifying the elution of compound mixtures, i.e. chromatography with steam mobile phase is experimentally based. The system of search for liquid-accumulating strains with combination of gas chromatography and also thin and liquid chromatography is widely used. On the analysis of triacylglycerol composition of plant oils and the products of their modification was shown that computing technique gives an opportunity of obtaining additional information by combination of different analy tical methods (including chromatography) with remified scheme of their applying. A number of other examples of applying of chromatogra phy for solving different problems in food and microbiolo gical industry are given.

367 P.1-53 DETERMINATION OF NITRATE AND CHLORIDE IONS IN FOOD BY SINGLE COLUMN ION CHROMATOGRAPHY ОПРЕДЕЛЕНИЕ НИТРАТ- И ХЛОРИД-ИОНА В ПИЩЕВЫХ ПРОДУКТАХ МЕТОДОМ ОДНОКОЛОНОЧНОЙ ИОННОЙ ХРОМАТОГРАФИИ J.Pentchuk, K.Ilmoja, Tartu, USSR U.Haldna, Tallinn, USSR Colorimetric and titrimetric methods for determination of nitrate and chloride ions in food have some drawbacks, such as interferences caused by other solutes, the need for silver salts and the necessity to use different proce dures for determination of the two ions. He have developed a single column ion chromatographic method for simultaneous determination of nitrate and chlo ride ions on a ppm level in potatoes, cabbages, etc. The influence of different factors (composition, concentration and flow rate of the eluent used) on the efficiency of separation and time of analysis has been estimated. The method developed has a detection limit of 0.2 and 2 ppm for chloride and nitrate ions, respectively. The relative root mean square error 0.03-0.05. The results of ion chtomatographic determination agree well with those obtained by potentiometric techniques using ion selective electrodes. The time for the simultaneous ion chromato graphic determination of chloride and nitrate ions is less than 15 min. A chromatogram for chloride and nitrate determination in beetroot is shown in figure. The operating conditions were: eluent 1.5 mM glyconic acid + 1.5 mM boric acid, pH - 8.6, flow rate 1.0 ml/min. Column (L - 15 cm, I.D.- 3 mm) contains anionic sorbent,BAKC-l (capa city - 0.031 mg-equiv/cm ).

247 368 P.1-54 PESTICIDE RESIDUES IN VEGETABLE DRUGS GAS CHROMATOGRAPHIC DETERMINATION OF RESIDUES OF DICHLORVOS, DIMETHOATE AND PARATHION-METHYL; EXAMPLE OF A TEST FROM THE GDR PHARMACOPOEIA (AB-DDR) ОПРЕДЕЛЕНИЕ ПЕСТИЦИДОВ З ОВОЩНЫХ КОНЦЕНТРАТАХ С ПОМОЩЬЮ ГЖХ Th.Gabrio, H.Schlenkrich, D.Ennet, Berlin, CDR Special problems of residue analysis in vegetable drugs are discussed and solutions presented for legal regula tions. For pesticides used to larger extent in medicinal plant cultivation or ubiquitous in the environment like triazines, chloro-organic and phosphoro-organic insectici des, standardized methods of residue analysis and tolerances are included in the AB-DDR. If residues occur for which there is no test provided in the AB-DDR, it should be de cided from toxicological aspects whether their type and amount admit the use of the drug tested. As an example of a standardized method, the test for residues of dichlorvos, dimethoate and parathion-methyl is presented. . The starting point for elaborating the test was the method of Becker /1/, modified as follows: The powered sample is treated with hydrochloric acid (0.01 mol/1) and then shaken with a mixture of chloroform- acetone (12:2). after purification on a silica gel-active carbon column (eluent mixture - chloroform-acetone), the purified extract is chromatographed )10% QF, on Chromosorb WHP deactivated with Cabowax 40O, temperature programme 130°C 4 min 4 C/min 2O0 C, phosphorus-specific detector). The recovery rate determined for different drugs lay around 60 to 100%. 1. G.Becker, Deutsche Lebensmlttel-Rundschau,67 (1971) 125.

369 P.1-55 MICROPACKED COLUMNS FOR THE GAS CHROMATOGRAPHIC CHARACTERIZATION OF DISTILLED ALCOHOLIC BEVERAGES МИКРОНАБИВНЫЕ КОЛОНКИ ДЛЯ ХАРАКТЕРИСТИКИ АЛКОГОЛЬНЫХ НАПИТКОВ С ПОМОЩЬЮ ГЖХ L.Wennrich, W.Engewald, Leipzig, GDR The direct gas chromatographic analysis of distilled alcoholic beverages requires the determination of higher alcohols, fatty acid ethyl esters, aldehydes, fatty acids and other substances in an aqueous ethanol medium. In addition to capillary columns packed columns filled with Carbowax 20M coated Carbowax В /1/ are applicable to this analytical problem. Using this packing material we studied the most favour able conditions for the analysis of alcoholic beverages.

24S These were obtained by; - use of micropacked columns /2/; - use of hydrogen as carrier gas; - optimization of the chromatographic parameters (carrier gas flow, temperatures, heating rate). A mixture of n-alcohols and 2- and 3-methylbutanol-l was selected for the optimization. The parameters were obtimized with respect to the sum of the separation num bers of the n-alcohols and to the resolution R of 2- and 3-methylbutanol-l. The efficiency, the polarity and the adsorption activity of the columns used were characterized by means of a test mixture containing n-alkanes, alcohols, esters, aldehydes, fatty acids and acetales. The results are compared with those obtained on capillary columns. 1. A.Di Corcia, R.Samperi, C.Severinit J.Chromatogr..,198 (1980) 347. 2. T.Welsch, W.Engewald, J.Poerschmann: J.Chromatogr.-148 (1978) 143.

370 P.1-56 INVESTIGATION INTO VOLATILE COMPONENTS OF RED CURRANT AND SOUR CHERRY ИССЛЕДОВАНИЕ ЛЕТУЧИХ КОМПОНЕНТОВ КРАСНОЙ СМОРОДИНЫ И ВИШНИ A.Sass-Kiss, Zs. Szabo-Forra's, Budapest, Hungary Authors investigated the composition of volatiles in red currant and sour cherry by means of chromatographic methods in order to find components probably contributing to the characteristic odour of the above fruits. The volatile carbonyl and acid composition of the fruits was studied. Carbonyl compounds were separated into groups by a multistep thin layer chromatographic procedure follow ing the formation of 2,4-dinitrophenylhydrazone deriva tives. Fractions were collected through preparative TLC and rechromatographed in different developing systems and thus separated into individual components. This way altogether 23 carbonyls were detected in red currant, of which 17 were identified, while in sour cherry 15 of the 19 separa ted components could be identified. Acid components were separated by a gas chromatographic method on FFAP stationary phase following steam distilla tion, isolation by salt formation and liberation by formic acid. Identification was performed by enrichment technique and by comparison of retention times of known compounds. Characteristic flavour compounds were studied by means of odour descriptive sensory analysis combined with gas chromatographic separation.

17.3ac.872 249 371 P.1-57 ATTEMPTS IN ORDER TO QUALIFY APPLE AROMAS PRODUCED IN INDUSTRIAL AROMA RECOVERY EQUIPMENTS ПОПЫТКИ ОЦЕНКИ АРОМАТА ЯБЛОК ПРИ ПРОИЗВОДСТВЕ ЭССЕНЦИИ E.Madarassy-Meraich, M.Petro-Turza, Gy.Teleky-Vamossy, F.Boross, I.Szarfoldi-Szalma, K.Fuzesi-Kardos, Budapest, Hungary Authors aimed at the objective instrumental qualifica tion of dilute aqueous industrial apple .aroma condensates, which gives a good correlation with sensory evaluation. Following the concentration of volatiles in organic sol vent in combination with salting-out, qualitative composi tion of the aroma extract was investigated prior to quan titative measurements.Separations were made by capillary gas chromatography. On the basis of previous examinations (isolation of the ester fraction, investigation into the effect of salting out on different types of compounds etc.) the identification war carried out by enrichment technique using two different stationary phases. This way 22 peaks were separated from the pentane extract, of which 17 could be identified. From the identified components the quantita tive determinations of four major acetate esters and two carbonyls were performed on СР-ИАХ-51 chemically bonded stationary phase, giving a better resolution. Parallel to measurment on capillary column, the amount of the alcohol constituents was determined directly from the aqueous so lution on a column packed with Chromosorb ЮЗ stationary phase. This way the instrumental analyses of 14 aromas of dif ferent quality was performed and it was tried to establish a relationship of chemical composition and detailed senso ry evaluations (determination of dilution threshold value and concentration-intensity functions).A relationship co uld be found for ranking different aromas into good, medium and week categories. Determination of the sequential order within each category and thus reliable substitution of sen sory examinations by instrumental measurments needs further investigations of a great number of samples, the evalua tion of which has to be carried out by mathematical methods. 372 P.4-27 ANALYSIS OF ROSE OIL BY GAS CHROMATOGRAPHY AND MASS-SPECTROMETRY АНАЛИЗ РОЗОВОГО МАСЛА МЕТОДАМИ ХРОМАТОГРАФИИ И МАСС-СПЕКТРОМЕТРИИ V.I.Tyutyunnik, V.N.Helnikov, Simferopol, USSR O.Teander, Uppsala, Sweden Using GC, MS and standard samples as methods of analy sis, the authors identified in rose essential oil 120 .components. GLC was carried out on Finnigan Model 9610 chromato-mass spectrometer, equipped with a glass capilla ry column 40 m x 0.3 mm i.d., filled with the stationary 250 phase [Icon 50-HB-5100, column temperature was programmed from 70°C to 280°C at 2°C/min. Rosoxide diastereoisomers were obtained by preparative GC. Isomeric cis- and trans-rosoxides were identified by N.M.R, spectrometry on Jeol Model РХ-ЭО Q. Geranyl acetate, geranial, elemol and ^f-cedinol were identified in the oil by high pressure LC on Varian Model 660, equipped with UV-detector (280 nm), stainless steel column 25 cm x 4,6 mm i.d., packed with Lichrosorb RP-8, elution gradient of aqueous solution of acetonitrile varied from 40% to 70%, solvent flow rate 2 ml/rain. 373 P.2-57 SAMPLE PREPARATION FOR GAS CHROMATOGRAPHIC ANALYSIS IN FLAVOUR RESEARCH - SOME EXPERIENCES НЕКОТОРЫЕ ДАННЫЕ О ПРИГОТОВЛЕНИИ ПРОБ ДЛЯ ГАЗОХРОМАТОГРАФИЧЕСКОГО АНАЛИЗА ЗАПАХОВ W.Rddel, Rehbrucke, GDR Gas chromatographic analysis in flavour research nor mally involves isolation and concentration of flavour compounds. These procedures have a great influence on accuracy and precision. In investigation of fats and oils we prefer molecular dictillaticn. Aqueous and partially solid samples also are mainly prepared by simultaneous distillation-extraction. The combination of Soxhlet-extraction and molecular distil lation gives good results in analysis of solid samples with a high amount of fat. The efficiency of mentioned methods is demonstrated by practical examples. 374 P.2-58 GAS CHROMATOGRAPHIC CHARACTERIZATION OF SESQUITERPENES FREQUENTLY OCCURRING IN ESSENTIAL OILS ГА30ХР0МАТ0ГРАФИЧЕСКАЯ ХАРАКТЕРИСТИКА СЕСКВИТЕРПЕНОВ, ЧАСТО ВСТРЕЧАЮЩИХСЯ В ЭФИРНЫХ МАСЛАХ E.Lemberkovics, G.Verzar-Petri, Budapest, Hungary A collection of the relative retention factors of ses quiterpenes, frequently occurring in essential oils have been prepared and presented in this paper. On account of the absence of sesquiterpene standards for the determination of factors we could't build up such standard series of sesquiterpenes like ones of monoterpe- nes /1/. Therefore the determination of sesquiterpene factors was carried out on the base of the data of iden tified sesquiterpene components in five different essen tial oils (Chamomile, Yarrow, Cannabis Basil and Wormwood oils) which were studied by identical GC parameters. The identification of the sesquiterpene components was made by GC-MS method and in each case with gas chromatographic standard addition, too. The retention factors were determined on two stationary phases of different polarity (3% OV-17 and 1.5% SP-2250 + 0.95% SP-2401) and they were referred to quaiazulene ses- 251 quiterpene standard. For assuring of reproducibility we have determined the retention factors of n-alkane series (CJC-CJQ) related to quaiazulene. The Standard deviation calculated from the relative retention factors of the same sesquiterpene comoonents in the different essential oils was not larger than 6 x 105. 1. E.Lemberkovics: J.Chromatogr., 286_, 293-300,(1984).

