DEVELOPMENT AND VALIDATION OF HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) METHOD FOR DETERMINATION OF PIPERINE IN PHYTOPHARMACEUTICAL PRODUCT CONTAIN CABE JAWA FRUCTUS EXTRACT (RETROFRACTII FRUCTUS)

• Main Author: Galih, Wening
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/73971
• Institution: Institut Teknologi Bandung

Cabe Jawa fruit (Retrofractii fructus) is a natural compound present in phytopharmaca preparations. Piperine, a secondary metabolite of Cabe Jawa has antipyretic, analgesic, anti-inflammatory, and central nervous system suppressing properties. Piperine has long-term and high-dose side effects, including impaired liver function. According to the EPA, the maximum daily use limit for piperine compounds is 0.300 to 0.540 mg/kg/person/day. There is currently no analytical technique for determining piperine levels in traditional phytopharmaca preparations, therefore this research was carried out to develop a piperine analysis method in phytopharmaca preparations. Optimizing the development of the piperine analysis method with the Shimadzu prominence HPLC SIL-20A Prominence autosampler using the One Factor at a Time (OFAT) approach to the mobile phase, sample preparation, and optimum conditions for analysis. Piperine was analyzed for solid and liquid preparation using High Performance Liquid Chromatography (HPLC) on a 5µm x 250mm x 4.6µm (X-Bridge C18) column with an acetonitrile:1% glacial acetic acid (48:52) mobile phase at a flow rate of 1 mL/min. The methanol dilution method was used for solid preparations.The liquid preparation was prepared using the liquid-liquid extraction (LLE) method, which included dichloromethane water, 1 gram of sodium sulfate anhydrate, and acetonitrile as a solvent. The analytical method was validated using parameters such as specificity, linearity, and range, detection and quantification limits, precision, accuracy, and robustness. A LOD value of 1.290 µg/mL was obtained in capsule preparations; LOQ 4.300 µg/mL respectively; precision with% RSD 0.157%; accuracy in the form of% recovery at three different concentrations, namely 94.232 ± 1.050%, 95.387% ± 0.176%, and 96.272 ± 1.043%. In liquid preparations, the LOD value of 1.300 µg/mL and LOQ were 4.000 µg/mL respectively; the precision with% RSD 0.174%; and the accuracy was 88.801 ± 0.428%, 89.473± 0.252% and 89.338 ± 0.052% at three different concentrations. The validation result showed that the analytical method met the requirements and could be used to analyze piperine in phytopharmaca capsule and liquid preparations. Testing of samples of phytopharmaca preparations using validated analytical methods showed that the sample content had met the requirement.