CHIRAL SEPARATION OF CITALOPRAM BY REVERSED PHASE HPLC USING SULFATED BETA CYCLODEXTRIN AS CHIRAL MOBILE PHASE ADDITIVE
Keywords:Enantiomeric resolution, Citalopram enantiomers, Citalopram enantiomeric purity, Chiral additive
Objective: To develop a simple and cost effective chiral HPLC method for the separation of citalopram (CIT) enantiomers using chiral mobile phase additives (CMPAs).
Methods: Sulfated beta cyclodextrin (S-Î²-CD) was synthesized in our laboratory and was evaluated as a CMPA. The parameters affecting the resolution were optimized. CIT enantiomers were resolved on an achiral Kromasil C8 column (150 mm Ã— 4.6 mm, 5 Âµm) using methanol and 20 mM KH2PO4 (pH 3) containing 12 mM S-Î²-CD (35:65) as the mobile phase with a flow rate of 1 ml/min at 240 nm. Chiral resolution capacity of synthesized S-Î²-CD was compared to the marketed product. The method using synthesized S-Î²-CD as CMPA was validated and applied for the quantitative determination of CIT enantiomers in bulk drug and tablet formulation.
Results: Synthesized S-Î²-CD gave a better resolution than the marketed form. This method was validated as per ICH guidelines and was found to comply with the standard norms. A good linearity was observed in the concentration range of 1-30 Âµg/ml with R2= 0.9993 for both enantiomers. The limit of detection and limit of quantification was 0.0272 and 0.0824 Î¼g/ml for the R-enantiomer and 0.0303 and 0.0920 Î¼g/ml for the S-enantiomer respectively.
Conclusion: A rapid and cost effective RP-HPLC method was developed and validated as per ICH guidelines to separate the CIT enantiomers. The method could be successfully applied for the quantitative determination of CIT enantiomers in bulk drug samples and pharmaceutical formulations.
Vaswani M, Linda FK, Ramesh S. Role of selective serotonin reuptake inhibitors in psychiatric disorders: A comprehensive review. Prog Neuropsychopharmacol Biol Psychiatry 2003;27:85-102.
MÃ¸rk A, Kreilgaard M, SÃ¡nchez C. The R-enantiomer of citalopram counteracts escitalopram-induced increase in extracellular 5-HT in the frontal cortex of freely moving rats. Neuropharmacol 2003;45:167-73.
Zhong H, Haddjeri N, SÃ¡nchez C. Escitalopram, an antidepressant with an allosteric effect at the serotonin transporterâ€”a review of current understanding of its mechanism of action. Psychopharmacol (Berl) 2012;219:1-13.
Leonard B, Taylor D. Escitalopramâ€”translating molecular properties into clinical benefit: reviewing the evidence in major depression. J Psychopharmacol 2010;24:1143-52.
Rauws AG, Groen K. Current Regulatory (draft) Guidance on chiral medicinal products: Canada, EEC, Japan, United States. Chirality 1994;6(2):72-5.
Branch SK. Guidelines from the International Conference on Harmonisation (ICH). J Pharm Biomed Anal 2005;38:798-805.
Guan J, Yang J, Bi Y, Shi S, Yan F, Li F. Determination of the enantiomeric impurity in S-(-)pantoprazole using high performance liquid chromatography with sulfobutylether-beta-cyclodextrin as chiral additive. J Sep Sci 2008;31(2):288-93.
Rocco A, Fanali S. Enantiomeric separation of acidic compounds by nano-liquid chromatography with methylated-beta-cyclodextrin as a mobile phase additive. J Sep Sci 2009;32(10):1696-703.
Tong S, Zhang H, Shen M, Ito Y, Yan J. Enantioseparation of mandelic acid derivatives by high performance liquid chromatography with substituted Î²-cyclodextrin as chiral mobile phase additive and evaluation of inclusion complex formation. J Chromatogr B: Anal Technol Biomed Life Sci 2014;962:44-51.
Rochat B, Amey M, Baumann P. Analysis of enantiomers of citalopram and its demethylated metabolites in plasma of depressive patients using chiral reverse-phase liquid chromatography. Ther Drug Monit 1995;17(3):273-9.
Carlsson B, Norlander B. Optimization and characterization of the chiral Separation of citalopram and its demethylated metabolites by response-surface methodology. Chromatographia 2001;53(5-6):266-72.
Haupt D. Determination of citalopram enantiomers in human plasma by liquid chromatographic separation on a chiral-AGP column. J Chromatogr B Biomed Appl 1996;685(2):299-305.
Kosel M, Eap CB, Amey M, Baumann P. Analysis of the enantiomers of citalopram and its demethylated metabolites using chiral liquid chromatography. J Chromatogr B Biomed Sci Appl 1998;719(1-2):234-8.
Rochat B, Amey M, Van Gelderen H, Testa B, Baumann P. Determination of the enantiomers of citalopram, its demethylated and propionic acid metabolites in human plasma by chiral HPLC. Chirality 1995;7(6):389-95.
Gupta VK, Ali I, Agarwal S. Enantiomeric analysis of citalopram in human plasma by SPE and chiral HPLC method. Int J Electrochem Sci 2011;6:5639â€“48.
El-Gindy A, Emara S, Mesbah MK, Hadad GM. Liquid chromatography determination of citalopram enantiomers using beta-cyclodextrin as a chiral mobile phase additive. J AOAC Int 2006;89(1):65-70.
Berzas NJJ, Guiberteau CC, VillaseÃ±or LMJ, RodrÃguez RV. Enantiomeric determination, validation and robustness studies of racemic citalopram in pharmaceutical formulations by capillary electrophoresis. J Chromatogr A 2005;1072(2):249-57.
Mandrioli R, Fanali S, Pucci V, Raggi MA. Enantiomeric separation of citalopram and its metabolites by capillary electrophoresis. Electrophoresis 2003;24(15):2608-16.
Kumar AP, Park JH. Azithromycin as a new chiral selector in capillary electrophoresis. J Chromatogr A 2011;1218(9):1314-7.
Unceta N, GÃ³mez-Caballero A, GarcÃa D, DÃaz G, Guerreiro A, Piletsky S, et al. Enantioselective extraction of (+)-(S)-citalopram and its main metabolites using a tailor-made stir bar chiral imprinted polymer for their LC-ESI-MS/MS quantitation in urine Samples. Talanta 2013;116:448-53.
Li M, Bao Z, Su B, Xing H, Yang Y, Ren Q. Enantiomeric separation of citalopram base by supercritical fluid chromatography. J Sep Sci 2013;36(18):3093â€“100.
Kenley KN, Qiqing Z, Kavita M, Nik C. Effect of sulfobutylether beta-cyclodextrin modifier on selectivity of reversed phase HPLC separations. J Liq Chromatogr Relat Technol 2012;35:2845â€“59.
Ameyibor E, Stewart JT. Resolution and quantitation of pentazocine enantiomers in human serum by reversed-phase high-performance liquid chromatography using sulfated Î²-cyclodextrin as chiral mobile phase additive and solid-phase extraction. J Chromatogr B Biomed Sci Appl 1997;703(1-2):273-8.
Wei M, Wang J, He J, Evans DG, Duan X. In situ FT-IR, in situ HT-XRD and TPDE study of thermal decomposition of sulfated Î²-cyclodextrin intercalated in layered double hydroxides. Microporous Mesoporous Mater 2005;78(1):53-61.