375 P.2-59 HPLC AS A METHOD FOR INVESTIGATING THE CHEMICAL MODIFICATION DYNAMICS OF PROTAMINES FROM STERGEON GONADS ВЭЖХ КАК МЕТОЛ ИССЛЕДОВАНИЯ ДИНАМИКИ ХИМИЧЕСКОЙ МОДИФИКАЦИИ ПРОТАМИНОВ ИЗ ГОНАД ОСЕТРОВЫХ V.K.Rybin, L.K.Moleva, N.V.Makarov, G.S.Katrukha, Moscow, USSR Protamines are the strongly basic polypeptide compo nents of sex cell chromatin with a molecular mass of about 4000. They are used in medicine for a sustained effect of various medicinal preparations and also as heparin antago nists. Protamines can also Inhibit the growth of some tumors. To enhance their antimicrobic and antitumor action, we have carried out a directed chemical modifica tion of two protamines from Caspian stugeon gonads - sturins A and В - and Immobilized on their NH^-groups certain antitumor and antimicrobic antibiotics. The direc ted modification was monitored by HPLC on a Du Pont 8800 model apparatus equipped with a Zorbax ODS column accor ding to our procedured. As a mobile phase we used a 0.15% solution of trifluoroacetic acid in water and an ethanol gradient at 1 ml/min with detection at 220 nm. The analy sis results were processed by a SP 4100 integrator. The elution conditions found by us make it possible to separa te sturin A from sturin B, as well intermediate products of their modification and covalent complexes of the type strain A - -«-antibiotic, sturin—•- antibiotic, and deter mine the number of the antibiotic residues covalently bound to the protamine. Assay time: 70 min.

376 P.1-62 CONCENTRATION OF MICROIMPURITIES OF WATER-ALCOHOL SOLUTIONS КОНЦЕНТРИРОВАНИЕ МИКРОПРИМЕСЕЙ ВОДНО-СПИРТОВЫХ РАСТВОРОВ D.M.Makeev, B.S.Tchernyaga, N.I.Slavutskaya, Moscow, USSR In water-alcohol solutions based on high quality alcohol ("Extra", "Luxe") micropurities are in the range of lO-4- 252 10~Ч. Such micropurities comprise aldehydes, esters, higher alcohols (components of fusel oil). Direct input into chromatographic equipment makes it possible to determine substances in the range of 10-10 That is way it is necessary to concentrate substances ly ing behind this threshold sensitivity. For this purpose vacuum-controlled concentrated columns are employed, that incorporate preconcentration column for sorption of analysed impurities, which then are directed through doser into analytical column.

253

i COMBINATION OE VARIOUS TYPES OF CHROMATOGRAPHY

377 P.2-45 DETERMINATION OF ENDOGENIC AND EXSOGENIC HYDROCARBONS IN SOME HYDROBIONTS BY USING HPLC, TLC AND GC ОПРЕДЕЛЕНИЕ СОДЕРЖАНИЯ ЭНДОГЕННЫХ И ЭКЗОГЕННЫХ УГЛЕВОДОРОДОВ В НЕКОТОРЫХ ГИДРОБИОНТАХ СОЧЕТАНИЕМ МЕТОДОВ ЖИДКОСТНОЙ, ТОНКОСЛОЙНОЙ И ГАЗОВОЙ ХРОМАТОГРАФИИ N.A.Pisareva, N.V.Sizova, B.P.Lapin, Moscow, USSR Determination of hydrocarbons in fish and other hydrobionts is important in connection with total environmental pollution. Analytical scheme for determination of total hydrocarbons content including n-alkanes, isoalkanes, cycloalkanes with monocyclic hexanes, bicyclic and tricyclic naphthenes, steranes and terpanes with different numbers of cycles, olefins (linear and branched alkenes, diterpe- nes and triterpenes among them), aromatics (including raono- di-, tri and polycyclic arenes) was developed with the aim of obtaining the safest and most informative method. The system developed for such determination is based on a com bination of HPLC, TLC, GLC, ultraviolet spectroscopy, fluorescence spectroscopy and GC-mass spectrometry. The analysis included extraction of total hydrocarbons, sepa ration into classes, identification and quantification. Natural background level of hydrocarbons was determined in some fish products and hydrobionts contamination with oil was also showr * some cases. The basic < was conducted by studying" the hydrocar bon content in different samples of sardine and small fish. Ultraviolet spectroscopy and a combination of TLC and GC assisted us in determining the presence of oil hydrocar bons in sea hydrobionts. Determination of aromatic hydro carbons' was conducted with the help of micro TLC on silica plates. The use of TLC in combination with UVespectrosco- py gives us rapid information about the presence or absen- .ce of contaminants. However the task of separation after identification by TLC, GC, UV spectroscopy, fluorescence spectroscopy can be simplified by using HPLC with diffe rent detectors. If standards are available, HPLC permits qualitative and quantitative determination of alkanes, alkenes and arenes. In particular, we used a spectrophotometric, a diode-array and a fluorometric detectors. The chromatographic analysis of a series of samples with in dication of oil contamination showed a characteristic dynamic of changes in concentration and number of compo nents. Some identifications were verified by GC-mass spectrometry. The data obtained can be used in practice for several rapid methods to determine individual classes of aliphatic and eromatic hydrocarbons in some hydxobionts and other products.

254 Analysis of hydrocarbons is necessary to evaluate the amount of endogenic alkanes, olefins, polyenes (including squalene), arenes and also possibly antropogenic contami nants of hydrobionts (in particular, PAU, mono-, di- and tricyclic aromatic hydrocarbons, short chain alkanes) which are not benign for organism from the point of view of food hygiene and also from the point of view of sale quality.

378 P.2-46 THE APPLICATION OF LIQUID COLUMN CHROMATOGRAPHY AND MOLECULAR MASS OF ENDO- ^-1,3-GLOCANASE FROM MARIN MOLLUSC SPISULA SACHALINENSIS ПРИМЕНЕНИЕ ЖИДКОСТНОЙ КОЛОНОЧНОЙ ХРОМАТОГРАФИИ И ЭЛЕКТРОФОРЕЗА ДЛЯ ОПРЕДЕЛЕНИЯ МОЛЕКУЛЯРНОГО ВЕСА ЭНДО~>а.,5-ГЛЮКАНАЗЫ ИЗ МОРСКОГО МОЛЛЮСКА SPISULA SACHALINENSIS A.A.Artukov, T.G.Svetasheva, M.B.Philitova, V.V.Sova, L.A.Elyakova, Vladivostok, USSR According to the gel filtration on Sephadex G-150 and P-lOO and thin-layer chromatography (Sephadex G-75) the molecular mass of & -*,3-gluconase is 22000. However, using polyacrylamid-gel electrophoresis in the presence of SDS the molecular mass 37000 was found. This discrepan cy may be explained either by the affinity of the enzyme to polymer matrix or by its very compact structure. To check the first assumption, the enzyme was chromatographed on Sephadex G-75 with and without high concentration of guanidine hydrochloride, the presence of guanidine hydro chloride blockaded interaction with the matrix. As stan dard substances myoglobin, chymotrypsinogen, bovine serum albumin were used. At chromatography with guanidine hydro chloride the molecular mass was 38OO0. although that of the "native" protein (at chromatography without guanidine hydrochloride) was only 22000. The comparison of the mole cular mass of "native" endo- -1,3-giuconase and that of irreversibly inactivated by the natural ingibithor cholis- tonosulphat (My = 778) by polyacrylamid-gel in the pre sence SDS did not indicate any differences.

379 P.2-47 CEREBROSPINAL FLUID LIPIDS ANALYZED BY MEANS OF TLC AND GLC. A COMPARISON TO SERUM LIPID PATTERN АНАЛИЗ ЛИПИДОВ ЦЕРЕБРАЛЬНОЙ ЖИДКОСТИ МЕТОДОМ ТОНКОСЛОЙНОЙ И ГАЗОЖИДКОСТНОЙ ХРОМАТОГРАФИИ J.Tichy, H.Ticha, Prague, &iSR In a group of 300 neurologic patients following techni ques were used to analyze lipid subtractions: the extrac tion procedure of Bligh and Dyer (1959) was adapted to separate lipids from 10 ml of CSF into 10 ml of chloroform (final volume) using three times a mixture chloroform: methanol:water (= csf) 2:4:4. In this system the proteins of CSF precipitate after centrifugation in the interphase 2SS between lower chloroform and upper water-methanolic phase. A) 3 ml of total lipid extract were used for two-step chromatography on silica gel plates (Merck No 5721) of po lar and neutral lipids on the plate. Fractions from-start: lysophosphatidylcholine, sphingomyelin» phosphatidylcho line, sulfatide, phosphatidylethanolamine, cerebroside, free fatty acids, free cholesterol, triglyceride, esterified cholesterol. Solvent l.s chloroform: methanol: water: acetic ac.: n--heptane=65:25:4 :1:10, solvent 11.:n-heptane: diethylether: acetic ac.=95:4:1 (v/v), 1. up to 14 cm, 11. up to 17 cm. Detection by charring at 140 С after cop per -acetate spray, evaluation by means of densitometry (Vitatron app.). B) 2 ml of total lipid extract were used for separation of cholesterol esters into five fractions according to the degree of unsaturation of their fatty acids. Detection and evaluation as under A). C) 5 ml of the totel lipid extract were separated on TLC and the fraction of phosphatidylcholine was scrapped off, trans- methylated by sodium-methoxide in methanol and fatty acid methylesters were analyzed by means of GLC on Perkin-El- mer F-30 apparatus (isothermal at 175 C, 15% DEGS on Chro- mosorb, carrier gas N,). Results t the lipia level in CSF is about 400 times lower than in the serum. In the lipid spectra there are some qualitative differences: in the serum cholesterol- linoleate (50%), in the CSF cholescerpl-oleate (35%) domi nate. Lysophophatidylcholine In the CSF used to be nearly absent. Phosphatidylethanolamine and free cholesterol are present in a higher percentage than in the serum. The amount of linoleic acid in serum phosphatidylcholine re presents about 22% of all FAMES, in CSF only 5%. In a smaller group (n=15) of senile dementia patients we have analyzed two subsequent portions of lumbar liquor. The second portion-proximal or more central ("cerebral") con tains a higher amount of phosphatidylethanolamine, sul- fatides and cerebrosides than the first, more distal one. There are differences in the CSF lipid composition between patients with blood-brain barrier damage (serum-like lipid pattern) and patients with various CSS structures degra dation (tissue-like lipid pattern ).

