BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF CLOPIDOGREL BISULFATE IN HUMAN PLASMA BY RP-HPLC
Objective: A simple, sensitive, rapid and precise bioanalytical RP-HPLC method was developed for estimation of clopidogrel bisulfate in human plasma.
Methods: The work was carried out on Shimadzu LC-2010 CHT HPLC system equipped with Waters C18 (250Ã—4.6 mm, 5Î¼) column with a mobile phase containing acetonitrile: methanol: water (75:20:05 v/v/v). The flow rate of mobile phase was 1 ml/min and the detection was carried out at 225 nm. The retention time of clopidogrel bisulfate was found to be 4.6 min.
Results: The developed bioanalytical method was found to be linear in the concentration range of 30.76-69.23 Âµg/ml. The simple regression analysis of chromatographic response showed the value of R2= 0.9917. The precision study revealed that the cumulative percentage variation was within acceptable limit and accuracy study showed the value of mean percent recovery between 103.60 to 109.80 %. The clopidogrel was stable in human plasma at different storage conditions.
Conclusion: The validation parameters of the method met the acceptance criteria. Sufficient stability of both LQC and HQC was shown to allow for completion of sample analysis in clinical trials. From the results, we can conclude that developed bioanalytical method is simple, rapid, accurate, and precise and can be used for routine analysis of clopidogrel bisulfate.Keywords: Bioanalytical Method, Clopidogrel Bisulfate, RP-HPLC, Validation
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