Bioanalytical Method for the Determination of Hydroxyproline in Mouse Kidney by High-Performance Liquid Chromatography with Tandem Mass Spectrometric Detection

Rika SHIOTA; Hirotoshi MORITA; Tomoko MATSUMOTO; Atsushi MORIMOTO; Jun HAYAKAWA; Masako OKA; Hiroshi KAMIMORI

doi:10.2116/analsci.33.719

Notes

Bioanalytical Method for the Determination of Hydroxyproline in Mouse Kidney by High-Performance Liquid Chromatography with Tandem Mass Spectrometric Detection

Rika SHIOTA, Hirotoshi MORITA, Tomoko MATSUMOTO, Atsushi MORIMOTO, Jun HAYAKAWA, Masako OKA, Hiroshi KAMIMORI

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Keywords: Hydroxyproline, mass spectrometry, mouse, kidney

JOURNAL FREE ACCESS

2017 Volume 33 Issue 6 Pages 719-722

DOI https://doi.org/10.2116/analsci.33.719

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Abstract

An analytical method was developed and validated for the measurement of hydroxyproline (Hyp) levels in mouse kidney by high-performance liquid chromatography with tandem mass spectrometric detection (LC/MS/MS) using an analytical column specially designed for the LC/MS/MS analysis for intact amino acids. Tissue hydrolyzed with hydrochloric acid could be directly injected into the LC/MS/MS, as well as separated and detected using the deuterium labelled Hyp as an internal standard. The calibration curve showed good linearity from 5 to 500 nmol/mg of tissue; the precision and accuracy, including within- and between-run, were less than 6% and within 100 ± 6%, respectively.

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