Determination of Psychotropic Phenylalkylamine Derivatives in Biological Matrices by High-Performance Liquid Chromatography with Photodiode-Array Detection

;,,,,.,alo/ChromatograS93p(199_a_hy, 7-94 /k_)__¢_._ Elsevier Science Publishers B.V., Amsterdam

CHROMSYMP. 2429

Determination of psychotropic phenylalkylamine derivatives in biological matrices by high-performance liquid chromatography with photodiode-array detection

Hans-J6rg Helmlin and Rudolf Brenneisen* Instituteof Pharmacy, University of Berne, Baltzerstrasse 5, CH~3012 Berne (Switzerland)

ABSTRACT

Several procedures using high-performance liquid chromatography with photodiode-array detection have been developed to create phytochemical and toxicological profiles of phenylalkylamine derivatives in biological samples (e.g. plant materials and urine). Mesca- line-containing cactus samples were extracted with basic methanol, using methoxamine as internal standard; the extraction and clean- up of urine samples were performed on cation-exchange solid-phase extraction columns. The extracts were separated on a 3-#m ODS column with acetonitrile-water-phosphoric acid-hexylamine as the mobile phase. Peak detection was performed a_ 198 or 205 nm; peak identity and homogeneity were ascertained by on-line scanning of the UV spectra from 190 to 300 nm. The detection limit of phenylalkylamine derivatives in urine and cactus material was 0.026-0.056 #g/ml and 0.04 t_g/mg, respectively. Following a single oral dose of 1.7 mg/kg methylenedioxymethylamphetamine (MDMA) the concentrations found in urine ranged from 1.48 to 5.05 #gml MDMA and 0.07-0.90 #g/mi methylenedioxyamphetamine (a metabolite of MDMA). The mescaline content of the cactus Trichocereus ?achanoi varied between 1.09 and 23.75 #g/mg.

INTRODUCTION South America, especially in the Andean regions and belongs, together with Lophophora williamsii Many natural and synthetic phenylalkylamine (Leto. ex Salm-Dyck) Coult. [4], to the mescaline- derivatives such as mescaline, amphetamine, meth- containing cactus species which are commercially ylamphetamine and 4-bromo-2,5-dimethoxyphen- available without legal restrictions in Switzerland ethylamine (see Table I) are known for their stim- and other European countries. ulant and/or hallucinogenic properties. Some very Considering the potential of abuse of phenyl- active ring-substituted amphetamines such as 3,4- alkylamine derivatives and mescaline-containing methylenedioxyamphetamine (MDA), 3,4-methyle- cactus species, it was the aim of this work to devel- nedioxymethylamphetamine (MDMA) and 3,4- op a selective, specific and sensitive analytical pro- methylenedioxyethylamphetamine (MDE) have cedureusinghigh-performanceliquidchromatogra- now appeared as "designer drugs" on the illicit pry with photodiode-array detection (HPLC- market, produced by clandestine laboratories [1]. DAD). This method should allow not only the iden- Owing to their high potential of abuse, most of tification of such compounds, but also the acquisi- these popular recreational substances are now in- tion ofphytochemical and pharmacokinetic profiles ternationally controlled. Despite this, MDMA in complex biological samples to estimate their toxi- I"Ecstasy", "XTC", "Adam") is used more fre- cological or therapeutic potency. The efficiency of quently as a controversial adjunct in psychotherapy HPLC-DAD in drug analysis, analytical toxicol- [2,3].Trichocereus pachanoi Britt. et Rose ("San Pe- ogy, forensic chemistry and phytochemistry of psy- dro") grows in subtropical and temperate areas of chotropic drugs has been shown previously [5-18].

"[_ TABLE I & -_._'_ STRUCTURES OF PHENYLALKYLAMINE DERIVATIVES I

s,",'_ Structure Peak no. Compound

[[__ NHa I Amphetamine t

2 3,4-Methylenedioxyamphetamine (MDA)

._/_ ! L_[[t'_YA_ N _ 3 Methylamphetamine '3 s

4 4 4-Methoxyamphetamine '" H.,,CO' v

[_"'_ NHa 5 Phentermine )

[J_ [ 6 3,4-Methylenedioxymethamphetamine (MDMA) °X..o/ ! CH30 i__ _,__ A. T NHa 7 5-Methoxy-3,4-methylenedioxyamphetamine (MMDA) '

H3COi_NH2 8 3,4,5-Trimethoxyamphetamine H3CO' T t OCH3 H I

' 9 3,4-Methylenedioxyethylamphetamine(MDE) !

