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Research Article

Quantitation of 2-hydroxyglutarate in human plasma via LC–MS/MS using a surrogate analyte approach

    Feng Yin

    *Author for correspondence: Tel.: +1 617 844 3306; Fax: +1 617 649 8618;

    E-mail Address: feng.yin@agios.com

    Agios Pharmaceuticals, Inc., 88 Sidney Street, Cambridge, MA 02139, USA

    ,
    Yonghua Ling

    PPD Inc., 3230 Deming Way, Middleton, WI 53562, USA

    ,
    Jennifer Keller

    PPD Inc., 3230 Deming Way, Middleton, WI 53562, USA

    ,
    Dennis Kraus

    PPD Inc., 3230 Deming Way, Middleton, WI 53562, USA

    ,
    Rohini Narayanaswamy

    Agios Pharmaceuticals, Inc., 88 Sidney Street, Cambridge, MA 02139, USA

    ,
    Heidi Mangus

    Agios Pharmaceuticals, Inc., 88 Sidney Street, Cambridge, MA 02139, USA

    ,
    Fumin Li

    PPD Inc., 3230 Deming Way, Middleton, WI 53562, USA

    ,
    Hua Yang

    Agios Pharmaceuticals, Inc., 88 Sidney Street, Cambridge, MA 02139, USA

    &
    Guowen Liu

    Agios Pharmaceuticals, Inc., 88 Sidney Street, Cambridge, MA 02139, USA

    Published Online:https://doi.org/10.4155/bio-2020-0131

    Aim: 2-Hydroxyglutarate (2-HG) is a target engagement biomarker in patients after treatment with inhibitors of mutated isocitrate dehydrogenase (mIDH). Accurate measurement of 2-HG is critical for monitoring the inhibition effectiveness of the inhibitors. Materials & methods: Human plasma samples were spiked with stable isotope labelled internal standard, processed by protein precipitation, and analyzed using LC–MS/MS. This method was validated following regulatory guidance and has been successfully applied in a clinical study for mIDH inhibition. Results: An LC–MS/MS method with a surrogate analyte approach was developed and validated to measure 2-HG in human plasma with acceptable intra- and inter-assay accuracy and precision. Conclusion: A sensitive and robust LC–MS/MS method was developed and validated for measuring 2-HG in human plasma.

    Papers of special note have been highlighted as: • of interest; •• of considerable interest

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