The following set of experiments introduce students to the applications of chromatography and electrophoresis. Experiments are grouped into five categories: gas chromatography, high-performance liquid chromatography, ion-exchange chromatography, size-exclusion chromatography, and electrophoresis.
Gas Chromatography
- Bishop, R. D., Jr. “Using GC–MS to Determine Relative Reactivity Ratios,” J. Chem. Educ. 1995, 72, 743–745.
- Elderd, D. M.; Kildahl, N. K.; Berka, L. H. “Experiments for Modern Introductory Chemistry: Identification of Arson Accelerants by Gas Chromatography,” J. Chem. Educ. 1996, 73, 675–677.
- Fleurat-Lessard, P.; Pointet, K.; Renou-Gonnord, M.-F. “Quantitative Determination of PAHs in Diesel Engine Exhausts by GC–MS,” J. Chem. Educ. 1999, 76, 962–965.
- Galipo, R. C.; Canhoto, A. J.; Walla, M. D.; Morgan, S. L. “Analysis of Volatile Fragrance and Flavor Compounds by Headspace Solid Phase Microextraction and GC–MS,” J. Chem. Educ. 1999, 76, 245–248.
- Graham, R. C.; Robertson, J. K. “Analysis of Trihalomethanes in Soft Drinks,” J. Chem. Educ. 1988, 65, 735–737.
- Heinzen, H.; Moyan, P.; Grompone, A. “Gas Chromatographic Determination of Fatty Acid Compositions,” J. Chem. Educ. 1985, 62, 449–450.
- Kegley, S. E.; Hansen, K. J.; Cunningham, K. L. “Determination of Polychlorinated Biphenyls (PCBs) in River and Bay Sediments,” J. Chem. Educ. 1996, 73, 558–562.
- Kostecka, K. S.; Rabah, A.; Palmer, C. F., Jr. “GC/MS Analysis of the Aromatic Composition of Gasoline,” J. Chem. Educ. 1995, 72, 853–854.
- Quach, D. T.; Ciszkowski, N. A.; Finlayson-Pitts, B. J. “A New GC-MS Experiment for the Undergraduate Instrumental Analysis Laboratory in Environmental Chemistry: Methyl-t-butyl Ether and Benzene in Gasoline,” J. Chem. Educ. 1998, 75, 1595–1598.
- Ramachandran, B. R.; Allen, J. M.; Halpern, A. M. “Air–Water Partitioning of Environmentally Important Organic Compounds,” J. Chem. Educ. 1996, 73, 1058–1061.
- Rice, G. W. “Determination of Impurities in Whiskey Using Internal Standard Techniques,” J. Chem. Educ. 1987, 64, 1055–1056.
- Rubinson, J. F.; Neyer-Hilvert, J. “Integration of GC-MS Instrumentation into the Undergraduate Laboratory: Separation and Identification of Fatty Acids in Commercial Fats and Oils,” J. Chem. Educ. 1997, 74, 1106–1108.
- Rudzinski, W. E.; Beu, S. “Gas Chromatographic Determination of Environmentally Significant Pesticides,” J. Chem. Educ. 1982, 59, 614–615.
- Sobel, R. M.; Ballantine, D. S.; Ryzhov, V. “Quantitation of Phenol Levels in Oil of Wintergreen Using Gas Chromatography–Mass Spectrometry with Selected Ion Monitoring,” J. Chem. Educ. 2005, 82, 601–603.
- Welch, W. C.; Greco, T. G. “An Experiment in Manual Multiple Headspace Extraction for Gas Chromatography,” J. Chem. Educ. 1993, 70, 333–335.
- Williams, K. R.; Pierce, R. E. “The Analysis of Orange Oil and the Aqueous Solubility of d-Limone,” J. Chem. Educ. 1998, 75, 223–226.
- Wong, J. W.; Ngim, K. K.; Shibamoto, T.; Mabury, S. A.; Eiserich, J. P.; Yeo, H. C. H. “Determination of Formaldehyde in Cigarette Smoke,” J. Chem. Educ. 1997, 74, 1100–1103.
- Yang, M. J.; Orton, M. L., Pawliszyn, J. “Quantitative Determination of Caffeine in Beverages Using a Combined SPME-GC/MS Method,” J. Chem. Educ. 1997, 74, 1130–1132.