HaCO j_NHa 10 2,5-Dimethoxyamphetamine '_ ["'_ /_"' 'OCH3

11 4-Bromo-2,5-dimethoxyphenylethylamine (DOBP,2-CB) HaCOj._/-_jNH2 Br" '"'cf -OCH3 H3CO j_NHa 12 2,5-Dimethoxy-4-methylamphetamine (DOM,STP) /' _ 'OCH_

· (Continued on p. 89)

4 LENNEISEN HPLC OF PHENYLALKYLAMINE DERIVATIVES 89

TABLE ] (continued)

Structure PeakNo. Compound

H3C_ [_NH2 13 4-Bromo-2,5-dimethoxyamphetamine (DOB)

'14 2,5-Dimethoxy-4-ethylamphetamine (DOET) V _ 'OCH3

15 Mescaline H3CO i_ ''._N H2 H_C0' T I., OH OCH3'1"

H3CO _NH2I,_,II 16 Methoxamine v 'OCH3

}

EXPERIMENTAL through a membrane filter (regenerated cellulose, 0.45 #m, Schleicher and Schuell) and degassed by Instrumentation sonication and during use with a constant flow of The HPLC-DAD system consisted of a Hewlett- helium. Methanol was used for washing the col- 0 Packard1090Mliquidchromatograph(Hewlett- umn. Packard, Waldbronn, Germany), an HP 1090L au- The separation of urine samples containing tosampler, an HP 1040M photodiode~array detec- MDMA and MDMA metabolites was performed _IMDA) tor, an HP 79994A Chemstation (software version isocratically at 40°C on a 125 x 4.0 I.D. column 1.05),an HP 7470A x/y plotter and an HP 2225A packed with 3-$tm Spherisorb ODS-1. The mobile Thinkjet printer, phase was acetonitrile-water (72:928, v/v; 57:943, w/w), containing 5.0 ml (8.5 g) orthophosphoric Chromatographic conditions acid (85%) and 0.28 ml (0.22 g) hexylamine per The separation of fourteen phenylalkylamine de- 1000 ml. The flow-rate was 0.8 mi/min. rivatives was performed at 40°C on a 125 x 4.0 mm The separation of the cactus samples was per- I.D. column packed with 3-tim Spherisorb ODS-1 formed isocratically at 25°C on a 150 x 4.6 mm (Phase Separations), filled by Stagroma (Wallisel- I.D. column with a 20 x 4.0 mm I.D. precolumn, len, Switzerland). The solvent gradient was devel- packed with 3-ttm Spherisorb ODS-1. The mobile oped by using the CARTAGO (computer assisted phase was acetonitrile-water (108:892, v/v), con- retention time prediction and gradient optimiza- taining 5.0 ml (8.5 g) orthophosphoric acid (85%) tion) software; details of this procedure are publish- and 0.28 mi (0.22 g) hexylamine per 1000 ml. The ed elsewhere [19,20]. Solvent A was water contain- flow-rate was I ml/min. ing 5.0 ml (8.5 g) orthophosphoric acid (85%) and BP,2-CB) 0.28 ml (0.22 g) hexylamine per 1000 mi; solvent B Chemicals and reagents was acetonitrile containing 100 mi water, 5.0 ml (8.5 Amphetamine sulphate was obtained from Sieg- g) orthophosphoric acid (85%) and 0.28 mi (0.22 g) fried (Zofingen, Switzerland) and methylampheta- _TP) hexylamineper 1000 ml. The gradient profile was as mine hydrochloride from Dr. Grogg Chemie follows: 0_10.6 min, 5.5% B in A (isocratic); 10.6- (Berne, Switzerland). Mescaline hydrochloride was 21.6 min, 5.5-39% B in A (linear gradient). The supplied by Laboratoires Plan (Geneva, Switzer- !inuedonp.89) flow-rate was 0.8 mi/min. The eluent was filtered land), methoxamine hydrochloride and MDA were 90 H.-J. HELMLIN, R. BRENNEISEN

