Identification Of The Species Source Of Collagen Based On Liquid Chromatography-Mass Spectrometry Technology Based On Characteristic Peptides
Keywords
Characteristic Peptides, Collagen, Liquid Chromatography-Mass Spectrometry Species Sources
Abstract
method for identifying the source of collagen species based on liquid chromatography-mass spectrometry technology based on characteristic peptides was established. After protein extraction, reduction, alkylation, and trypsin digestion, the sample was separated using an Eksigent C18 chromatographic column (75 ¦Ìm ¡Á 150 mm, 3 ¦Ìm), using mobile phase 0? ? 1% formic acid water-acetonitrile solution (98:2) and 0? ? 1% formic acid acetonitrile-water solution (98:2) gradient elution, in the positive ion mode, detected by nanoelectrospray quadrupole time-of-flight mass spectrometry, the data was analyzed by ProteinPilotTM software and blast, and potential characteristic peptides were screened out. . The digested sample was then separated using an Eclipse Plus C18 chromatographic column (2?? 1 mm ¡Á 100 mm, 1?? 8 ¦Ìm). The mobile phase was gradient eluted with acetonitrile and 1% formic acid aqueous solution. In the positive ion mode, the column was electrolytically separated. The multiple reaction monitoring of the spray quadrupole/linear ion trap tandem mass spectrometer triggered the enhancer ion scanning mode for detection to further confirm the specificity of the peptide. Finally, 3 characteristic peptides of porcine-derived collagen, 4 characteristic peptides of bovine-derived collagen, and 1 characteristic peptide of sheep-derived collagen were screened and confirmed. The selected characteristic peptides have good heat resistance and can provide a highly specific, accurate and reliable detection method for the identification of animal-derived collagen.
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Original research was done by Liu Yixiong, Zhu Yuwei, Li Yi, Du Yegang, Yang Guowu, Wang Weida, Yang Hongbo
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