INFORMATION TO USERS The most advanced technology hak been used to photo- ' graph and reproduce this manuscript from the microfilm master. UMI films the original text directly from the copy submitted. Thus, some dissertation copies are in typewriter face, while others may be from a computer printer. ! In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyrighted material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are re produced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Each oversize page is available as one exposure on a standard 35 mih slide or as a 17" x 23" black and white photographic print for an additional charge. Photographs included in the original manuscript have been reproduced xerographically in this | copy. 35 mm slides or 6" x 9" black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order.
Accessing the Worldls Information since 1938
300 North Zeeb Road. Ann Arbor. Ml 4B106-1346 USA
Order Number 8804000
Chromatographic behavior of anions and the first row transition metal ions
Arbogast, James Kevin, Ph.D.
The Ohio State University, 1987
UMI 300 N. Zeeb Rd. Ann Aibor, MI 48106 ! PLEASE NOTE:
In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark_
1. Glossy photographs or pages _____
2. Colored illustrations, paper or print ______
3. Photographs with dark backgrou nd.
4. Illustrations are poor copy______
5. Pages with black marks, not original copy
6. Print shows through as there is text on both sides of p age ______
7. Indistinct, broken or small print on several pages ______
8. Print exceeds margin requirements_____
9. Tightly bound copy with print lost In spine______
10. Computer printout pages with indistinct print ______
11. Pagefs) lacking when material received, and not available from school or author.
12. Page(s) seem to be missing In numbering only as text follows.
13. Two pages numbered . Text follows.
14. Curling and wrinkled pages _____
15. Dissertation contains pages with print at a slant, filmed a s received
16. Other______
CHROMATOGRAPHIC BEHAVIOR OF ANIONS AND THE FIRST ROW
TRANSITION METAL IONS
DISSERTATION
Presented in Partial Fulfillment of the Requirements for
the Degree Doctor of Philosophy in the Graduate
School of the Ohio State University
By
James K. Arbogast, B.S.
*****
The Ohio State University
1987
Dissertation Committee: Approved by
Dr. T. Sweet
Dr. S. Olesik Advise Dr. P. Dutta Department of Chemistry ACKNOWLEDGMENTS
I would like to thank my adviser, Dr, T.R. Sweet, for his help
throughout this work and particularly for his patience over the years.
I want to thank my fellow researchers, especially Jorge Zayas,
Atchana Wongchaisuwat, Kent Dougherty, Mary Davis, and the undergraduate researchers for their efforts.
I also would like to express my appreciation to family and friends whose support, understanding, and prayers kept this from being an
impossible task. I especially appreciate the loving support of my wife, who over the last few months, has tolerated more than her share
and has contributed directly to the preparation of this document.
Finally, 1 would like to thank Jane Zblnden for her professional * typing.
ii VITA
July 4, 1956 Born - Bridgeport, Ohio
1978 . . . . B.C. in Chemistry, Wheeling College, Wheeling, West Virginia
1978-1984 . Teaching Associate, Department of Chemistry, The Ohio State University, Columbus, Ohio
1986-Present Research Assistant, Byrd Polar Research Center, The Ohio State University, Columbus, Ohio
PUBLICATION
"High Performance Liquid Chromatography of Metal Ions on a Bonded Stationary Phase" (With K. T. DenBleyker and T. R. Sweet), Chromatographia, 17, 8 (1983) p. 449.
iii TABLE OF CONTENTS
ACKNOWLEDGMENTS...... ii
VITA ...... ill
LIST OF T A B L E S ...... v
LIST OF FIGURES...... vii
INTRODUCTION ...... 1
CHAPTER PAGE
1. THE ANALYSIS OF ANIONS IN ICE SAMPLES BY ION CHROMATOGRAPHY ...... 2
Introduction...... 2 T h e o r y ...... 4 Experimental ...... 18 Results and Discussion ...... 22
2. THIN LAYER CHROMATOGRAPHY OF THE FIRST ROW TRANSITION METAL I O N S ...... 81
Introduction ...... 81 Theory ...... 85 Experimental ...... 88 Results and Discussion...... 92
3. ELECTROPHORESIS OF THE FIRST ROW TRANSITION METAL IO N S ...... 114
Introduction ...... 114 T h e o r y ...... 117 Experimental...... 123 Results and Discussion ...... 125
SUMMARY...... 133
LIST OF REFERENCES ...... 135
iv LIST OF TABLES
TABLE PAGE
1. Slple Chemistry Core B ...... 24
2. Siple Chemistry Core A ...... 30
3. Siple Chemistry Pit 1 Profile A ...... 32
4. Siple Chemistry Pit 1 Profile B ...... 33
5. Slple Chemistry Pit 3 Wall B ...... 34
6. Dunde Chemistry xx C o r e ...... 35
7. Dunde Chemistry Pit 2 36
8. Dunde Chemistry Pit 5 ...... 37
9. Comparison of Efficiencies ...... 77
10. Comparison of Injection Methods ...... 79
11. Solvent Systems Used For TLC ...... 100
12. Comparison of Stationary Phase and Pretreatment Techniques ...... 101
13. Behavior of Metal Ions in Normal Phase TLC ...... 102
14. Behavior of Metal Ions in Normal Phase T L C ...... 103
15. Behavior of Metal Ions in Reverse Phase T L C ...... 104
16. Behavior of Metal Ions in Reverse Phase T L C ...... 105
17. Effects of pH on Rf Values (Normal P h a s e ) ...... 106
18. Effects of pH on Rf Values (Aluminum O x i d e ) ...... 107
19. Effects of pH on Rf Values (Reverse P h a s e ) ...... 108
v TABLE PAGE
20. Behavior of Metals on Reverse Phase HPTLC ...... 109
21. Developing Reagents for Locating Cations on TLC Plates . 110 •» 22. Colors of Metal Complexes Formed With Developing Reagents...... Ill
23. Colors of Metal Complexes Formed With Developing Reagents (after 1 hour) ...... 112
24. Colors of Metal Complexes Formed With Developing Reagents (after 2 hours) ...... 113
25. Solvent Systems Used for T L E ...... 126
26. Behavior of Metal Ions In T L E ...... 127
27. Behavior of Metal Ions in T L E ...... 128
vi LIST OF FIGURES
FIGURES PAGE
1. Important Parameters In Retention ...... 4
2. Hj - Longitudinal Diffusion ...... 9
3. Hc - Multipath Effect ...... 9
4. H . - Resistance to Mass Transfer...... U ml 5. HETP Curve for a Liquid Chromatography Column ...... 15
6. Important Parameters in Resolution ...... 16
7. Location of Dunde Ice Cap and Siple Station ...... 19
8. Location of Cores and Pits at Siple Station, Antarctica 20
9. Chloride in Core Samples, Siple ...... 39
10. Sulfate in Core Samples, Siple ...... 40
11. Chloride in Wall B Profile A, Slple ...... 41
12. Chloride in Wall B Profile B, Siple ...... 42
13. Chloride in Fit 3 Wall B, Slple ...... 43
14. Sulfate in Wall B Profile A, Siple ...... 44
15. Sulfate in Wall B Profile B, Slple ...... 45
16. Sulfate in Pit 3 Wall B» S l p l e ...... 46
17. Nitrate in Wall B Profile A, Siple ...... 47
18. Nitrate in Wall B Profile B, Siple ...... 48
19. Comparison of Sample Loading Methods ...... 51
20. 100 ppb Standard in Mobile Phase (or 30) ...... 54
Vii FIGURES PAGE #> 21. 100 ppb Standard in Mobile Phase (or 1 0 ) ...... 54
22. 1 ppb Standard; 8.5 ml on Column Loading (or 30) . . . 55
23. 1 ppb Standard; 17 ml on Column Loading (or 30) .... 55
24. 10 ppb Standard; 8.5 ml on Column Loading (or 30) . . . 56
25. 10 ppb Standard; 17 ml on Column Loading (or 30) . . . 57
26. 1 ppb Standard; 0.64 ml Injection (or 3 0 ) ...... 58
27. 1 ppb Standard; 0.64 ml Injection (or 1 0 ) ...... 58
28. 100 ppb Standard; 0.64 ml Injection (or 30 ) ...... 59
29. 100 ppb Standard; 0.64 ml Injection (or 10 ) ...... 60
30. 1 ppb Standard; 17 ml Using Concentrator Column (or 30) 61
31. 1 ppb Standard; 8.5 ml Using Concentrator Column (or 3 0 ) ...... 62
32. 10 ppb Standard; 17 ml Using Concentrator Column (or 3 0 ) ...... 63
33. ,1 ppb Standard; 17 ml on Column Loading (or 10) .... 64
34. Sample Containing Chloride, Nitrate, and Sulfate at Approximately 300 ppt, 17 ml on Column Loading (or 1 0 ) ...... 65
35. Milli-Q Water, 17 ml on Column Loading (or 30) .... 65
36. 20 ppt Sample; 51 ml on Column Loading (or 10) .... 66
37. Ideal Working Curve ...... 69
38. Working Curve for Fluoride ...... 70
39. Detector Response vs Loading for 10 ppb SO ^ ...... 71
40. Working Curve for Chloride ...... 72
41. Working Curve for Nitrite ...... 73
42. Working Curve for Ni t r a t e ...... 74
43. Working Curve for H2P 0 ^ ...... 75
viii FIGURES PAGE
44. Working Curve for Sulfate ...... 76
45. Pit Sample 56,'0.64 ml Injection (or 10) ...... 80
46. Pit Sample 56, 17 ml on Column Loading (or 10) .... 80
47. Factors Important in Plate Efficiency ...... 86
48. Resolution on a TLC P l a t e ...... 87
49. Effectiveness of Cooling in T L E ...... 131
50. Effectiveness of Cooling in TLE ...... 132
ix INTRODUCTION
This work was undertaken with the intention of analyzing ice samples from ice cap regions of the earth and to develop Improvements in analytical methods and techniques used for this purpose• The need for such analysis is great since thousands of years of meteorological history of our planet is locked in ice caps in the polar regions and some mountain areas of our world. Trace analyses in the ppm and often in the ppb or ppt range are necessary. Ion chromatography* thin layer chromatography, and electrophoresis will be considered.
1 CHAPTER 1
THE ANALYSIS OF ANIONS IN ICE SAMPLES BY ION CHROMATOGRAPHY
Since its introduction in the early 1970's, ion chromatography has been used for a wide variety of analysis. Analysis of trace anions is one application for which this method is very well suited.
Delmas and coworkers published several articles (1,2,3) on the chemical analysis of Antarctic snow and ice. In the first article, they analyzed for sulfate by a titration with lead perchlorate. In the next paper, they determined both sulfate and nitrate by ion chromatography using a preconcentration column. Analysis time was about 1/2 hour for a detection limit of 1 ppb. The article published in 1984 reported the results of analysis for chloride, nitrate, and sulfate. They employed a large volume injection loop (5 ml) for the
Bulfate and nitrate analyses. Chloride analyses were run separately with a 1.5 ml injection volume. This method was used for analysis of volcanic deposits in Antarctic snow and ice.
Jenke et al. (4) measured the chloride, nitrate, and sulfate content of snow samples from the Rocky Mountains using both suppressed and non-suppressed ion chromatography. They found good agreement between the two methods. The non-suppressed Method was leBS sensitive and required either sample preconcentration or large sample volume. They found no system which could be used for the analysis of both chloride and sulfate within a reasonable time (less than 30 minutes).
Herron (5) reported the determination of fluoride, chloride, nitrate, and sulfate in Greenland and Antarctic precipitation. In his work, he mentioned the use of large sample volumes (10 ml) but gave no details as to the analytical procedure (i.e. whether sample loop or concentrator column was used).
Spencer and coworkers (6,7) used ion chromatography for the analysis of bromide, fluoride, chloride, sulfate, and nitrate in samples from Heard Island (South Indian Ocean) and Greenland. They used a 0.2 ml sample loop. Samples were Bpiked with concentrated eluant to eliminate the water dip. Precision was 5X or better for each anion.
The previous work was Improved to allow for determination of fluoride, chloride, sulfate, nitrate, nitrite, and dihydrogen phosphate at the ppb level using suppressed ion chromatography. This method was used to analyze Bamples from China and Antarctica. This method was further Improved to permit more accurate determination in the ppb range and detection in the ppt range. Theory
Retention
The purpose of the analytical column is to preferentially
retain the components of a sample
s i &
Figure 1. Important Parameters In Retention
The retention time (tr) is related to the flow rate of the mobile
phase (F) by:
(1) tr - Vr/F
where Vr is the retention volume, which is the volume of mobile
phase required to elute the solute peak maximum.
The equation for the retention time can also be expressed in
terms of the linear solvent velocity (p in cm/sec) as derived by
the following: 5
(2) p s - ji(Xm)
where u is the average velocity of the sample (or solute) and Xtn S is the average fraction of solute molecules in the mobile phase.
The partition coefficient (Kr) is defined as the ratio of the concentration of solute in the stationary phase (Cs) to the concentration of solute in the mobile phase (Cm).
(3) Kr - Cs/Cm
The phase ratio £ is simply the ratio of the volumes of the two phases
(4) * - Vm/Vs
I Since Xra is the fraction of solute molecules in the mobile phase
(5) Xm « CmVm/(CmVm + CsVs).
By combining equations, equation (2) becomes
(6) ps - p l l / O + Kr/*)]
The retention time is related to u by S
(7) *ia - L/Tr 6 where L is Che column length. Combining equations (6) and (7) we get
(8) L/Tr ■ git 1/(1 + Kr/*)]
or
(9) Tr - (L/*i)(l + Kr/jJ)
Since to ■ L/p, equation (9) becomes
(10) Tr ■ To (I + Kr/jg )
The solute capacity factor (k1) is the ratio of moles of solute in the stationary phase and mobile phase
(11) k' - CsVs/CraVm - Kr/*
Combining equations (10) and (11):
(12) Tr - To(l + k') or
(12a) k' - (Tr - To)/To
Equation (12) can be expressed in volume terms by multiplying by the flow rate 7
(13) Vr - Vo(l + It') or
(13a) k' - (Vr - Vo)/Vo
where Vo is Che dead volume of the system Vo * (to)F .
