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Method development and validation for estimation of Felodipine in human plasma by LCMS/MS
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After extraction from plasma by liquid - liquid extraction method, Felodipine and FelodipineD5, (IS), were separated on a Reverse phase chromatography using the mobile phase mixture of ammonium acetate and methanol at a flow rate of 1.0 ml/min. The analytes were detected in API 4000 Mass spectrometer in the positive atmospheric pressure chemical Ionization (APCI) mode with multiple reactions monitoring (MRM). The MRM transitions were monitored by following m/z for parent ion 384.0&daughter ion 338.0 (Felodipine), and m/z 389.1&daughter 338.1 (Felodipine D5, IS). A linear calibration plot of Felodipine was achieved in the concentration ranges of 0.105 ng/ml to 20.000 ng/ml. Mean recovery was 69.4%.This method was fully validated for specificity, precision, accuracy, reproducibility and other criteria as per regulations.
Keywords
Felodipine, Liquid liquid extraction, LCMS/MS, Human plasma, Validation
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