380 P.2-48 QUANTITATION OF AMINOPHENAZONE, PHENACETIN AND ETHYLMORPHINE HYDROCHLORIDE IN PHARMACEUTICAL DOSAGE FORMS BY HICH PRESSURE LIQUID CHROMATOGRAPHY AND CAPILLARY GAS CHROMATOGRAPHY КОЛИЧЕСТВЕННОЕ ОПРЕДЕЛЕНИЕ АМИН0ФЕНА30НА, ФЕНАЦЕТИНА И ГИДРОХЛОРИДА ЭТИЛМОРФИНА В ФАРМАЦЕВТИЧЕСКИХ ПРЕПАРАТАХ ВЭЖХ И КАПИЛЛЯРНОЙ ГЖХ A.Szemes, L.Gemesi, Budapest, Hungary A reversed phase HPLC method is reported to quantita tion aminophenazone, phenacetin and ethylmorphine hydro- chlorid in pharmaceuticals. Stock solutions were prepa red In MeOH/ 0.05 M KH-PO. aqueous solution. An aliquot 256 of sample solution was injected into a Nucleosil C„ co lumn, eluted with aqueous KH.PO./MeOH (pH 2.5) , containing 0.05 H heptansulfonic acid Ha in aqueous phase. The capa city factors of the three drug molecules were measured as a function of pH. The method was also applied to the deter mination of drug dissolution. Our assay was compared with a GC method developed for quantitation of the three mole cules . 381 P.2-49 GLC AND HPLC DETERMINATION OF ISOSORBIDE DINITRATE (ISDN) AND ITS MONONITRATES IN PHARMACEUTICAL PREPARATIONS ОПРЕДЕЛЕНИЕ И30С0РБИД ДИНИТРАТА /ИСДН/ И ЕГО МОНОНИТРАТОВ В ФАРМАЦЕВТИЧЕСКИХ ПРЕПАРАТАХ МЕТОДАМИ ГЖХ И ВЭЖХ L.Gemesi, A.Szemes, Budapest, Hungary ISDN is widely used as coronary vasodilator for the treatment of angina pectoris, thus many methods have al ready been published for the determination of ISDN and its mononitrates (ISMN) by TLC, polarography, spectropho tometry, GLC and HPLC. We present GLC and HPLC methods which are specific for the determination of ISDN, ISMN and their impurities. As ISDN and ISMN are thermally labile molecules, we have studied the following parameters which affect the decom position of these molecules in the GLC column: temperature of the injector and the coluirn, residence time and surface effects. This work presents the optimum experimental con ditions for quantitative determination of ISDN, ISHU and the impurities both in crude preparations and in fresh and aged pharmaceutical end-products, with packed-column and capillary GLC. Finally the results of the complex formation of ISDN with 3-cyclodextrins were studied. The GLC results were checked by HPLC technique.

382 P.2-50 THE IDENTIFICATION AND DETERMINATION OF N-SUBSTITUTED PIPERAZINES BY TLC AND GC ИДЕНТИФИКАЦИЯ И ОПРЕДЕЛЕНИЕ ЗАМЕЩЕННЫХ ШПЕРАЗИНОВ МЕТОДОМ ТСХ И ГЖХ L.Dimofte, Bucharest, Romania The results of the separation, identification and deter mination of some N-substituted piperazines by TLC and GC are presented. The compounds studied were: piperasine (I), N-hydroxy- ethylpiperazine (II), N,N'-bis(hydroxyethyl)piperasine (III) , N-p-chlorobenzhydryloxyethoxyethylpiperasine (IV), N-phenylbenzyl-N'-hydroxyethoxypiperazine (V), N-p-chlo- rophenylbenzyl-N"-hydroxyethoxypiperazine (Hydroxizin) (VI) and N,N'-bis(p-chlorobenzhydryloxyethoxyethyl)pipe- razine (VII).

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I The literature data are concerned only with the identi fication and determination by TLC and GC of Hydroxizin which is used as a drug /1-6/. The separation and identification of the above-mentio ned compound» have been performed by TLC using the elution system methyl alcohol t ammonia (100:1.5) and spraying with Dragen-dorff reagent. The Rf values of the separated compounds have been measured. For the quantitative determination of the above-mentio ned compounds a new GC method was applied by using a column packed with Silicone SE-ЗО 5% on silanized Gas- Chrom Q (80-100 mesh). He have worked with a Carlo Erba Fractovap 2400 V with FID detector connected to a computer integrator Spectra Physics Minigrator. He have worked with temperature programmation from 120° to 250°C at Ю C/min and a final isotherm of 20 min. The retention times of the above-mentioned compounds have been measured and there after used for the identification of the compounds in various reaction mixtures. The method was used for the determination of the quali tative and quantitative composition of the reaction mix tures in order to follow a given reaction, helping thus to find the best reaction conditions. 1. Analytical Profiles of Drug Substances, vol.7, Ed.Klaus Florey, Academic Press, N.Y.-San Francisco-London, 197B, p.330. 2. C.Cardini, V.Querela, A.Carlo, J.Chromatogr., 37, (1968) 190. 3. Б. van der Kleijn, G.C.Beelen, M.A.Frederink, Clin.Chem. Acta, 34, (1971) 345. 4. B.Caddy, F.Fisch, J.Tranter, Analyst, 99 (1974) 555. 5. P.Rartvig, H.llandl, Acta Phara.Swed., T? (1975) 349. 6. H.G.Fanda, D.C.Hohhrand, J.E.Stamhough, J.Parm. Sci., 68 (1979) 1456. 383 P.4-46 APPLICATION OF FUSED SILICA CAPILLARY MICRO-PACKED COLUMNS IN GAS CHROMATO-MASS-SPECTROHETRY ПРИМЕНЕНИЕ КВАРЦЕВЫХ КАПИЛЛЯРНЫХ МИКРОНАСАДОЧНЫХ КОЛОНОК В ГАЗОВОЙ ХРОМАТО-КАСС-СПЕКТРОМЕТРИИ V.S.Gavrichev, A.R.Jumaev, A.Malik, Yu.M.Borisov, I.A.Lvov, Yu.H.Lyubitov, V.I.Mikhailov, V.D.Sokovikh, V.N.Khokhlov, Moscow, USSR The dependence of separation power of fused silica capillary micro-packed columns (CMPC) has been investiga ted in comparison with open tubular capillary'columns. It has been shown that fused silica CMPC may be used for the analysis of a greater sample than that for open tubular columns. The results obtained from the basis for the application of CMPC in gas chromatograph in combination with maes- spectrometer (in order to incrsase the sensibility of trace analysis). The chromato-mass-spectrometer included

2S8 a monopole mass-spectrometer as the detector and a turbo- molecular pump. Some practical examples of analytical application have been given. 384 P.4-14 CHROMATOGRAPHIC METHODS FOR DETERMINATION OF PHOSPHOLIPIDS FROM RAT BRAIN ХРОМАТОГРАФИЧЕСКИЕ МЕТОДЫ ОПРЕДЕЛЕНИЯ ФОСФОЛИПИДОВ ИЗ МОЗГА КРЫС A.T.Dischkelov, Sofia, Bulgaria Using different chromatographic methods, phospholipids from rat brain in pre and postnatal period of development were studied. Separation of phospholipids from the total lipid fraction was done using column chromatography on "Florisil". The next separation of phospholipids was car ried out on TLC plates "Merck" covered with silica gel- H and fluorescence indicator. The plate was developed in chloroform-methanol-water (65:25:4). Identification of the separated phospholipids was made using Bartlet's method. The fatty acids from the phospholipids were separated and identified as methyl esters on a 5* DEGS column using "Termochrom-23 P" gas chromatograph provided with FID at 190°C. Qualitative evaluation of the separated fatty acids was done using a standard mixture from "Sigma". All results show quantitative and qualitative differen ces in the phospholipids and their fatty acids content of the brain from rat in pre and postnatal stage of develop ment but the highest differences are in phosphati-dilcho- line and phosphatidilethanolamine. 385 P.4-47 DTLC AND GC-MS INVESTIGATION OF (-) DEPRENYL METABOLISM AND FRAGMENTATION ИССЛЕДОВАНИЕ МЕТАБОЛИЗМА И ФРАГМЕНТАЦИИ (-) ДЕПРЕНИЛА МЕТОДОМ ВЫТЕСНИТЕЛЬНОЙ ТОНКОСЛОЙНОЙ ХРОМАТОГРАФИИ И ХРОМАТО-МАСС-СПЕКТРОМЕТРИИ H.Kalasz, J.Knoll, I.Hollo's!, J.Pucsok, Budapest, Hungary (-) Deprenyl (N-methyl-N-propargyl-(2-phenyl-l-methyl) ethyl ammonium hydrochloride), JUHEXR, is a highly selec tive and potent inhibitor of the В form of monoamine oxi dase enzyme. Its metabolism and in vitro fragmentation were followed by displacement thin-layer chromatography (DTLC) and gas chromatography-mass spectrometry (GC-MS). Radiolabeled (-)deprenyl was injected to rats , sun- cutane, the urine was collected for 24 hours, and the radiolabeled metabolites were separated by DTLC. The "in vivo" metabolism of (;) deprenyl was also fol lowed by GC-MS. Fragmentation of (-) deprenyl by oxidizing agent (H-O,) was monitored by DTLC and the decomposition products were identified by GC-MS. Some of the products were the same as