Column Efficiency
The efficiency of a column is expressed in terms of the number of theoretical plates (N). Martin and Synge (8) showed that N could be calculated by:
(14) N - (tr/ where N is related to the height equivalent to a theoretical plate (H) by (15) H - L/N. Glddings (9) and Huber (10) extended the theory first developed by van Deemter and coworkers (11) for CC. The theory assumes that each individual component to band spreading can be considered independent and, when summed with the other components, will give the overall band dispersion. There are three processes which contribute to the band spreading: longitudinal diffusion, convective or coupled mixing, and resistance to mass transfer at the mobile phase/stationary phase interface. Mathematically this can be expressed as <16> "total ■ "l + "c + "t.1 where Is the contribution from longitudinal diffusion, Hc is the contribution from mixing, and Hmi is the contribution from mass transfer The longitudinal diffusion is a result of the solute not filling the entire chromatographic system while being eluted. Axial concentration gradients therefore exist along the column. This results in diffusion of solute molecules in the longitudinal direction. Theoretically, diffusion occurs in both stationary and mobile phases; however, the mobile phase diffusion rate is much greater. Band-broadening is proportional to the solute dlffusivity (Dm) and the reciprocal of the mobile phase linear velocity (u). The broadening is related to the column packing geometry (A) or interparticle tortuosity factor (Tm). (17) ■ 2^Dm/u or (18) Hx - 2DM/Tmu 9 indicates mobile phase Jdirection solute $ ■ stationary phase Figure 2. - Longitudinal Diffusion The second term in equation (16), Hc, was for convective or coupled mixing. Both eddy diffusion, or multipath effects, and mass transfer kinetics in the mobile phase contribute to this term. The multipath effect is a result of the difference in path lengths taken by solute molqcules. m 6 © Figure 3. Hc - Multipath Effect 10 This spreading depends on the physical nature of the column (Y) and the diameter of the column packing particle (dp). (19) He -Ydp Convective mixing also results from the lateral diffusion of solute molecules from one streamline to another. These velocity unequalities are less important at higher diffusion rates. (20) Hid - Wjdp^u/Dm where is a function of packing structure (12). Giddings (13) combined the eddy diffusion and lateral diffusion terms. The result is (21) H - 1/(1/He + 1/Hld) or c (22) H - 1 Dm c (l//dp) + Wjdp^ The resistance to mass transfer is a result of the non- instantaneous transfer of solute molecules between the mobile phase and the stationary phase. 11 Figure 4. Hmi - Resistance to Mass Transfer Humber and Hulsman (14) derived the following equation for 1 j « ml (23) B mi » 0 < * « * *rf> d as where € is the volumn volume occupied by flowing fluid. € is' m s the volume occupied by static fluid, is the column volume occupied by the mobile phase, 0 Is the ratio of total surface area of stationary phase to column volume, Ks is the solute partition coefficient, tJ is the mobile phase viscosity, Da is the mass of solute transported across the interface per unit concentration difference, Ds is the solute diffusibility in the stationary phase. Das 1b the combination of Da and Ds, and Ts is an intraparticle tortuosity factor. 12 Combining equations (16), (18), (22), and (23) we get (2‘> "total ' "l + "c + "at f f M + 1______+ J _ ( T.u lA»d ♦ *u 5.7 V + * , ♦ j _ yyggfr y»-yy,,* t (l-^)DM« 5 0 6. »AS Assuming that the difference between ^ and €m is very small, and recognizing that the capacity factor is related to the solute partition coefficient by (25) - KsAs/Vm - Ks/fi m (where As is the stationary phase surface area and Vm is the mobile phase volume), equation (24) becomes (26) „ . J ! h _ + ------1 + ; » ’ T.u l/v»dp ♦ DM/Wjdp2u 5.7Dm*(1-€b) 1+k' 1 k ' (X-€ ) T . d 2u ^ a ____ ■ ® P 3 0 Equation (26) is often simplified to (27) H - Au0,33 + B/u + Cu where A, B» and C are constants for a given column and correspond to convective mixing* longitudinal diffusion* and resistance to mass transfer* respectively. Figure 5 shows a typical plot of how H and its components vary with u. At high linear velocity* resistance to mass transfer in the stationary phase is the major contribution to H. At low linear velocity* longitudinal diffusion predominates. At intermediate velocities, the H term is most affected by convective mixing and resistance to mass transfer in the mobile phase. Frequently columns are evaluated in terms of reduced plate height (h) and reduced linear velocity (v). These are defined as: (28) h - H/dp and (29) v ■ dp u/Dm where Dm is the diffusion coefficient which can be calculated by: (30) Dm - 7.A x 10-8 (froMm)*5!/!*v where v is the solute molar volume* Mm is the molecular weight of the mobile phase,fm is an association factor for the solvent, and lit T is the temperature in eK. The reduced plate height is composed of axial diffusion (B), anisotropic flow (A), and slow mass transfer (C). (31) h - B/v + Av0,33 + Cv Composite HETP Curve H Mass Transfer in the Mobile Phase/Stationary Phase Interface Longitudinal diffusion Coupled Mixing /» Figure 5. HETP Curve for a Liquid Chromatography Column Da teeter retpease Resolution resolution. Resolution is determined by the distance between the peak maxima maxima peak the widths between peak and distance the by determined is Resolution Purnell (15) derived an equation relating the resolution to the the to resolution the relating equation an derived (15) Purnell ounefcec, eetvt, n te ouecpct factor. capacity solute the and selectivity, efficiency, column (32) Rs - (Tr2 - T r j ) / ^ + U’2 + ) ^ / ) j r T - (Tr2 - Rs (32) 3) s- ,5*(J1+ 1/4*) + * ( ) j K + 0,25N*S(KJ/1 - Rs (33) The separating power of a system is referred to as the the as to referred is system a of power separating The Figure 6. Important Parameters in Resolution in Parameters Important 6. Figure TI m 16 17 From equation 33, we can see that the resolution can be improved by: 1) increasing the column efficiency, usually done by optimizing flow rate or increasing column length, 2) varying the solute capacity factor (kf)» accomplished by changing the mobile phase (k' is reduced with increasing solvent strength) and, 3) varying the selectivity factor (•<.), which can be done by changing the stationary phase or controlling temperature. (Temperature can also affect solute capacity factors and column efficiency.) 18 Experimental Analyses of ice cores have been shown to contain records of climatic changes dating back hundreds of years (16,17,18). The interpretation of these records can be enhanced by the study of pit Bamples. Research teams from the Byrd Polar Research Center drilled ice cores and excavated pits on the Dunde Ice Cap, China, and at Siple Station, Antarctica (see Figure 7). The cores obtained were as deep as 302 meters which would contain a 500 year record ,of precipitation (19). The pit depths are on the order of 2 to 3 meters and give a detailed record of the meteorological conditions of the last few years. The accumulation rate at Siple Station has been estimated at 1.3 meters per year (20). Thus a 2 meter pit would contain about 1.5 years of precipitation. In the deeper cores, the ice becomes compressed and; therefore, an annual accumulation is leBs than 1 meter thick. Figure 8 is a map showing the location of the cores and pits in Antarctica. Chemical analysis was conducted on a 9 meter section of core B (132 meter core or 130 years) and 2 meters of core A (302 meter core or 500 years). Two profiles from wall B of pit 1 were also analyzed. These profiles were approximately 38 cm apart in the center of the four meter wide wall. The profile from pit 3 wall B was collected 68 cm from the corner. All samples were collected in pre-cleaned containers which were tested prior to shipment to the field. Samples were kept frozen until just prior to analysis. CURRENT SITES FUTURE SITES * COMPLETED PROJECTS O PRELIMINARY STUDIES COMPLETED • PROPOSALS SUBMITTED Q ' N O DATA AVAILABLE Figure 7. Location of Dunde Ice Cap and Siple Station 20 N\S iple Station, Antarctica, 1985/86 Drilling and Pit Program \2 B Mitt Antenno Skiway Siple Station \ V \ S. Summer Comp True b .) Drill Site \ 4 Pit I. Science Quadrant b.) ^Prevailing wind: Dec. 3 - Jan. B, 150* true North & .Drilling Trench 0 4 4 km vPit I . • 302 m core O ^ Z/ k m 0.5km • 132 m core Pit 2 ^ V ^ p | t 3 o 2 0 m core Figure 8. Location of Cores and Pits at Siple Station, Antarctica 21 Anion analyses were performed on a Dionex 20101 ion chromatograph with a fiber suppressor. Separation was achieved on an AS-4A column using an eluant of 0.0006 moles/liter sodium bicarbonate and 0*.017 moles/liter sodium carbonate. The eluant flow rate was maintained at 1.7 ml/min and the regenerant flow was set as 3.0 ml/min. Samples were injected through a 0.25 m Millipore filter into a 0.65 ml sample loop. The size of this sample loop permitted detection in the PPB range without the need to preconcentrate the sample. The volume of the injected sample (usually 4 ml) was large enough to flush the sample loop with at least 5 times the loop volume. Additionally, between each sample the loop was flushed with at least 5 ml of lab water. The chemicals used for standards and eluants were of reagent grade. The standards were made and all analyses were conducted in a class 100 clean room. The clean room water is reagent-grade deionized and filtered water provided by a Millipore Milli-Q3 and Milli-Q2 system. Standard stock solutions in the PPM range were remade monthly. Dilute standards in the PPB range were remade weekly. Standards and blanks were run at the beginning and end of each day. In addition the standards were run periodically during the day - usually after every fifth sample. Duplicate or triplicate runs were performed on samples to insure reproducibility. Approximately 1/4 of all samples were duplicated. Any sample which showed extraordinarily high or low values was also reanalysed. Results and Discussion •. Antarctic Samples Tables 1 through 5 give the results for analyses done* on core and pit samples from Antarctica. Table 1 shows the results for a 9 meter section of core B. The data for a 2 meter section at the same starting depth Is given in Table 2. The average chloride concentration for core A is 62.5 ppb. The average for the first 2 meters of core B was 66.2 ppb. The sulfate levels are in even closer agreement. The average in core A is 35.9 ppb while the average for 2 meters of core B is 36.3 ppb. Two profiles of wall B in pit 1 were analyzed and the results are given In Tables 3 and 4. The chloride averages for profile A and B are 52.4 ppb and 54.6 ppb respectively. For sulfate the average in profile A is 31.9 ppb and in profile B is 29.9 ppb. Table 5 presents the data from a wall oriented in the same direction as wall B in pit 1, but from a different pit. The chloride average here is somewhat higher than in the other pit samples (66.8 ppb) and the sulfate average is lower than In any other set of samples (20.7 ppb). The difference here is likely due to the location of the pit. It 1b 0.4 kilometer from pit 1 and 1.0 kilometer from the core drilling site. The ratio between sulfate and chloride is also consistent in the two cores and two profiles from pit 1. Core A has a ratio of 0.57 and core B a ratio of 0.58. Profile A of pit 1 has a ratio of 0.61 and profile B has a ratio of 0.55. 23 China Samples Tables 6, 7, and 8 give Che results obtained from the analyses of two pits and one core from China. The levels of chloride and sulfate are generally one to two orders of magnitude larger than Chose found in Antarctic samples. In the samples from the xx core* the chloride and sulfate levels rise and fall together whereas in Antarctic samples* the rise of one corresponded more closely to the fall of the other. For both chloride and sulfate* the highest concentration (almost double the second highest) is found in the last sample. Pit 2 also shows the highest concentrations in the last samples. Similarly* this pit has changes in sulfate levels corresponding to the changes in chloride. Pit 5 again shows the general pattern of sulfate and chloride levels varying simultaneously except in the last 3 samples where chloride values Increase but sulfate values do not. The variation is much more pronounced in the sulfate values where the range is from 0.06 to 1.73 ppm. The similarity of results between the two cores from Antarctica can best be seen graphically. Figure 9 shows the chloride concentration as a function of depth for both core A and B. The same trends appear in both cores with yearly cycles occuring at approximately every 0.5 meter. An offscale chloride peak is found at a depth of 105.7 meters in both cores. The sulfate levels versus depth also show a yearly cycle which can be clearly seen in each core (see Figure 10). It is Table 1. Siple Chemistry Core n o " Sample Number Section Number Depth F Cl n o " H 2P04 9 105 105.420 0.0 61.0 27.0 6.0 30.0 41.0 10 105 105.470 0.0 55.0 63.0 3.0 35.0 22.0 11 105 105.520 0.0 42.0 89.0 5.0 53.0 20.0 12 105 105.570 0.0 73.0 53.0 2.0 54.0 60.0 13 105 105.620 0.0 85.0 31.0 2.0 50.0 76.0 14 105 105.670 0.0 197.0 35.0 0.0 41.0 66.0 15 105 105.720 0.0 144.0 23.0 1.0 42.0 40.0 16 105 105.770 0.0 89.0 20.0 1.0 64.0 109.0 17 105 105.820 0.0 62.0 14.0 2.0 42.0 50.0 18 105 105.870 0.0 82.0 23.0 2.0 36.0 50.0 19 105 105.920 0.0 59.0 38.0 4.0 40.0 45.0 20 105 105.970 0.0 35.0 36.0 12.0 30.0 31.0 1 106 106.020 0.0 53.0 45.0 5.0 40.0 13.0 2 106 106.070 0.0 39.0 42.0 3.0 51.0 61.0 3 106 106.120 0.0 39.0 44.0 5.0 49.0 42.0 4 106 106.170 0.0 39.0 37.0 5.0 25.0 0.0 5 106 106.220 0.0 47.0 18.0 3.0 36.0 76.0 6 106 106.270 0.0 87.0 24.0 2.0 59.0 93.0 7 106 106.320 0.0 85.0 19.0 6.0 58.0 39.0 8 106 106.370 0.0 50.0 20.0 18.0 50.0 87.0 9 106 106.427 0.0 37.0 15.0 28.0 37.0 47.0 10 106 106.487 0.0 16.0 23.0 19.0 16.0 37.0 11 106 106.544 0.0 32.0 45.0 21.0 32.0 24.0 12 106 106.594 0.0 63.0 84.0 21.0 63.0 16.0 13 106 106.644 0.0 44.0 66.0 21.0 44.0 37.0 14 106 106.694 0.0 34.0 37.0 19.0 34.0 70.0 15 106 106.744 0.0 29.0 33.0 16.0 29.0 45.0 16 106 106.794 0.0 26.0 20.0 15.0 26.0 59.0 17 106 106.844 0.0 42.0 21.0 17.0 30.0 89.0 18 106 106.894 0.0 39.0 18.0 15.0 39.0 52.0 25 ooooooooooooooooooooooooooo a*o CM m m o\ *o — CMCMOoicncMfMcnrMOrMOiCMOioifficntMcn©pHeo n n N n d < o o o o O O o o o o o o o o o o o o o O o o o O i cn « • • • •• • . . . .