High-Performance Liquid Chromatography
- Batchelor, J. D.; Jones, B. T. “Determination of the Scoville Heat Value for Hot Sauces and Chilies: An HPLC Experiment,” J. Chem. Educ. 2000, 77, 266–267.
- Beckers, J. L. “The Determination of Caffeine in Coffee: Sense or Nonsense?” J. Chem. Educ. 2004, 81, 90–93.
- Betts, T. A. “Pungency Quantitation of Hot Pepper Sauces Using HPLC,” J. Chem. Educ. 1999, 76, 240–244.
- Bidlingmeyer, B. A.; Schmitz, S. “The Analysis of Artificial Sweeteners and Additives in Beverages by HPLC,” J. Chem. Educ. 1991, 68, A195–A200.
- Bohman, O.; Engdahl, K.-A.; Johnsson, H. “High Performance Liquid Chromatography of Vitamin A: A Quantitative Determination,” J. Chem. Educ. 1982, 59, 251–252.
- Brenneman, C. A.; Ebeler, S. E. “Chromatographic Separations Using Solid-Phase Extraction Cartridges: Separation of Wine Phenolics,” J. Chem. Educ. 1999, 76, 1710–1711.
- Cantwell, F. F.; Brown, D. W. “Liquid Chromatographic Determination of Nitroanilines,” J. Chem. Educ. 1981, 58, 820–823.
- DiNunzio, J. E. “Determination of Caffeine in Beverages by High Performance Liquid Chromatography,” J. Chem. Educ. 1985, 62, 446–447.
- Ferguson, G. K. “Quantitative HPLC Analysis of an Analgesic/Caffeine Formulation: Determination of Caffeine,” J. Chem. Educ. 1998, 75, 467–469.
- Ferguson, G. K. “Quantitative HPLC Analysis of a Psychotherapeutic Medication: Simultaneous Determination of Amitriptyline Hydrochloride and Perphenazine,” J. Chem. Educ. 1998, 75, 1615–1618.
- Goodney, D. E. “Analysis of Vitamin C by High-Pressure Liquid Chromatography,” J. Chem. Educ. 1987, 64, 187–188.
- Guevremont, R.; Quigley, M. N. “Determination of Paralytic Shellfish Poisons Using Liquid Chromatography,” J. Chem. Educ. 1994, 71, 80–81.
- Haddad, P.; Hutchins, S.; Tuffy, M. “High Performance Liquid Chromatography of Some Analgesic Compounds,” J. Chem. Educ. 1983, 60, 166–168.
- Huang, J.; Mabury, S. A.; Sagebiel, J. C. “Hot Chili Peppers: Extraction, Cleanup, and Measurement of Capscaicin,” J. Chem. Educ. 2000, 77, 1630–1631.
- Joeseph, S. M.; Palasota, J. A. “The Combined Effect of pH and Percent Methanol on the HPLC Separation of Benzoic Acid and Phenol,” J. Chem. Educ. 2001, 78, 1381–1383.
- Lehame, S. “The Separation of Copper, Iron, and Cobalt Tetramethylene Dithiocarbamates by HPLC,” J. Chem. Educ. 1986, 63, 727–728.
- Luo, P.; Luo, M. Z.; Baldwin, R. P. “Determination of Sugars in Food Products,” J. Chem. Educ. 1993, 70, 679–681.
- Mueller, B. L.; Potts, L. W. “HPLC Analysis of an Asthma Medication,” J. Chem. Educ. 1988, 65, 905–906.
- Munari, M.; Miurin, M.; Goi, G. “Didactic Application to Riboflavin HPLC Analysis,” J. Chem. Educ. 1991, 68, 78–79.
- Orth, D. L. “HPLC Determination of Taurine in Sports Drinks,” J. Chem. Educ. 2001, 78, 791– 792.
- Remcho, V. T.; McNair, H. M.; Rasmussen, H. T. “HPLC Method Development with the Photodiode Array Detector,” J. Chem. Educ. 1992, 69, A117–A119.
- Richardson, W. W., III; Burns, L. “HPLC of the Polypeptides in a Hydrolyzate of Egg-White Lysozyme,” J. Chem. Educ. 1988, 65, 162–163.
