Capillary electrophoresis methods

There are several different forms of capillary electrophoresis, each of which has its particular advantages. Several of these methods are briefly described in this section. 

A form of capillary electrophoresis in which separations are based on differences in the solutes electrophoretic mobilities. 

Capillary Zone Electrophoresis The simplest form of capillary electrophoresis is capillary zone electrophoresis (CZE). In CZE the capillary tube is filled with a buffer solution and, after loading the sample, the ends of the capillary tube are placed in reservoirs containing additional buffer solution. Under normal conditions, the end of the capillary containing the sample is the anode, and solutes migrate toward 

UVA/is absorbance solute must have UVA/is absorbing chromophore 10-5-10- yes 

Source Adapted from Baker, D. R. Capillary Electrophoresis. Wiley-Interscience New York, 1995. Concentration depends on the volume of sample injected. 

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H. Engelhar dt, W. Beck and T. Schmitt, Capillary Electrophoresis Methods and Potentials, Vieweg, Wiesbaden, Germany (1996). 

Frazierm, R.A. et ah. Development of a capillary electrophoresis method for the simultaneous analysis of colours, preservatives and sweeteners in soft drinks, J. 

Frazier, R. A. (2001). Recent advances in capillary electrophoresis methods for food analysis. Electrophoresis 22,4197-4206. 

Engelhardt, H., Beck, W., Kohr, J., and Schmitt, T., Capillary electrophoresis methods and scope, Angewandte Chemie, 32, 629, 1993. 

H. Watzig, M. Degenhardt, and A. Kunkel, Strategies for capillary electrophoresis Method development and validation for pharmaceutical and biological applications, Electrophoresis, 19, 2695 (1998). 

Fan et al. [106] developed a high performance capillary electrophoresis method for the analysis of primaquine and its trifluoroacetyl derivative. The method is based on the mode of capillary-zone electrophoresis in the Bio-Rad HPE-100 capillary electrophoresis system effects of some factors in the electrophoretic conditions on the separation of primaquine and trifluoroacetyl primaquine were studied. Methyl ephedrine was used as the internal standard and the detection was carried out at 210 nm. A linear relationship was obtained between the ratio of peak area of sample and internal standard and corresponding concentration of sample. The relative standard deviations of migration time and the ratio of peak area of within-day and between-day for replicate injections were <0.6% and 5.0%, respectively. 

Bortocan and Bonato [112] presented a capillary electrophoresis method for the enantioselective analysis of primaquine and its metabolite carboxyprimaquine. The method is used for the simultaneous determination of the drug and its metabolite, in... 

P. Schmitt Kopplin, Capillary Electrophoresis. Methods and Protocols, Humana Press, Neuherberg (2008). 

G. Lunn (Ed.), Capillary Electrophoresis Methods for Pharmaceutical Analysis, Wiley-Interscience, New York, 2000. 

R.A. Frazier, E.L. Inns, N. Dossi, J.M. Ames and H.E. Nursten, Development of a capillary electrophoresis method for the simultaneous analysis of artifical sweeteners, preservatives and colours in soft drinks. J. Chromatogr.A, 876 (2000) 213-220. 

Tim Wehr is Staff Scientist at Bio-Rad Laboratories in Hercules, California. He has more than 20 years of experience in biomolecule separations, including development of HPLC and capillary electrophoresis methods and instrumentation for separation of proteins, peptides, amino acids, and nucleic acids. He has also worked on development and validation of LC-MS methods for small molecules and biopharmaceuticals. He holds a B.S. degree from Whitman College, Walla Walla, Washington, and earned his Ph.D. from Oregon State University in Corvallis. 

Zhou, L., Thompson, R., Song, S., Elhson, D., Wyvratf J.M. A strategic approach to the development of capillary electrophoresis methods for pharmaceutical basic compounds nsing sulfated cyclodextrins. J. Pharm. Biomed. An. 2002, 27, 541-553. 

Table 12 gives an orientation help for CE separations sorted by pharmaceutical substances published in review articles. As this chapter focuses on the technical development of drug substances and products, only drug substances and drug formulations are covered. A useful compendium of CE applications in the pharmaceutical environment can be found in the book Capillary Electrophoresis Methods for Pharmaceutical Analysis written by G. Lunn. The book covers more than 700 active pharmaceutical ingredients and contains short method descriptions, sample preparation steps, and references. 

Hammitzsch, M., Rao, R. N., and Scriba, G. K. E. (2006). Development and validation of a robust capillary electrophoresis method for impurity profiling of etomidate including the determination of chiral purity using a dual cyclodextrin system. Electrophoresis 27(21), 4334—4344. 

Marini, R. D., Servais, A.-C., Rozet, E., Chiap, P., Boulanger, B., Rudaz, S., Crommen, J., Hubert, P., and Fillet, M. (2006). Nonaqueous capillary electrophoresis method for the enantiomeric purity determination of 5-timolol using heptakis(2,3-di-0-methyl-6-0-sulfo)-p-cyclodextrin validation using the accuracy profile strategy and estimation of uncertainty. /. Chromatogr. A 1120(1—2), 102-111. 

Muijselaar, P. G., hammers, N. G. E M., and Gerding, T. K. (2004). Development and validation of a capillary electrophoresis method for the enantiomeric purity determination of SLV307, a basic potential antipsychotic compound. Electrophoresis 25(16), 2854—2859. 

Sanger-van de Griend, C. E., Wahlstrom, H., Groningsson, K., and Widahl-Nasman, M. (1997). A chiral capillary electrophoresis method for ropivacaine hydrochloride in pharmaceutical formulations validation and comparison with chiral liquid chromatography. /. Pharm. Biomed. Anal. 15(8), 1051-1061. 

Toro, L, Dulsat, J. E, Fabregas, J. L., and Claramunt, J. (2004). Development and validation of a capillary electrophoresis method with ultraviolet detection for the determination of the related substances in a pharmaceutical compound. /. Chromatogr. A 1043(2), 303 — 315. 

Jamali, B., and Lehmann, S. (2004). Development and validation of a high-resolution capillary electrophoresis method for multi-analysis of ragaglitazar and arginine in active pharmaceutical ingredients and low-dose tablets. /. Pharm. Biomed. Anal. 34(3), 463—472. 

Wynia, G. S., Windhorst, G., Post, P. C., and Maris, F. A. (1997). Development and validation of a capillary electrophoresis method within a pharmaceutical quality control environment and comparison with high-performance liquid chromatography. /. Chromatogr. A 773(1—2), 339—350. 

Novatchev, N., and Holzgrabe, U. (2003). Capillary electrophoresis method for determination of related substances in glutathione reduced drug substance. Chromatographia 57(5—6), 345—349. 

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