•. .. • . •. . .. ** o CM m CM O s inm pi » o m o o cno cn|M p H CM p H cnCM m «»fM z n co CO •»n cnCM CM CM cn m cncnMT cn*»CM m cn ooooooooooooooooooooooooooo 'cT* ON«COO[N^00'^NOH[^*Hiri^OQOOOONOOlOOlH z N M ^1 »4 H H ^ ^ Pi M HHNMpIMmN o o o o o o o o o o O o o o o o o o o o o o o o R 'T 1 . .. * «« . . . . . • . • •* * * . . **. O oooooooooooooooooooooooooooo o o o 1 • ••• p H NasnA<}pinoisvocoininvOionioniMOp ooooooooooooooooooooooooooo oooooooooooooooooooooooooooliattllflllllllllllllltlll* •j L0J i B 3 lOlOC'l*>NNNNNf'r*MNNIsr>«Nr't>NMCBnoOC008 Z ooooooooooooooooooooooooooo 108 108 108 108 108.272 108.321 0.0 108.421 0.0 146.0 0.0 143.0 36.0 90.0 27.0 1.0 27.0 1.0 36.0 16.0 38.0 48.0 49.0 35.0 38.0 C 108 108.371 0.0 154.0 26.0 21.0 43.0 36.0 ■poH U TS 6J 01 V) 3 C c o u u u 1 z ffiO^Nn-juiiOP«oo »OpHNcn>rirtiOAeooipiNn>»in*Neoo>o at pH CM £ n cs H w Table 1. (continued) Sample Number Section Number Depth F Cl~ 2 O S0I i ro N03 H2P°/ LI 108 108.471 0.0 36.0 12.0 17.0 31.0 59.0 12 108 108.520 0.0 29.0 17.0 23.0 31.0 44.0 13 108 108.570 0.0 18.0 16.0 21.0 36.0 48.0 14 108 108.620 0.0 29.0 77.0 26.0 53.0 43.0 15 108 108.670 0.0 47.0 113.0 13.0 58.0 35.0 16 108 108.720 0.0 45.0 72.0 18.0 50.0 49.0 17 108 108.770 0.0 35.0 22.0 . 16.0 33.0 57.0 18 108 108.820 0.0 49.0 30.0 15.0 28.0 36.0 19 108 108.870 0.0 53.0 22.0 2.0 43.0 48.0 20 108 108.920 0.0 283.0 20.0 0.0 30.0 37.0 1 109 108.970 0.0 178.0 25.0 14.0 34.0 59.0 2 109 109.020 0.0 273.0 41.0 19.0 27.0 49.0 3 109 109.070 0.0 267.0 32.0 22.0 28.0 39.0 4 109 109.120 0.0 27.0 30.0 30.0 35.0 37.0 5 109 109.170 0.0 26.0 33.0 9.0 28.0 45.0 6 109 109.220 0.0 25.0 32.0 24.0 26.0 44.0 7 109 109.270 0.0 43.0 90.0 24.0 45.0 70.0 8 109 109.320 0.0 38.0 101.0 7.0 47.0 39.0 9 109 109.370 0.0 33.0 33.0 17.0 59.0 102.0 10 109 109.420 0.0 45.0 31.0 17.0 55.0 62.0 11 109 109.470 0.0 102.0 31.0 14.0 65.0 48.0 12 109 109.520 0.0 144.0 53.0 8.0 47.0 62.0 13 109 109.570 0.0 89.0 25.0 13.0 41.0 56.0 14 109 109.620 0.0 46.0 21.0 20.0 30.0 67.0 15 109 109.670 0.0 63.0 22.0 10.0 24.0 36.0 16 109 109.720 0.0 86.0 39.0 9.0 31.0 53.0 17 109 109.770 0.0 45.0 74.0 13.0 56.0 36.0 18 109 109.820 0.0 60.0 46.0 10.0 55.0 61.0 19 109 109.870 0.0 65.0 35.0 11.0 42.0 33.0 20 109 109.920 0.0 80.0 44.0 20.0 69.0 19.0 Table 1. (continued) Sample Number Section Number Depth F Cl“ s°: N02 N03 H2POi 1 110 109.970 0.0 130.0 36.0 13.0 35.0 0.0 2 110 110.020 0.0 132.0 44.0 14.0 26.0 0.0 3 110 110.070 0.0 104.0 45.0 19.0 25.0 0.0 4 110 110.120 0.0 82.0 83.0 17.0 39.0 0.0 5 110 110.170 0.0 45.0 70.0 17.0 34.0 7.0 6 110 110.220 0.0 50.0 184.0 11.0 38.0 0.0 7 110 110.270 0.0 41.0 112.0 12.0 40.0 0.0 8 110 110.320 0.0 50.0 46.0 3.0 24.0 0.0 9 110 110.370 0.0 185.0 103.0 2.0 8.0 0.0 10 110 110.420 0.0 41.0 126.0 25.0 27.0 0.0 11 110 110.470 0.0 37.0 133.0 23.0 34.0 0.0 12 110 110.520 0.0 34.0 159.0 19.0 16.0 0.0 13 110 110.570 0.0 20.0 136.0 21.0 35.0 0.0 14 110 110.620 0.0 53.0 149.0 14.0 24.0 0.0 15 n o 110.670 0.0 50.0 142.0 19.0 21.0 0.0 16 110 110.720 0.0 28.0 112.0 21.0 32.0 0.0 17 110 110.770 0.0 32.0 118.0 15.0 25.0 0.0 18 110 110.820 0.0 35.0 231.0 14.0 45.0 0.0 19 110 110.870 0.0 44.0 349.0 16.0 56.0 0.0 20 no 110.920 0.0 44.0 220.0 18.0 46.0 0.0 1 111 110.970 0.0 65.0 164.0 16.0 25.0 0.0 2 111 111.020 0.0 34.0 197.0 23.0 26.0 0.0 3 111 111.070 0.0 90.0 231.0 12.0 16.0 0.0 4 111 111.120 0.0 22.0 206.0 14.0 15.0 0.0 5 111 111.170 0.0 25.0 223.0 30.0 22.0 0.0 6 111 111.220 0.0 12.0 179.0 26.0 32.0 0.0 7 111 111.270 0.0 14.0 170.0 21.0 37.0 0.0 8 111 111.300 0.0 33.0 79.0 14.0 38.0 0.0 9 111 111.230 0.0 50.0 55.0 9.0 27.0 0.0 10 111 111.390 16.0 61.0 33.0 30.0 21.0 0.0 M H V to -aB a*m t- re HO«OODM»Ul« o p m H* p m ►-* N- M 9 P* P-* N* ►■* H* H- N- TO TO ro ro ro to ro ro ro ro ro ro ro ro ro IO ro ro ro ro ro cas §■ IP o NNNNMNNNNNMIOMNMNNNN h m h m h n n - w ^ U h IP 1 3 vCOOOO'»J‘«JC'i(J'LnUl.£'-C'LOU>fONJ>—j>vO£»voi>vox>'ua£'voi>»a.C'vo^>vO£''«o^vo.C'vaf'vo£»o.ovOi>vox'►— O O l'OVOCOOO-J«vlC'CJ'U1t‘/«.c'-*> oaoooooooaooooooooooooooooooooo H^H*«jroioioM(ji*«^u>»-oo%O'ON)£>O'OOa'<^v0uiotow£*oo ooooooooooooooooooooooooooaoooo VO VO •—> Ul UUB't>0'CD^i-N)X'Um)lO0Iffvi>vJUtv)NNlnlJlLJUlvJ UUMM«HfrOOvU M £'LL3X'CTv'^J-C'U)^-^-UJl*JU1i>UJNJ£'»-*K)H-N-t-JVjlOvi— ►-NJfOMfsJUJ in MHUiwooNUova«a» »>* ro M ro ro ro p * ro c j ro ro ro ro ro ro 00 O s ro ro m ro ro o ro CO Ui w o pm Ui o o s 00 M s C 09 ro C/1 •t* co ro u s 00 ** § • • • • • « « • • ■ V • t * » •• * • • •«•• « •* * •• • ro I o o o o o o o o o o O o o o o o o o o o o o o o o o o o o o o NNMKINN h N»UMNUU iN h n n h K1MM{>^x h n MUWK) ss X'MO-C'OV'»JVOOU>^-0-OOlNj-C'f>OOCvOOOOH-K>|sJLJ‘-H-C7'-C'-C>C7>'^U) Wo | ooooooooooooooooooooooooooooooo X oooooooooooooooooooooooooaooooo ro ...... *9 ooooooooooooooooooooooooooaoooo o ** I 8Z Table 1. (continued) Sample Number Section Number Depth F Cl S0I N°; NO- h 2p °; 1 113 112.990 0.0 44.0 12.0 9.0 30.0 0.0 2 113 113.040 1.0 86.0 30.0 7.0 14.0 0.0 3 113 113.090 5.0 72.0 33.0 10.0 20.0 0.0 4 113 113.140 4.0 62.0 73.0 7.0 33.0 0.0 5 113 113.190 3.0 33.0 68.0 7.0 34.0 0.0 6 113 113.240 1.0 32.0 56.0 6.0 52.0 0.0 7 113 113.290 3.0 155.0 108.0 5.0 46.0 0.0 8 113 113.340 10.0 169.0 115.0 16.0 42.0 0.0 9 113 113.390 11.0 99.0 47.0 18.0 58.0 0.0 10 113 113.440 12.0 175.0 34.0 19.0 29.0 0.0 11 113 113.490 10.0 135.0 44.0 18.0 23.0 0.0 12 113 113.540 20.0 37.0 60.0 18.0 21.0 0.0 13 113 113.590 10.0 22.0 . 89.0 19.0 26.0 0.0 14 113 113.640 20.0 24.0 93.0 20.0 33.0 0.0 15 113 113.690 11.0 34.0 69.0 15.0 19.0 0.0 16 113 113.740 10.0 76.0 57.0 17.0 23.0 0.0 17 113 113.790 10.0 114.0 69.0 12.0 20.0 0.0 18 113 113.840 12.0 49.0 79.0 17.0 31.0 0.0 19 113 113.890 17.0 27.0 89.0 20.0 37.0 0.0 20 113 113.940 5.0 74.0 68.0 16.0 24.0 0.0 1 114 113.990 5.0 98.0 77.0 5.0 18.0 0.0 2 114 114.040 20.0 41.0 112.0 17.0 22.0 0.0 3 114 114.090 15.0 24.0 172.0 22.0 37.0 0.0 4 114 114.140 25.0 33.0 151.0 20.0 28.0 0.0 5 114 114.190 10.0 34.0 152.0 14.0 21.0 0.0 6 114 114.240 7.0 42.0 151.0 17.0 22.0 0.0 7 114 114.290 0.0 74.0 117.0 0.0 0.0 0.0 N «o Table 2. Siple Chemistry Core Sample Number Section Number Depth F Cl SOl N02 N03 v ° ; 5 105 105.220 0.0 79.0 19.0 1.0 26.0 0.0 6 105 105.270 0.0 27.0 11.0 5.0 28.0 0.0 7 105 105.320 0.0 22.0 17.0 4.0 16.0 0.0 8 105 105.370 0.0 39.0 25.0 1.0 17.0 0.0 9 105 105.420 0.0 42.0 120.0 1.0 35.0 0.0 10 105 105.470 0.0 28.0 83.0 0.0 44.0 0.0 11 105 105.520 0.0 45.0 50.0 0.0 34.0 0.0 12 105 105.570 0.0 72.0 28.0 0.0 24.0 0.0 13 105 105.620 0.0 255.0 36.0 0.0 20.0 0.0 14 105 105.670 0.0 217.0 25.0 0.0 20.0 0.0 IS 105 105.720 0.0 120.0 19.0 3.0 22.0 0.0 16 105 105.774 0.0 44.0 12.0 9.0 29.0 0.0 17 105 105.827 0.0 72.0 20.0 7.0 31.0 0.0 18 105 105.881 0.0 45.0 39.0 9.0 20.0 0.0 19 105 105.935 0.0 23.0 35.0 13.0 23.0 0.0 1 106 105.989 0.0 42.0 41.0 14.0 35.0 0.0 2 106 106.043 0.0 29.0 52.0 23.0 48.0 0.0 3 106 106.096 * 0.0 24.0 47.0 22.0 42.0 0.0 4 106 106.150 0.0 26.0 34.0 5.0 23.0 0.0 5 106 106.200 0.0 41.0 16.0 1.0 28.0 0.0 6 106 106.250 0.0 84.0 21.0 0.0 32.0 0.0 7 106 106.300 0.0 63.0 17.0 0.0 31.0 0.0 8 106 106.350 0.0 36.0 11.0 0.0 16.0 0.0 9 106 106.400 0.0 62.0 14.0 0.0 12.0 0.0 10 106 106.450 0.0 38.0 21.0 0.0 10.0 0.0 11 106 106.500 0.0 18.0 71.0 0.0 33.0 0.0 12 106 106.550 0.0 20.0 92.0 0.0 39.0 0.0 13 106 106.600 0.0 22.0 61.0 0.0 29.0 0.0 14 106 106.650 0.0 46.0 32.0 4.0 24.0 0.0 15 106 106.700 0.0 94.0 35.0 2.0 20.0 0.0 Table 2. (continued) Sample Number Section Number Depth F Cl“ n o " h 2p °. S O l N°3 16 106 106.750 0.0 109.0 24.0 1.0 16.0 3.0 17 106 106.800 0.0 36.0 13.0 2.0 20.0 0.0 18 106 106.850 0.0 64.0 16.0 0.0 15.0 0.0 19 106 106.900 0.0 99.0 20.0 1.0 17.0 0.0 20 106 106.950 0.0 91.0 36.0 1.0 18.0 0.0 1 107 107.000 0.0 140.0 58.0 0.0 13.0 0.0 2 107 107.050 0.0 46.0 45.0 0.0 25.0 0.0 3 107 106.100 0.0 15.0 50.0 2.0 30.0 0.0 cn 9 UIWNNNNNNmnhnhpnkki- _ (D U)»UtC cn n o 3 Z C al 3 Spe hmsr Pt Poie A Profile 1 Pit g* Chemistry Siple 3. Table n •^•■^••Mra^HMMHMMOOOOOOOOOOOOOOO a n 00"-JO'a,'Vn£'.£'U>tO>-‘ *-OvOvOOO,«J'-JCT'Cn.e-.C'liJrOfO>-'00 •a ► *WOO>-f-slOWO'VOSJUlCSMi.sJOtJIJ'iONU'IXMX'vJO CnOOOOOQOOOOOOQQOOOOOOOOOOOO OOOVOCnCnOOOOOOOOOOOUlWW.e'CJroU).~>-W ooooooooooooooooooooooooooo CT\LiJH-l— i- - h - r o liJ U '0 0 V O liJ * - .e 'O 'tO liJ - O 'O V O 'O H -r o U > i- » U '.£ « HO\Q)OMUiOOOM09>'JMHUiMa\UUat^090i>0(atN ooooooooooooooooooooooooooo O ' vn O'u incn rororo ro t—* 44 M ro ro ro cn to M CO O' cn co o * o o cn vo vo u N*cn vo o 00 vo O' Ov o w ro «o cn VO 00 cn O' 00 e • • 4 * » • » 4 4 4 • 4 4 4 * 4 4 4 4 4 4 4 4 4 4 • o o o o o o oo o o o o o o o o o o o o o o o o o o o z ooooooooooooooooooooooooooo Wo I ooooooooooooooooooooooooooo M^U^^MUMMUfWUN^UUMMNNMOOUlHMts} z i0HWMt0t>Mln0«U s oooooooooooooooooo 0-0 o o o o o o o ro*0 ooooooooooooooooooooooooooo o •B* I z t Table 4. Siple Chemistry Pit I Profile B Sample Number Section Number Depth F~ CL n o 2 S04 N°3 h 2p o 26 1 0.000 27.0 53.0 84.0 0.0 27.0 0.0 27 1 0.073 8.0 35.0 35.0 0.0 33.0 0.0 28 1 0.146 5.0 16.0 16.0 0.0 26.0 0.0 29 1 0.218 5.0 19.0 29.0 0.0 47.0 0.0 30 1 0.291 5.0 10.0 16.0 0.0 22.0 0.0 31 1 0.364 0.0 144.0 51.0 0.0 25.0 0.0 32 1 0.437 0.0 138.0 34.0 0.0 21.0 0.0 33 1 0.510 0.0 70.0 22.0 0.0 25.0 0.0 34 1 0.582 0.0 85.0 33.0 0.0 28.0 0.0 35 1 0.655 0.0 61.0 14.0 0.0 29.0 0.0 36 1 0.728 0.0 52.0 19.0 0.0 36.0 0.0 37 1 0.801 0.0 45.0 14.0 0.0 40.0 0.0 38 1 0.874 0.0 85.0 17.0 0.0 24.0 0.0 39 I 0.946 0.0 54.0 0.0 0.0 22.0 0.0 40 1 1.019 0.0 55.0 6.0 0.0 20.0 0.0 41 1 1.092 0.0 44.0 13.0 0.0 38.0 0.0 42 1 1.165 0.0 109.0 28.0 0.0 44.0 0.0 43 1 1.238 0.0 80.0 22.0 0.0 22.0 0.0 51 1 1.310 0.0 37.0 27.0 0.0 18.0 0.0 45 I 1.383 0.0 29.0 17.0 0.0 12.0 0.0 46 1 1.456 0.0 50.0 73.0 0.0 32.0 0.0 47 1 1.529 0.0 16.0 54.0 0.0 17.0 0.0 48 1 1.602 0.0 16.0 29.0 0.0 15.0 0.0 52 1 1.674 0.0 22.0 45.0 0.0 35.0 0.0 53 1 1.747 0.0 41.0 41.0 0.0 43.0 0.0 ICA NNNNNNNNN n m n w HH id OSMO'lnf'WN^O'OOSMOMni' MMQVOOOMIJ'Uie-UNM Z nid (A n n o x« O' JJ*O'O' O' o*O' o*O' O' O' O' O' O-O' O' o»O* o*O' O' O' o*O' 9 o o o o o o o o o o o o o o o O o o e o O o o o o o a a CO 00 00 00 oo 00 00 00 Oo 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 z a c 8- ft *1 H*o* ID VA I—*** N— M M O o o a o a o o o o a • * * **••»• *•* •*«••• ■ « ■ « • * ID vo 00 00 -4 O'O' VAo* VO u»ro M o o vo 0000 O' VAVAO' VOro •o -o *a to VA•>4o VO O' 00 O'O' vo ro O* -o o vo VA00 VO O' vo ft tA u *4o> •—*00 ro -4 00 VA ro 3* M. 00 VA toVO O' o VA VO O' VO o O' •o K* ID 9 OQOOOOVa QOOQOOQVAOOOOOOO O'VA tn W O' Ui oooooooooooooooooooooooooooo i prn VO •— t* WmUlUS)WN>U)UlU'JO'Ul'JOBO ro ro vo vo to ro vo o* ro CA O' 00 VA O ' vo vo 00 O' o • « » • •* ••• op. o o o o o o o o o z oooooooooooooooooooooooooooo n P. oooooooooooooooooooooooooooo z oooooooooooooooooooooooooooo o VO I oooooooooooooooooooooooooooo s oooooooooooooooooooooooooooo IO*o 000*0000000000000000000000000 o «* I *lZ 35 Table 6. Dunde Chemistry xx Core Sample Number Cl SO^ 1 1.08 1.46 2 1.41 1.35 3 1.11 0.78 4 1.47 1.55 5 0.78 0.71 6 1.84 1.72 7 1.30 1.16 8 0.80 0.61 9 1.70 1.87 10 3.29 3.70 36 Table 7. Dunde Chemistry Pit 2 Sample Number Cl” SO^" Sample Number Cl” SO^" 1 0.55 0.50 23 0.67 0.59 2 0.32 0.55 2A O.AA 0.33 3 0.15 0.17 25 0.69 0.53 A 0.59 0.79 26 0.52 0.A2 5 2.62 3.52 27 0.51 0.35 6 0.17 0.13 28 0.A6 0.27 7 0. AO 0.19 29 0.38 0.37 8 O.AA 0.26 ‘ 30 0.26 0.22 9 0.50 0.33 31 0.35 0.59 10 0.35 0.26 32 0.51 0.56 11 0.36 0.17 33 0.55 0.67 12 0.2A 0.10 3A 1.21 0.A0 13 0.33 0.11 35 0.26 0.32 1A 0.50 0.19 36 0.31 O.AA 15 0.2A 0.1A 37 1.17 0. AO 16 0.A1 0.05 38 0.69 1.01 17 0.3A 0.06 39 0.52 0.72 18 0.78 0.30 AO 0.A5 0.1A 19 0.A5 0.31 A1 0.95 0.25 20 0.80 0.71 A2 2.58 1.56 21 0.69 0.29 A3 3.A5 7.96 22 0.77 0.69 37 Table 8. Dunde Chemistry Pit 5 Sample Number Cl” SO^" Sample Number Cl” SO^" 1 0.77 0.62 15 0.75 0.7A 2 0.78 1.26 16 0.38 0.A2 3 O.AA 0.A2 17 0.A1 0.3A A 0.31 0.1A 18 0.18 0.26 5 0.35 0.13 19 0.20 0.2A 6 0.A0 0.15 20 0.39 0.31 7 0.A2 0.13 21 0.35 0.30 8 0.32 0.06 22 0.51 0.37 9 0.36 0.08 23 0.6A O.A6 10 0. A3 0.11 2A ' 0.76 0.33 11 0.90 O.AA 25 1.76 0.21 12 1.51 1.A3 26 1.60 0.27 13 1.62 1.23 27 1.50 0.63 1A 1.28 1.A6 38 interesting to note that the cycle still occurs at 0.5 meter* but the sulfate cycle is offset. The highs in sulfate levels correspond to lows in the chloride level. Figures 11* 12* and 13 show the chloride levels in the two profiles of pit 1 and one profile of pit 3 from Antarctica. Although the averages are different (see earlier discussion), the same trend of highs and lows are clearly seen in each of the three profiles. The sulfate levels in the two profiles of pit 1 (Figures 14* 15, and 16) show good agreement at each depth. In the samples from pit 3* the same trend can generally be seen. However, between 0.4 and 0.5 meter in depth* the high sulfate levels seen in pit 1 samples are not as distinct as in pit 3 samples. Figures 17 and 18 show the nitrate levels as a function of depth. The same pattern is exhibited in both profiles although at 0.3 meter the high value seen in profile A is not seen in profile B. Overall, the nitrate averages are in good agreement. The difference between the two averages is only 3.3 ppb. The use of ice core samples to uncover records of events of climatic importance can be seen in Figure 10. The yearly cycle exhibited by sulfate in core B is disrupted between 110.5 and 111.5 meters. This is likely due to volcanic activity in the southern hemisphere approximately 100 years ago (21). 39 U•J f* ID W DEFTH 1*1 com INI Core A «. H V C flB C t lit M D I M M I k COKE (HI Core B Figure 9. Chloride in Core Samples, Siple S PVERnnE = 5«. J *0.0- _J PPB 110.O' 1 100.0. 1 1*0,0 •o.o. *0.0. JO.O * * 0 0 . . 0 0 ' ' iue 3 Clrd nPt alB Siple B» 3 Mall Pitin Chloride 13. Figure A3 AA at 1.00 Sulfate VailIn B Profile ASiple t DEPTH IN DEPTH CORE IM) Figure 1A. ' 0.00 0 . 0.0' 10.0 to.o- 10 Jf.O II.O' 21.0. s.o Qdd has 4 5 4 1.00 LD UJ UJ Cl 1.09 DEPTH IN DEPTH CORE (HI Figure 15. Sulfate Uallin B Profile B, Siple 0.00 . 0.& 10.0 li.O t.O 31.0 10 «0.0 30.0- «i.O' add hos soil PPB - O . M w.o- n.o- li.O- 10 . 