- Silveira, A., Jr.; Koehler, J. A.; Beadel, E. F., Jr.; Monore, P. A. “HPLC Analysis of Chlorophyll a, Chlorophyll b, and \(\beta\)-Carotene in Collard Greens,” J. Chem. Educ. 1984, 61, 264–265.
- Siturmorang, M.; Lee, M. T. B.; Witzeman, L. K.; Heineman, W. R. “Liquid Chromatography with Electrochemical Detection (LC-EC): An Experiment Using 4-Aminophenol,” J. Chem. Educ. 1998, 75, 1035–1038.
- Sottofattori, E.; Raggio, R.; Bruno, O. “Milk as a Drug Analysis Medium: HPLC Determination of Isoniazid,” J. Chem. Educ. 2003, 80, 547–549.
- Strohl, A. N. “A Study of Colas: An HPLC Experiment,” J. Chem. Educ. 1985, 62, 447–448.
- Tran, C. D.; Dotlich, M. “Enantiomeric Separation of Beta-Blockers by High Performance Liquid Chromatography,” J. Chem. Educ. 1995, 72, 71–73.
- Van Arman, S. A.; Thomsen, M. W. “HPLC for Undergraduate Introductory Laboratories,” J. Chem. Educ. 1997, 74, 49–50.
- Wingen, L. M.; Low, J. C.; Finlayson-Pitts, B. J. “Chromatography, Absorption, and Fluorescence: A New Instrumental Analysis Experiment on the Measurement of Polycyclic Aromatic Hydrocarbons in Cigarette Smoke,” J. Chem. Educ. 1998, 75, 1599–1603.
Ion-Exchange Chromatography
- Bello, M. A.; Gustavo González, A. “Determination of Phosphate in Cola Beverages Using Nonsuppressed Ion Chromatography,” J. Chem. Educ. 1996, 73, 1174–1176.
- Kieber, R. J.; Jones, S. B. “An Undergraduate Laboratory for the Determination of Sodium, Potassium, and Chloride,” J. Chem. Educ. 1994, 71, A218–A222.
- Koubek, E.; Stewart, A. E. “The Analysis of Sulfur in Coal,” J. Chem. Educ. 1992, 69, A146–A148.
- Sinniah, K.; Piers, K. “Ion Chromatography: Analysis of Ions in Pond Water,” J. Chem. Educ. 2001, 78, 358–362.
- Xia, K.; Pierzynski, G. “Competitive Sorption between Oxalate and Phosphate in Soil: An Environmental Chemistry Laboratory Using Ion Chromatography,” J. Chem. Educ. 2003, 80, 71–75.
Size-Exchange Chromatography
- Brunauer, L. S.; Davis, K. K. “Size Exclusion Chromatography: An Experiment for High School and Community College Chemistry and Biotechnology Laboratory Programs,” J. Chem. Educ. 2008, 85, 683–685.
- Saiz, E.; Tarazona, M. P. “Size-Exclusion Chromatography Using Dual Detection,” Chem. Educator 2000, 5, 324–328.
Electrophoresis
- Almarez, R. T.; Kochis, M. “Microscale Capillary Electrophoresis: A Complete Instrumentation Experiment for Chemistry Students at the Undergraduate Junior or Senior Level,” J. Chem. Educ. 2003, 80, 316–319.
- Beckers, J. L. “The Determination of Caffeine in Coffee: Sense or Nonsense?” J. Chem. Educ. 2004, 81, 90–93.
- Beckers, J. L. “The Determination of Vanillin in a Vanilla Extract,” J. Chem. Educ. 2005, 82, 604– 606.
- Boyce, M. “Separation and Quantification of Simple Ions by Capillary Zone Electrophoresis,” J. Chem. Educ. 1999, 76, 815–819.
- Conradi, S.; Vogt, C.; Rohde, E. “Separation of Enatiomeric Barbiturates by Capillary Electrophoresis Using a Cyclodextrin-Containing Run Buffer,” J. Chem. Educ. 1997, 74, 1122–1125.
- Conte, E. D.; Barry, E. F.; Rubinstein, H. “Determination of Caffeine in Beverages by Capillary Zone Electrophoresis,” J. Chem. Educ. 1996, 73, 1169–1170.
