Development and Validation of a Bioanalytical Method for the Quantification of Aceclofenac in a Small Volume of Human Serum by RP-HPLC

In this study, a simple, accurate, precise, and cost-effective method based on reversed-phase high-performance liquid chromatography with diode array detection for the quantification of aceclofenac in a small volume of human serum (100 µL) was developed and validated according to US-FDA guidelines. Acetanilide was used as an internal standard. The analyte and internal standard were spiked in human serum and extracted using a simple protein precipitation technique. The analysis was carried out on a reversed-phase C 18 HPLC column (5 μm, 4.6 × 250 mm) using 0.1% formic acid and acetonitrile in a ratio of 40 : 60 (v/v) as the mobile phase with UV detection at 275 nm at a flow rate of 1.2 mL/min. The developed method was found to be sensitive, selective, and linear over the concentration range of 0.8–20 µg/mL with good precision (RSD ≤ 15%) and accuracy (relative error ≤15%). The limit of detection and lower limit of quantification were found to be 0.3 and 0.8 µg/mL, respectively.