0 0.00 . iue 6 Slaei i 3Wl B Siple B, 3 Wall Pit in Sulfate 16. Figure E T I CR (HI COREDEPTH IN 1.00 o 46 o z PPB 100.0 60 eo.C' so.o. 30 >10.0' 70.0 0.0- . . 0 0 - ' iue 7 Ntae nWl BPoieA Siple A, Profile B in Wall Nitrate 17. Figure E T I CR IH) COREDEPTH IN 1.05 UJ L3 47 kOj PP8 20 to. o. 10 10.0- 0.0. . . 0 0 O.OD ' ' iue 8 Ntaei al rfl » Siple B» Profile B in Wall Nitrate 18. Figure E T I CR IMJ COREDEPTH IN o 48 49 Injection Methods In 1981, Broquaite and Guinebault (22) showed that large volumes (up to 1 ml) could be Injected into a normal phase chromatographic column without loss of column efficiency if the sample was prepared in a non-eluting solvent. Buchholz and co-workers (23) showed that this technique worked in ion chromatography for samples of up to 2 ml. Large volume injection in ion chromatography has also been studied by Okada and Kuwamoto (24) and by Heckenberg and Haddad (25). The application of this technique to microbore was investigated by Silver and co-workers (26) and by van der Wal (27). The technique of large volume injections relies on the use of a non-eluting solvent for sample preparation. When the sample is forced from the sample loop onto the column, the non-eluting solvent allows the sample to be concentrated on the top of the analytical column. The limit for injection volumes in the above studies was 6 ml; however, most work involved less than 2 ml. This is due to the difficulty of working with large loops on Injectors (I.e. large back pressures and the pressure difference between injection and load positions). Early in this study, it was found that the use of large volume injection (0.64 ml) allowed detection in the ppb range without use of a concentrator column. When the sample was in a non-eluting solvent, no loss of efficiency occured. As a sample in a non- eluting solvent enters the analytical column, the sample is 50 retained on the top of the column until moved by the mobile phase. When a large volume'of sample is UBed, the retention on and elution from the top of the column will result in a more concentrated sample (on column concentration). As mentioned earlier, previous work with large sample volumes were limited to 6 ml or less due to problems of working with large sample loops. In thiB study, the sample loop was eliminated and very large volumes (up to 51 ml-) were loaded directly onto the analytical column (see Figure 19). Figure 20 shows a 100 ppb standard of fluoride, chloride, and sulfate using a 0.64 ml injection loop (output range 30). The standard was made in the eluant. This shows the broad peaks and low peak heights which result from using an eluting solvent which does not take advantage of the on column concentration. The 100 ppb standard is shown again in Figure 21 with the output range decreased 10 fold (sensitivity increased 10 fold). These peak heights are about what a 10 ppb standard with the previous output range (30) shows when done by on column concentration (Figure 25). Figure 22 shows a 1 ppb standard of fluoride, chloride, nitrite, nitrate, dlhydrogen phosphate, and sulfate with a 5 minute (8.5 ml) on column loading. The low base line for the first two minutes is due to the water from the sample eluting from the column. The same standard is shown in Figure 23 after a 10 mln (17 ml) on column loading. 51 Standard Method Mobile PhaBe 3 - C Analytical Fiber Column Suppressor Injection Detector Loop Very Large Volume on Column Concentration Mobile Phase Valve Sample Pump Analytical Fiber Column Suppressor Detector Figure 19. Comparison of Sample Loading Methods 52 A 10 ppb standard, 5 minute (8.5 ml) on column loading is shown in Figure 24, The 10 ppb minute (17 ml) on column loading is shown in Figure 25. The 1 ppb standard was rerun using an injection loop- (0.64 ml) in Figure 26. The only peak noticeable is for chloride. The dip in the baseline at one minute is due to water from the sample eluting from the column. Figure 27 shows the 1 ppb standard at a higher sensitivity (output range 10). The fluoride, chloride, nitrate, and sulfate peaks are detectable but much smaller than those obtained by on column concentration. Figure 28 shows a chromatogram of a 100 ppb standard at an output range of 30 using a 0.64 ml injection loop. These peaks are substantially smaller than those for 10 ppb (Figure 25) using on column concentration. Even with the sensitivity increased 3 fold (output range 10), the 100 ppb peaks (Figure 29) are only about the same as the 10 ppb peaks using on column concentration (Figure 25). Figure 30 shows the 1 ppb standard using a concentrator column loaded with 17 ml. The peaks are not as high or gausslan as those done by the on column concentration (Figure 23), Figure 31 is a chromatogram of a 10 ppb standard with 8.5 ml loaded onto a concentrator column. This should be comparable to Figure 24 of 10 ppb with a 5 minute loading, but the peaks from the concentrator column are larger. This will be discussed under the linearity of response section. 53 £n Figure 32, a 17 ml sample of 10 ppb was loaded onto the concentrator column. The fluoride peak is much smaller than that seen for the 17 ml of 10 ppb loaded by on column concentration (Figure 25). Other peaks are about the same peak height. Figure 33 shows a 1 ppb standard (17 ml) on column loading with an output range of 10. At the same output range and volume (on column loading), a sample was run (Figure 34) showing 300 ppt of chloride, nitrate, and sulfate. This shows that chloride, nitrate, and sulfate can be detected in the ppt range. Figure 35 is a chromatogram of a typical 17 ml sample of Milli Q water analyzed by on column concentration (output range 30). Small amounts of fluoride, chloride, nitrite, and sulfate can be seen. Figure 36 is a 20 ppt sample with a 30 minute (51 ml) loading. The output range was 10. Fluoride, nitrate, and sulfate can be detected at this level. The high chloride peak is due to contamination which 1b a serious problem at this level. Contamination from airborne chloride was a consistent problem with the longer loading times. Figure 20. 100 ppb Standard in Mobile Phase (or 30) J___I Figure 21. 100 ppb 'Standard in Mobile Phase (or 10) 55 NO SO, I i "■" I 0-11 Min. Figure 22. 1 ppb Standard; 8.5 ml on Column Loading (or 30) NO, m ■"V so Figure 23. 1 ppb Standard; 17 ml on Column Loading (or 30) 56 Cl i i i" —■ i i ■ i . . . i i \ 0-11 Min. Figure 24. 10 ppb Standard. 8.5 ml on Column Loading (or 30) 57 H 2P04 0-11 Min. Figure 25. 10 ppb Standard, 17 ml on Column Loading (or 30) 58 \rCl \v i i ■ " i i i r~ r i ■ ■ ■ t i i 0-11 Min. Figure 26. 1 ppb Standard, 0.64 ml Injection (or 30) Cl NO. SO. ) fJ h 0-11 Min. Figure 27. 1 ppb Standard, 0.64 ml Injection (or 10) 59 F NO NO, r ■" i i i i 111 ■ i i 1 ■ i i 0-11 Min. Figure 28. 100 ppb Standard, 0.64 ml Injection (or 30) 60 F I I 9 I no: NO H.PO SO 0-11 Min. Figure 29. 100 ppb Standard* 0.64 ml Injection (or 10) 61 < i i ■ i ■ "i — ■ i . . i i i \ 0 - U Min. Figure 30. 1 ppb Standard, 17 ml Using Concentrator Column (or 30) 62 Cl NO H-PO SO i > ""I i ■ i i I i i 0-11 Min. Figure 31. 1 ppb Standard, 8.5 ml Using Concentrator Column (or 30) 63 NO I----- 1 I 1 I 1 ■ I — i— | ■ > ' —r— — —r — — .'f ! 0-11 Min. Figure 32. 10 ppb Standard, 17 ml Using Concentrator Column (or 30) 64 Cl NO, SO NO, 0-11 Min. Figure 33. 1 ppb Standard, 17 ml on Column Loading (or 10) 65 Cl NO soT 0-11 Min. Figure 34. Sample Containing Chloride, Nitrate, and Sulfate at Approximately 300 ppt, 17 ml on Column Loading (or 10) Cl 0-11 Min. Figure 35. Milli-Q Water, 17 ml on Column Loading (or 30) 66 Cl NO 0-11 Min. Figure 36. 20 ppt Sample; 51 ml on Column Loading (or 10) 67 Linearity of Response An ideal set of working curves is shown in Figure 37. Figure 38 shows the working curve for fluoride. The 5 minute loading for on column concentration shows the best linearity but lowest detector response. For a 5 minute loading, only 8.5 ml (as compared with 17.5 ml) Is loaded onto the column which results in the lower detector response. It is interesting to note that in most cases the response of the 5 minute on column concentration is less than 1/2 of the 10 minute loading though it is the same technique but with 1/2 the volume. This probably is a result of the finite time needed to flush the column. As the loading time is increased, the time needed to flush becomes less significant and the response vs. time curve (Figure 39) becomes more linear. Figure 40 presents the working curve for chloride. From this figure it can be seen that the concentrator column and on column concentration (10 minutes) give comparable results for chloride. The time needed for re-equilibration resulted in peak broadening for the chloride when using on column concentration. This broadening resulted in the loss of base line resolution between chloride and nitrite. As a result, the on column concentration gave a lover apparent peak height for nitrite than the concentrator column (Figure 41). The working curve for nitrate (Figure 42) shows the similarity in response of the concentrator column and on column concentration (10 minute loading). In this case the concentrator 68 column gave slightly better linearity and detector response. Figure A3 shows the working curve for HjPO]*. Here the 10 minute loading for on column concentration gave the exact, same results as the concentrator column. Table AA shows the working curve for sulfate. The on column concentration shows better linearity than the concentrator column. The non-zero y-intercept for most lines is a result of the impurities found in the Milli Q water in which the standards were made (see Figure 35). { PPB 5 tain, loading on column concentration 10 min. loading on column.concentration concentrator column Figure 37. Ideal Working' Curve 0 5 10 PPB + ” 5 mln. loading on column concentration ■ 10 min. loading on column concentration x ” concentrator column Figure 38. Working Curve for Fluoride ° Detector Response i 10 - 0 Figure 39. Detector Response vs Loading for 10 ppb S0^ 10 ppb for Loading vs Response Detector 39. Figure 5 10 odn (tain.) Loading 15 20 25 m 30 Detector Response Q * 10 min. loading on column concentration concentration column on loading 10min. * Q + ■ 5 rain, loading on column concentration concentration 5column rain, on loading ■ + x « concentrator column concentrator « x iue4. okn uv for Chloride Curve Working 40. Figure PPB 0 5 10 PPB + = 5 min. loading on column concentration 0=1- min. loading on column concentration x ■ concentrator column N l W Figure 41. Working Curve for Nitrite Detector Response .8 .6 .4 .2 1. 0 0 concentrator column concentrator 10 min. loading on column concentration concentration column on loading 10 min. 5 min. loading on column concentration concentration column on loading 5min. Figure 42. Working Curve for Nitrate Curve Working 42. Figure PPB 5 10 .c~ Detector Response 5 0 □ « 10 min. loading on column concentration concentration column on loading 10 min. « □ + ■ 5 min. loading on column concentration concentration column on loading 5 min. ■ + x x 3 concentrator column concentrator Figure 43. Working Curve for for H^PO^ Curve Working 43. Figure PPB in _ 10 Detector Response 6 5 4 3 2 0 1 + ** 5 min. loading on column concentration column on loading 5 min. **+ x = concentrator column concentrator = x • 10 min. loading on column concentration concentration column on loading 10 min. • Figure 44. Working Curve for Sulfate for Curve Working 44. Figure PPB 10 O' 77 Efficiency Table 9 shows che efficiency of the system with the four methods examined during this study. Efficiencies were measured from the sulfate (longest retained) peak. Table 9. Comparison of Efficiencies ______Method______No. of Plates 10 Ml loop-sample in eluting solvent 650 0.64 ml loop-sample in non-eluting solvent 900 concentrator column 900 large on column concentration (8.5 ml) 1020 large on column concentration (17 ml) 1020 large on column concentration (51 ml) 1038 The 10 m 1 loop with sample in an eluting solvent showed the poorest efficiency. The 0.64 ml loop gave the same results as the concentrator column. The large on column concentration gave the best results. Pit Samples Several pit samples from profile A of pit 1 from Siple Station, Antarctica, were run by both large volume (0.64 ml) injection loop and very large volume (17 ml) on column concentration. Figure 45 shows the chromatogram of a pit sample using the injection loop. Figure 46 is a chromatogram of the same sample using a 17 ml sample. It can be seen that the peaks are 78 much more obvious for the larger sample volume. This makes peak height measurements much easier and more accurate. Table 10 shows the results of 4 pit samples using both methods. The values obtained from the larger sample volumes are slightly lower for chloride and sulfate. Generally, there is good agreement between the samples and the same trends are definitely present. These four pit samples were chosen to be run by both methods because they had sufficient volume for multiple analyses and had low concentrations of all anions such that none would be offscale using the more sensitive method. 79 Table 10. Comparison of Injection Methods 0.64 Injection Loop Sample Number F Cl NO^ S0^ 56 - 16 28 36 58 - 25 29 36 59 - 27 28 25 69 - 18 42 33 17ml on Column Concentration Sample Number F Cl N0^ S0^ 56 1 11 31 58 8 22 33 30 59 6 25 23 23 69 8 14 47 29 Figure 45. Pit Sample 56,. 0.64 ml Injection (or 10) Cl I t i t SO Figure 46. Pit Sample 56, 17 ml on Column Loading (or 10) CHAPTER 2 THIN LAYER CHROMATOGRAPHY OF THE FIRST ROW TRANSITION METAL IONS Introduction The beginning of TLC can be found back as far as 1889 with the work of Beyerinek (28) who used a thin layer of gelatin to separate hydrochloric acid from sulfuric acid. In 1898, Wijsman (29) used the same technique to separate enzymes from malt diastase. Ismailov and Schraiber (30) were the first to use TLC as a screening tool for column chromatographic systems. They used a thin layer of aluminum oxide on a glass plate and developed the plate by first placing a drop of the solution to be separated in the middle of the plate, followed by drops of solvent, resulting in concentric zones. Their procedure was called "drop chromatography". In 1948, Meinhard and Hall (31) used a binder to adhere alumina to microscope slides. These plates were used to separate inorganic ions using drop chromatography. Further details of the history of inorganic TLC will be given later. In the 1903*a, Kirchner jet al. developed the techniques of TLC as we know it today (32,33,34,33,36). In 1938,' Stahl (37) coined the term "thin layer chromatography". Detailed histories of TLC have been provided by Stahl (38), Heftmann (39), Kirchner (40,41,42) and Pelick (43). 