A combination of qualitative and quantitative methods was developed and validated to elucidate the chemical constituents in Ya-tong-yi-li-wan (YTYLW), a traditional Chinese medicine formula used for the treatment of toothache in China. Few reports are available on the chemical analysis of YTYLW, so far. For qualitative analysis, a method based on high-performance liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry (HPLC/DAD/ESI-MSn) was established. A total of 48 compounds in YTYLW were characterized, including 10 bufadienolides, 9 isoflavones, 7 chalcones, 7 saponins, 6 flavanones, 4 flavonols, 2 flavones, 2 coumarins, and 1 pterocarpan. For quantitative analysis, 12 major compounds in YTYLW were simultaneously determined by HPLC-UV at 270 nm in 9 batches of commercial products. Precision, reproducibility and accuracy of the method were fully validated. The method established in this study could be used for the quality control of YTYLW.
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