- Demay, S.; Martin-Girardeau, A.; Gonnord, M.-F. “Capillary Electrophoretic Quantitative Analysis of Anions in Drinking Water,” J. Chem. Educ. 1999, 76, 812–815.
- Emry, R.; Cutright, R. D.; Wright, J.; Markwell, J. “Candies to Dye for: Cooperative, Open-Ended Student Activities to Promote Understanding of Electrophoretic Fractionation,” J. Chem. Educ. 2000, 77, 1323–1324.
- Gardner, W. P.; Girard, J. E. “Analysis of Common Household Cleaner-Disinfectants by Capillary Electrophoresis,” J. Chem. Educ. 2000, 77, 1335–1338.
- Gruenhagen, J. A.; Delaware, D.; Ma, Y. “Quantitative Analysis of Non-UV-Absorbing Cations in Soil Samples by High-Performance Capillary Electrophoresis,” J. Chem. Educ. 2000, 77, 1613–1616.
- Hage, D. S.; Chattopadhyay, A.; Wolfe, C. A. C.; Grundman, J.; Kelter, P. B. “Determination of Nitrate and Nitrite in Water by Capillary Electrophoresis,” J. Chem. Educ. 1998, 75, 1588–1590.
- Herman, H. B.; Jezorek, J. R.; Tang, Z. “Analysis of Diet Tonic Water Using Capillary Electrophoresis,” J. Chem. Educ. 2000, 77, 743–744.
- Janusa, M. A.; Andermann, L. J.; Kliebert, N. M.; Nannie, M. H. “Determination of Chloride Concentration Using Capillary Electrophoresis,” J. Chem. Educ. 1998, 75, 1463–1465.
- McDevitt, V. L.; Rodríguez, A.; Williams, K. R. “Analysis of Soft Drinks: UV Spectrophotometry, Liquid Chromatography, and Capillary Electrophoresis,” J. Chem. Educ. 1998, 75, 625–629.
- Palmer, C. P. “Demonstrating Chemical and Analytical Concepts in the Undergraduate Laboratory Using Capillary Electrophoresis and Micellar Electrokinetic Chromatography,” J. Chem. Educ. 1999, 76, 1542–1543.
- Pursell, C. J.; Chandler, B.; Bushey, M. M. “Capillary Electrophoresis Analysis of Cations in Water Samples,” J. Chem. Educ. 2004, 81, 1783–1786.
- Solow, M. “Weak Acid pKa Determination Using Capillary Zone Electrophoresis,” J. Chem. Educ. 2006, 83, 1194–1195.
- Thompson, L.; Veening, H.; Strain, T. G. “Capillary Electrophoresis in the Undergraduate Instrumental Analysis Laboratory: Determination of Common Analgesic Formulations,” J. Chem. Educ. 1997, 74, 1117–1121.
- Vogt, C.; Conradi, S.; Rhode, E. “Determination of Caffeine and Other Purine Compounds in Food and Pharmaceuticals by Micellar Electrokinetic Chromatography” J. Chem. Educ. 1997, 74, 1126– 1130.
- Weber, P. L.; Buck, D. R. “Capillary Electrophoresis: A Fast and Simple Method for the Determination of the Amino Acid Composition of Proteins,” J. Chem. Educ. 1994, 71, 609–612.
- Welder, F.; Colyer, C. L. “Using Capillary Electrophoresis to Determine the Purity of Acetylsalicylic Acid Synthesized in the Undergraduate Laboratory,” J. Chem. Educ. 2001, 78, 1525–1527.
- Williams, K. R.; Adhyaru, B.; German, I.; Russell, T. “Determination of a Diffusion Coefficient by Capillary Electrophoresis,” J. Chem. Educ. 2002, 79, 1475–1476.
The following texts provide a good introduction to the broad field of separations, including chromatography and electrophoresis.
- Giddings, J. C. Unified Separation Science, Wiley-Interscience: New York 1991.
- Karger, B. L.; Snyder, L. R.; Harvath, C. An Introduction to Separation Science, Wiley-Interscience: New York, 1973
- Miller, J. M. Separation Methods in Chemical Analysis, Wiley-Interscience: New York, 1975.
- Poole, C. F. The Essence of Chromatography, Elsevier: Amsterdam, 2003.
