Determination Of Hydroxyproline In Collagenous Proteins By High Performance Liquid Chromatography-Mass Spectrometry Determination Of Hydroxyproline In Collagenous Proteins By High Performance Liquid Chromatography-Mass Spectrometry
Keywords
High Performance Liquid Chromatography-Mass Spectrometry (Hplc-Ms),Hydroxyproline,Collagenous Proteins High Performance Liquid Chromatography-Mass Spectrometry (Hplc-Ms),Hydroxyproline,Collagenous Proteins
Abstract
simple and rapid high-performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI/MS) method for the determination of hydroxyproline (HYP) in collagen was established. After acid hydrolysis, collagen was separated on a C8 reversed-phase column at a flow rate of 1.0 mL/min using acetonitrile-0.05% trifluoroacetic acid aqueous solution (volume ratio 5:95) as the mobile phase. Using theanine as the internal standard, mass spectrometry was used to determine HYP qualitatively and quantitatively. Selected ion monitoring was performed on m/z132 and m/z175 ions in electrospray positive ion mode. In the range of 11.7 to 117 mg/L, the peak area ratio of HYP to the internal standard theanine and the mass concentration of HYP showed a good linear relationship, with a correlation coefficient of 0.9993. The relative standard deviation of the content determination was 1.87%, and the recovery rate was 97.85% to 101.76%. This method has a simple mobile phase, short analysis time and no need for derivatization. It has strong anti-interference ability and can accurately and quickly determine the HYP content in collagen.
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Original research was done by XIA Jingen,CHEN Bo,YAO Shouzhuo
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