'l provided by Sigma (St. Louis, MO, USA). MDMA, x 0.5 mi of methanol-hydrochloric acid (7.3%; ,._ MDE, 5-methoxy-3,4-methylenedioxyampheta- 97.5:2.5) at a flow-rate of about 0.5 ml/min. Ali- mine (MMDA), 4-methoxyamphetamine, 2,5-di- quots of 10 #1 of the defined volume of the eluates methoxyamphetamine, 2,5-dimethoxy-4-methylam- were injected into the HPLC-DAD system for the 5 phetamine (DOM,STP), 2,5-dimethoxy-4-ethylam- determination of MDMA and MDA. For !ow phetamine (DOET), 3,4,5-trimethoxyampheta- MDMA and MDA levels a concentration stepmay mine, 4-bromo-2,5-dimethoxyamphetamine (DOB) be necessary. An aliquot of 1.5 ml ofthe eluates was and phentermine were donated by the Division of added to 68 #1 of I M KeHPO4, concentrated to ":; Narcotic Drugs, United Nations (Vienna, Austria). about 100 #1 under a stream of nitrogen and recon- 4-Bromo-2,5-dimethoxyphenethylamine (DOBP,2- stituted to 150.0 #1 with methanol-water (50:50). _ CB) was a gift of the Swiss Association for Psycho- The resulting solution, with a pH of about 4-5, was lyric Therapy. All other chemicals and reagents then sonicated for 2 min and filtered through the tip ';7, were of HPLC or analytical-reagent grade and were of a Pasteur pipette filled with cotton wool. Aliqu- , purchased from Merck (Darmstadt, Germany) or ots of 10/al were used for determination by HPLC- Fluka(Buchs,Switzerland). DAD. The cactus samples were cut in half, lyophilized ._ Urine and cactus samples and stored in a desiccator under vacuum and pro- The urine samples were obtained from patients tected from light until used for analysis. A repre- treated with MDMA by psychiatrists of the Swiss sentative sample of the cactus specimen was pulver- Association for Psycholytic Therapy. Urine sam- ized with a grinder. An accurately weighed amount pies were collected approximately 6 h after the ad- of the powdered sample (about l0 mg) was then ministration of 1.7 mg/kg MDMA. The specimens washed four times with 1 mi of diethyl ether by so- of Trichocereus pachanoi Britt. et Rose and Lo- nication (5 min) and filtration through a 0.2-#m re- phophora diffusa (Croizat) Bravo (Cactaceae) were generated cellulose filter (Spartan 13/30, Schleicher bought at flower shops and shopping centres in & Schuell). The defatted sample was extracted four Switzerland, or obtained from private collections, times with 0.5 ml of methanol-ammonia (33%; 99:1), containing 150.0 #g/ml methoxamine hydro- ., Sample preparation chloride as internal standard (I.S.), by sonication (5 The extraction and clean-up of urine samples min) and filtration through a 0.2-#m regenerated , (real, spiked, blank) were carried out on Adsorbex cellulose filter. Aliquots of 5 #1 were injected into

