Identification of Colla Corii Asini and Examination of Six Animal Heterologous Components Based on UPLC-MS/MS Characteristic Peptide Detection Technology
Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method to detect the six animal heterologous components such as horse hide glue,cattle hide glue,pig hide glue,camel hide glue,sheep hide glue,deer hide glue in Colla Corii Asini.Methods Colla Corii Asini powder was dissolved with 1%ammonium bicarbonate solution for ultrasonic dissolution,filtration,and enzymolysis with trypsin for 15 h.UPLC-MS/MS method was adopted,with the Colla Corii Asini characteristic peptide[mass charge ratio(m/z)539.8 → 924.3],donkey characteristic peptide(m/z 570.5 → 698.1),horse characteristic peptide A(m/z 386.4 → 377.3),cattle characteristic peptide A(m/z 641.2 →726.3),pig characteristic peptide(m/z 774.5 → 1 035.7),camel characteristic peptide(m/z 603.8 → 281.8),sheep characteristic peptide(m/z 553.0 → 450.8),and deer characteristic peptide(m/z 733.1 → 962.5)as detection ion pairs.Multi-reaction monitoring(MRM)was adopted with apositive ion mode of electric spray ion source(ESI).The quality of 10 batches of medicinal Colla Corii Asini samples and six batches of food-grade Colla Corii Asini samples were evaluated.Results The selected characteristic peptide segments of each animal were only detected in the corresponding animal hide glue,and the specificity of the method was strong.The sensitivity of the method met the quality control requirements.The repeatability,injection precision,and sample stability of enzymolysis all met the requirements of methodological validation,and the RSDs of the results were all lower than 9. 0％(n=6). In six batches of food-grade Colla Corii Asini samples,no Colla Corii Asini component was detected and the main components were determined to be cattle hide glue,camel hide glue,horse hide glue and pig hide glue. The Colla Corii Asini was detected in eight batches of medicinal Colla Corii Asini samples,of which the content of Colla Corii Asini in one batch was low,and horse hide glue was detected in one batch. Two batches of medicinal Colla Corii Asini were judged as counterfeit products because no ingredient of Colla Corii Asini was detected. Conclusion The established method for the detection of six animal hide glue has strong specificity and high sensitivity,which can be used for the identification of Colla Corii Asini and the detection of adulterated hide glue.
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