259

1. that of the metabolism of (-) deprenyl suggesting the possibility of similar oxidative dealkylation in vivo and in vitro. 386 P.4-48 ANALYSIS OF HYDROCARBON COMPOUNDS IN EXTRACTS OF HUNGARIAN COALS BY HIGH RESOLUTION GAS CHROMATOGRAPHY/ MASS SPECTROMETRY АНАЛИЗ УГЛЕВОДОРОДОВ В ЭКСТРАКТАХ ЕЕНГЕРСКИХ УГЛЕЙ МЕТОДОМ ГАЗОВОЙ ХРОМДТОГРАФИИ-МАСС-СПЕКТРОМЕТРИИ G.Alexander, I.Hazai, Budapest, Hungary J.Albaiges, J.Grimalt, Barcelona, Spain Diagenetic changes during coalification are recogniza ble through determination of some compounds (molecular markers), which are derived from ancient biota. Even tho ugh these compounds are present in the sample in low con centrations they can serve for characterization. In the present work a set of Hungarian subbituminous coals and related rock samples were investigated. The sam ples were extracted and the hydrocarbons after being fra ctionated were analyzed by high resolution capillary gas chromatography and gas chromatography/mass spectrometry. Analyzing the aliphatic hydrocarbon fractions of coals we found an unusual predominance of even numbered alkanes in the C16-20 region, while the alkanes-with higher mole cular masses showed the usual odd predominance with the maximum at 029. The even dominance in the low molecular region in rock extracts was present even more expressively. This unique alkane distribution is probably caused by the different origin of the alkanes in these regions. The al kanes with low molecular masses (C16-20) are derived from marine sources and may formed from glycerides in the early stage of diagenesis. In consequence of the reducing envi ronment the even dominance (characteristic for fatty acids in living organisms) has saved. On the other hand the alka nes with higher molecular masses are possibly derived from cuticular waxes terrestial plants by a defunctionalization reaction. Many other molecular markers (aliphatic, hydroaromatic and aromatic hydrocarbon compounds) were identified on the basis of their retention behaviour and mass spectra. 387 P.4-49 CHARACTERIZATION OF COALS BY PYROLYSIS - CAPILLARY GAS CHROMATOGRAPHY - MASS SPECTROMETRY ХАРАКТЕРИСТИКА УГЛЕЙ МЕТОДОМ ПИРОЛИТИЧЕСКОИ КАПИЛЛЯРНОЙ ГАЗОВОЙ ХР0МАТ0ГРА<ШИ - МАСС-СПЕКТРОМЕТРИИ M.Blazso, .E.Jakab, Budapest, Hungary It is believed that for coals, the tar consists of mini mally distrubed fragments of the original organic matrix. The functional group composition of tars for a bituminous coal is remarquably similar to the functional group compo sition of the parent material according to their elementar 260 composition IR spectra and NMR spectra /]./. Thus the low temperature pyrolysis,a process which appears to be cont rolled by combined effects of pyrolysis and vaporization, results in products characterizing th-э coal structure. The mass spectrometric analysis of coal pyrolysis products for coal characterization was extensively used by Meuzelaar /2/. However, in the case of sulfur containing coals and of low rank coals, the separating efficiency of a capilla ry column is needed, in order to distinguish between cer tain fragment ions, since ions of some characteristic mass numbers may originate equally from aromatic hydrocar bons, oxygen or sulfur containing compounds. The selective ion monitoring of the capillary column pyrograms at mass numbers of basic importance acquires data for the characterization of coals. A set of selective ion pyrograms will be presented for coals of different rank and different sulfur content at mass numbers repre senting alkanes, isoalkanes, cycloalkanes, alkyl aromatics, alkyl indenes, alkyl phenols, fluorenes thiols and thiophenes. 1. P.R.Solomon, K.R.Squire, R.M.Carnagelo, ЛСБ Div. or Fuel Chemistry Preprints, 2_9 (1984) 10. 2. H.L.C.Meuzelaar, Q.M.Carper, G.R.Hill, P.H.Given, Fuel, £3 (1984) 640. 388 P.4-50 CONFIRMATION OF IDENTITY OF DDT-TYPE RESIDUES IN MEDICINAL PLANTS BY CHEMICAL TRANSFORMATION AND GC/MS ИДЕНТИФИКАЦИЯ СОЕДИНЕНИИ ТИПА-ДДТ В ЛЕКАРСТВЕННЫХ РАСТЕНИЯХ МЕТОДОМ ХИМИЧЕСКОЙ ТРАНСФОРМАЦИИ И ХРОМАТО-МАСС-СПЕКТРОМЕТРИИ R.Beneeke, Leipzig, GDR R.A. de Zeeuw, Groningen, The Netherlands For the gas chromatographic analysis of DDT isomers and metabolites in extracts of Flores Chammomiilae and Radix Valerianae, the separation on a packed QF-1/0V-17 column was compared with various capillary columns of the CP-Sil type. Identification of the individual compounds could be achieved by comparing the retention behaviour, chemical transformation of DDT and DDE by treatment with KOH and chromium (VI)-oxide, respectively, as well as by capilla ry GC-MS using single ion monitoring of substance-charac teristic ion masses. It this way, residues of p, p'-DDT; o, p'-DDT; p,p'-DDE and p,p*-TDE could be identified and the quantitative character of the transformation could be established. Based on these results a routine method for the quality control analysis of medicinal plants utilizing packed columns before and after chemical transformation was deve loped. The method worked well in both the qualitative and quantitative analysis of DDT residues in all medicinal plants tested.

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i 389 P.4-51 QUADRUPOLE MASS SPECTROMETRY APPLIED IN NATURAL PRODUCTS INVESTIGATION КВАДРУПОЛЬНАЯ МАСС-СПЕКТРОМЕТРИЯ В ИЗУЧЕНИЙ ПРИРОДНЫХ СОЕДИНЕНИИ F.De Малек, B.Pollet, M.Lesleur, C.Claude, Rueil-Malmaison, Prance Natural products analysis is still considered as a wide area in analytical chemistry. The problems encounted range from light products to rather heavy, polar and thermolabi- le ones. Therefore the analytical techniques vary accor ding to the nature of the investigated products. In our poster presentation we will emphasize the capabilities offered by a quadrupole mass spectrometer. According to the inlet systems, a gas chromatograph, a high performance liquid chromatograph, different classes of natural com pounds can be pertinently investigated. Additionally, we will also show that a mass spectrometer used as an inlet system, therefore—the use of a MS/MS instrument is nowa days a tool which offers new step in selectivity, the most outstanding feature in complex mixture analysis.

390 P.4-52 INVESTIGATION OF THE BRANCHED POLYETHYLENE IHIDE AS A STATIONARY LIQUID PHASE AND A REAGENT IN THE SOBSTRACTION METHOD ИЗУЧЕНИЕ РАЗВЕТВЛЕННОГО ПОЛИЭТИЛЕНИМИДА В КАЧЕСТВЕ НЕПОДВИЖНОЙ ЖИДКОЙ ФАЗЫ И РЕАГЕНТА В МЕТОДЕ ВЫЧИТАНИЯ M.Gancheva, Sofia, Bulgaria - V.G.Berezkin, P.A.Gembitsky, T.P.Popova, Moscow, USSR The chromatographic properties of the branched polyethy lene imide as a stationary liquid phase in the capillary gas chromatography were investigatei The retention charac teristics for different kinds of hydrocarbons - alcohols, esters and ethers and other compounds were measured. The application of the polyethylene imide as a reagent in the substraction method was studied. The polyethylene imide substracts selectively aldehydes, glycols. Practical instances testifying about adventages of use of the polyethylene imide as SLP and a reagent for substraction in gas chromatography are shown.

262

I. 391 P 4~"53 THE INVESTIGATION OF NATURE AND THE DEVELOPMENT OF THE DETERMINATION METHOD OF GROUP COMPOSITION OF SULPHIDES OF GASOIL FRACTIONS BY CHROMATO-MASS SPECTROMETRIC METHOD ИССЛЕДОВАНИЕ СОСТАВА И РАЗРАБОТКА МЕТОДИКИ ОПРЕДЕЛЕНИЯ ГРУППОВОГО СОСТАВА СУЛЬФИДОВ ГАЗОИЛЕВЫХ ФРАКЦИИ НЕФГЕЙ МЕТОДОМ ХРОМАТО-МАСС-СПЕКТРОМЕТРИИ E.A.Kruglov, Yu.N.Popov, T.V.Portnova, S.I.Ezhova, Ufa, USSR The present mass spectral method of determination of sulphides group composition have essential drawbacks. These drawbacks are due to the superioposition on the molecular masses fragments of high molecular weight com pounds and fragments of other classes of compounds, for example, hydrocarbons. The use of the chromato-mass spectromatric method with highly efficient column allcws to avoid these difficul ties and obtain more reliable and valid data. The procedure is based on the fragments analysis of peaks masses of chromatogram of each molecular mass of sulphides separation of sulphides molecular ions and determination of the average number of hydrocarbon atom of each group. Coefficient of sensitivity fos each group have been taken from the literature. The use of SO m capillary column allowed to separate the mixture to be analysed essentially into individual compounds and un ambiguously isolate sulphides. Comparison of the data obtained by chromato-mass spectral method and the common mass spectral method showed that the group composition of sulphides including alkylsulphides, mono-, di-, trl-, tetra- and pentathio- cyclanes have essential differences (in some cases up to 50%).

392 P. 4-54 INVESTIGATION OF THE COMPOSITION OF CONDENSATION PRODUCTS OF ACETONE BY THE METHOD OF CHROMATO-MASS SPECTROMETRY ИССЛЕДОВАНИЕ СОСТАВА ПРОДУКТОВ КОНДЕНСАЦИИ АЦЕТОНА МЕТОДОМ ХРОМАТО-МАСС-СПЕКТРОМЕТРИИ K.M.Meisberg, Ii.F.Korzhova, Y ; ."n opov, T.V.Portnova, Ufa, USSR To investigate the composition of petrochemical syn thesis complex product with different classes of organic compounds GC/MS system with highly efficient capillary columns, computer and library mass spectra were used. But in some instances the results were not successful. The reason is that the mass spectra of cyclic hydrocar bons and cyclic ketone are very similar. A combination of two methods was used for the complete identification of all components. At first, the products were investi gated as a whole, then they were separated on silica gel 263 by the method of preparative elution chromatography. In the next step, the separated fractions of hydrocarbons and oxygen-containing compounds were studied by the chromato- maes spectrometer under the same conditions as in the case of the original fraction. AB a result of this procedure more than 50 compounds were detected: CQ-C.- alkyl cyclohexenes and alkyl cyclo- hexanes, unsaturated and saturated ketones both aliphatic and cyclic series from acetone to C,, and some other com pounds as well. In terms of identification data we can judge of the mechanism of acetone condensation and hydrocondensation processes. The obtained data were applied to the chromatograms of capillary gas chromatographs and were used while studying kinetics and activity of different catalyst samples.

393 P.2-44 GLC AND HPLC OF STERIC AND STRUCTURAL ANALOGS OF ESTRANE ГЖХ И ВЭЖХ ПРОСТРАНСТВЕННЫХ И СТРУКТУРНЫХ АНАЛОГОВ ЭСТРАНА I.E.Pakhomova, A.V.Platonova, M.V.Dodonov, S.N.Ananchenko, Moscow, USSR The chromatographic behaviour of steric and structural analogs of estrane and 18-methylestrane was studied by GLC and HPLC. These compounds synthesized in the laboratory of steroid chemistry of Shemyakin Institute of Bioorganic Chemistry show high biological activity in regulating the vital processes in human and animal organisms. These ana logs were chromatographed both as unprotected estrones and estradiols and in the form of 3-Me-, 3-Ac-, 3-TFac- ethers. It is shown that some pairs of steric analogs (8j8 and BtL ; BeL, 14/3 and 9£ , 14jg) can be identified only with the help of the combination of GLC and HPLC. The substituent at C17-atom (ketone or hydroxyl) can be de fined from the shape of absorption isotherm. The influence of ether-group at C3-atom on the retention time (RT) was shown. The additivity of log(RT) for the analogs was obser ved. This permits to predict the RT of new analogs of estrane and 18-methylestrane. The liquid phases of various polarity were used in GLC capillary and packed columns. For HPLC "DuPont-830" was used in semi-preparative and analytical regimes on. Lichrosorb-Sl 60-7 ;u and ultrasphe- re-Si 5 AI columns.