A more recent discussion of peak capacity is presented in the following papers.
- Chester, T. L. “Further Considerations of Exact Equations for Peak Capacity in Isocratic Liquid Chromatography,” Anal. Chem. 2014, 86, 7239–7241.
- Davis, J. M.; Stoll, D. R.; Carr, P. W. “Dependence of Effective Peak Capacity in Comprehensive Two-Dimensional Separations on the Distribution of Peak Capacity between the Two Dimensions,” Anal. Chem. 2008, 80, 8122–8134.
- Li, X.; Stoll, D. R.; Carr, P. W. “Equation for Peak Capacity Estimation in Two-Dimensional Liquid Chromatography,” Anal. Chem. 2009, 81, 845–850.
- Shen, Y.; Lee, M. “General Equation for Peak Capacity in Column Chromatography,” Anal. Chem. 1998, 70, 3853–3856.
The following references may be consulted for more information on gas chromatography.
- Grob, R. L., ed, Modern Practice of Gas Chromatography, Wiley-Interscience: New York, 1972.
- Hinshaw, J. V. “A Compendium of GC Terms and Techniques,” LC•GC 1992, 10, 516–522.
- Ioffe, B. V.; Vitenberg, A. G. Head-Space Analysis and Related Methods in Gas Chromatography, Wiley-Interscience: New York, 1982.
- Kitson, F. G.; Larsen, B. S.; McEwen, C. N. Gas Chromatography and Mass Spectrometry: A Practical Guide, Academic Press: San Diego, 1996.
- McMaster, M. C. GC/MS: A Practical User’s Guide, Wiley-Interscience: Hoboken, NJ, 2008.
The following references provide more information on high-performance liquid chromatography.
- Dorschel, C. A.; Ekmanis, J. L.; Oberholtzer, J. E.; Warren, Jr. F. V.; Bidlingmeyer, B. A. “LC Detectors,” Anal. Chem. 1989, 61, 951A–968A.
- Ehlert, S.; Tallarek, U. “High-pressure liquid chromatography in lab-on-a-chip devices,” Anal. Bioanal. Chem. 2007, 388, 517–520.
- Francois, I.; Sandra, K.; Sandra, P. “Comprehensive liquid chromatography: Fundamental aspects and practical considerations—A review,” Anal. Chim. Acta 2009, 641, 14–31.
- Harris, C. M. “Shrinking the LC Landscape,” Anal. Chem. 2003, 75, 64A–69A.
- Meyer, V. R. Pitfalls and Errors of HPLC in Pictures, Wiley-VCH: Weinheim, Germany, 2006.
- Pozo, O. J.; Van Eenoo, P.; Deventer, K.; Delbeke, F. T. “Detection and characterization of anabolic steroids in doping analysis by LC–MS,” Trends Anal. Chem. 2008, 27, 657–671.
- Scott, R. P. W. “Modern Liquid Chromatography,” Chem. Soc. Rev. 1992, 21, 137–145.
- Simpson, C. F., ed. Techniques in Liquid Chromatography, Wiley-Hayden: Chichester, England; 1982.
- Snyder, L. R.; Glajch, J. L.; Kirkland, J. J. Practical HPLC Method Development, Wiley-Interscience: New York,1988.
- van de Merbel, N. C. “Quantitative determination of endogenous compounds in biological samples using chromatographic techniques,” Trends Anal. Chem. 2008, 27, 924–933.
- Yeung, E. S. “Chromatographic Detectors: Current Status and Future Prospects,” LC•GC 1989, 7, 118–128.
The following references may be consulted for more information on ion chromatography.
- Shpigun, O. A.; Zolotov, Y. A. Ion Chromatography in Water Analysis, Ellis Horwood: Chichester, England, 1988.
- Smith, F. C. Jr.; Chang, R. C. The Practice of Ion Chromatography, Wiley-Interscience: New York, 1983.
The following references may be consulted for more information on supercritical fluid chromatography.
- Palmieri, M. D. “An Introduction to Supercritical Fluid Chromatography. Part I: Principles and Applications,” J. Chem. Educ. 1988, 65, A254–A259.
- Palmieri, M. D. “An Introduction to Supercritical Fluid Chromatography. Part II: Applications and Future Trends,” J. Chem. Educ. 1989, 66, A141–A147.
