Development and validation of a methodology for HPLC analysis of clopidogrel

Abstract

Aim to develop a methodology for the chemical and toxicological analysis of clopidogrel using HPLC.
 Material and methods. The study was performed on Clopidogrel bisulfate substance-powder, manufacturer: PJSC "Farmak", Ukraine, corresponding to the European Pharmacopoeia with an active substance content of 99.31% (LM2504208 series). For the chromatographic analysis we used a micro-column liquid chromatograph "Milichrome A-02" (CJSC "Econova", Russia). The analysis was done according to the unified HPLC technique developed by G.I. Baram.
 Results. The highest spectral ratio by HPLC was observed at 280 nm / 210 nm (maximum clopidogrel peak area R = 1.0787). The calibration curve for the HPLC method was described by the equation: у = 0.0005912х 0.001385 (r = 0.9998; S02 = 25.2910-7); linearity within 1400 mcg/ml, which corresponded to the content of clopidogrel in the sample from 2 ng to 800 ng, respectively. Based on the values of the calibration curve, the LOD and LOQ were calculated: 2,334 mcg/ml and 7,074 mcg/ml, respectively.
 Conclusion. A technique for the quantitative determination of clopidogrel by HPLC has been developed that allows for determining the substance in a mixture. The methodology was validated according to the following criteria: specificity, limit of detection, limit of quantitative determination, linearity, correctness and precision.