DEVELOPMENT AND VALIDATION OF UPLC-TQD/MS METHOD TO DETERMINE ACTIVE COMPOUND IN THAI HERBAL HARAK FORMULA

Abstract

This study aimed to develop an analytical method for determining four anti-inflammatory active compounds in the Harak herbal formula, an herbal formula in Thailand's national list of essential medicines with antipyretic properties. The targeted analytes were pectolinarigenin, gallic acid, ferulic acid, and (+)-catechin. The analysis utilized UPLC-TQD/MS, and chromatographic separation was achieved on an HSS T3 column (1.8 μm, 2.1 × 100 mm) using a gradient elution with 0.1% formic acid in water and 0.1% formic acid in acetonitrile. This method completes the separation of all four active compounds within 8 minutes. The method was validated by assessing linearity, precision, accuracy, recovery, LOD, and LLOQ. The linearity range for pectolinarigenin and gallic acid was 1.5-200 ng/mL, while for ferulic acid and (+)-catechin, it was 1-200 ng/mL. All active compounds exhibited excellent linearity, with coefficients of determination (R²) ≥ 0.999. Precision, accuracy, and recovery were within the acceptance criteria for all active compounds. Finally, the validated method was successfully applied to assess the quality of 14 marketed Harak herbal formula samples. Due to the variations in the active compounds found, quality control of herbal raw materials is crucial for ensuring the quality of herbal medicines.