Validated High Performance Liquid Chromatographic (HPLC) Method for Anti-inflammation Activity of Lom-Am-Ma-Preuk Remedy

Abstract

Lom-Am-Ma-Preuk (LAP) remedy is a Thai traditional medicine procedure for muscle pain relief. Active ingredients from LAP remedy are eugenol, myristicin and piperine. These standard markers also have been proven to have anti-inflammatory activity. The aim of this study was to develop a validated HPLC method for determination of markers in the LAP remedy to assess the validation parameters. Results show that the validation parameters were specificity, limit of detection, limit of quantification, linearity, accuracy and precision. The developed method was specific to these markers with the retention times of 45.7, 56.6 and 53.9 min for eugenol, myristicin and piperine, respectively. The coefficient of determination (R²) was more than 0.999. The accuracy and precision were in an acceptable range with good recovery verified by coefficient of variation of less than 2%. The limits of detection (LOD) of eugenol, myristicin and piperine were 0.5, 0.2 and 1.3 μg/mL and limits of quantitation (LOQ) were 1.9, 0.6 and 5 μg/mL, respectively. Total analysis time was approximately 70 min. The contents of eugenol,myristicin and piperine in LAP could be obtained in one analysis for the first time. The present method is useful for routine quality control of the LAP remedy.