81 82 Metal Ion Separation Frache (44,45,46) reported the TLC of up to 61 metal Ions on silica gel plates. They used mobile phases of nitric acid, sulfuric acid, or phosphoric acid in acetone. Fe(IIl), Nl(II), and Cu(II) were separated using pure acetone. Varshncy (47) also studied the TLC of metal ions on silica gel plates. Separation of 46 metal ions was attempted using methanol/ nitric acid as the mobile phase. Qureshi and Thakur (48) examined the chromatographic behavior of 47 metal ions on silica plates. The mobile phase was 1M aqueous sodium chloride/acetone. Ni was separated from Cu and Zn, but large spot sizes prevented further separation. Baffi, Dadone, and Frache (49) examined the TLC of Zn, Nl, and Cu on cellulose. As a mobile phase they tried various concentrations of tartaric acid and ammonium hydroxide. Soljic and Grba (50) separated Ni, Mn, Co, Cu, Fe, and Zn with cellulose as the stationary phase. The mobile phase was 86% acetone, 8% hydrochloric acid, and IX water. The same mobile phase could not effect a separation with a stationary phase of silica gel. Diethylaminoethyl-cellulose was used by Kuroda et al. (51) while examining the behavior of 47 metal ions using various concentrations of sodium thlosulfate in water. Little separation was achieved. 83 Separation of Metal Complexes Rao and Sheker (52) used silica plates to separate the xanthates of Cu, Ni, and Zn. Toluene/benzene was used as the mobile phase. Oksala and Krause (53) also uBed silica plates to separate metal ion complexes. The acctylacetonates of Fe(Ill), Cu(II), Ni(II), Co(ZII), and Cr(lll) were separated uBing a mobile phase of diethyl ether. Cu and Ni were easily resolved; however, Fe showed an elongated spot. Silica plates were also used by Singh (54) to separate metal- morphollne-4-carbodithloate complexes. Using n-propanol, Ni and Cu could be separated. Fc, Ni, and Cu were separated with chloroform. Lohmueller and coworkers (55) examined the use of silica and alumina for TLC of dithlzonateB of Zn(II), Cu(II), and Ni(II). They tried a variety of eluants including: benzene, carbon tetrachloride, tetrahydrofuran, acetonitrlle, toluene, and chloroform. Complete separation was not achieved due to spot elongation. Cagliardi and Deutschmann (56) used alumina plates with a chloroform mobile phase for the separation of Fe, Cu, Zn, Co, and Pd complexes of x-methyl-N-2(pyridyl)methylene dithlocarbazate. No system was found in the literature which could separate all ten first rwo transition metal ions. Generally, the available literature reports only the chromatographic behavior of a few of 84 the transition metal Ions in any one system. In this work, twenty mobile phases were Investigated, three different types of stationary phases were used, and two methods of pretreatment of the stationary phases were examined. Thirteen complexlng agents were studied for use in locating the metal ion spots. The results for all ten metal ions are reported for each system. The goal of this project was to develop in a systematic way a separation and detection method which could be used for the first rwo transition metal ions in environmental samples. The TLC methods developed here could be adopted for HPLC which would be useful in the analysis of trace metals in ice samples. 85 Theory Retention In thin layer chromatography* the results are usually reported as an Rf value. This retention factor is defined as the ratio of the migration distance. (34) Rf ■ distance traveled bysolute distance traveled by solvent The distance traveled by the solute is determined by the amount of time spent in the mobile phase relative to the total time. The capacity factor of the stationary phase (K) is defined as (35) K ■ time in stationary phase ■ ts time in mobile phase tm Since (36) (36) Rf ■ ts/tm + ts then (37) Rf - 1/K + I 86 Plate Efficiency Another important parameter is the efficiency of a chromatographic separation measured by the height equivalent to a theoretical plate (H) (38) H -«r2/Rf(L - ZQ) where L is the total distance moved by the solvent front, Zq is the distance from the solvent reservoir to the original solute spot, and CT is the variance as calculated from the length of the final solute spot. original final solvent .spot spot front 4 * Figure 47. Factors Important in Plate Efficiency The number of theoretical plates (N) is given by (39) N - L/H 87 The selectivity of a chromatographic system is defined s b the ratio of the Rf values. (AO) Selectivity ■ Rf solute 1 > 1 Rf solute 2 Resolution The degree of separation is referred to as the resolution. Figure 48. Resolution on a TLC Plate The resolution Is calculated from equation (41): (41) R - D2 - DlAOfj + W2) 88 Experimental Stationary Phase For normal phase chromatography, both Adsorbosil-Plus (Applied Science) and Silica Gel G (EM Reagents) were used. The plates were coated using a Desaga-Brinkman variable thickness spreader. The silica was made into a slurry using isopropyl alcohol (reagent grade from Mallinckrodt). Generally, coating thickness was 0.25 mm. For reverse phase plates, plates of Adsorbosil-PluB were placed into a developing tank containing a 10% solution of octadecyltrichlorosilane (ODS) (PCR research chemicals or Fluka) in toluene (reagent grade from Fisher). These plates were allowed to stand and react until 10 min. after the ODS solution had reached the top. Baker-flex sheets (J.T. Baker Chemical Co.) of silica gel 182 were also used in this study by reacting by the aforementioned method. This method of reacting however proved unsuitable for use with larger plates. Plates in which the width was greater than 5 cm would only partially react. These plates could be used for two-dimensional chromatography since they would have an approximately 5 cm strip of reverse phase with the rest of the plate normal phase. For work requiring larger plates, manufactured plates of silica gel 60 on plastic (Alltech Association) were used. These plates were reacted by placing 100 ml of \% ODS in toluene in a 19 cm x 30 cm pyrex baking dish (or 150 ml of solution in a 22 cm 89 x 34 cm dish). The baking dish was tipped so that the solution was all on one side* and the plateB were placed on the bottom of the dish. The dish was then quickly set to a level position, allowing the ODS solution to completely cover the plates. A cover was placed over the dishes to prevent reaction of the ODS with atmospheric moisture. The plates were allowed to react for 20 minutes. For HPTLC, Whatman Linear-K High Performance Silica Gel plates (LHP-KD) were used. The plates were 20 cm x 10 cm with a 200 micrometer thickness. When used for reverse phase work, these plates were reacted using the second method. Spotting 100 mg/ml standard solutions of metals were prepared according to Standard Methods for Examination of Water and Wastewater (57). A 1000 ug/ml Scandium standard was obtained from Alfa Products. For metals which were not easily detected at 1.0 mg/ml, more concentrated solutions of the metal chlorides were used. For reverse phase work, the metal ion solutions were prepared in either 50% isopropyl alcohol (reagent grade from Mallinckrodt) or acetone (reagent grade from Mallinckrodt). The aqueous metal ion solutions were made from zinc(II) chloride, copper(II) nitrate, nickel(II) nitrate, cobalt(II) chloride, iron(III) nitrate, manganese(II) sulfate, chromium(lll) chloride, and scandium(III) chloride. Vanadlum(V) oxide was dissolved in sodium hydroxide. Titanium(IV) dioxide was dissolved 90 In hydrofluoric acid. The transition metals ions in solution can form a variety of complexes. For example Titanium dioxide _2 dissolved in aqueous hydrofloric acid is likely to form TiF^ , but +2 can also form complexes with water such as TifOH^d^O)^ . Therefore* only the oxidation states of the metals will be listed. For spots of more than 1 microliter, a Hamilton 701-N syringe was used. A Hamilton 7001 or 7101 syringe was used for sub- mlcroliter spots. Development Glass tanks of approximately 5 L volume were used. Approximately 70-100 ml of solvent was added. Whatman no. 1 filter paper was placed inside the tank to help equilibrate the air Inside the tank. The tank was then covered with a glass lid and allowed to equilivrate for at least 20 minutes. Next, the plate was placed Inside and allowed to develop. Detection When the solvent reached an appropriate point (usually 11 cm), the plate was removed and immediately dried on an L & R Ultrasonic Dryer.' This was done to limit futher diffusion of the spots. The plates were then sprayed with a color forming complex. The chemicals for detection will be discussed in the results and discussion section. General Other chemicals used but not specified above were of reagent grade. Water used was either distilled or Nanopure. 92 Results and Discussion Spotting Techniques On several of the types of plates used, In particular the plates which had been pre-equilibrated with the mobile phase, the Initial spot diffused rapidly. Heating the plate during the spotting evaporated the solvent so that smaller Initial spots could be obtained. Separation Table 11 gives the list of mobile phases which were used for TLC. Several stationary phases were examined. These include: Adsorbosil (Silica Gel), Silica Gel G, C-18 bond Adsorbosil, and Aluminum Oxide G. In addition, experiments were conducted to determine the effect of pre-treatments of the stationary phases. Table 12 shows the difference in the two types of silica gel used and the effect of pre-treatments. There was no major difference between Adsorbosil and Silica Gel G. The most noticeable difference Is for Ti(IV) where the Rf Is conslstantly higher for the Silica Gel G. The V(V) spot was visible on the Silica Gel G, but was undetectable on Adsorbosil. Cr(IIl) which did not move on the pre-heated Adsorbosil, spread over almost the entire plate on the pre-heated Silica Gel G. [Note: Rf values were not reported for spots which were very diffuse.] Ni(II) moved better on the Adsorbosil than on silica gel when the plate was either heated or not pre-treated. The pre-equilibrated 93 Silica Gel G; however, ahowed a higher Rf value for Ni(II) than the pre-equilibrated Adsorboail. The particle size difference accounts for the difference in behavior between the two gels. Adsorbosil has a smaller particle size and, therefore, a larger surface area. The larger surface area would result in a lower Rf value for an ion which was attracted to the OH groups on the surface [Ti(XV)]. The smaller particle size will lead to more compact spots. The pre-equilibrated plates generally showed lower Rf values than non-equillbrated plates. Ti(IV) is the only metal which showed equal or higher. Rf values for the pre-equilibrated plates. Several metals [V(V), Sc(IlI), Fe(IIl), Co(II), and Cu(II)] showed comparable or slightly larger spot widths when pre- equilibrated. The other metals showed much narrower spots' on the pre-equilibrated plates. Cr(lll), for example, had a spot width of 2.5 cm on the Silica Gel G, but only a width of 0.7 cm on the pre-equilibrated plate. The pre-heating of the plates made little difference in either the Rf value or the spot shape. Several sets of metals can be separated using this mobile phase. Ti(IV) [Rf - 0.1], Mn(II) [0.6J, Ni(II) [0.7], and Fe(lII) [1.0} can be separated on Adsorbosil. On the pre- equilibrated Adsorbosil plates, Mn(II) can be separated from Sc(IIl), Cr(III), Fe(III), Cu(Il), and Zn(II). Fe(III) can be separated from Ti(IV), Cr(III), Mn(II), Co(II), and Ni(II). Table 13 shows the behavior of metals on silica gel (normal phase) plates with 3 different solvent systems. With chromotropic 94 acid (solvent G) as the mobile phase, all the metals were detectable and each metal had a lower Rf on the pre-equilibrated plate. With 8-Quinolinol (solvent G), Sc(III) and Cr(ZXZ) showed higher Rf values on the pre-equilibrated plates. All other metals followed the trend of a low Rf values with pre-equilibration. The pre-heating resulted in approximately the same Rf but up to 3 times the spot width. Several metals can be separated using 8-Quinolinol on Silica G: Cr(III) [OJ. Sc(ZII) [0.11, Ti(IV) [0.7], and V(V) [0.9]. The other mobile phase/stationary phase combinations did not look promising. For 0.1M Formic Acid (H), all metals travelled very close to the solvent front; therefore, this system was considered for further work in electrophoresis. Table 14 shows the normal phase TLC with five different mobile phases. In the last four, pre-equilibration is the only method reported as it gave the narrower spot widths. The silica plate run in pure acetone (N) could not be considered pro-equilibrated because the acetone evaporated during the spotting procedure. The acetone/silica gel G system does permit the separation of Cr(III) [0.1] from Sc(III) [0.4] or Ti(IV) [0.3] from Fe(III) [0.6]. Several other mobile phases were capable of separation of binary mixtures. Diphenylcarbohydrazide (L) could separate Fe(III) from Sc(III), Ti(IV), Cr(lll), Mn(II), and Ni(II). Chromotropic acid (F) could separate Sc(III) from Ti(IV) and Cr(III) and Fe(IIl) from Ti(IV), Cr(III), Co(II), and Ni(II). 