The following references may be consulted for more information on capillary electrophoresis.
- Baker, D. R. Capillary Electrophoresis, Wiley-Interscience: New York, 1995.
- Copper, C. L. “Capillary Electrophoresis: Part I. Theoretical and Experimental Background,” J. Chem. Educ. 1998, 75, 343–347.
- Copper, C. L.; Whitaker, K. W. “Capillary Electrophoresis: Part II. Applications,” J. Chem. Educ. 1998, 75, 347–351.
- DeFrancesco, L. “Capillary Electrophoresis: Finding a Niche,” Today’s Chemist at Work, February 2002, 59–64.
- Ekins, R. P. “Immunoassay, DNA Analysis, and Other Ligand Binding Assay Techniques: From Electropherograms to Multiplexed, Ultrasensative Microarrays on a Chip,” J. Chem. Educ. 1999, 76, 769– 780.
- Revermann, T.; Götz, S.; Künnemeyer, J.; Karst, U. “Quantitative analysis by microchip capillary electrophoresis—current limitations and problem-solving strategies,” Analyst 2008, 133, 167–174.
- Timerbaev, A. R. “Capillary electrophoresis coupled to mass spectrometry for biospeciation analysis: critical evaluation,” Trends Anal. Chem. 2009, 28, 416–425.
- Unger, K. K.; Huber, M.; Hennessy, T. P.; Hearn, M. T. W.; Walhagen, K. “A Critical Appraisal of Capillary Electrochromatography,” Anal. Chem. 2002, 74, 200A–207A.
- Varenne, A.; Descroix, S. “Recent strategies to improve resolution in capillary electrophoresis—A review,” Anal. Chim. Acta 2008, 628, 9–23.
- Vetter, A. J.; McGowan, G. J. “The Escalator—An Analogy for Explaining Electroosmotic Flow,” J. Chem. Educ. 2001, 78, 209–211.
- Xu, Y. “Tutorial: Capillary Electrophoresis,” Chem. Educator, 1996, 1(2), 1–14.
The application of spreadsheets and computer programs for modeling chromatography is described in the following papers.
- Abbay, G. N.; Barry, E. F.; Leepipatpiboon, S.; Ramstad, T.; Roman, M. C.; Siergiej, R. W.; Snyder, L. R.; Winniford, W. L. “Practical Applications of Computer Simulation for Gas Chromatography Method Development,” LC•GC 1991, 9, 100–114.
- Drouen, A.; Dolan, J. W.; Snyder, L. R.; Poile, A.; Schoenmakers, P. J. “Software for Chromatographic Method Development,” LC•GC 1991, 9, 714–724.
- Kevra, S. A.; Bergman, D. L.; Maloy, J. T. “A Computational Introduction to Chromatographic Bandshape Analysis,” J. Chem. Educ. 1994, 71, 1023–1028.
- Rittenhouse, R. C. “HPLC for Windows: A Computer Simulation of High-Performance Liquid Chromatography,” J. Chem. Educ. 1995, 72, 1086–1087.
- Shalliker, R. A.; Kayillo, S.; Dennis, G. R. “Optimizing Chromatographic Separations: An Experiment Using an HPLC Simulator,” J. Chem. Educ. 2008, 85, 1265–1268.
- Sundheim, B. R. “Column Operations: A Spreadsheet Model,” J. Chem. Educ. 1992, 69, 1003– 1005.
The following papers discuss column efficiency, peak shapes, and overlapping chromatographic peaks.
- Bildingmeyer, B. A.; Warren, F. V., Jr. “Column Efficiency Measurement,” Anal. Chem. 1984, 56, 1583A–1596A.
- Hawkes, S. J. “Distorted Chromatographic Peaks,” J. Chem. Educ. 1994, 71, 1032–1033.
- Hinshaw, J. “Pinning Down Tailing Peaks,” LC•GC 1992, 10, 516–522.
- Meyer, V. K. “Chromatographic Integration Errors: A Closer Look at a Small Peak,” LC•GC North America 2009, 27, 232–244.
- Reid, V. R.; Synovec, R. E. “High-speed gas chromatography: The importance of instrumentation optimization and the elimination of extra-column band broadening,” Talanta 2008, 76, 703–717.