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Rapid analysis of pyridinoline and deoxypyridinoline in biological samples by liquid chromatography with mass spectrometry and a silica hydride column
Author(s) -
Naffa Rafea,
Watanabe Seiichiro,
Zhang Wenkai,
Maidment Catherine,
Singh Preet,
Chamber Paul,
Matyska Maria T.,
Pesek Joseph J.
Publication year - 2019
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201801292
Subject(s) - pyridinoline , chromatography , deoxypyridinoline , chemistry , mass spectrometry , column (typography) , hydride , high performance liquid chromatography , organic chemistry , alkaline phosphatase , osteocalcin , enzyme , structural engineering , connection (principal bundle) , engineering , hydrogen
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.