A method was established for detecting the ingredients of horse, ox, sheep, pig, camel and deer skin in colla corii asini using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).Theoretical marker peptides were selected by comparing the collagen sequences in the skins of donkeys, horses, oxen, sheep, pigs, camels, and deer.Marker peptides were identified by protease cutting techniques and high resolution mass spectrometry and then analyzed by UPLC-MS/MS.The separation was performed on a UPLC system with a BEH C18 column via the gradient elution of acetonitrile containing 0.1%(v/v) acetic acid and water containing 0.1%(v/v) acetic acid.The marker peptides were identified in the modes of electrospray positive ionization (ESI+) and multiple reaction monitoring (MRM).The marker peptides for the horse, ox, sheep, pig and camel skin were detected in 15 samples.The method is simple, highly reproducible, and suitable for the identification of mixed skin ingredients in colla corii asini.It has been successfully used to detect the authenticity of colla corii asini.
Keywords: colla corii asini; collagen; marker peptide; ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).