394 S.5-5 POLISH LIQUID-CRYSTALLINE STATIONARY PHASES ПОЛЬСКИЕ СТАЦИОНАРНЫЕ ФАЗЫ НА ОСНОВЕ ЖИДКИХ КРИСТАЛЛОВ Z.Witkiewicz, Warsaw, Poland Liquid crystals are finding ever wider application in gas chromatography. They present highly selective statio nary phases and are therefore particularly useful for se- 244 parating isomers. At the same time gas chromatography pre sents a method for testing the properties of liquid crys tals. for some years studies are conducted in Poland in which new liquid-crystalline stationary phases are seeked and their physico-chemical and separating properties tested. In the course of these studied several dozen new liquid crystals were obtained with properties suitable for their application in gas chromatography. These liquid crystals cover a wide applicability temperature "range from low temperature to high temperature ones. Some of these high temperature liquid crystals may be used even at temperatu res exceeding 300°C. Apart from conventional rod-like liquid crystals also their polymers as well as disc-like ones (hexasubstituted triphenylenes and benzenes) have been synthesized. The physico-chemical tests concentrated on the interac tions of liquid crystals with different supports as well as on the effects of the support on the phase transition temperature in dependence on the amount of the liquid crystal deposited. The polarity of the tested stationary phases was also determined. Alongside individual phases the properties of phase mixtures were also tested. A relationship was observed between the structures of the molecules of liquid crystals and the properties of the latter as stationary phases. It was found that in the case of mixed phases the increased separating ability with respect to the chromatographed mixtures is related to the lowered melting temperature of the liquid crystal mixture. The tested phases were used chiefly in conventional analytical columns but they also found application in micropacked ;-.nd capillary columns. Good separations of mixtu res belonging to different groups of compounds were obtai ned. Aliphatic through alkylaromatic (including disubstituted benzene isomers) to polycyclic hydrocarbons were tested as well as esters, ethers, et al. The tests carried out as well as the analyses of the results allowed us to propose a set of stationary phases with the most advantageous properties. These phases are manufactured and offered on a commercial scale.

395 P.4-28 CHANGE IN FATTY ACID LIPID COMPOSITION OF REPRODUCTIVE ORGANS OF ALMOND AND PEACH AS A RESULT OF FERTILIZATION ИЗМЕНЕНИЕ ЖИРНОКИСЛОТНОГО СОСТАВА ЛИПИДОВ РЕПРОДУКТИВНЫХ ОРГАНОВ МИНДАЛЯ И ПЕРСИКА В РЕЗУЛЬТАТЕ ОПЛОДОТВОРЕНИЯ A.A.Rikhter, Yalta, USSR During the plants fertilization, the lipid complex plays an important role. Thus, in the process of plants pollination, papillae of stigma secret a liquid which con tains lipids. In this work we have studied the change in composition

of free fatty acids and those found in lipids 'Ся -C22 )

18.3«к.872 ,-c in developing anthers and pistils of almond and peach. The ]ipids were extracted with the mixture chloroform-methanol (2:1). The fatty acid composition of lipids was determined after saponification by means of GLC, under conditions of temperature programming from 120 till 185 С, with column length 2 m, diameter 2 mm, filled with 15% degs on the Chromosorb W AW. It was stated that in tissues of anthers in the stage of one-celled pollen. In the lipids palmitic, oleic, linoleic and linolenic acids prevail, whereas in the anthers in the stage of two-celled pollen, palmitic, linoleic and linolenic acids predominate in lipids (16,5-19,1%, 20.4- 25.2%2and 19.2-22.5%, respectively). These acids mainly present in pistils. In ovules of almond and peach palmitic, oleic, linoleic and linolenic acids were found after fertilization. Lino lenic acid content in the ovules reaches 12-14% which is somewhat lower than in anthers and pistils. The fatty acids conversion in tissues of developing anthers is directed on accumulation of linoleic and lino lenic acids which is probably connected with desaturation of oleic acid. Linolenic acid formation in lipids of ovule tissues is evidently blockaded as the oleic acid accumula tion continues. The data show that the structural organi zation of reproductive tissues in fruit plants is closely related to fatty acid composition of lipids, and the fer tilization process is accompanied with a deep change in fatty acid metabolism.

396 P.4-29 COMPARATIVE ANALYSIS OF VOLATILE ORGANIC PLANT CONSTITUENTS BY MEANS OF GAS LIQUID CHROMATOGRAPHY СРАВНИТЕЛЬНЫЙ АНАЛИЗ ЛЕТУЧИХ ОРГАНИЧЕСКИХ ВЕЩЕСТВ РАСТЕНИЙ МЕТОДАМИ ГА30ЩЦК0СТН0Н ХРОМАТОГРАФИИ Yu.A.Akimov, Yalta, USSR То study dynamics of accumulation and emission of vola tile organic substances (VOS) by plants, a diagram of the comparative analysis using various methods of GLC has been developed. The diagram includes consecutive or parallelic determi nation of VOS contents and composition in plants emitted by plant» in air under natural growth conditions and in laboratory, analysis of products obtained by means of wa ter distillation of plant material. Determination of VOS in a plant is carried out by means of rapid heating of the plant material in special devices and introducing the vapors into the chromatographic column. Vos emitted by a plant in air were determined Con centrating them on polymeric sorbents of kinds of Polysorb and Porapak, with subsequent temperature desorption, intermediate concentration and chromatography. Under natu ral conditions, the determination was carried out with isolation of certain plant parts, and in laboratory - using the "bucket* method. For obtaining and analysing the 266 water distillation products, conventional methods have been used. Given diagram allow to estimate efficiently the quan titative and qualitative parameters of VOS accumulation and emission, influence of ecological conditions on them, presence of bound fonts of volatile compounds and simpli fies the identification of constituents. For quantitative estimation of VOS of plants a simpli fied method with recording the compounds analysed as one peak is proposed.

397 P.5-66 MULTIPLE MEASUREMENT AND HIGH RESOLUTION GAS CHROMATOGRAPHY BASED ON "ZICHROMAT" WITH A DOUBLE OVEN ГАЗОВАЯ ХРОМАТОГРАФИЯ С МНОГОКРАТНЫМ ИЗМЕРЕНИЕМ И ВЫСОКОРАЗРЕЫАЮЩЕЙ СПОСОБНОСТЬЮ НА БАЗЕ "ЗИХРОМАТ" С. ДВОЙНОЙ ПЕЧЫО F.Muller, Karlsruhe, FRG Abstract not arrived on time for printing.

398 SEPARATION OF URINARY PEPTIDES BY HPLC РАЗДЕЛЕНИЕ ПЕПТИДОВ МОЧИ МЕТОДОМ ВЭЖХ J.Macek, L.tliterova, M.Adam, Prague, CSSR Enhancement degradation of connective tissue compounds takes place in almost all diseases. Products of this degra dation are partially excreted in urine. The monitoring of these fragments may be an important method for studying of some pathological processes. In order to isolate fraction of urinary peptides having molecular weight larger than 4000, gel permeation chroma tography on Bio-Gel P 4 was used. Further separation was accomplished by reversed phase HPLC. The Beckmann Ultrapo- re RPSC column (75 mm x 4.6 mm I.D.) with 30 nm mean pore size was used. The i.iobile phase A was deionized water a adjusted with trifluoroacetic acid to pH 2.2, the mobile phase В was acetonitrile. After 4 min isocratic step (100%) a linear gradient to 65% A 35S В was applied for 5 min followed by isocratic elution for 6 min. In 15 min we were able to obtain 30 peaks on a chromatogram. The mobile phase flow-rate was 0.8 ml/min, column temperature 40°C. The column eluent was monitored at 230 nm. 49 samples were analysed - 18 from healthy controls and 31 from patients with rheumatoid arthritis. Areas of indi vidual peaks normalized for 24 hours urine volume were compared. It was found that studied groups differed signi ficantly in four peaks with retention times about 10 rain. These peaks were isolated and further characterized for amino acids and sugars content. 267 399 HPLC SEPARATION OF OLIGONUCLEOTIDES РАЗДЕЛЕНИЕ ОЛИГОНУКЛЕОТИДОВ МЕТОДОМ ВЭЖХ A.H.Csarnyi, M.Vajda, Budapest, Hungary The synthesis of alternating oligonucleotides is get ting more and more important, as oligonucleotides of pre scribed sequence are potentially useful in genetic engi neering. For this purpose a very high purity of the compounds is essential.. Compounds of sufficient purity can only be obtained if the preparative and the analytical separation methods are sufficiently accurate and selective. Seveial HPLC methods were tried (experiments on an APS column and phosphate buffer; reverse-phase chromatogra phy on an ODS column with a buffer and methanol) till optimal separation of d(AT),, d(TAT), d(TATA), d

400 STABILITIES AND ION EXCHANGE PROPERTIES OF SOLVENT IMPREGNATED SULFONATE ION EXCHANGERS WITH LOW AND EXTRA LOW CAPACITIES СТАБИЛЬНОСТЬ И ИОНООБМЕННЫЕ СВОЙСТВА ПРОПИТАННЫХ РАСТВОРИТЕЛЕМ СУЛЬФОНАТНЫХ И0Н00БМЕННИКОВ С НИЗКОЙ И КРАЙНЕ НИЗКОЙ ЕМКОСТЬЮ D.N.Muraviev, Moscow, USSR E.Hdgfeldt, Stockholm, Sweden Samples of sulfonate ion exchangers with widely vari able capacities (from 0.0014 to 0.11 mgequiv/g) have been obtained by a wet impregnation method using toluene solu tions of dinonylnaphthalene sulfonic acid (HD) for modi fying beads of polystyrene-divinylbenzene copolymer (Am- berlite XAD-2). Capacities of impregnated resins are directly proportional to the concentration of the modi fying solution. Stabilities of resins impregnated with HD depend on the initial values of their capacities. Samples with high specific capacity (0.08 mgequiv/g dry polymer and more) are less stable than those with low capacities. Absolute capacity losses depend mostly on the number of ion exchange cycles that have been carried out. The results of studying ion exchange equilibria in the system Na+ - H+ on resins with different capacities have shown that the-ion exchangers under investigation ha'e similar properties for both organic solutions of HD and for conventional sulfonated resins. Increasing capacities of resins lead to a decreasing selectivity towards sodium ions. It has been shown that, the simple model used for 268

L treating ion exchange data obtained with solid and liquid ion exchangers may also be used in the case of solvent im pregnated resins. It has been shown that beads of resins with different capacities have different values of surface charge density, that may be considered as one of the possible ways for interpreting the dependency between ion exchange equilib rium constant and capacity of the resin.

401 REVERSED-PHASE LIQUID CHROMATOGRAPHY OF HYDROPHILIC PROTOLYTES BY USE OF ION-PAIRING PROBES 0БРАЩЕНН0-ФА30БАЯ ЖИДКОСТНАЯ ХРОМАТОГРАФИЯ ГИДРОФИЛЬНЫХ ПРОТОЛИТОВ С ИСПОЛЬЗОВАНИЕМ ИОН-ПАРНЫХ ПРОБ E.Arvidsson, G.Schill, D.Westerlund, Uppsala, Sweden Injection of a solute, without inherent detectable pro perties, in reversed-phase liquid-solid systems using de tectable ions in the mobile phase, will give respons by affecting the distribution of the detectable ion between the mobile and solid phase /1-5/. The chromatogram con tains peaks with constant retention characteristic for the system (system peaks) and one sample peak for each solute injected. The background absorbance is fairly high, and the sample peaks can be positive and negative depending on the retention relative to the system peak. The respons, and thereby the detectable amount, is dependent on the relative retention of the peaks. This sensitivity has a maximum when the sample peak comes close to the retained system peak. The sensitivity and the number of solutes detected can be improved by using more than one detectable ion. It is also, in these systems, possible to follow more than one UV-absorbing ion and the concentration changes of the detectable ions in different zones can be observed. The systems were used for the detection of mono- and polyvalent hydrophilic protolytes. A hydrophobic polystyre ne phase, was used as the solid phase. Its hydrophobic cha racter facilitate the retention of the hydrophilic solutes and its stability over a large pH-range makes it possible to exploit the protolytic properties of the compounds to a full extent. The effect of changes in the concentration of the ions included in the mobile phase has been investigated, and retention of mono- and divalent ions was compared. In these systems the retention is affected by changes of most ions included in the mobile phase. 1. M.Denkert, L.Hackzell, G.Schill, E.Sjogren, J.Chromatogr., 218 (1981) 31. 2. L.Hackzell, G.Schill, Chromatographia, 15 (1982) 437. 3. L.Hackzell, T.Rydberg, G.Schill, J.Chromatogr., 282 (1983) 179. 4. J.Crommen, J.Pharm. Biomed. Anal., 1 ,(1983), 549. . 5. J.Crommen, P.Heme, J. Pharm. Biomed. Anal., 2 (1984) 241.

