Development and Validation of Analytical Method for Piperine in Phytopharmaceutical Product Containing Cabe Jawa Fructus by HPLC

Cabe Jawa fruit (Retrofractii fructus) has empirically known to have effects such as anti-inflammatory and analgesic due to the presence of piperine. The maximum daily dose for piperine is 0.300–0.540 mg/kg/person/day. A reliable analytical method is required to ensure that phytopharmaceutical preparation is safe. This study developed and validated a piperine analysis method using phytopharmaceutical samples containing Cabe Jawa. A Shimadzu prominence HPLC SIL-20A Prominence autosampler and an X-Bridge C18 column (5 m x 250 mm x 4.6 mm) were used in the research devices. The OFAT method optimised the mobile phase, instrument systems, and sample preparation. The instrument system was optimized for acetonitrile:1% glacial acetic acid (48:52), 40°C, and a 1 mL/min flow rate. Each solid and liquid sample was prepared using the multilevel methanol dilution and water-dichloromethane liquid-liquid extraction (LLE) method. The results of validating the analytical procedures for solid and liquid samples were LOD and LOQ of 1.290 g/mL and 4.300 g/mL, 1.310 g/mL and 4 g/mL, respectively. Precision: 0.157% and 1.174%. Accuracy for three different concentrations were 94.232±1.050%, 95.387%±0.176%, and 96.272±1.043%; 88.801±0.428%, 89.473 ±0.252%, and 89.338 ±0.052% respectively. The analysis of the sample's piperine revealed that this method met the criteria for the validation parameters.