95 Dimethyl glyoxlme (H) could separate Ti(IV) and Fe(lII) from Cr(III), Mn(II), and Co(II). Table 15 shows the reverse phase behavior of metals on reverse phase plates. Each of 3 solvents were run with and without pre- equlllbratlon. The pre-equilibration did not affect the Rf values as much as in normal phase TLC. Chromotropic acid (F), pre-equilibrated, was able to separate Ni(II) [0.1], Cr(lII) [0.2], Co(II) [0.4], Fe(III) 10.7], and V(V) [0.9]. Dimethyl glyoxlme, pre-equilibrated, separated Ni(II) [0.2], Cr(III) [0.4], V(V) [0.5], Fe(III) [0.7], and Sc(IXl) [0.9]. The Dlphenylcarbohydrazide (L) could separate Ni(II) [0.1], Cr(IIl) [0.3], Zn(II) [0.4], and Fe(III) [0.5]. Table 16 shows the reverse phase TLC with 3 other solvent systems. Alizarin (K), without pre-equilibration, could separate V(V) [0.1], Cr(III) [0.5], and Cu(II) [0.6], For Dithizone (I), all the metals moved with the solvent front. Ti(IV) could be separated from V(V), Cr(IlI), Fe(III), Co(II), and Cu(ll) by a mobile phase of PAN (J). Further separation with PAN could not be achieved due to the large spot widths. Buffered EDTA solutions were used as mobile phases for 3 different stationary phases. The buffers of pH 1.81, 4.10, 6.09, 7.96, and 9.91 were made by adding 0.0 ml, 25.0 ml, 42.5 ml, 60.0 ml, and 77.0 ml respectively of 0.2 N HaOH to a 100 ml mixture of phosphoric, acetic, and boric acids (0.04 M). The EDTA solution was 0.1 M. 96 Table 17 shows the results when these solutions were used with silica gel plates as the stationary phase. Except for Cr(IIX), Rf values were the sane or less for the pre-equilibrated plate. Ci(Il) showed the sane Rf volues for each solution. Ni(II) had a higher Rf for a pH of 6.09. Cr(Ill) and Co(II) also showed higher Rf values at higher pH'B. V(V), however, showed less migration at higher pH values. On an aluminum oxide stationary phase all metals showed substantially less movement (see Table 16). Co(Il) and Zn(ll) both showed higher Rf values when the plates were pre- equilibrated. The low Rf values made differences harder to detect. For V(V) and Cr(III), Rf values were the same for all buffer systems. Cu(II), Co(II), and Fe(III) moved less in the pH of 4.10 system. The reverse phase plates (Table 19) shoved a marked Increase in movement for all ions, except Fe(III), when the pH was Increased from 1.81 to 4.10. The movement of V(V) dropped off as pH was further increased. Except for Zn(II) at pH 7.96, all other values remained about the same as the pH increased. Both Cu(ll) and Co(II) separated into two spots with each system. The Rf value given is that of the larger, more concentrated spot. There are several factors which can influence the retention of metal ions in TLC. In fact, any one TLC separation is likely to be a result of several different mechanisms acting simultaneously. It is believed that the major interaction in normal phase TLC 97 is competition between the OH groups on the surface of the silica gel and the complexing agent in the mobile phase. For the pre- equilibrated platesi this changes to a partitioning between the complexing agent adsorbed onto the surface and the complexing agent remaining in the mobile phase. This would account for the lowering of the Rf values for pre-equilibration of metals which Interact strongly with the complexing agent (Rf ^ 0.4). For ions which Interacted more strongly with the OH groups of the silica gel, an increase in Rf value was seen after pre-equilibration due to the lower availability of OH groups which were now occupied by complexing agent. In reverse phase chromatography the predominant Interaction is likely to be a partitioning of the metal complex between the stationary and mobile phases. This partitioning would not be greatly affected by pre-equilibration. This is shown in the data. HPTLC Use of the HPTLC plates (Table 20) resulted in smaller spot widths. As a result, smaller differences in Rf values are required for separation. The spot sizes in this study were reduced by approximately 1/2 with HPTLC. Chromotropic acid (F) mobile phase on a reverse phase plate could separate N1(X1) [0.1], Tl(IV) [0.4], Zn(II) [0.5], and Fe(XlI) [0.7] without pre-equilibration. With pre-equilibration, only binary solutions could be separated. 98 Dimethyl glyoxlme (M) without pre-equilibration was able to separate Cr(lll) [0.2], Zn(II) [0.4], and Fe(lll) [0.5]. With pre-equlllbratlon, Nl(II) [0.3], V(V) [0.4], and Fe(lII) [0.6] or Co(II) [0.5] showed the best separation. Dlphenylcarbohydrazlde (L) showed good separation of Ni(II) [0.2], Zn(II) [0.4], and Fe(III) [0.6] with and without pre- equilibration. Detection Several different spray reagents were used to locate the spots after development. Table 21 gives the list of reagents and their method of preparation as used in this study. Table 22 shows the colors formed between the complexing reagents and the 10 metals immediately after the application of the reagent. The relative visibility Is also given. With time, some spots became more visible while others nearly disappeared. Table 23 gives the colors and Intensities after 1 hour and Table 24 gives the colors and intensities after 2 hours. V(V) with PAR is a good example of a spot fading with time. By the end of 2 hours it was little more than colorless. After 3 hours it had completely faded. Alizarin had several spots which were barely visible immediately, but became very noticeable within 1 hour after spraying. PAR ahows the best overall color forming ability; therefore, it was chosen as the main spray reagent. Dlphenylcarbazide was also used because it reacted with Cr(III) (PAR did not). 100 Table 11. Solvent Systems Used For TLC Letter Code Chemical Composition A Water B 1.5% Citric Acid C 1.5% Lactic Acid D Acetone/HCl/Acetlc Acid (90/10/2) E Chromotropic Acid (1% aqueous) F Chromotropic Acid (0.2% in acetone) ♦ G 8-Quinolinol (0.2% In water) H Formic Acid (0.1 M aqueous) 1 Dithlzone (0.2% In acetone) J PAN (0.2% In acetone) K Alizarin (saturated in methanol) L Diphenylcarbohydrazlde (1% In ethanol) M Dimethyl Gloxime (1% in methanol) N Acetone Table 12. Comparison of Stationary Phase and Pretreatment Techniques Stationary Mobile Phase Phase V______Sc_____ Ti_____ Cr_____ Mn Fe C o ____ Ni Cu Zn Adsorbosil D 8.1-9.0 0-4.5 5.5-8.1 9.1-10 7.4-8.9 6.2-8.5 8.4-9.5 (10cm) (-) (0.8) (0.1) (-) (0-6) (1.0) 10.8) (0.7) (0-9) (-) Adsorbosil D - A.8-6.5 0-4.5 4.6-5.3 2.2-4.0 5.7-6.9 3.1-5.1 3.2-4.6 4.3-5.7 5.0-6.0 Pre- (10cm) (-) (0.6) (0.1) (0.5) (0.3) (0.6) (0.4) (0.4) (0.5) (0.5) equilibrated Adsorbosil D — 9.2-10 0-2.3 0 7.2-10 9.9-10 8.9-10 6.9-8.0 9.2-10 9.9-10 Heated (10cm) (-) (0.9) (0.1) (0) (0.8) (1.0) (0.9) (0.7) (0.9) (1.0) Silica G D 9.1-9.8 8.0-9.2 I.8-3.7 7.0-9.5 4.5-9.0 9.2-10 7.2-9.4 4.0-8.8 8.5-10 6.6-7.5 (10cm) (0.9) (0.9) (0.3) (0.8) (0.7) 0 . 0 ) (0.8) (0.6) (0*9) (0.7) Silica G D 6-5.7 4.7-5.7 3.0-4.0 4.8-5.5 3.2-4.7 5.5-7.5 4.3-5.3 3.8-4.6 4.5-5.8 5.7-6.4 Pre- (10cm) (0.5) (0.5) (0.4) (0.5) (0.4) (0.6) (0.5) (0.4) (0.5) (0.6) equilibrated Silica G D B.1-9.5 8.4-9.4 1.9-3-3 0-9.3 4.6-8.4 9.2-10 8.2-9.9 4.6-7.7 7.9-10 8.7-10 Heated (10cm) (0.9) (0.9) (0.3) (-) (0.6) (0.9) (0.9) (0.8) (0.9) (0.9) Distance traveled in cm ( ) • Rf value Table 13. Behavior of Metal Ions in Normal Phase TLC Stationary Mobile Phase Phase V______Sc_____ Ti_____ Cr_____ Mn_____ Fe_____ Co_____ Ni Cu Zn Silica G G 3.5-9.0 2.0-4.9 7.3-10 0-8.5 7.6-10 5.2-10 8.2-10 8.6-10 2.2-10 3.0-9.0 (10cm) (-) (0.4) (0.9) (-) (0.9) (0.8) (0.9) (0.9) ( ) ( ) Silica G E 0 0-6.0 3.2-5.4 4.5-5.8 3.6-5.5 2.5-6.0 3.3-5.8 3.5-5.7 1.3-5.8 3.6-6.1 ‘ Pre- (10cm) (0) (0.3) (0.4) (0.5) (0.5) (0.4) (0.5) (0.5) (0.4) '0.5) equilibrated 0 Silica G E 0 0-6.5 7.2-8.8 6.4-9.5 2.0-10 0-10 4.0-10 1.5-10 1.5-10 7.5-10 Heated (10cm) (0) (0.3) (0.8) (0.8) (-) (-) (0.9) (0.7) (-) (0.8) Silica G G (r 8.5-9.3 0-2.5 6.0-8.0 0 6.0-9.0 0-8.4 6.5-9.1 6.9-9.1 3.3-8.3 6.5-9.2 (10cm) (0.9) (0.1) (0.7) (0) (0.8) ( ) (0.8) (0.8) (0.6) (0.8) Silica G G 0-5.0 5.5-6.5 5.0-6.0 5.6-6.6 4.2-6.0 0-6.0 5.3-6.5 4.3-5.8 0-4.0 4.8-7.2 Pre- (10cm) (0.3) (0.6) (0.6) (0.6) (0.5) (0.3) (0.6) (0.5) (0.2) (0.6) equilibrated 00 0 1 * Silica G H 7.7-10 8.2-9.8 8.2-10.! 8.4-11.5 7.7-10.5 8.0-10 • 9.3-10. 7.0-9.1 0-8.8 (11.Sea) (0.8) (0.8) (0.8) (0.9) (0.8) (0.8) (0.8) (0.8) (0.8) (-) Distance traveled In cm { ) ■ Rf value Table 14. Behavior of Metal Ions In Normal Phase TLC Stationary Mobile Phase Phase V______Sc_____ Ti_____ Cr_____ Mn_____ Fe_____ Co_____ NI Cu Zn Silica G . N 3.0-4.1 3.3-3.5 0-2.0 0-4.0 6.0-7.0 2.5-4.5 _ 3.2-6.2 0-6.8 (10cm) (-) (0.4) (0-3) (o.i) (0-2) (0.6) (0.4) (-) (0.5) (0.3) Silica G H 8.0-9.5 7.7-8.5 5.4-7.9 8.0-8.8 6.4-8.6 6.0-7.5 6.6-7.6 7.4-8.3 6.0-7.1 6.2-7.2 Pre- (10cm) (0.9) (0.8) (0.7) (0.8) (0.7) (0.7) (0.7) (0.8) (0.7) (0.7) equilibrated Silica G L — 0-4.5 0-3.0 0-2.0 0-4.0 5.8-7.0 0-5.5 0-2.4 4.5-6.5 0-7.5 Pre- (lOca] (-) (0.2) (0.2) (0.1) (0.2) (0.6) (0.3) (0.1) (0.6) (-) equilibrated Silica G F p - 5.4-6.5 3.1-5.2 3.0-4.6 5.7-7.1 3.3-5.5 3.5-6.0 0-8.0 Pre- (10cm) (-) (0.6) (0.4) (0.4) (-) (0.6) (0.4) (0.5) (0.4) (-) equilibrated Silica G M — 4.0-7.6 5.5-7.0 3.5-4.9 1.5-3.9 6.2-7.2 2.2-4.0 3.5-7.2 5.0-7.0 Pre- (10cm) (-) (0.6) (0.6) (0.4) (0.3) (0.7) (0.3) (0.5) (-) f 0.6) equilibrated Distance traveled In cm ( ) - Rf value Table 15. Behavior of Metal Ions in Reverse Phase TLC Stationary Mobile Phase Phase V______Sc Ti_____ Cr_____ Mn_____F e _____ Co Ni Cu Zn C-18 F _ 0-3.3 1.8-5.9 7-2-11 4.9-8.0 1.0-4.2 5.6-10.2 6.5-7.5 (11.5cm] (-) (-) (0.1) (-) (0.3) (0.8) (0.6) (0-2) (0*7) (0.6) C-18 F 9.3-11. < —— 2.3-3.3 — 6.2-9.4 3.3-6.0 .8-1.9 4.2-7.5 Pre- (11.5cm] (0-9) (-) (-) (0-2) <-) (0.7) (0.4) (0.1) (0.5) (-) equilibrated C-18 M 6.0-8.0 — - 2.5-4.7 — 7.0-7.8 4.2-6.6 1.7-4.5 6.2-7.3 (10.5cm; (0.7) (-) (-) (0.3) (-) (0.71 (0.5) (0.3) (0.6) (-) C-18 M 5.2-6.4 9.2-9.8 — 4.0-4.9 — 6.6-7.3 4.3-5.9 2.2-3.0 6.5-7.3 Pre- (10.5cm] (0.6) (0.9) (-) (0-4) (-) (0.7) (0-5) (0-2) (0.7) (-) equilibrated C-18 L — 4.2-5.8 — 2.0-4.2 — 5.5-6.0 3.5-5.0 .3-1.5 5.0-5.9 4.2-5.0 (10.5cm) (-) (0.5) (-) (0-3) (-) (0.5) (0-4) (o.D (0*5) (0.4) C-18 L — 0-3.1 1.5-2.0 .7-4.0 6.4-7.2 4.0-4.6 0-2.6 5.5-6.6 5.5-6.2 Pre- (9.5cm) (-) (0.2) (0.2) (0.2) (-) (0.7) .(0.4) (0.1) (0-6) (0.6) equilibrated Distance traveled in cm ( ) ■ Rf value Table 16. Behavior of Metal Ions In Reverse Phase TLC Stationary Mobile Phase Phase V______Sc_____ Ti_____ Cr_____ Mn_____ Fe_____ Co_____ Ni Cu Zn C-18 K .5-3.0 4.5-6.4 5.2-6.5 4.8-6.5 6.5-8.5 _ (12cm) (0.1) (-) (-) (0.5) (-) (0.5) (0.5) (-) (0.6) (-) C-18 K — 7.0-8.0 5.5-6.5 — 5.5-6.9 6.3-7.5 0-7.0 5.0-6.0 » Pre- (10cm) (“ ) (o.a> (0.6) (-) (0.6) (0.7) (-) (-) (0.6) (-) equilibrated C-18 I 9.5-10 9.5-10 9.5-10 9.5-10 9.5-10 9.5-10 9.5-10 9.5-10 9.5-10 9.5-10 (10cm) (1.0) (1.0) (1.0) (1.0) (1.0) (1.0) (1-0) (1-0) (1.0) (1.0) C-18 J 7.0-9.0 - 2.5-3.5 7.8-9.7 « 7.5-9.0 6.0-7.5 0-8.5 4.5-10 1.0-8.0 (0-8) (-) (0.3) (0.9) (-) (0.8) (0.7) (0.4) (0.7) (0.4) Distance traveled in cm ( ) * Rf value Table 17. Effects of pH on Rf Values (Normal Phase) Sta tion ary Mobile Phase Phase V___ Sc Ti Cr Mn Fe Co Ni Cu Zn Silica EDTA _ 7.5-8.7 _ 7.1-8.1 . 6.5-8.0 6.5-8.2 Gel pH 1.81 ( - ) ( - ) ( - ) (0.8) ( - ) ( - ) (0.8) ( - ) (0.7) (0.7) K301R pre- equil EDTA 9.5-9.8 — — 6.7-8.2 6.7-7.3 5.8-8.5 6.8-9.0 — 6.0-10.0 6.5-10.0 pH 1.81 (1.0) ( - ) ( - ) (0.7) (0.7) (0.7) (0.8) ( - ) (0.8) (0.8) EDTA 9.2-10.0 _ — 8.4-9.8 — 8.8-10.0 6.4-9.5 8.6-10.0 5.5-10.0 7.8-10.0 pH 4.10 (1.0) ( - ) ( - ) (0.9) ( - ) (0.9) (0.8) (0.9) (0.8) (0.9) EDTA 9.5-10.0 9.1-10.0 8.4-10.0 9.5-10.0 8.0-9.0 8.8-10.0 9.0-10.0 9.2-10.0 5.4-9.6 5.2-9.5 pH 6.09 (0.9) (1.0) (0.9) (1.0) (0.8) (0.9) (1.0) (1.0) (0.8) (0.7) EDTA 8.2-8.8 8.5-10.0 9.2-10.0 9.5-10.0 6.8-9.1 9.0-10.0 8.0-10.0 6.3-10.0 8.0-10.0 pH 7.96 (0.8) (0.9) (1.0) (1.0) ( - ) (0.8) (1.0) (0.9) (0.8) (0.9) EDTA 8.6-10.0 _ — 9.2-10.0 — 6.7-8.1 8.0-10.0 8.7-10.0 7.0-10.0 4.7-10.0 pH 9.91 (0.9) ( - ) ( - ) (1.0) ( - ) (0.7) (0.9) (0.9) (0.8) (0.7) Table 18. Effects of pH on Rf Values (Aluminum Oxide) Sta tion ary Mobile Phase Phase V Sc Ti Cr Mn Fe Co Ni Cu Zn Alum EDTA 0.5-0.7 0.8-1.7 _ 0-5.1 0-5.2 1.0-1.9 0-4.6 0-5.5 inum pH 1.81 (0.1) ( - ) ( - ) (0.1) ( - ) (0.3) (0.3) (0.1) (0.2) (0.3) Oxide pre- G equil EDTA 0.5-1.0 —— 0-1.5 3.2-5.0 1.0-2.0 0.5-1.4 0-4.5 0-5.0 pH 1.81 (0.1) ( - ) ( - ) (0.1) ( - ) (0.4) (0.2) (0.1) (0.2) (0.2) EDTA 0-2.5 —— 0-1.5 0-1.0 0-2.2 0-1.5 0-2.3 0-10.0 pH 4.10 (0.1) ( - ) ( - ) ( - ) (0.1) (0.1) (0.1) (0.1) (0.1) ( - ) (4.5cm) EDTA 0-1.3 —_ 0-1.8 — 2.0-6.0 1.3-4.8 0-3.2 1.8-5.3 1.6-5.0 pH 6.09 (0.1) ( - ) ( - ) (0.1) ( - ) (0.4) ( ) ( ) ( ) ( ) EDTA — _ — 1.2-6.0 1.0-5.0 0-3.0 1.2-5.2 0.8-4.8 pH 7.96 ( - ) ( - ) (- ) ( - ) ( - ) (0.4) (0.3) (0.2) (0.3) (0.3) EDTA 0-1.2 — 1.7-6.0 3.5-5.2 1.0-2.8 1.0-5.2 1.2-4.2 pH 9.91 ( - ) ( “ ) ( - ) (0.1) ( - ) (0.4) (0.4) (0.2) (0.3) (0.3) o Table 19. Effects of pH on Ef Values (Reverse Phase) Sta tion ary Mobile Phase Phase_____ V______Sc______Ti______C r ______Mn Fe Co______Ni Cu Zn Silica EDTA 0.4-0.7 _ 0.2-1.6 _ 5.4-6.3 0.3-5.0 0.2-2.5 0.3-2.1 0.2-2.3 Cel pH 1.81 (0.1) ( - ) ( - ) (0.1) ( - ) (1.0) (0.4) (0.2) (0.2) (0.2) K301R (6.6cm) Reverse Phase EDTA 2.0-5.5 5.5-6.8 - 5.4-6.1 5.5-6.8 5.4-6.2 5.2-6.6 5.3-6.8 5.3-6.8 6.0r6.8 pH 4.10 (0.6) (0.9) ( - ) (0.8) (0.9) (0.8) (0.9) (0.9) (0.9) (0.9) (6.8cm) EDTA 1.6-3.1 5.3-6.8 5.2-6.2 5.3-6.8 5.3-6.8 5.4-6.8 5.6-6.8 0.6-6.8 6.2-6.8 pH 6.09 (0.3) (0.9) ( - ) (0.8) (0.9) (0.9) (0.9) (0.9) ( - ) (1.0) (6.8cm) EDTA 0.5-0.8 7.0-8.0 * 6.6-7.2 6.5-8.0 6.5-8.0 6.2-8.0 6.3-8.0 6.0-8.0 0.2-1.2 pH 7.96 (0.1) (1.0) ( - ) (0.9) (0.9) (0.9) (0.9) (0.9) (0.9) (0.1) (8.0cm) EDTA 0.3-1.5 4.6-6.0 4.5-5.5 5.0-5.6 5.0-6.0 5.0-6.0 5.0-6.0 5.0-6.0 4.8-6.0 5.2-6.0 pH 9.91 (0.2) (0.9) (0.8) (0.9) (0.9) (0.9) (0.9) (0.9) (0.9) (0.9) (6.0cm) Table 20. Behavior of Metals on Reverse Phase HPTLC Stationary Mobile Phase Phase V Sc T1_____ Cr Hn_____ Fe_____ Co_____ HI Cu Zn C-18 F 0-2.0 3.8-4.2 5.9-6.9 4.7-5.7 0-1.7 4.6-5.8 3.5-5.2 (9cm) (0.1) (-) (0.4) (-) (-) (0.7) (0.6) (0.1) (0-6) (0-5) C-18 F 0-3.6 3.3-5.5 — — 2.4-2.6 4.5-7.1 3.7-5.3 0-3.0 3.9-5.1 3.1-4.5 Pre- (8cm) (0.2) (0.6) (-) (-) (0-3) (0.7) (0.6) (0.2) (0.6) (0.5) equilibrated C-18 M ——— I.2-2.8 — 5.4-6.0 4.4-5.0 2.5-2.9 4.3-5.2 3.1-5.0 (9cm) (-) (-) (-) (0-2) (-) (0.6) (0.5) (0.3) (0.5) (0.4) C-18 H 3.8-4.0 ——— — 4.3-6.8 4.1-6.2 2.1-3.6 1.5-6.0 3.4-5.0 Pre- (9cm) (0.4) (-) (-) (-) (-> (0.6) (0-6) (0-3) (0-4) (0.5) equilibrated C-18 L - — — 3.8-4.7 5.4-6.0 5.3-5.8 4.3-6.5 0-3.3 4.5-7.7 3.5-4.2 (10cm) (-) (-) (-) (0.4) (0.6) (0.6) (0.5) (0.2) (0.6) (0.4) C-18 L 0-4.4 4.5-4.7 — 4.2-4.7 — 5.1-5.9 4.1-5.8 0-3.8 4.1-5.3 3.4-4.9 Pre- (10cm) (0.2) (0.4) (-) (0.4) (-) (0.6) (0.5) (0.2) (0.5) (0.4) equilibrated Distance traveled In cm ( ) * Rf value 110 Table 21. Developing Reagents for Locating Cations on TLC Plates Developing Reagents Abbreviations Preparations Alizarin AL Saturated solution in methanol Benzidine BZ 12 in glacial acetic acid Chromotropic Acid CA 12 aqueous Dimethyl Glyoxlme DC 12 in methanol Diphenylcarbizide DP 12 In ethanol Diphenylcarbazone DC 12 in methanol Dithizone DZ 0.12 in chloroform Erlochrome black t EB 0.042 in 0.1M ammonia solution l-(2-Pyridylazo)-2-Naphthol PAN 12 in methanol 4-(2-Pyridylazo)-Resorcinol PAR 12 in methanol Quinolinol Q 12 in methanol Stannous Chloride/ Potassium Iodide SC/KI S grams SnCl- in 10ml conc. HC1, dilute to 100ml, add 0.5 grams KX Tannic Acid TA 102 aqueous Table 22. Colors of Metal Complexes Formed With Developing Reagents (immediate) V Sc Ti Cr Mn Fe Co Ni Cu Zn Alizarin Colorless Light Yellow Blue Pink Yellow Purple Purple Yellow Yellow (saturated Yellow solution in - 1 1 1 1 2 1 3 2 2 methanol) Dithizone Yellow Yellow Pink Yellow Yellow Yellow Purple Yellow Yellow Yellow (.0022 in 1 2 2 1 2 2 1 I 1 2 chloroform) Diphenyl- Pink Pink Pink Pink Purple Carbohydrazide 3 1 2 3 1 (12 in ethanol) PAR (saturated Pink Yellow Yellow Pink Yellow Pink Pink Pink Yellow solution in 2 3 3 3 3 3 3 3 1 methanol) Benzidine *• Blue Blue ““ 3 2 Chromotropic Acid ------Dimethyl Glyoxlme ■“ Pink ** 3 Diphenyl- Yellow Yellow Pink Pink Yellow Purple Purple Purple Yellow Carbizlde 1 1 1 2 2 3 34- 3 2 Diphenyl- • Yellow/Blue Pink Pink Pink Yellow Purple Purple Yellow Yellow Carbazone 1 1 1 1 1 3 34- 1 1 PAN • Yellow Yellow Pink Yellow Purple Purple Purple Purple 1 2 1 2 3+ 34- 3 1 Qulnollnol • Yellow Yellow * 1 1 t SC-KI * Gray ** • • * ** 2 Key: I - 3 ■ light to dark Table 23. Colors of Metal Complexes Formed With Developing Reagents (after 1 hour) V Sc Ti Cr Mn Fe Co Ni Cu Zn Alizarin Pink Yellow Pink Pink Purple Yellow Purple Purple Yellow Yellow 1 1 1 1 2 3 2 2 3 3 Benzidine Gray • Gray “ Gray 1 1 1 Chronotropic Gray Gray •• ——• Acid 1 1 Dimethyl- • ** Pink GLyoxime 3 Diphenyl- Pink Pink Red Purple Pink Yellow Pink Purple Yellow Yellow Carblzide 2 2 3 3 1 2 3 34- 1 1 Diphenyl- Yellow Red Purple Pink Yellow Red Purple Gray Gray Carbazone 1 3 3 1 1 . 