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Research Article

Measuring NAD+ levels in mouse blood and tissue samples via a surrogate matrix approach using LC–MS/MS

    Xiaorong Liang

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Lulu Yang

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Ann R Qin

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Justin Ly

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Bianca M Liederer

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Kirsten Messick

    Department of Safety Assessment, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Shuguang Ma

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Mark Zak

    Discovery Chemistry, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Peter S Dragovich

    Discovery Chemistry, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Brian J Dean

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    ,
    Cornelis ECA Hop

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    &
    Yuzhong Deng

    Drug Metabolism & Pharmacokinetics, Genentech Inc., South San Francisco, CA 94080, USA

    Published Online:https://doi.org/10.4155/bio.14.8

    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.

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