269

/ 402 DIRECT SEPARATION OP ENANTIOMERS BY USE OF LIQUID- LIQUID CHROMATOGRAPHY ПРЯМОЕ РАЗДЕЛЕНИЕ ЭНАНТИОМЕРОВ МЕТОДОМ ЖИДКОСТНО- ЖИДКОСТНОЙ ХРОМТОГРАФИИ E.Heldin, C.Pettersson, H.W.Stuurman, Uppsala, Sweden Separation of enantiomers is of great importance espe cially in the field of biomedical science. Many drug mole cules are optically active and the two enantiomeric forms can have quite different p; jSiological activity /1/. Liquid - liquid chromatographic methods for separation of enantiomeric aminoalcohols, e.g., ephedrine and practolol, with DBT (dibutylester of L-tartaric acid) as the chiral complexing agent is presented. The aminoalcohols are distributed with hexaflourophosphate as counter ion to the organic phase (ion-pair chromatography). In reversed phase systems the chiral ester, DBT, was applied as a liquid stationary phase or. the support, Phe nyl Hypersil. Symmetrical peaks with separation factors of 1.1 to 1.2 were obtained for the enantiomers of ephedrine and analogues. The system was stable for more than two months /2/. Retention was regulated by the counter ion con centration. In straight phase systems DBT was added to the organic phase (dichloromethane). In these systems the ephedrine analogues gave somewhat higher stereoselectivity (separa tion factors 1.2 to 1.3 ) than the reversed phase systems and furthermore it was possible to separate the enantiouers of some /'-receptor blocking agents (e.g. practolol) .In creasing the concentration of DBT in the mobile phase gave no significant effect on the stereoselectivity, while the retention of the enantiomers decreased. 1. E.J.Ariens et al. Stereochemistry and Biological Acti vity of Drugs, 31acwell Scientific Publications, Oxford, 1983. 2. C.Pettersson and H.W.Stuurman, J. Chromatogr. Sci. 22 (1984),441,

270 AUTHOR INDEX ADAM M., 089, 398 BENECKE R. , 388 AGHATHONOS P., 022 BENKOVITCH Y-J.I. , 364 AGITSKY G.Yu., 298 BENEV D.K., 327 AHUJA S., 066 BERANEK I., 191 AKIMOV Yu.A., 396 BEREK D., 088,110,120,322, ALBAIGES J., 386 323 ALEKSEEVA L.B., 194 BEREZKIM V.G., 151,289,383, ALEXANDER G., 386 390 ALEXANDROV M.L., 061 BEREZKINA L.G., 236 ALEXANDROV S.D., 244 BEREZVAI E.Vincze-Pal, 205 ALEXEEVA K.V., 176 BERG van den J.H.M., 067 ALIEV V.K., 111 BERLIZOV Yu.S., 172, 273 ALYMOV A.M., 259 BEYRICH Th., 092 ANANCHENCO S,N., 393 BEZZUBOV A.A., 202 ANDERS G., 266 BIACS P.A., 360 ANDERS K., 266 BIDLO-IGLOY M., 205 ANDERSON A.A., 165 BIEGANOWSKA M., 145 AJJDRIANOVA I. P., 101 ANDRONIKASHVILI T.G., 318, BIENERT M., 098 ANDROSHINA I.A., 081 1 BLAZSO M., 387 ANDRYSIAK A., 335 BLEDZKI A.. 104 ANH-TOAN N., 095 BODHAR J. 360 ARATSKOVA A.A., 341 BOGATYRYOV .L. , 041 ARTUKOV A.A., 378 BOGDANOVA O.A., 327 ARUTUNYAN A.M., Ill BOGHER A.M., 126 ARUTUNYAN Z.A. , 221 BOIKO A.I., 234 ARVIDSSON E., 401 BOJARSKI J., 083 ATABEKYAN T.A., 038 BOLOTINA N.E., 347 AYRAPETYAN S.S., 311 BONDARENKO I.V., 350 BONDAREV V.B. , 248 BONDARYUK O.M., 211 BABAYAN S.G., 111,311 BORDAS В., 058 BABICH P.A., 302 BORISOV Yu.M., 383 BABSKI V.G. , 297 BORISOVA S.I., 236 BAKAEVA T.I., 034 BORODAI L.V., 190 3ALLSCHMITER K., 016 BOROSS F,, 371 EALOGH L. , 043 BOROSSAY J., 181 BANAKH O.S., 342 BORISENKO S.N., 299 BANNIKOVA G.E., 081 BOUfiOET G., 226 BARAM G.I., 051, 145 BOZEK P., 120, BARANOVA N.I., 201 BRANDSTETEROVA Ё*., 096 , 361 BARLA F., 106, 138 BRATSLAVSKAYA A.L., 322 BARANSKY R.I., 342 BREZGUNOV V.N., 306 BARLOW G.H., 277 BROWN P.R. , 057 BAYER E., 0O9 BRZOBOHATA H., 321 BATHORI M., 127 BRUGMANN L., 196 BEBRIS N.K., 324 BUDOVICH V.L., 350 BEGLARYAN A.A., 311 BUDTOV V.P., 071 BELYAEV N.F. , 338 BURCK D., 2B5 BELENKII B.C., 0O4, 143 BURYAN P., 273 BELODED L.N., 212 BUTKIEWICZ K., 099 BEL'NOV V.K., 049 BOTOV Yu.S., 056 BELOV V.V., 042 BUTZKE P., 048 BELYAKOVA L.D., 346 BOZINKAIOVA Т., 192

271 САР К., 180 ECKNIG W., 146 CELLAR P., 269 EFENDIEV A.A., 320 CHARNETSKAYA L.M., 342 EGOROV I.A., 202 CHAYLYAN S.G., 111 EGOROVA K.V., 338 CHERESHNYA O.P., 174 EL'KIN G.E., 038, 039 CHIANTORE 0., 088 ELOMAA M.A., 179 CHIKIN G.A., 032, 03S ELTEKOV Vu.A., 131, 136 CHIZHKOV V.P., 007, 261 ELTEKOVA N.A. , 325 CHIZOVA H.B.,, 039 ELYAKOVA L.A., 073, 378 CHLENOV M.A., 079, 132 EMETS L.V., 046 CHUPKAKOV Yu.D. , 035 ENGEWALD W., 170, 369 CHURA6EK J., 048, 265 ENNET D., 368 CHUVELEVA E.A., 042 ERDELMEIER C.A.S. , 355 CHUVILYAEVA N.I., 186 ERISTAVI V.D., 033 CIGANKOVA M., 087 CLAUDE C., 389 ERMOSHKIN A.E., 078 CLAUSS H., 317 ERNST W., 232 CLEVELAND C., 277 EROSHENKOVA G.V., 015 COUPEK J., 309 ESIN M.S., 338 Van CRUCHTEN J.H.M., 252 EVREINOV V.V. , 068 CSAKVARI В., 210 EVSEEVA Z.A., 331 CSAPd &., 103 EZHELEVA A.E., 244 CSARNYI A.H., 129, 399 EZHOVA S.L., 391 CSEH G., 208 CSERHATI ?., 058, 204 FARKAS F., 138 CZELAXOWSKI И., 335 FARXAS P., 271 CZERWINSKI W., 231 FAZAXAS L., 178 FEDOTOVA G.I., 302 FEILER D., 199 FELL A.F., 050 FELT V., 203 DACHEVA M., 293 FELTL L., 319 DALLEHBACH-TOLKE K., 356 FESENKO V.P., 301 DAVANKOV V.A. , 017 FIEDLER G., 124 DAVYDOV V.Ya., 119 FILATOVA Z.I., 247 DAVYDOVA S.L., 238 FILIPPOV Уu.S., 193 DAWES E.F., 257 FILONENKO G.V., 168 DAWES Г.А., 257 FISCHER W., 294 DEMENTYEV A.A. , 073 FIRSOVA L.A. , 042 DEMYANOV P.I., 197 FODOR-CSORBA K., 307 DIETRICH P., 108 FRANZ P., 196 DIMITROV Chr., 326, 328 FREIDGEIM L.I., 211 DIMOFTE L. , 382 FRIDMAN E.A., 353 DISCHKELOV A.T. , 384 FOGGERTH E., 283 DISKINA D.E., 185 FONTOV V.N. , 246 DMITRIKOV V.P., 084,172,273 FUZESI-KARDOS K. , 371 DODONOV M.V., 393 DOLEZALOVA И. , 128 GABP.IO Th., 368 DOLPHIN J., 217 GAIBAKYAN D.S. , 290 DOMBROVSKY V.A. , 132 GALACZ J., 210, 226 DRESSLER II. , 214 GALEV E.E., 242 DUBROVA H.A. , 241 GALITSKAYA N.B., 331 DUFEK P., 075 GANCHEVA M., 390 DOFKA 0., 267, 272 GANKINA E.S., O04 DOTKA P. , 307 GANZLER K., lOO DZHABAWOV D.N., 153 GARTZKE J., 285 DZHAfARIDZE M.T. , 245 GARAJ J"., 096,158,251,322,361 272 GARCIA-RASO A., 166, 167 HELFFERICH F.G., 013 GARCIA-RASO J., 166, 167 HENDEL P. , 199 GASPARIC J., 282 HENKE M., 329 GAVRICHEV V.S., 383 HERMENAU H., 199 GAVRILOVA T.B., 326, 328, 348 HERZSCHUH R. , 170 GAVRILYUK I.I. , 292 HOFFMANN H., 329 GAVRYUSHKIN A.V., 306 HOHN H., 285 GAZDA K., 3S1 HOLLAN S.R., 095 GEBRUNE A.B., 353 HOLLOSI I., 385 GELMAN B.G., 162 HORNA A., 267 GEMBITSKY P.A., 390 HORVXTH M 051 GEMESI L., 380, 381 H0SCHKE A 360 GERASIMENKO V.A., 172 HRIVNAK J. 262, 263, 268 GERGELY S., 251 HROUZEK J 269 GIDDINGS J.C., Oil HUBER J.F.K., 001 GLADYSHEV P.P., 303 HOBL W., 072 GLOCKNER G., 067 HULM M., 118 GOBER В., 122 HUSEK P., 203 GOLDINA V.G., 338 GOLOS I.Ya., 342 ILAVSKY J., 262 GOLOVNYA R.V., 008,076,163,229 ILMOJA K. , 367 GONCHARYONOK V.M., 194 ILIYIN N.P., 289 GORBUNOV A.A., 069,107,134 ILNITSKY Yu.S., 036 GORDETZOV A.C., 174 IOFFE B.V., 010 GORSHKOV A.V., 068 1SACK0V A.A., 111 GORSHKOV V.I., 029, 049 ISENBEKG A., 270 GRACHEV M.A. , 061 IVANOV St., 326, 328 GRAFHETTEROVA J., 12 3 IVANOV V.A., 049 GRAMOLIN S.L., 366 IVANOVA M.M., 245 GREBENYUK V.D. 030 GREGOIRE J., 159,184,239 JAKAB E., 387 GRIGORYEVA D.N., 076, 163 JANAK J., 026 GRIGORYEVA V.D., 077 JANOERA P., 048, 053 GRIMALT J., 386 JANOS E., 204 GRIMM U., 092 JARCNIEC J.A., 054 GROBLER A., 178 JIRA Th., 092 GOBITZ G., 121 JIRSA M. , 090, 286 GURYANOVA V.V. , 133 JOST W., 294 JUMAEV A.R., 383 HABERLANO D., 225 JURKOVA M., 116 HAEUPKE K., 329 JURASEK A., 322 HAIS I.M., 206 JUVANCZ Z., 129 HAJDU F., 360 HALASZ A., 296 KAISER R.E., 002 HALDNA V., 367 KALABINA L.I., 213 HALFPENNY A.P. , 057 KALACHEVA N.I., 240 HANSEN S.H., 060 KALASHNIKOVA E.V. , 156 HARA S., 147 KALASZ H., 097,280,281,385 HARITONOV O.V. , 042 KALCHENKO O.I., 198 HARTINGER A. , 121 KALININ A.I., 212 HAUCK H.E., 294 KALININ N.Ph., 313 HAZAI I., 386 KALINITCHEV A.I., 024, 037 HEINSOO E.J. , 334 KALJURAND M. , 175 HEISE K.H., 114 KALMANOVSKY V.I., 164 HELBOE P., 060 KAL6Y K., 094 HELDIN E., 402 KAMENIKOVA I., 195 273 KANEV A.S., 113, 291 KOVAR P. , 142 KARAISKAKIS G., 022 KOWALSKI W.J., 223 KARAPETYAN A.A., 111 KRAFT R., 109 KARGMAN V.B., 044, 045, 3: KRASOTSKII S.G., 244, 254 KARLINA L.V., 034 KKATCHANOV Ch.G., 359 KARLOV N.V., 019 KRAUS G., 270, 316 KASYHOVA G.A., 299 KRAUS§ G.-J., 080 KATRVKHA G.S., 375 KRAXNER H. , 106 KATRUKHA S.P., 112 KREjfil M., 093 KATSANOS H.A. , 022 KRIVOLAPOV S.S., 348 KATYKHIN G.S., 113, 291 KRIZ J., 265 KAZAKEVITCH Yu.V. , 136 KROKHIN V.A., 274 KAUFMANN K.-D. , 055 KROTENKO V.E. , 301 KELEMEN-HALLER A., 138 KRUGLOV E.A. , 391 КЕМША W., 086 KRUPCIK J., 192, 251 KENNDLER E., 001 KRYLOV V.A., 244, 254 KERECSEN L. , 097 KRYUCHKOVA O.V., 034 KERSTEN D. , 122 KUDRYASHOV L.I., 079 KERTSCHER U., 055, 098 KUDRYAVTSEVA S.P., 042 KERY A., 284, 357 KUKBS V.G., 112 KEVBR E.E., 143 KULLIK E., 175 KHALILOV K.F., 015, 020 KUNINGAS K.R,, 171 KHAZEL M.Yu., 031 KUNZ P.A., 249, 258 KHITAROV A.V., 220, 221 KUNZE M.E., 277 KHOKHLOV V.N.,117,207,213, KUPRIENKO V.I., 30O