' SCX (100 mg) cation-exchange extraction columns the HPLC-DAD system, g.i (Merck),usingan AdsorbexSPUsampleprepara- tion unit. Frozen urine samples(stored at -20°C) Quantitation were warmed to room temperature in an ultrasonic Quantitation of the urine samples was performed [ bath and centrifuged, if necessary (2000 g for 5 at 198 nm by measuring the peak areas of MDMA ' min). An aliquot of 1.0 mi was added to 0.5 ml of and MDA and using the external standard method. i ] 0.05 M KH2PO4 and then sonicated for 1 min in a The calibration graphs for MDMA and MDA (lin- _' stoppered 2.5-ml vial. The Adsorbex columns were ear regression analysis) were obtained by analyzing f preconditioned with 2ml of methanol, I ml of water twice pooled blank urine spiked with MDMA and i and I ml of 0.017 M KH2PO4. The sample was then MDA in the concentration ranges 0.5-17 and 0.08- i applied to the preconditioned extraction column, 1.6 _g/ml, respectively. The extraction was per- which was not allowed to dry out at the end of the formed as described earlier. Quantitation of the i the preconditioning step. The vial was rinsed with cactus samples was performed by measuring the 0.05 ml of 0.05 M KH2PO4. After drying the extrac- peak areas of mescaline and the I.S. at 205 nm. The tion column for about 1 min, urine interferences calibration graph was obtained by measuring three were removed by washing the cartridge with 3 x times standard solutions in the concentration range il 0.5 ml of 0.017 M KH2PO4 and I mi of methanol, 20-75 _g/ml mescaline with an addition of 128/_g/ 'i followed by drying the column for about 1 rain un- ml I.S. (aqueous solution, calculated as base).

dervacuum.Theelutionstepwascarriedoutwith4 i ...... 91 loric' acid (7'3%, Precision Recovery study 0.5 ml/m.m. Ali,. Theinter-day precision of MDMA in urine was The recoveries of MDMA and MDA from urine [me of the eluates determinedby analyzing two pooled urine samples specimens were measured with the spiked samples iD system for the spikedwith 1.8 and 8.8 #g/mi MDMA. The inter- used for the determination of the precision. The re- iMDA. For low day precision of MDA in urine was determined by covery of mescaline from cactus material was deter- atration step may analyzingtwo pooled urine samples spiked with 0.4 mined with a dried and pulverized mescaline-free 0ftheeluateswas and 0.8 #g/ml MDA. The intra-day precision of cactus sample (Lophophora diffusa) spiked with a ; concentrated to mescalinein the cactus material was determined by solution of 1 mg/ml mescaline (corresponding to 10 irogen and recon- analyzinga dried and pulverized mescaline-free cac- #g/rog dried material). The efficiency of the extrac- ol-water (50:50). lus specimen (Lophophora diffusa) spiked with a tion and clean-up procedures was calculated by r>fabout 4-5, was methanolic solution of 1 rog/mi mescaline (corre- comparing the peak areas of MDMA, MDA and :d through the tip sponding to 10#g/mg dried material). The solvent mescaline with those of similar aqueous standard Iton wool. Aliqu- wasevaporated before analysis. All analyses were solutions. All analyses were performed three times ration by HPLC- repeatedthree times on two different days during a following the described procedures, but without one-week period using the described procedures, adding I.S. for the cactus material. half, lyophilized vacuumandpro- 10 12 nalysis. A repre- 3oe. a :linen was pulver- weighed amount 10rog)wasthen 8 ethyletherby so- 2ee. )ugha0.2-#mre- 3 14 13/30, Schleicher = 2 11 as extractedfour E _45 ammonia (33%; 1oe- hoxamine hydro-

!-t_mregenerated _ereby sonicationinjectedinto(5 -_ _ i__ _ 00 _ Jb _s 2b 2s Time (mln ) 10o- _ b _'swas performed areas of MDMA tandardmethod. 80. kandMDA(lin- g

dthMDMAand ; 2, 6

'actionwasper- 0.5-17and0.08-mtitation of the _ 40-'______x_ 1,3 y measuring the ew v-_'x ;.measuringat 205 nm.threeThe e _ _ -_'_-'__ _centration range 2_ 220 2,/e _60 2ee 3_0 :!ition of 128pgi _q_vele,',a:h (,,m) ed as base). Fig. 1. (a) Chromatogram recorded at 198 nm and (b) on-line UV spectra of a standard mixture of phenylalkylamine derivatives. For peaks° see Table I. Chromatographic conditions as described under Experimental. 92 H.-J. HELMLIN, R. BRENNEISEN l

