Thai herbal Sahatsatara formula (STF) has been used for more than three decades to relieve musculoskeletal pain, numbness and anti-flatulence in Thai traditional medicine (TTM). STF composed of 21 ingredients. Black pepper is the major constituent in this formula. Previous studies revealed that piperine was used for quality control assessment of STF. However, only one biomarker is not enough to assure quality determination of the poly-herbal medicine. In this work we established untargeted metabolomic profiling of STF using Liquid Chromatography-Mass Spectrometry (LC-MS) and identification compounds with Traditional Chinese medicine (TCMs) library in UNIFI software. STF was extracted three concentrations. The untargeted metabolomics profiles of STF were represented A total of 21 natural products were detected in in all STF extract concentrations both positive and negative ionization modes Additionally, there were discovered new chemical markers for quality control. Moreover, the method was simplifying and reproducibility.