3 3+ 1 1 Dithizone Gray ■" 1 PAN Yellow Gray ■“ Pink Yellow Blue Pink Pink Gray 1 1 1 2 3 3 1 1 PAR Pink Yellow Yellow Pink Yellow Pink Pink Pink ■" 1 1 1 2 1 2 2 2 Quinolinol ■“ Gray • •• Gray Gray 1 1 1 SC-KI • Gray “ • 1 Key: 1 - 3 • light - dark Table 24. Colors of Metal Complexes Formed With Developing Reagents (after 2 hours) V Sc Ti Cr Mn Fe Co Ni Cu Zn Alzarin Pink Yellow Yellow Pink Pink Yellow Purple Purple Yellow Yellow 1 1 1 1 1 3 1 2 3 3 Benzidine • Gray White * White Gray Red Black Gray 1 2 1 2 1 2 1 Chronotropic Yellow White Gray White • Pink White White Acid 1 2 1 3 1 2 2 Dimethyl- • ■* • • ** Pink • Glyoxime 3 Diphenyl- Red Pink Red Purple Pink Yellow Pink Purple Yellow Yellow Carbizlde 3 3 3 3 1 2 3 3+ 2 2 Diphenyl- Gray Pink Purple Pink Yellow Red Purple Gray Gray Carbizone 1 3 3 1 1 3 3+ 1 1 Dithizone • Gray • ■" — 1 PAN • Yellow Gray • Pink Yellow Blue Pink Pink Gray 1 1 1 2 3 3 2 1 PAR Pink Yellow Yellow/ Pink Pink Yellow Pink Pink Pink Gray 1 1 1 1 1 1 2 2 2 Quinolinol • • Yellow * 2 SC-KI Gray “ * 2 Key: 1 - 3 = light - dark CHAPTER 3 ELECTROPHORESIS OF THE FIRST ROW TRANSITION METAL IONS Introduction The origins of electrophoresis are found as early as 1809 when F.F. Reuas described the migration of clay particles under an electric field (58). Strain and coworkers (59) separated inorganic radioactive mixtures using thick sheets of paper 3-6 ft. long and 20 in. wide. The sheets were held between thin flexible plastic sheets with the ends dipping into separate electrode vessels. Potentials of 5 V/cm were applied for up to 2 days. Radloautographical detection was used. Co, Zn, Cu, and Sc, could be separated using 0.1 M lactic acid. Lederer and Ward (60) separated Co, Nl, and Fe using KCNS in ethyl alcohol. More recently Bhatnagar and Yusufi (61) used aqueous and mixed water - alcohol media with NaNO^ as complexing electrolyte for the separation of Mn, Fe, Co, Ni, Cu, Zn, Ag, Cd, Hg, and Pb. Potentials of 5 V/cm were used. Separation of binary and ternary mixtures were accomplished. Yadava et al. (62) used paper electrophoresis in the study of formation constants of mixed ligand complexes in solution. Formation constants for Cu, Zn, and Co with glutaric and nitrolotriacetic acids 114 were determined. These studies were later extended to include NTA, tartaric acid, and adlplc acid complexes with Cu, An, Ni, Co, VO^, and Th(V). (63,64) Thin Layer Electrophoresis Thin layer electrophoresis has many advantages over paper electrophoresis. Thin layer plates allow for better cooling, higher applied voltages, and shorter analysis times. Moreover, use of thin layer plates allows a greater choice of separation media such as agar, ion exchange resins, and silica gel. Cellulose Shukla et al. (65) studied the separation of ferritin and transferrin complexes of the group I11A metals on paper and cellulose acetate. Ermolenko and coworkers (66) studied the separation of Cu, Ni, Co, and Ag on paper and cellulose derived supports. Yamazaki (67) and coworkers used cellulose acetate to determine the dissociation constants of Group VIB oxoacids. Silica Gel Frache and Dadone (68, 69) used buffered EDTA solutions, citric acid, and various electrolytes containing hydrochloric acid, water, methanol, ethanol, propanol, acetone, and dioxane. These electrolytes were tried on different materials such as silica gel, cellulose, and ion exchange resins. Silica gel gave 116 Che best results. Separation of up to six transition metals was obtained. Mori and coworkers (70) attempted the separation of 34 common metallic ions. A 0.1 M citric acid solution was used as the electrolyte. Experiments were run for only 5 minutes at 300 V. The separation of all ten first rwo transition metal ions by thin layer electrophoresis has not yet been reported. The purpose of this study was to use results from the TLC work to develop a system for the electrophoretic separation of the first row transition metal ions. The results for all ten metal ions are given for each of the ten systems studied. With each of these techniques, band broadening by diffusion is a limiting factor in determining the number of metals which can be separated. High performance thin layer chromatography plates have been shown to greatly reduce the amount of spot elongation in thin layer chromatography. The same advantage should be obtainable in thin layer electrophoresis. 117 Theory The term electrophoresis Is used to describe the migration of electrically charged particles in solution under the influence of an electric field. Migration depends on the timet the strength of the applied electric fieldt properties of the medium (pH of the electrolytic solution* ionic strength( temperature, viscosity, etc.), and properties of the particle (charge, size, hydration, tendency to dissociate, etc.). Ionic mobilities (u) can be calculated using the equation: (42) u ■ d/t x 1/e where d (cm) is the distance traveled by the ion during electrophoresis, t (sec) is the time of electrophoresis, 1 (cm) is the distance between the electrodes, and e (volts) the applied voltage. Factors Influencing Mobility: a) Applied voltage and heating effect- When voltage is applied across the plate, current will flow. The current will be determined by the equation: (43) I - V/R where I (amps) is the current, V (volts) is the applied potential and R (ohms) is the resistance. 118 The passage of current generates heat in the plate according to the equation: (44) calories - VIt/4.18 where t (sec) is the time. This heating effect can cause temperature differences in the field (see section (h) and evaporation (see section J>. b) Size of partlcle- Tho larger the particle, the slower it will migrate. c) Charge on particle- Thc greater the charge the faster it will move in the electric field. d) Ionic strength- In general, Increasing ionic strength results in decreasing ion mobility. Also increasing ionic strength means increasing conductivity and thereby increasing the heating effect (see section a). e) pH- The pH influences the type of complexes which form and the amount of hydration. Buffer solutions are used to keep the pH constant for a given run. Adjusting the pH can allow further control over the separation. 119 f) Adsorption- If a substance to be separated is adsorbed on the thin layer plate, only the non-adsorbed part of the substance migrates resulting in a reduction of migration speed and an increase in tailing. g) Diffusion- Substances in a highly concentrated area tend to migrate to less concentrated areas. Migration paths also differ for particles of the same substance causing further diffusion. Diffusion is controlled by selecting support materials of highly uniform grain, allowing minimum time after spotting of the sample to application of voltage, and drying the support immediately after the run. h) Zone flaws- Zone front irregularities result from inhomogcneity of the support, non-uniform moisture content, electrolytic concentration gradients and temperature differences in the electric field. These irregularities can be kept to a minimum with careful work. i) Zeta-potential and electroosmosis- In all electrophoresis, the solid phase is charged with respect to the surrounding solution to an extent which depends on the pH. (This is the zeta-potential.) Application of an electric field causes movement of the liquid with respect to the stationary solid support (electroosmosis). The electroosmotlc flow can be so large under certain conditions that the particles appear to be going in reverse. 120 The electroosmotic effect increases with increase in the structural fineness of the supporting material and in the field strength. Increase in pH renderB a negative zeta-potential smaller. Higher pH increases a positive zeta-potential. j) Suction effects- As evaporation takes place from the thin layer plate, buffer is pulled onto the plate to replace it. The buffer is moving from the edge to the center directly opposite the migration of the species to be separated. This flow of buffer reduces the migration speed. If the electrode compartments are not filled to equal heights, or the chamber is not level, electrolyte will be siphoned into the lower level. Suction effects can be reduced by supplying coolant to the chamber or plate to reduce evaporation, filling the electrode compartments equally, and leveling the chamber. Application of the Double-Layer Theory to Electrophoresis Earlier in this chapter it was stated that the larger molecules migrated slower. This is a very simplified description. In actuality, one must not only consider the size of the molecule but also the ionic atmosphere. Any charged molecule in solution will be surrounded by a layer of oppositely charged particles. Such a situation requires a more detailed treatment using the theory of double-layers. Reviews of the double-layer theory can be found elsewhere and will not be included here. Instead we will 121 concentrate on the usage of these theories. Huckel (71) presented an equation for the mobility (u) of a sphere surrounded by an electrical double-layer In an applied field (45). (45) u - 2€0 €r* /3n where V is the Burface potential, n is the viscosity of the liquid, and tr is the dielectric constant. Onsager (72) showed a derivation of that equation. The ionic atmosphere around a particle of radius a can be divided into concentric spherical shells of radius r and thickness dr. The 2 applied field exerts a force equal to 4 r pEdr. The velocity, v, of the shell can be found using the equation 2 (46) v - 4nr pEdr/fcrrnr where p is the space charge density. The total ionic atmosphere gives the innermost layer a velocity which can be found from the equation: 4lfr pEdr/6fTnr The force acting on the particle with charge Q will result in a velocity given by 122 (48) vpartt - QE/*TTna. The net velocity is the sum of these two opposing forces: <49) V - QE/Wna + y*4ffr^pEdr/i/rnr. Using electroneutrality and applying Poisson's law for spherical symmetry this is transformed into equation (50). (50) Q - It is worth noting that when expressing the mobility as a function of the surface potential, neither the size of the particle nor the thickness of the double-layer appears explicitly in the equation. Smoluchowskl derived an equation for particles surrounded by a relatively thin double-layer. This equation, which applies to particles of arbitrary shape, is similar to Huckel's except that the denominator is 4 rather than 6 (73). Later, Henry (74) showed that Smoluchowskl was correct for thin double-layers and Huckel's equation for thick double-layers. 123 Experimental All electrophoresis vork was done using a Gelman Instrument Co. voltage control and a Gelman Deluxe Electrophoresis Chamber. The chamber was cooled using tap water. Both inlet and outlet temperatures of cooling water were monitored to assure constant temperature. Developing solutions were placed in the chamber at least 6 hours before a run, allowing for saturation of the air in the chamber to reduce evaporation. During this 6 hour period, a piece of filter paper connected the two compartments of the chamber. This was done to eliminate any siphoning effects during the run itself. Paper electrophoresis was performed using Whatman ill filter paper or chromatography paper. Thin layer plates were made as described in the experimental section of Thin Layer Chromatography. Whatman LHF-K thin layer plates were used for the high performance thin layer electrophoresis. All solutions were made from reagent grade chemicals and Nanopure water unless otherwise specified. Solution was wicked onto the plate using Whatman ffl filter paper. Plates were allowed to equilibrate for at least 1 hour before a run. For spots of more than 1 microliter, a Hamilton701-N syringe was used. A Hamilton 7001 or 7101 syringe was usedfor sub-microliter spots. The spots were applied Immediately prior to applying the voltage. Time was set by timer control on a voltage control unit and checked by a "time-it" timer down to a hundredth of a minute. After a run, the plates were quickly dried using a L&R Ultrasonic Dryer. Metals were located by spraying with 0.5% 4-(2-Pyridylazo)-resorcinol (PAR) In methanol (unleBB otherwise indicated.). The metal Ion solutions were prepared as discussed in the Experimental section of Chapter 2. 