C/_ RESULTSAND DISCUSSION 210 nm) where the phenylalkylamine derivatives ex- ___ hibitthehighestabsorption.At theoptimumdetec- HPLC-DAD tion wavelengths of 198 and 205 nm the sensitivity Among the reversed-phase materials tested (Cs, is approximately ten times greater for MDMA and g .?,4 Cls), with particle sizes of 3, 4 and 5/_m and from MDA (log _los 4.557 and 4.569) and more than 60 t different manufacturers, only the 3-#m spherical times greater for mescaline (log _2o5 4.675) com- C_s phase with a minimum plate number of pared with the absorption maxima and detection : 120 000/m showed the efficiency necessary to obtain wavelengths reported previously (233-234 nm for j_ the HPLC profiles of a complex mixture of fourteen MDMA and MDA [25,26] and 268 nm for mesca- structurally related phenylalkylamine derivatives line [27]). The detection limit for MDA at 198 nra t (Fig. la) and to produce peak shapes which were and a signal-to-noise ratio of 5:1 was about 0.03 , '_-] generally sharp and symmetrical. It is well known /_g/ml in urine (Table II). The excellent sensitivity, 7' that basic compounds may show a pronounced tail- the wide !inearity range and the good overall repro- ing effect on certain reversed-phase columns due to ducibility allow the detection and reliable determi- interactions with the residual polar silanol groups nation ofphenylalkylamine derivatives in biological of the stationary phase [21,22]. The addition of an matrices, even at very iow concentrations. , aminemodifierto the mobilephaseas a masking I agent for the silanol groups improves the peak Determination of MDMA and MDA in urine shape and changes the capacity factor (k') of basic The use of solid-phase extraction as an alterna- substances [23,24]. It has to be noted that the selective to liquid-liquid extraction for the isolation of r tivity of the chromatographic system can be widely xenobiotics from body fluids is recommended be- ! lX influenced by changing not only the ratio of aceto- cause of excellent recoveries and ease of operation _ ' nitrile-water but also the concentration of hexyl- [5,18]. The sample clean-up of small volumes of t_ amine. With the addition of orthophosphoric acid urine containing MDMA and its main metabolite lX to the mobile phase, an acidic eluent with a pH of MDA can be performed rapidly and effectively by ti approximately 2 is obtained, so that the compo- the use of short cation-exchange solid-phase extrac- h nents of interest, such as MDMA, MDA, mescaline tion columns. Fig. 2 shows that MDMA and MDA $ e and other basic phenylalkylamine derivatives, are are well resolved and separated from the endoge- e ti '-r' protonated and eluted as associates with phosphate ]' ti . ions. The gradient was designed by the application TABLE11 of the CARTAGOsoftware,a computer-based e_ method development tool which can be extremely VALIDATIONDATA FOR MDMA AND MDA useful in the selection and optimization of chro- i matographic conditions for complex mixtures. The Parameter MDMA MDA details of CARTAGO are reported elsewhere _ [19,20]. Linearity range01gJml) 0.5-17 0.08-1.6 Correlationcoefficient 0.999 0.999 ' The selectivity of HPLC is significantly improved Mean recovery(%: n = 3): ; by coupling with a photodiode-array detector, al- 0.4 #g/ml sample - 97.7 lowing peak identification through the retention 0.8/_g/mlsample - 100.7 ?, time and UV spectrum as well as peak purity checks 1.8/_g/mlsample 98.5 - E through up-slope, apex and down-slope spectra 8.8 #g/misample 99.2 - matching. Fig. lb, with the UV spectra (190-300 Mean0.4 #gprecision/mi sample(#g:ml:C.V.,%; n = 6) - 0.39;11.6 nm) of amphetamine sulphate, methylamphetamine 0.8 #g/mlsample - 0.78;6.t hydrochloride, MDMA, MDA and 3,4,5-trimeth- 1.8/_g/mlsampe 1.77:0.9 - oxyamphetamine (see Table I) shows that only 8.8 #g/mi sample 8.69:0.8 - phenylalkylamine derivatives with a distinct ring- DetectionAbsolutelimiff(ng) 2.8 1.3 substitution pattern can be differentiated. The low Relative(#g/ml) 0.056 0.026 UV cut-off of the water-acetonitrile modifier phase ': gives the possibility of measuring in a range (190- _ Signal-to-noiseratio = 5:1. Fig

