Validation of HPLC Method for Determination of Hydroxychavicol in the Extract from The Thai Traditional Remedy “Ruean-Mhoon-Nok” to Treat Psoriasis Disease
Keywords:
Hydroxychavicol, Ruean-Mhoon-Nok Remedy, Method validationAbstract
Ruean-Mhoon-Nok (RMN) remedy is listed in the National Thai Traditional Medicine Formulary in Honour of His Majesty the King B.E. 2562. The RMN remedy consists of 9 herbs used for the treatment of psoriasis disease (the stage characterized by white scars, scaly, raised edges, itching, and spreading all over the body). Previous research has shown that hydroxychavicol is an active compound of RMN remedy, but the analytical method was not validated. Therefore, this research aimed to develop and validate a method for analyzing the amount of hydroxychavicol in RMN remedy extract using high-performance liquid chromatography (HPLC). The results indicated that standard hydroxychavicol was eluted at a retention time of 33.927 min. Similar chromatogram was also found in the RMN remedy extract with the same retention time (33.797 min), confirming that it is hydroxychavicol. Moreover, the standard calibration curve of hydroxychavicol expressed a good linearity within the range of 25 – 400 µg/mL (R2 = 0.9996). The LOD and LOQ values were 4.31 and 13.05 µg/mL, respectively. Besides, the method demonstrated excellent intra-day and inter-day precisions of hydroxychavicol at 50, 100, and 200 µg/mL, with a relative standard deviation (%RSD) of 1.12-1.45% and 1.25-1.42%, respectively. In addition, the percentage recovery of hydroxychavicol was found to be 94.44 ± 0.58, 98.30 ± 1.11, and 100.79 ± 1.38%, respectively, indicating a high degree of accuracy of the method. Additionally, the quantification of hydroxychavicol in three different batches of RMN remedy extract was found to be within the range of 21.24 ± 0.16 to 22.80 ± 0.80 %w/w of extract. This method, therefore, will be very useful for standardization and quality control of RMN remedy extract, ensuring its efficiency and safety for future applications.
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