A simple chiral separation method using capillary electrophoresis with UV detection for analysis of enantiomers of drug compounds

  • Nadia Mira Kusumaningtyas, Prapin Wilairat and Rattikan Chantiwas
Keywords: Capillary electrophoresis, Chiral separation, Chiral selector, Drug chemical structure


A rapid and simple method for separation of enantiomers of a variety of compounds is desirable as information regarding the enantiomeric composition may not be provided by the chemical suppliers especially chemical drug compounds. These drugs are commercially available with no information regarding whether they are pure enantiomers or racemic mixtures. Each enantiomer may have different pharmacological activity. Thus the information of chirality is important. This work aims to employ a capillary electrophoresis with UV detection as a simple and rapid method for separation of enantiomers to determine the compound chirality. The developed method has been applied for separation of different functional chemicals of drugs, such as β-blockers and neurotransmitter, by using a single capillary electrophoresis method. The buffer conditions for the capillary method were optimized with regards to the chiral selector type and concentration for improving the efficiency of enantiomer separation. The optimum condition of buffer was phosphate buffer (20.0 mM, pH 2.7) containing CM-β-CD (4.0 mM). The single method was applied to the separation of four different drug compounds, viz. atenolol, propranolol, octopamine, and ibuprofen, to demonstrate the effectiveness of the developed method.


1. Lloyd DK, Ahmed A, Pastoré F. A quantitative relationship between capacity factor rand selector concentration in capillary electrophoresis and high performance liquid chromatography: evidence from the enantioselective resolution of benazoin using human serum albumin as a chiral selector. Electrophoresis 1997;18(6):958-64.
2. Le Potier I, Tamisier-Karolak SL, Morin P, Megel F, Taverna M. Comparison of native, alkylated and charged cyclodextrins for the chiral separation of labetalol stereoisomers by capillary electrophoresis. J Chromatogr A 1998;829(1-2):341-9.
3. Paik MJ, Nguyen DT, Kim KR. N-Menthoxycarbonylation combined with trimethylsilylation for enantioseparation of β-blockers by achiral dual-column gas chromatography. J Chromatogr A 2006;1103(1):177-81.
4. Lv L, Wang L, Zou Y, Chen R, Yu J. Chiral separation by nonaqueous capillary electrophoresis using L-sorbose-boric acid complexes as chiral ion-pair selectors. RSC Advances 2016;6(106):104193-200.
5. Su Y, Mu X, Qi L. Development of a capillary electrophoresis system with Mn(ii) complexes and β-cyclodextrin as the dual chiral selectors for enantioseparation of dansyl amino acids and its application in screening enzyme inhibitors. RSC Advances 2015;5(36):28762-8.
6. Mavroudi MC, Kapnissi-Christodoulou CP. Combined use of L-alanine tert butyl ester lactate and trimethyl-beta-cyclodextrin for the enantiomeric separations of 2-arylpropionic acids nonsteroidal anti-inflammatory drugs. Electrophoresis 2015;36(19):2442-50.
7. Lebedeva MV, Prokhorova AF, Shapovalova EN, Shpigun OA. Electrophoretic enantioseparation of profens in water-methanol solutions using eremomycin as a chiral selector. Mosc Univ Chem Bull 2013;68(5):215-8.
8. Maruszak W, Trojanowicz M, Margasińska M, Engelhardt H. Application of carboxymethyl-β-cyclodextrin as a chiral selector in capillary electrophoresis for enantiomer separation of selected neurotransmitters. J Chromatogr A 2001;926(2):327-36.
9. Kong D, Yang W, Duan K, Cui Y, Xing S, Zhang X, et al. Capillary electrophoretic enantioseparation of m-nisoldipine using two different beta-cyclodextrins. J Sep Sci 2009;32(18):3178-83.
10. Machín R, Isasi JR, Vélaz I. β-Cyclodextrin hydrogels as potential drug delivery systems. Carbohydr Polym 2012;87(3):2024-30.
11. Weinberger R. Practical capillary electrophoresis. Second ed. New York: Academic Press; 2000. 459 p.
12. Fanali S. Identification of chiral drug isomers by capillary electrophoresis. J Chromatogr A 1996;735(1-2): 121-77.
13. Podar A, Suciu S, Oprean R. Review-Recent enantiomer separation strategies of nonsteroidal anti-inflammatory drugs (NSAIDs) by capillary electrophoresis. Farmacia 2016;64(2):159-70.
14. Tophan P. Simple capillary electrophoresis system for chiral analysis of illicit drugs. M.Sc. Thesis, Mahidol University. Thailand; 2014.
15. Saz JM, Marina ML. Recent advances on the use of cyclodextrins in the chiral analysis of drugs by capillary electrophoresis. J Chromatogr A 2016;1467:94-79.
16. Fang L, Du Y, Hu X, Luo L, Guo X, Yu J. Carbocymethyl-β-cyclodextrine as chiral selector in capillary electrophoresis: enantioseparation of 16 basic chiral drugs and its chiral recognition mechanism associated with drugs’s structural features. Biomed Chromatogr 2017;31:e3991.
17. Zhu X, Lin B, Epperlein U, Koppenhoefer B. Enantiomeric resolution of some nonsteroidal antiinflammatory and anticoagulant drugs using β‐cyclodextrins by capillary electrophoresis. Chirality 1999;11(1):62-5.
18. Schmitt T, Engelhardt H. Derivatized cyclodextrins for the separation of enantiomers in capillary electrophoresis. J High Resolut Chromatogr 1993;16(9):525-9.
19. Zhu Q, Scriba GKE. Advances in the use of cyclodextrins as chiral selectors in capillary electrokinetic chromatography: fundamentals and applications. Chromatographia 2016;79(21):1403-35.
20. Ranzuglia GA, Manzi SJ, Gomez MR, Belardinelli RE, Pereyra VD. An analytical model for enantioseparation process in capillary electrophoresis. Physica A 2017;487:153-63.
21. Lancioni C, Keunchkarian S, Castells CB, Gagliardi LG. Enantiomeric separations by capillary electrophoresis: Theoretical method to determine optimum chiral selector concentration. J Chromatogr A 2018;1539:71-7.
22. Pang N, Zhang Z, Bai Y, Liu H. A study of the interaction between enantiomers of zolmitriptan and hydroxypropyl-beta-cyclodextrin by capillary electrophoresis. Anal Bioanal Chem 2009;393(1):313-20.
23. Al Azzam KM, Saad B, Aboul‐Enein HY. Determination of the binding constants of modafinil enantiomers with sulfated β‐cyclodextrin chiral selector by capillary electrophoresis using three different linear plotting methods. Electrophoresis 2010;31(17):2957-63.