! IRENNEISEN HPLC OF PHENYLALKYLAMINE DERIVATIVES 93 rivatives ex- 6 imum detec- 120: _e sensitivity dDMA and 1zo. lore than 60 4.675) com- ez. id detection -234 nm for _ sz. 1 for mesca- E I [ about 0.03 t sensitivity, 2_- verall repro- bleat determi-198 nm 4_.z -----J _, 2 · in biological a 2 ,/ 6 fi Ih 1 ,ns. Time (m{n ) Fig. 2. Chromatogram of a human urine sample obtained after oral administration of 1.7 mg/kg MDMA. For peaks, see Table 1. urine an alterna- isolation of n0us matrix. The results of the recovery study for identified was MDA, formed by N-demethylation mended be- MDMA and MDA, listed in Table II with further and excreted in concentrations of 0.07 to 0.90 #g/ )f operation validation data, show the efficiency of the extrac- ml. The pharmacokinetic data will be published in volumes of ti0n procedure. The volatility of MDMA and detail elsewhere. t metabolite MDA may cause sample loss during the concentra- _ectively by ti0n step. This can be avoided by the addition of Determination of mescaline in cactus plants hase extrac- hydrochloric acid to the eluent and K2HPO4 to the Solvent extraction was chosen for the extraction and MDA extract before evaporation. After oral administra- of cactus material. Mescaline is almost quantita- the endoge- ti0n, MDMA is mainly exreted unchanged in urine, tively extracted from the finely powdered and defat- The concentrations found in the urine of four pa- ted cactus matrix by sonication with alkalized tients who received an oral dose of 1.7 mg/kg rang- methanol (solvent/sample, 200:1, v/w). The selected ed from 1.48 to 5.05/_g/ml. The main metabolite I.S. (methoxamine) is a synthetic ring-substituted )A 60o, 15 lA MDA 500' t 0.08-1.6 0.999 400- 97.7

100.7 _ 300' -- E

).9 - IOO: ).8 - 0.78;0.39;11.66.1 200' _1 _ fl li 1.3 0 _ - · 0.026 T; me (m i n )