125 Results and Discussion Table 25 shows the solvent systems which were examined in TLE for metal ion separation. Table 26 shows the results of the 5 different buffer systems. Lactic acid (E) showed the best separation and longest migration distances. This system could separate Ti(IV), Mn(II), Zn(II), and Co(ll). The HPTLE using the same buffer system could separate Cr(lll), V(V), Mn(Il), and Fe(lll) or Ti(IV), Zn(Il), MN(II), V(V) and Cr(lII). The results of four more solvent systems arc given In Table 27. These systems showed much slower migration than the citric, lactic, and formic acid solutions. The 1.5% oxalic acid solution had a high conductivity even at low voltages (125 V), which resulted in increased temperature during the run. The siphoning effects which result from the higher temperatures account for the lack of movement with this system. As mentioned in the theory section, there are several factors which influence the mobility of ions in electrophoresis. The effect of many of these factors were reduced considerably during the experiments (see High Performance Thin Layer Electrophoresis). The mobilities reported here are largely a result of two factors: a) true electrophoretic mobility and b) electroosmosis. These factors are not easily resolved and can work in opposing directions. The opposing action of electroosmosis could explain the little movement in the tartaric acid and oxalic acid (0.7%) systems. Tabic 25. Solvent Systems Used for TLE Letter Code Chemical Composition A Citric Acid(1.5% aqueous) B Lactic Acid(1.5% aqueous) C Formic Acld(0.1M aqueous) D Formic Acid(1.0M aqueous) E Formic Acid/Sodium Formate(0.02M/0.02M) F Tartaric Acid(1.5% aqueous) G Oxalic Acid(1.52 aqueous) H Oxalic Acld(0.7% aqueous) I Tartaric Acid/Sodium Tartrate(0.1M/0.1M) Table 26. Behavior of Metal Ions in TLE Stationary Mobile Phase Phase V Sc T1_____ Cr_____ to_____ Fe_____ Co_____ NI Cu Zn . Silica G A 5.5-6.2 5.6-6.5 5.5-6.0 5.0-6.1 4.1-4.7 4.0-7.0 20cm (13.0) ( - ) (13.4) ( - ) ( - ) ( - ) (12.8) (12.3) (9.8) (12.2) Silica G B 6.4-6.9 - .9-1.3 6.1-6.3 1.8-2.3 6.9-7.3 6.8-7.3 6.5-7.5 6.0-6.4 5.8-6.5 20cm (14.8) ( - ) (2.4) (13.8) (4.6) (15.8) (15.7) (15.6) (13.8) (13.7) Silica G C .5-1.0 .8-1.0 — .6- ? .6-1.3 .5-1.6 .7-1.7 .3-1.6 * 20cm (1.7) (2.0) ( - ) ( ) (2.1) (2.3) (2.7) ( - ) (2.1) ( - ) Silica G D 0-1.0 —— 0-4.0 0-1.8 1.0-1.7 1.5- ? 1.4-2.0 • 25.4cm (1.4) ( - ) ( - ) (5.6) (2.5) (3.8) ( ) (4.8) ( - ) ( - ) Silica G E —— 2.8-4.5* — 0-4.3 0 0-4.6 0-4.8 0-3.8 0-4.9 20.4cm ( - ) ( - ) (8.3) ( - > (4.9) ( 0 ) (5.2) (5.4) (4.3) (5.5) HPTLE B I.0-1.2 1.1-1.3 2.3-2.7 .5-1.0 1.3-1.8 2.0-2.5 1.3-2.2 1.2-2.2 1.2-3.1 1.5-2.1 (1.2) (1.3) (2.8) (0.8) (1.7) (2.5) (1.9) (1.9) (2.4) (2.0) *Moved toward anode Distance traveled in an- ( ) ■ mobilities x 10 Table 27. Behavior of Metal Ions in TLE Sta tion ary Mobile Phase Phase V Sc Ti Cr Mn Fe Co Ni Cu Zn Silica F 0 0 0 0 0 0 0.5-1.4 0.2-0.8 0 0 Gel ( 0 ) ( o ) ( 0 ) ( 0 ) ( 0 ) ( 0 ) (2.1) (1.1) ( 0 ) ( 0 ) (20cm) Silica G 0 0 0 0 0 0 0 0 0 0 Gel ( 0 ) ( o ) ( o ) ( 0 ) ( 0 ) ( 0 ) ( 0 ) ( 0 ) ( 0 ) < 0 ) (10cm) Silica H 0 0 0 0 0 0-0.3 0.6-1.2 0.1-0.5 0 0.5-0.6 Gel ( 0 ) ( 0 ) ( 0 ) ( 0 ) ( 0 ) (0.2) (1.0) (0.3) ( 0 ) (0.6) (10cm) Silica I 0-0.4 0.2-0.4 0-0.6 0.5-1.5 0.3-0.8 0-0.6 0 Gel 2 hours (0.1) (0.2) (0.2) (0.6) (0.3) (0.2) ( o ) (10cm) Distance traveled in cm, ( ) * nobilities x 10' 129 HlRh Performance Thin Layer Electrophoresis In developing the technique of HPTLE, several factors that influence nobility were addressed (see theory for the list of factors influencing nobility). First, it was realized that HPTLE would enploy high voltages resulting in larger heating effects. A cooling systen was developed which Involved a "Cold Comfort" Cold Pack (3 N) cooled to 0°C before the run began. The cool pack was placed in the chamber and the thin layer plate was set on top of it. If the cold pack was placed in the freezer and cooled below 0°C, the electrolyte would freeze on the plate resulting in no movement of the ions of interest. The effectiveness of the cooling technique was demonstrated by measuring the movement of indicators in an acetic acid/ammonium acetate buffer on silica gel plates. Figure 29 shows the distance travelled by the indicators at 300, 400, and 500 volts with no cooling system. Figure 50 shows the mobility of the indicators with the cold pack in place. The movement of the Indicators continued to Increase between 400 and 500 volts when cooled, but showed a marked decrease for the uncooled system. Adsorption will adversely affect the mobility. In this study, potential buffer systems were first screened by TLC in order to choose only systems in which no adsorption would occur (i.e. Rf near 1.0). Diffusion was controlled by limiting the time that the ions were in solution. Voltage was applied as soon as the electrolyte covered the plate and high voltages were used to reduce the run time. Plates were heated to evaporate the electrolyte Immediately after the run. These techniques reduced the time Ions were in solution by up to 90%. The zone flaws were minimized by UBing HPTLC plates instead of regular TLC plates. Suction effects were reduced by cooling the plate (see previous discussion). The siphoning effect was eliminated by placing several pieces of filter paper across the divider between the two electrode compartments. In this way, any siphoning which was going to happen occurred prior to the electrophoretic run. Distance (cm) 300 Q - Bromophenol Blue Blue Bromophenol - Q X - Phenol Red Phenol - X Figure 49- 49- Figure oCo*^ System Cool*"^ Wo Effectiveness Effectiveness Voltage of Cooling in XLE in Cooling of 500 Distance (cm) A . 300 Q Peo e w Red Phenol - X Figure 50.. Effectiveness of Cooling in TLE in Cooling of Effectiveness 50.. Figure ~ Bromophenol Blue Bromophenol ~ Cold Pack Cooling System Cooling Pack Cold Voltage A00 500 SUMMARY Ion chromatography was used to analyze ice samples from Dunde Ice Cap, China and Siple Station* Antarctica. Results are given for F", Cl", NO", NO", H2PO", and SO*. Separation of the first row transition metal ions was attempted by both thin layer chromatography and thin layer electrophoresis. The chromatographic behavior of all ten of the metal ions is reported for each of the thirty mobile phases, three stationary phases, and the two pretreatment techniques studied. All ten of the metal ions were also studied in ten electrophoretic systems. No one system was capable of separating all ten ions. However, a number of useful separations were suggested and many others could be devised based on the given data. Thirteen complexing agents were studied for locating the metal ion spots. During these experiments, novel approaches to solving various problems were developed: The use of on column concentration for the chromatographic analysis of large volumes of very dilute solutions was explored. A new method was developed which eliminates the need for a sample loop or concentrator column. The result is that much larger columns can be concentrated quite easily. To load volumes as large as 17 ml by an injection loop or through a concentrator column would require the use 133 134 of a second pump to overcome the back pressure of either of these two. The method developed In this work does not require a second pump, as It uses the primary pump to load the sample. In addition, it has been shown that the new method enhances the efficiency of the system. During the work on the electrophoresis of metal Ions, spot diffusion was a consistent problem. The technique of high performance thin layer electrophoresis was developed by careful attention to each of the factors contributing to spot spreading. The time a spot has to diffuse during an experiment can be reduced by use of higher voltages. Higher voltages in turn result in heating which reduce migration speeds. This problem was overcome by the development of an " “ inexpensive, easy to use, cooling system. The effectiveness of the cooling system was demonstrated. The techniques developed here should be applicable to a wide variety of other systems. LIST OF REFERENCES 1. Delmas, R., Boutron, C., J. of Geophysical Research, 85 (1980) p. 5645. 2. Arlstarain, A.J., Delmas, R.J., Briat, M., J. of Geophysical Research, 87 (1982) p. 11,004. 3. Legrand, M., De Angel is, M., and Deltnas, R.J., Analytics Chltnica Acta, 156 (1984) p. 181. 4. Jenke, D.R., Mitchell, P.K., and Paenkopf, G.K., J. of Chromatography Science, 21 (1983) p. 487. ~y: Herron-, *M.M.;-JT~of Geophysical Research, 87 (1982) p. 3052. 6. Spencer, M.J., Mayevskl, P.A., and Lyons, U.B., J. of Glaciology, 31 (1985) p. 233. 7. Mayewski, P.A., Lyons, W.B., Spencer, M.J., Twickler, M . , Dansgaard, W., Koci, B., Davidson, C.I., and Honrath, R.E., Science, 232 (1986) p. 975. 8. Martin, A.J.P., and Synge, R.L.M., J. Blochem., 35 (1941) p. 1358. 9. Giddlngs, J.C., Dynamics of Chromatography, Part I, Marcel Dekker, New York (1965). 10. Huber, J.F.K., and Hulsman, J.A.R., Anal. Chim. Acta, 38 (1967) p. 305. 11. van Deemter, J.J., Zviderweg, F.J., and Kllnkenberg, A., Chem. Eng. Sci., 5 (1956) p. 271. 12. Karger, B.L., Snyder, L.R., and Horvath, C., An Introduction to Separation Science, Willey-Interscience, New York (1973). 13. Giddlngs, J.C., Dynamics of Chromatography, Part I, Marcel Dekker, New York (1965). 14. Huber, J.F.K., and Hulsman, J.A.R., Anal. Chim. Acta, 38 (1967) p. 305. 135 136 15. Purnell, J.H., J. Chem. Soc., (1960) p. 352. 16. Mosley-Thompson, E., Krusa, P.D., and Thompson, L.G., Annals of Glaciology, 7 (1965) p. 26. 17. Mosley-Thompson, E., and Thompson, L.G., Quaternary Research, 17 (1982) p. I. 18. Thompson, L.G., Mosley-Thompson, E., and Arnao, B.M., Science, 226 (1984) p. 50. 19. Mosley-Thompson, E., Mountain, K.R., and Pasklevitch, J.F., Antarctic J. of the United States, XX (1986) in press. 20. Schwander, J. and Stauffer, B., Nature, 311 (1984) p. 45. 21. Mosley-Thompson, E., personal communication. 22. Broqualre, M., and Gulnebault, P.R., J. of Liquid Chrom., 4 (1981) p. 2039. 23. Buchholz, A.E., Verplough, C.I., and Smith, J.L., J. of Chromatogr. Sci., 20 (1982) p. 499. 24. Okado, T., and Kuwanoto, T., J. of Chromatogr., 350 (1985) p. 317. 25. Heckenberg, A.L., and Haddad, P.R., J. of Chrom., 299 (1984) p. 301. 26. Silver, M.R., Trosper, T.D., Gould, M.R., Dickerson, J.E., and Desotelle, G.H., J. of Liquid Chrom., 7 (1984) p. 559. 27. van der Wal, S., J. of Liquid Chrom., 9 (1986) p. 1815. 28. Beyerinek, M.W., Z. Phys. Chem., 3 (1889) p. 110. 29. Wijsman, H.P., DeDiastase, beschouwd als mengsel van Mattase en Dextrlnase, Amsterdam, (1898). 30. Izmailov, N.A., and Schralber, M.S., Farmatsiya (Sofia) 1938, p. 1. 31. Meinhard, J.E., and Hall, N.F., Anal. Chem., 21 (1949) p. 185. 32. Miller, J.M., Kirchner, J.G., Anal. Chem. 23 (1951) p. 428. 33. Kirchner, J.G., and Keller, G.J., J. Am. Chem. Soc., 72 (1950) p. 1867. 137 34. Miller, J.M., Kirchner, G.J., Anal. Chem., 24 (1952) p. 1480. 35. Miller, J.M., Kirchner, G.J., Anal. Chem., 25 (1953) p. 1107. 36. Miller, J.M., Kirchner, G.J., Anal. Chem., 26 (1954) p. 2002. 37. Stahl, E., Chem. Ztg., 82 (1958) p. 323. 38. Stahl, E., Thin Layer Chromatography, 2nd ed., Springer Verlag, Berlin (1969). 39. Heftmann, E., Chromatography - A Laboratory Handbook of Chromatographic and Electrophoretic Methods, 3rd ed., Van Rostrand Reinhold, New York (1975). 40. Kirchner, J.G., J. Chromatogr. Sci., 13 (1975) p. 558. 41. Kirchner, J.G., Thin Layer Chromatography, 2nd ed., Wiley, New York (1978). 42. Kirchner, J.G., Thin Layer Chromatography - Quantitative Environmental and Clinical Applications, Wiley - Interscience, New York (1980). 43. Felick, N., Bolllger, H.R., Mangold, H.K., Advances in Chromatography, Vol. 3, Marcel Dekker, New York (1966). 44. Frache, R., Dadone, A., Chromatographla, 5 (1972) p. 449. 45. Frache, R., Dadone, A., Chromatographla, 6 (1973) p. 433. 46. Frache, R., Dadone, A., Baffl, F., Chromatographla, 9 (1976) p. 83. 47. Varshney, R.G., Curr. Sci., 45 (1976) p. 54, 48. Qureshi, M., Thakur, J.S., Sep. Sci., 11 (1976) p. 467. 49. Baffl, F., Dadone, A., Frache, R., Chromatographla, 9 (1976) p. 280. 50. Soljlc, Z., Grba, V., Z. Anal. Chem., 278 (1976) p. 278. 51. Kuroda, R., Matushe, H., Oguma, K., Radioanal. Chem., 36 (1977) p. 119. 52. Rao, A.L.J., Shekar, C », Fresenlus Z . Anal. Chem., 277 (1976) p. 126. 53. Oksala, R.H. Jr., Krause, R.A., Anal. Chlm. Acta, 85 (1976) p. 351. 138 54. Singh, N., Kumar, P., Kansal, B.D., Chromatographla, 11 (197B) p. 408. 55. Lohmueller, M., Herrmann, P., Ballschmlter, K., J. Chromatogr., 137 (1977) p. 165. 56. Gagliardl, E., Deutschmann, G., Mikrochim. Acta, 2 (1976) p. 23. 57. Franson, M.A., editor, Standard Methods for the Examination of Water and Waste Water, 14th Edition, American Public Health Association, Washington, D.C. (1976). 58. Chin, H.P., Cellulose Acetate Electrophoresis, Ann Arbor - Humphrey Science Pub., Ann Arbor (1970) p. 1. 59. Strain, H.H., and Sullivan, J.C., Anal. Chem., 23 (1951) p. 816. 60. Lederer, M., and Ward, F.L., Anal. Chim. Acta, 6 (1952) p. 355. 61. Bhatnager, R.P., and Yusufi, F.A.K., Indian Chem. Soc., 59 (1982) p. 10. 62. Yadava, J.R., and Yadava, K.L., Electrochem. India, 30 (1981) p. 250. 63. Sircar, J.K., and Yadava, K.L., J. Chem. Eng. Data, 27 (1982) p. 231. 64. Singh, R.K.P., and Yadava, K.L., Trans Saest, 16 (1981) p. 163. 65. Shukla, S.K., Blotta, I., and Masella, R., Anal. Chem. Symp. Ser., 14 (1983) p. 179. 66. Ermolenko, I.N., and Lazareva, T.G., Vesrsi. Akad. Navuk BSSR, Ser. Khim., 6 (1983) p. 30. 67. Yamazakl, H., Tsujimoto, K., Gohda, S., Hiraki, K., and Nishikawa, Y., Bunseki Kagaku 28 (1979) p. 424. 68. Frache, R., and Dadone, A., Chromatographla, 6(6), (1973) p. 266. 69. Frache, R., and Dadone, A., Chromatographla, 6(10), (1973) p. 430. 70. Mori, I., Tsunematsu, N., and Shinogl, M., Yakugaku Zasshi, 89 (1969) p. 1669. 71. Huckel, E., Physik Z., 25 (1924) p. 204. 72. Onsager, L., Physik Z., 27 (1926) p. 388. 139 73. Rlghetti, P.G., Van Oss, C.J., and Vanderhoff, J.W., Electrokinetic Separation Methods, Elsevier/North-Holland Biomedical Press, New York (1979). * 74. Henry, D.C., Proc. Roy. Soc. London, A133 (1931) p. 106.