Analytical Sciences

Abstract − Analytical Sciences, 33(6), 719 (2017).

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,*** and Hiroshi KAMIMORI***
*Shionogi TechnoAdvance Research Co., Ltd., 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan
**Biomarker R&D Department, Shionogi & Co. Ltd., 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan
***Pharmaceutical Research Division, Shionogi & Co. Ltd, 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan
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.