Method Development and Validation of 9α‐Fluoro‐16β‐Methyl‐Prednisolone‐17‐Valerate by HPLC

In this study, the development and validation of an analytical method for the assay of 9α‐fluoro‐16β‐methyl‐prednisolone‐17‐valerate (FMPV) using reversed‐phase high performance liquid chromatography (HPLC) is reported. The chromatographic separation is achieved with methanol‐water (63:37, v/v) as mobile phase, a C18 column, and UV detection at 240 nm. The method was critically validated to demonstrate its selectivity, linearity, precision, accuracy, specificity, limit of detection and quantitation. The calibration curve showed good linearity (r 2=0.9999) over the concentration range 20 to 150 µg/mL. The mean percent relative standard deviation values for precision studies were less than 0.37%. Mean recoveries were 99.73–100.02%. The limit of detection was 3.0 µg/mL and limit of quantitation was 20.0 µg/mL. This method represents a useful protocol for routine testing of 9α‐fluoro‐16β‐methyl‐prednisolone‐17‐valerate drug substance.