Benjakul is a Thai Traditional medicine preparation, used for balanced health. From selective interviews of folk
doctors in southern Thailand, it was used as the adaptogen drug for cancer patients. In our previous study, the ethanolic
extract of Benjakul preparation exhibited high cytotoxic activity against lung cancer cell lines (COR-L23). Piperine has been
identified as the main compound in the extract. In addition, plumbagin was found as the most cytotoxic compound. In this
study, a reversed-phase high performance liquid chromatography (HPLC) method for quality control such as chemical
fingerprint, quantification and stability of the ethanolic extract of Benjakul preparation was developed. The reversed-phase
HPLC was performed with a gradient mobile phase composed of water and acetronitrile, and peaks were detected at 256 nm.
Based on validation results, this analytical method is precise, accurate and stable for quantitative determination of piperine
and plumbagin which are cytotoxic compounds isolated from the ethanolic extract of Benjakul preparation. This method could
be suitable for analysis of Benjakul extract.

Keywords: Benjakul preparation, HPLC, Quantification, piperine, plumbagin