Chinese Journal of Chromatography

Chinese Journal of Chromatography ›› 2017, Vol. 35 ›› Issue (12): 1251-1256.DOI: 10.3724/SP.J.1123.2017.09020

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Determination of hydroxyproline in liver tissue by hydrophilic interaction chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry

LIU Wei1,2, QI Shenglan4, XU Ying1,2, XIAO Zhun1,2,3, FU Yadong1,2,3, CHEN Jiamei1,2, YANG Tao5, LIU Ping1,2,3   

  1. 1. Institute of Liver Disease, Shuguang Hospital Affiliated with Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
    2. Key Laboratory of Liver and Kidney Disease of the Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
    3. E-Institute of Shanghai Municipal Education Commission, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
    4. Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
    5. Institute of Cardiovascular Disease, Shuguang Hospital Affiliated with Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • Received:2017-09-09 Online:2017-12-08 Published:2017-12-15
  • Supported by:

    Key Project of National Natural Science Foundation of China (No. 81530101); Shanghai Sailing Program (No. 17YF1419800); Project Funded by China Postdoctoral Science Foundation (No. 2016M601643).

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Abstract:

A method for the determination of hydroxyproline (Hyp) in liver tissue of mice by hydrophilic interaction chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (HILIC-HRMS) was developed. The liver tissue samples of normal mice and liver fibrosis mice induced by carbon tetrachloride were hydrolyzed by concentrated hydrochloric acid. After filtrated and diluted by solution, the diluent was separated on an Hypersil GOLD HILIC column (100 mm×2.1 mm, 3 μm). Water-acetonitrile (28:72, v/v)were used as the mobile phases with isocratic elution. Finally, the target analytes were detected in positive model by HRMS equipped with an electrospray ionization source. The linear range of hydroxyproline was from 0.78 to 100.00 μg/L with the correlation coefficient (R2) of 0.9983. The limit of quantification was 0.78 μg/L. By detecting the spiked samples, the recoveries were in the range of 97.4%-100.9% with the relative standard deviations (RSDs) between 1.4% and 2.0%. In addition, comparison of the measurement results by this method and the chloramine T method was proceeded. It was found that the linear correlation between the two methods was very good, and the Pearson correlation coefficient was 0.927. And this method had simpler operation procedure and higher accuracy than chloramine T method. This method can be used for the quick determination of hydroxyproline in liver tissue samples.

Key words: hydrophilic interaction chromatography (HILIC), hydroxyproline (Hyp), liver fibrosis, liver tissue, quadrupole/orbitrap high resolution mass spectrometry (HRMS)

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