Characterization and quantification of chemical constituents in Angong Niuhuang Pill using ultra-high performance liquid chromatography tandem mass spectrometry

Material basis researches and quality evaluation of traditional Chinese medicine preparations (TCMPs) face great challenges due to the complex composition and abundant unexpected interference. Angong Niuhuang Pill (ANP), one of the most famous TCMPs in China, containing 11 crude drugs has been commonly used for the treatment of febrile diseases. However, previous literatures of comprehensive chemical constituents in ANP were still limited. Herein, Ultra-high performance liquid chromatography-ion mobility-quadrupole time-of-flight mass spectrometry (UHPLC-IM-QTOF MS) method was established to effectively recognize the chemicals in Angong Niuhuang Pill (ANP). A total of 205 compounds, containing 72 confirmed with reference standards, were characterized from ANP with the application of a systematic strategy integrated polygonal mass defect filtering (MDF) and diagnostic fragment ion filtering (DFIF)/neutral loss filtering (NLF). Additionally, 68 major constituents in 8 batches of ANP samples were simultaneously determined by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QQQ MS/MS) within 40 min. The quantitative method was validated regarding linearity, precision, repeatability, stability, and accuracy. These findings proved the established liquid chromatography-mass spectrometry (LC-MS) method was efficient and dependable for qualitative and quantitative chemical profiling of ANP. Besides, this research could provide the material basis for further pharmacological researches and quality control of ANP.