Development of Reverse Phase Liquid Chromatographic Method for Determination of (+)-(S)-(o-Chlorophenyl)-6,7-Dihydrothieno [3,2-c] pyridine-5(4H)-acetic acid,Hydrochloride and Methyl (+/-) – (o-Chloro phenyl)-4,5-Dihydrothieno[2,3-c]pyridine-6(7H)-acetate, Hydrochloride from Clopidogrel Besylate

Author(s)

Pawaskar, P. S., Dighe, V. V., Adhyapak, S. S., Shambhu, N. S., Mestry, D. Y.

Author's Affiliation

Abstract

Clopidogrel besylate contain single stereogenic center and has impurities like ((+) - (S) - (o-chlorophenyl)-6,7-dihydrothieno[3,2-c] pyridine-5(4H)- acetic acid, hydrochloride) which is known as impurity A and (Methyl (+/-) - (o-chloro phenyl)-4,5-dihydrothieno[2,3-c] pyridine-6(7H)-acetate, hydrochloride) which is known as impurity B. They are introduced during production. A simple, sensitive, precise and high performance liquid chromatographic (HPLC) method has been developed and validated for quantitative determination of impurity A and impurity B from clopidogrel besylate in bulk drug using uv detector at 220 nm. The developed method was able to separate impurity A and impurity B of clopidogrel besylate from its bulk drug within 50 min. The chromatographic separation was carried out by reverse phase chromatography using C₈ column (Zorbax SB C₈ 250 mm x 4.6 mm x 5 µm), with mobile phase comprising of buffer solution and acetonitrile in the gradient composition, at a flow rate of 1.0 ml/min, at 25°C temperature. The limit of detection  and limit of quantitation  of impurity A were found to be 0.07 µg/ml and 0.20 µg/ml and of impurity B were found to be 0.10 µg/ml and 0.30 µg/ml respectively. The linearity of response of impurity A was in the range of 0.20 µg/ml to 3.0 µg/ml with r > 0.9999. The linearity of response of impurity B was in the range of 0.30 µg/ml to 4.5 µg/ml with r > 0.9995. The method was validated and found to be suitable for determination of impurity A and impurity B from clopidogrel besylate bulk drug.

Keywords

Clopidogrel besylate, ((+)-(S)-(o-chlorophenyl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-acetic acid,          hydrochloride)   i.e. (impurity A), (Methyl (+/-) - (o-chloro phenyl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-acetate, hydrochloride) i.e. (impurity B), High performance liquid chromatography, Method validation and quantitation.Nephrotoxicity, Anti-tubercular drugs, Thymoquinone

Cite This Article

Pawaskar, P. S., Dighe, V. V., Adhyapak, S. S., Shambhu, N. S., & Mestry, D. Y. (2013). Development of Reverse Phase Liquid Chromatographic Method for Determination of (+)-(S)-(o-Chlorophenyl)-6,7-Dihydrothieno [3,2-c] pyridine-5(4H)-acetic acid,Hydrochloride and Methyl (+/-) - (o-Chloro phenyl)-4,5-Dihydrothieno[2,3-c]pyridine-6(7H)-acetate, Hydrochloride from Clopidogrel Besylate. International Journal for Pharmaceutical Research Scholars (IJPRS), 2(1), 16-23.