Development and validation of an LC-MS/MS method for quantifying NAD + and related metabolites in mice sciatic nerves and its application to a nerve injury animal model.

Autor: Ma Y; College of Chemistry, Beijing Key Laboratory of Environmentally Harmful Chemical Analysis, Beijing University of Chemical Technology, Beijing 100029, China; DMPK Department, Sironax (Beijing) Co., Ltd, Beijing 102206, China., Deng L; DMPK Department, Sironax (Beijing) Co., Ltd, Beijing 102206, China., Du Z; College of Chemistry, Beijing Key Laboratory of Environmentally Harmful Chemical Analysis, Beijing University of Chemical Technology, Beijing 100029, China. Electronic address: duzx@mail.buct.edu.cn.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Apr 26; Vol. 1721, pp. 464821. Date of Electronic Publication: 2024 Mar 15.
DOI: 10.1016/j.chroma.2024.464821
Abstrakt: Recent studies highlight the pivotal roles of Nicotinamide adenine dinucleotide (NAD + ) and its metabolites in aging and neurodegeneration. Accurate quantification of NAD + and its metabolite levels in cells or tissues is crucial for advancing biochemical research and interventions targeting aging and neurodegenerative diseases. This study presents an accurate, precise, and rapid LC-MS/MS method using a surrogate matrix to quantify endogenous substances NAD + , nicotinamide mononucleotide (NMN), nicotinamide (NAM), adenosine diphosphate ribose (ADPR), and cyclic adenosine diphosphate ribose (cADPR) concentrations in mice sciatic nerves. Considering the properties of the phosphate groups in the analytes, the column and mobile phase were systematically optimized. These five polar analytes exhibited excellent analytical performance and baseline separation within 5 min on an Atlantis Premier BEH C18 AX column, with methylene phosphonic acid as a mobile phase additive. Enhanced sensitivity addressed the challenges posed by the small sample size of mice sciatic nerve and low NMN and cADPR detection. The method was fully validated, with linear correlation coefficients exceeding 0.992, precision (%relative standard deviation, RSD) values within 8.8%, and accuracy values between 92.2% and 107.3%, suggesting good reproducibility. Analytical recoveries in spiked and diluted matrix ranged from 87.8% to 104.7%, indicating the suitability of water as a surrogate matrix. Application of the method to quantify NAD + and its metabolite levels in normal and injured mice sciatic nerve identified cADPR as a sensitive biomarker in the nerve injury model. This method is anticipated to deepen our understanding of the connections between NAD + and its metabolites in health and disease, potentially improving diagnoses of various neurological disorders and aiding drug development for aging and neurodegenerative diseases.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
Externí odkaz: