Identification of Calculus Bovis and its mixed varieties by ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) combined with the principal component analysis (PCA) method

•The mixed solvent system can be used to extract the chemical composition of the sample to the maximum extent.•The run time of each sample was 52 min by UHPLC-Q-TOF/MS.•UHPLC-Q/TOF-MS, Progenesis QI and Makerlynx XS software were integrated to clarify the difference of chemical components from Calculus Bovis and its mixed varieties.•The appropriate multi-response monitoring(MRM) and the characteristic marker components can be used integrately for the qualitative identification. The method of ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) was established and combined with principal component analysis (PCA) to identify natural Calculus Bovis, in vitro cultured Calculus Bovis and artificial Calculus Bovis. PCA, which was particularly powerful in dealing with multicollinearity and variables that outnumber the samples, was used to analyze the UHPLC-MS data of the processed samples, and potential markers were analyzed and described based on orthogonal partial least-squares discriminant analysis. According to the results in this study, the approach of combining UHPLC-QTOF-MS with PCA was proven to be credible and could be used to identify Calculus Bovis from in vitro cultured Calculus Bovis and artificial Calculus Bovis and to determine if there is Calculus Bovis in patented Chinese medicines that should contained Calculus Bovis medicinal materials.