Fig. 3. Chromatogram of a Trichocereus pachanoi cactus specimen. For peaks, see Table I. 94 H.-J.HELMLIN,R.BRENNEISENr Journalof Chrorr Elsevier Science 1 TABLE III REFERENCES VALIDATION DATA FOR MESCALINE _ CHROMSYMP. I R. M. Baum, Chem. Eng. News, 63 (1985) 7. 2 D. E. Nichols, in K. K. Redda, C. A. Walker and G. Barnett ! Parameter Mescaline (Editors), Cocaine, Marijuana. Designer Drugs.' Chemistry, Pharmacology. and Behavior, CRC Press, Boca Raton, FL. Hi _ll-l,)(_ Linearity range (,ug/ml) \ 20-75 1989, p. 175. Correlation coefficient 0.999 3 S. J. Peroutka (Editor), Ecstasy.' The Clinical, Pharmacolog- of 0_'toc Mean recovery (%; n = 3) 99.1 ical and Neurotoxicological Effects of the Drug MDMA, t Mean precision _g/mg; C.V., %; n = 6) 9.9; 3.3 Kluwer, Dordrecht, 1990. Detection limit' 4 H.-J. Helmlin and R. Brenneisen, in preparation. A. aaruffini Absolute (ng) I 5 B. K. Logan, D. T. Stafford, I. R. Tebbett and C. M. Moore, Relative (gg/mg) 0.04 J. Anal. Toxicol., 14 (1990) 154. Department of Pr 6 K. Zech and R. Huber, J. Chromatogr., 282 (1983) 161. Signal-to-noise ratio = 5.1. 7 A. Fell, B. J. Clark and H. P. Scott, J. Chromatogr., 316 (1984) 423. ABSTRACT 8 R. O. Fullinfaw, R. W. Bury and R. F. W. Moulds, J. Chro- matogr., 415 (1987) 347. phenylalkylamine derivative with chemical proper- 9 E. I. Minder, R. Schaubhut, C. E. Minder and D. J. Von- _ _-Tocopheryl n ties very similar to those of mescaline but not in- derschmin, J. Chromatogr., 419 (1987) 135. broad:_.TNinrangecosmetic°fco terfering chromatographically (Fig. 3). The efficien- 10 377.D. W. Hill and K. J. Langner, J. Liq. Chromatogr., 10 (1987) cy of extraction was tested by spiking mescaline- 11 I. s. Lurie, J. M. Moore, D. A. Cooper and T. C. Kram, J. free cactus material with mescaline hydrochloride. Chromatogr., 405 (1987) 273. INTRODUCTIO] The recovery of mescaline (Table III) was 399%, 12 R. Brenneisen and S. Geisshfisler, Pharm. Acta Heir., 60 0985)290. Variousderi showing that the loss of mescaline during the defat- 13 R. Brenneisen and S. Borner, Pharm. Acta Heir., 60 (1985) ting process, which is necessary to remove interfer- 302. drugs as perip lng !ipids and waxes, is not significant. The mesca- 14D. Bourquin and R. Brenneisen,Anal.Chim.Acta,198 nicotinic acid e line content of six Trichocereuspachanoi specimens (1987)183. usedin cosmetJ ranged from 1.09 to 23.75 #g/mg dried cactus mate- 15s. BornerandR.Brenneisen,J.Chromatogr., 408 (1987)402. ators. _-Tocop rial, showing the extreme variability of the psycho- 16 neisen,H,H.Helmlin,Pharm. ActaD. Bourquin,Helv., 64M.(1989)de Bernardini178. andR.Bren- effects of its ct tropic potency. 17 D. Bourquin and R. Brenneisen Proceedings of the 41th _ acid [2], withol AAFS Meeting, Las Vegas, NV, American Academy of Fo- shown by othei ACKNOWLEDGEMENTS rensic Sciences, Colarado Springs, CO, 1989, p. 143. and heat sensa 18 R. Brenneisen, K. Mathys, S. Geisshfisler, H. U. Fisch, U. tion and stimu Koelbing and P. Kalix, J. Liq. Chromatogr., 14 (1991) 271. This work was supported by grants from the E. 19 H.-J. Helmlin, L. Szajek, D. Bourquin and J. T. Clerc, Pro- active ingredk Steinegger Foundation for Medicinal Plant Re- ceedings of the 15thInternationalSymposium on Column products [3-12 search and the Swiss Federal Office of Public Health Liquid Chromatography (HPLC '91), Basel,199l,Poster and creams) a (Foundation of Narcotics Research). We thank Dr. ll8/t, Abstractbookp. 76. and shampoosl 20 H.-J. Helmlin, D. Bourquin and J. T. Clerc, in preparation. Some reven J. W. Roth, Chairman of the Swiss Association for 21 D. Chan Leach, M. A. Stadalius, J.S. Berus and L. R. Psycholytic Therapy, for providing the urine sam- Snyder, LC. GC Int., I (1988) 22. chromatograp[ pies. We also thank the Division of Narcotic Drugs 22 H. Engelhardt and M. Jungbeim, Chromatographia,29 tion of tocoph of the United Nations for the donation of phenyl- (1990)59. [13-16]. Only t alkylamine derivatives and Mr. J. Liithy, Institute 23 R. Gill, S. P. Alexander and A. C. Moffat, J. Chromatogr.. tion of _-TN i: 247 (1982) 39. suitable for mo of Botany, University of Berne, for the identifica- 24 I. S. Lurie and S. M. Carr, J. Liq. Chromatogr., 6 (19831 tion and supply of Trichocereuspachanoi speci- 1617. This paper d mens. 25 Analytical Profiles of Substituted 3,4-Methylenedioxyamphe' for the determi tamines.' Designer Drugs Related to MDA, CND Analytica]. tions. Auburn, AL, 1988.

26 C. R. Clark, A. K. Valaer and W. R. Ravis, J. Liq. Chroma- EXPERIMENTAl togr., 13 (1990) 1375. 27 A. C. Moffat (Editor), Clarke's Isolation andldentificationq Drugs in Pharmaceuticals, Body Fluids, and Post-Mortern ._laterials Material, Pharmaceutical Press, London, 1986. _-TocopheD

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