Measuring Nad + Levels in Mouse Blood and Tissue Samples Via A Surrogate Matrix Approach Using LC–MS/MS

Background: NAD+ is an endogenous analyte and is unstable during blood sample collection, both of which present obstacles for quantitation. Moreover, current procedures for NAD+ sample collection require onsite treatment with strong acid to stabilize the NAD+ in mouse blood cells. Results: NAD+ can be stabilized by addition of acid before the frozen mouse blood sample was thawed. A simple sample collection procedure was proposed to facilitate the analysis of NAD+ in mouse blood and tissue samples. A LC–MS/MS method was developed for quantifying NAD+ in mouse blood and various tissue samples. The described method was used to measure endogenous NAD+ levels in mouse blood following oral administration of the nicotinamide phosphoribosyltransferase inhibitor GNE-617. Conclusion: This study presents a suitable assay and sample collection procedure for high throughput screening of NAD+ samples in preclinical discovery studies.