A Stability Indicating Lc-Ms/Ms Method for Quantification of a Nox Inhibitor R14 in Its Bisulfite Adduct Form for Pharmacokinetic Studies

Authors

Yang Wang
Jing Ma
Shiaw-Yih Lin
Huan Xie
Dong Liang

Publication Date

5-10-2023

Journal

Journal of Pharmaceutical and Biomedical Analysis

Abstract

R14, also known as NOX Inhibitor VII, is a potent inhibitor of NADPH oxidases (NOX) which has recently been identified as a novel agent targeting to triple-negative breast cancer. It is also rapidly degraded in collected pharmacokinetic plasma and blood samples even stored under - 70 °C. The purpose of this study was to develop a stability indicating LC-MS/MS assay that would be suitable for quantification of R14 in plasma and blood. In the presence of sodium sulfite under acidic pH, R14, an aryl lactam compound which is not a typically reactive compound for bisulfite addition, readily and completely converted to R14 bisulfite adduct, which was more stable in plasma and blood. The adduct has MRM transition at m/z 340.1-127.0 in negative mode and showed high sensitivity in LC-MS/MS quantification. Thus, monitoring the adduct provided a suitable way of quantitating R14 concentrations in mouse whole blood. The reacting conditions were optimized based on detecting R14 bisulfite adduct, and the assay was established and validated on a SCIEX 6500+ Triple Quad LC-MS/MS System. The method was then successfully adapted to pharmacokinetic studies after oral administration of R14 to mice.

Keywords

Mice, Animals, Chromatography, Liquid, Tandem Mass Spectrometry, Sensitivity and Specificity, Reproducibility of Results

Comments

Data Availability Statement

PMID: 36924633