Rapid analysis of pyridinoline and deoxypyridinoline in biological samples by liquid chromatography with mass spectrometry and a silica hydride column

Publication Date

April 2019

Document Type

Article

Publication Title

Journal of Separation Science

Volume

42

Issue

8

DOI

10.1002/jssc.201801292

First Page

1482

Last Page

1488

Abstract

Pyridinoline and deoxypyridinoline crosslinks are biomarkers found in urine for collagen degradation in bone turnover. For the first time, a rapid, sensitive, and ion‐pairing free method is described for the analysis of pyridinoline and deoxypyridinoline using ultra‐high performance liquid chromatography with Cogent Diamond Hydride column and detection by Q Exactive hybrid quadrupole‐orbitrap high resolution accurate mass spectrometry. The separation was achieved using both isocratic and gradient conditions and run time <5 min under isocratic conditions of 20% acetonitrile in water containing 0.1% formic acid. Pyridoxine was used as an internal standard and relative standard deviation of the retention times of both pyridinoline and deoxypyridinoline were <1%. The limit of detection was 0.082 ± 0.023 μM for pyridinoline and 0.118 ± 0.052 μM for deoxypyridinoline. The limit of quantitation was 0.245 ± 0.070 μM for pyridinoline and 0.354 ± 0.157 μM for deoxypyridinoline. The method was validated by the detection and quantitation of both pyridinoline and deoxypyridinoline in skin and urine samples.

Keywords

deoxypyridinoline, mass spectrometry, pyridinoline, silica hydride column, urine analysis

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