KURGANOV A.AW 017 222 383 KURKINA V.V., 218 KHOKHLOVA T.D., 074 XUSMINYKH V.A., 035 KIRYUKHIN I.F., ЗОО, 301 KUSZ P., 335 KLIMA J., 123 KOTKINA O.P., 126 KLINTSOVA S.P., ЗОО, 301 KUZNETSOV A. A. , 014 KLOPOV V.I., 338 KUZNETSOVA N.B., 338 KLYACHKO Yu.A., 364 KVASNICKOVA E., 206 KNAUSZ D., 129, 210, 226 KNOLL J., 097, 280, 385 KOBAC V.V., 259 LAGUCHEV A.S., 019 KOLCHIN Yu.A., 259, 276 LAKSZNER K., 157 KOLESNIC G.B., 366 LANDER A.V., 211 KOLEVA R.S., 139 LANHAM J.W., 277 KOLIKOV V.M., 354 LANSKOVA T.G., 177 KOLOMIETS L.N., 213 LAPERASHVILI L.Ya., 318 KOLOTINSKAYA E.V. , 037 LAPIN B.P. , 140, 377 KOL'TSOV P.A., 056 IARIKOV V.V. , 227 KOMAREK, K., 265 LARIN A.V. , 02 3 KOPYLQVA V.D., 044 LARSKY E.G., 115 KORDAC V., 090, 286 LAVRENOVA A. S . , 190 KOROL A.N., 168 LAZARIS A.Ya., 202 KOROLEV A.A. , 260 LAZAREV V.A. , 254 KOROLYOVA E.M. , 143 LEHOTAY J., 096, 361 KORZHOVA E.F., 392 LEIKIN Yu.A., 333 KOSIK O.G., 3O0, 301 LEISZTNER L., 095 KOTAL P., 090, 286 LEITOLD A. , 065 KOTOVA I.A. , 117 LEMBERKOVICS E., 374 KOTSIS E., 094 LENCHIC N.V., 1J3 KOUfclLOVA D., 093 LEONT'EVA S.A., 228 KOVACHEV G., 189, 293 LESIEDR M., 389 KOVALEVA N.V. , 336 LE^NIAK K., 141

274 LEUKBFELD W., 224 MINAEV V.A., 259, 276 LEVINA N.B., 125 MINCHEVA Т., 293 LEWIS M.L., 277 MINDLIN L.O., 233, 241 LICHTMAN T.V., 201 MIRGORODSKAYA O.A., 061 LXNDBERG J.J., 179 MISHARINA T.A. , 163 LIPAVSKY V.N., 219 MISHINA E.A., 174 LISICHKIN G.V., 310 MITEROVA L., 089, 398 LITCHEV V.I.» 202 MLADENOV I., 162 LITVIN E.F., 007 MOCAK J., 251 LOBAZOV A.F., 143 MOHNKE M., 196 LOMOTSEVA T.I., 247 MOKEEVA R.N., 232 LOPATIN S.A., 081 MOLEVA L.K., 375 LUKYANOV A.E. , 107 MOLLER E., 225 LUNSKII M.Kh., 186 MORAVKOVA V., 272 LVOV I.A., 383 MOROZ P.A., 222 LYUBITOV YU.M., 383 MORRISON D., 277 MOSTOVNIKOV V.A., 143 MAACK de F., 389 MUCH H. , 108 MACAK J., 273 MUCHIN L.M., 154 MACEK J., 089, 398 MULLER F., 397 MACHOVA J., 304, 305 MULLER K.-D., 317 MACKO Т., 088, 110 MUNARI F., 161 MADARASSY-№RSICH E.,371 MURADOV H.H., 125 MAKARENKO N.P., 078 MUSILEK V., 116 MAKAROVN.V., 302, 375 MYAGKOV E.A., 350 MAKAROVA S.B., 314,330,332 MYLYNSKAYA I.N., 247 MAKEEV D.M., 376 ИАКиСН В., 351 NABIVACH V.I1. , 172, 273 MALIK A., 383 NADIRADZE N.A., 118 MALTSEV V.G., 004 NAGEL M., 199 MALYKH Ya.N., 049 NAGY G., 181 MANN G., 170 NAGY J., 280, 281 MARES P., 256 NAGYVARADI E.M., 2 37 MARTASEK P., 286 NAVARRO X., 288 MARTIN-AYUSO S., 288 NAVRATIL J.D., 028 MARTIROSSYAN J.G., 311 NAZAROV P.P. , 042 MARTON J., 262 NAZAROVA V.I. , 156 MASLOVA G.B., 042 NAZIMOV I.V. 125 MATISOVA E., 255 NECHAEV S.V. , 143 MATORINA N.N., 034 NEMETS V.M. , 246 MATOUSEK P., 267, 272 NETSCH W. , 108 MATYSIK G., 287 NEUMANOVA E., 319 MCHEDLISHVILI B.V., 254 NICLAS H.-J., 108 MEHLIS В., 055, 098 NICULITCHEVA S.I., 117 MEIER В., 355, 356 NIKITIN Yu.S., 074, 324 MEISNER I.S., 137 NIKOLOV R.N., 070, 243 MEKSS P.A., 165 NIKONOROV E.A., 228 MELENEVSKY А.Т., 039, 349 NILSSON D., 064 MELNIKOV V.N., 372 NIOTIS A. , 022 MELNIKOVA S.V. , 236 NOBILIS M., 206 MERTEN H., 215 NONDEK L., 322 MESZTICZKY A., 210 NOVAK I., 120, 322, 323 MICHALKOVA A., 268 NOVAK M., 128 MIKHAILOV V. I. , 383 NOVIKOVA Т.Е., 056 MIKICHUR N.I., 144 NOVOTNY M., 005 MIKITA K., 355 NYREDY Sz., 355, 356

