Quantitative Detection Method Of Collagen Based On Biological Mass Spectrometry
Keywords
Collagen, Quantitative Detection, Characteristic Peptides, Liquid Chromatography Tandem Mass Spectrometry
Abstract
This study aims to establish a quantitative detection method for collagen based on characteristic peptides, screen characteristic peptides of collagen through sequence comparison, use trypsin to enzymatically hydrolyze bovine type I collagen standard, and use liquid mass spectrometry technology ( HPLC-MS) is used to detect characteristic peptides, establish the corresponding relationship between the abundance of characteristic peptides and collagen concentration, and use it for actual sample analysis. The results show that 6 characteristic peptides were detected in bovine type I collagen. Among them, the peptide geagpsgpagptgar is suitable as a characteristic peptide for quantitative detection due to its high abundance and stable secondary mass spectrometry. The signal intensity of the peptide is closely related to the protein concentration (0.1-3.0 mg/ml ) shows a good linear relationship. The developed method was applied to actual sample analysis. The collagen content of bovine Achilles tendon was 90.2%, and the collagen content of collagen sponge was 93.4%. The detection results were consistent with the results based on amino acid composition analysis. This study shows that it is feasible to quantitatively detect collagen based on HPLC-MS characteristic peptide analysis method. This method has application prospects in the quality control of biological products such as collagen-containing medical devices.
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Original research was done by Sun Kun, Yang Fan, Kong Yingjun, Kang Jiyao, Cao Wei, Yang Xiaoyan, Zha Shenghua, Zhang Guifeng, Wang Minglin
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