275 OBERDORFER E., 225 POLLET В., 389 OBSEPYAN A.M., 221 POLSTER H.-J., 146 OEHLKE J. , 055 POLYAKOVA E.A., 135 OGLOBLINA I.P., 245 POMAZANOV V.V., 259,276 OKHOTNIKOV B.P., 350 PONOMAREVA E.L., 071 ONORIN S.A., 313 PONOMAREVA T.M. , 017 OPPERMANN G. , 160,215 POPOV Yu.N., 391,392 ORLOV A.M., 019 POPOVA M.I1., 359 OROS G.Yu., 032 POPOVA T.P., 260,390 OSTROVSKY I., 269,270 POPP P., 160,215 OVSEPYAN A.M., 220 PORSCH B., 090 PORSHNEV N.V. , 248 PADALKINA V.S., 364 PORTNOVA T.V., 391,392 PAIZAHSKAYA I.L., 186 POSHKUS P.P., 345 PAKHOMOV V.P., 302 POTTER H. , 118 PAKHOMOVA B.I., 338 PRAT J., 288 PAKHOMOVA I.E., 393 PRILEPSKAYA K.K., 233 PANINAL.I., 314,330,331 PROKHOROV A.M., 019 PANTSURKIN V.I., 227 PRYAKHINA V.M. , 234 PAPTSOVA I.I. , 046 PRZYBOROWSKA M., 150 PARSHINA I.V., 326 PSCHEIDL H., 225 PASECHNIK V.A., 107,135,35 POSCOK J., 385 PASTUSHENKO V.G., 348 PUNNING K., 040 PATRYKIEJEW A., 315,340 PYATYGIN A.A., 324 PAVLENKO N.U., 275,363 RABE H., 102 PAVLOV A.V. 133 RADDA E., 124 PAYER K. , 129 RAISZ I., 025 PEKOV G.I., 343 RAKHOVSKAYA S.M., 347 PENTCHUK J., 367 RANG S.A., 171 PERELROYZEN M.P., 145 RAPOPORT L.M., 238 PERL I., 209 RASO M.A., 166 PETTERSSON C, 402 RATONYI I., 094 PETRENKO V.F., 298 RAY C.A., 258 PETROSYAN V.S., 197 REICH G. , OOl PETRO-TURZA И. , 371 REICHMANN G., 102 PETROV Yu. N. , 019 REINHARO J., 266 PETROVA M.L., 332 REINHAEDT K., 092 PETROVA R.S. , 156 PETSEVN., 326,328,343 REMEN J,,268 PHILITOVA M.B., 378 REPKA D., 251 PICK J., 095 RETL V., 195 PIMENOV A.M., 055,137 REZNIKOV V.I., 020 PINTER-SZAKACS M. , 209 RIEDER R.J., 002 PIRKLE W.H., 063 RIGIN V.I., 230 PIROGOV V.S., 349 RIJKS J.A., 252 PISAREVA N .A. , 140,337 RIKHTER A.A., 395 PISARIKOVA A. , 304,305 RODEL W., 373 PISHEV D.I., 243 ROGOVIK V.I., 342 PLANK L.D., 277 ROGOZHIN S.V.- 081 PLATONOVA A.V. , 393 ROHDS K.-H., 196 PLHACKOVA K. , 304 ROMANOV N.A., 247 PODMANICZKY L., 157 ROSHCHIN S.A., 135 PODOSYENOVA N.G., 071 ROSHCHINA T.M., 328 POGOSBEKYAN G.V., 247 ROTIN V.A., 162 POLANUER 3.M., 229 ROTZSCHE H., 317 POLYANSKAYA N.Y., 353 P6ZYL0 J.K.,054,290 RUDOLFI T.A., 233,241 276 RYAZANOV V.I., 227 SHUKUN S.A., 306 RYBIN V.K., 375 SHUSHUNOVA A.F., 235 RUBTSOVA Т.Е., 140 SHYMANSKA M.V., 165 RUDENKO B.A., 153, 201 SIDOROV R.I., 169 RUSIN0V L.A., 218 SIMON J. , 138 ROSTOV В., 102 SIMON R.K., 258 RZEPA J. , 105 , 250 SIRONI A., 161 SIZOVA N.V., 377 SAFONOV M.S., 49 SKORNYAKOV V.V. , 045 SAHARTOVA O.V. , 77 SKVORTSOV A.M., 069, 134 SAJEWICZ И. , 250, 105 Sl.AIS K. , 093 SAKODYNSKII K.I.,254,314,О SLAVUTSKAYA N.I., 376 SALDADZE K.M., 27,44,45 SLEDZINSKA В OSS SALO S., 179 SLIWIOK J., 105 SAMOLYUK S.I., 190 SMIRNOV A.V. , 332 SAMSONOV G.V., 38,39,349 SMIRNOV V.A., 338 SAPOZHNIKOV У.М., 117 SMOLKOVA-KEOLEMANSOVA E.,319 SARAVALLE C.A. , 161 SMOLYANINOV G.A., 193 SARKISSYAN E.H. , 311 SMRZ* M. , 048 SARSUNOVA M. , 268 SOCZEWINSKI E., 279, 287 SASS-KISS A., 370 SOJAK L., 269, 270, 271 SAURA-CALIXTO F., 166, 16 7 SOKOLOVA G.V., 244 SAZONOV M.L., 186 SOKOLOVA L.I., 076 SCHILL G., 62, 401 SOKOLOVA L.P., 045 SCHIRUTSCHKE M., 215 SCHLENKRICH H., 368 SOKOVIKH V.D., 383 SCHNEIDER G., 91 SOLDATOV V.S., 312 SCHWACHULA G., 329 SOLOMATINA L.S., 176 SCOBELEVA S.E., 174 SOLOMONOV A.P. , 115 SEBEKOVA K., 120 SOLOV'EV A.A., 246 SEDIVY J. , 123 SOLTES L., 323 SEGURA R., 288 SOLYOM J., 129 SELEMENEV N.F., 032 SOMER T., 040 SEMENOV V.A., 274 SOTNIKOV E.E., 173, 177 SEMENOVA V.A. , 274 SOVA V.V., 378 SEMINA G.N. , 337 SPYCHAJ T., 104 SEMENOVSKAYA T.D., 37 SROLER A., 206 SEMIONOV S.N., 014 STAROVEROV S.M., 310 SENIN N.N., 193 STEFANOVA A.D., 24 3 SERDAN A.A., 310 STEIMANN H., 317 SERGIEVSKY V.V. , 245 STEIN J., 098 SHAPOSHNIK V.A., 47 STERKHOV N.V., 007, 261 SHARAEVA Т.К., 298 STICHER D., 355, 356 SHATIRISHVILI I.SH., 365 STOEV G., 182 SHATZ V.D., 77 STOLYAROV B.V., 242 SHCHEDRINA M.M. , 233, 241 STOTTMEISTER E., 199 SHCHERBAKOVA K.D., 156 STOYANOVA G.P., 188 SBEEHAN T., 249 STRENZE T.V., 171 SHEPETYUK L.V., 34 STROKINA L.M., 346 SHIRYAEVA V.E., 260 STRUCHALINA T.I., 126 SHIROKOV V.P., 352 STRUPPE H.G., 266 SHKLYAEV V.S., 227 STUORMAN H.W., 402 SHMIDEL E.B., 213 STYSKIN E.L., 115 SHONIA N.K., 324 SULTANOVICH Yu.A. 366 SHTARK M.B., 144 SOPINA W.R. 253 SUPRYNOWICZ Z. 264, 340

277 SVETASHEVA T.G., 378 UJSZASZI K., 129, 226 SVETLOVA N.I., 076 UNGER K.K., 308 SVITELSKI V.P., 198 URALETS U.P., 274 SWANN D.F.K., 216 SYBILSKA D., 083, 187 VAJDA J., 095 SZABO-FORRAS Zs., 370 VAJDA W., 399 SZAKACS-DOBOZI M., 296 VALKO K., 059 SZANTO J., 051 VARADI L., 280 SZARFOLDI-SZALMA I., 371 VARGEK И.,208 SZCZEPANIAK W., 183, 344 VARLAMOV V.P., 081 SZEDERKENYI F. , 205, 210 VEISSERIK J., 040 SZEMES A., 380, 381 VEITH J., 224 SZENDREI K., 127 VELEV V.N., 202 SZEPESY L., 051, 157 VERESS T., 051 VERZAR-PETRI G., 103,284,374 TARABA B. , 180 VESPALEC R., 087, 321 TCHERNYAGA B.S., 376 VETROVA Z.P., 341 TEANDER D., 372 VIGDERGAUZ U.S., 012 TELEKY-VAMOSSY Gy., 371 VIGH fTy. , 065 TELKOVA T.N., 132 VINOGRADOV B.A., 275, 363 TEREKHOVA G.P., 330 VINTINA O.G., 117 TER-STEPAHYAN E.C., 220,22 VISKA J., 265 TESARIK K., 265 VITENBERG A.G., 240 TETENYI P., 357 VLASENKO E.V., 328 THOMSEN M., 060 VLASOVSKAYA O.N., 330 TICHA H., 379 VOITOVICH V.B., 031 TICHY J., 379 VOLF L.A., 046 TIHANYI E., 205 VOLKHIN V.V., 313 TIKHOMIROVA L.P., 117 VOLKOV S.A. , 015,020,173.193 TIKHONOV Yu.V., 018,056,1 VOLMUT J., 200 TIMOFEEVA A.N. , 228 VOLODIN V.S., 303 TITOVA E.V., 079 VOLOVENKO V.O. , 198 TODD P., 277 VOLYNETS M.P., 289 TOGUZOV R.T., 018,056,137 VONDRACEK M., 305 TOLIKINA N.F. , 193 VOSKRESENSKY N.M., 049 TOMORI E., 208 VRCHIABSKY H., 321 TOPALOVA I., 326, 328 TORKOS K., 181 WAINER I.W., 062 TORtlASI E., 208 HEISBERG K.H., 392 TOROSYAN G.K., 177 HEISSFLOG W., 270 ТбТН A., 094 WENNRICH L. , J69 TOTH Т., 237, 25.2 WESTERLUND D.., 064, 401 TRESTIANU S., 161 NIGAND G., 224 TROITSKY G.V., 278, 298 WINTERSTEIGER R. , 121 TRUBNIKOV V.I., 117 WITKIEWICZ Z., 021,264,^35, TRYBITSINA E.S., 364 394 TSARFIN Y.A., 232 WOLFBEIS O.S., 121 TSODIKOV M.V. , 260 WOLFENBERCER A., 253 TOLUPOVE.E., 152, 194 WOLFF M., 122 TURIAK Gy., 284, 357 WORONTSOV A.M., 113, 291 TYIHAK E., 003, Or8 WRONSKI 3., 021 TYMCZYNSKI R. , 148 WULFSON A.N.,- 130 TYUTYUNNIK V.I. , 372 WURST M., 116 UGOLEV D.A., 362 YAKIMOV S.A. , 130 UHLIR M., 191 YAKUBOVA M.A., 019 278 YAKUSHIN M.A., 274 ZEEUW de R.A., 388 YANOVSKII S.M., 155 ZELVENSKY V.Yu., 190 YASHIN Ya.I., 052, 341 ZEMAN I., 082 YEGOROVA S.A., 347 ZEMANOVA I., 116 YUDINA I.P., 337 ZEMEK J., 120 YUDOVITCH V.B., 117 ZHAKEVICH M.L., 140 YORIN O.A. , 132 ZENKOV A.P., 145 YUSTIN V.S., 247 ZHILTSOVA L.I., 186 ZHUKHOVITSKII A.A., 006 ZHUKOV M.Yu., 297 ZABOKRITSKY И.Р., 007 ZHURAVLEVA I.L., 229 ZAGOREVSKAYA E.V. , 336 ZILBERMAN H.V., 313 ZALINYAH V.P., 220, 221 ZOLOTAREV P.P., 014 ZAMANSKII S.M., 015 ZOTIKOV E.C., 071 ZAFOROZHETS A.I., 362 ZSOLDOS L. , 360 ZASCHKE H., 270 ZURABASHVILI Z.A., 207 ZAVYALOV V.P., 306 ZVYAGINTSEVA T.N., 073

279 CONTENTS

GENERAL PROBLEMS OF CHROMATOGRAPHY 3 ION-EXCHANGE CHROMATOCPAPHY 20 LIQUID CHROMATOGRAPHY 33

GAS CHROMATOGRAPHY 105

CAPILLARY CHROMATOGRAPHY 16g

THIN-LAYER CHROMATOGRAPHY AND ELECTROPHORESIS 18< SORBENTS 206 PREPARATIVE CHROMATOGRAPHY 235 CHROMATOGRAPHY OF NATURAL PRODUCTS 243

COMBINATION OF VARIOUS TYPES OF CHROMATOGRAPHY 254 AUTHOR INDEX 271

Издше осуществлено с оршпоши, подготовлеявого к петая Иясяпутом фкапкша хами АН СССР

V ДУНАЙСКИЙ СИМПОЗИУМ ПО ХРОМАТОГРАФИИ Тюкы wHCjaan « гтшдоша сообаяшЛ (и «гл. шп)

Утверждено к печати Иистюутом фктасков хшшш АН СССР

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