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Research Article

Development of an LC–MS/MS assay for quantification of intact INSL3 in rat plasma

    Nicole A Schneck

    *Author for correspondence:

    E-mail Address: nicole.x.schneck@gsk.com

    Bioanalysis, Immunogenicity & Biomarkers, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    ,
    Lela Mortezavi

    Bioanalysis, Immunogenicity & Biomarkers, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    ,
    Alan R Olzinski

    Novel Human Genetics Research Unit, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    ,
    Diane Posavec

    Novel Human Genetics Research Unit, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    ,
    Larry J Jolivette

    Drug Metabolism & Pharmacokinetics, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    ,
    Timothy W Sikorski

    Bioanalysis, Immunogenicity & Biomarkers, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    ,
    Shan-Shan Zhang

    Therapeutics Division, 23andMe, 349 Oyster Point Blvd, South San Francisco, CA 94080, USA

    ,
    Christine G Schnackenberg

    Novel Human Genetics Research Unit, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    &
    Hermes Licea-Perez

    **Author for correspondence:

    E-mail Address: hermes.2.licea-perez@gsk.come

    Bioanalysis, Immunogenicity & Biomarkers, GSK, 1250 S. Collegeville Rd, Collegeville, PA 19426, USA

    Published Online:https://doi.org/10.4155/bio-2023-0120

    Background: Relatively large disulfide-linked polypeptides can serve as signaling molecules for a diverse array of biological processes and may be studied in animal models to investigate their function in vivo. The aim of this work was to develop an LC–MS/MS assay to measure a model peptide, INSL3, in rat plasma. Results: A dual enrichment strategy incorporating both protein precipitation and solid phase extraction was utilized to isolate INSL3 from rat plasma, followed by targeted LC–MS/MS detection. The method was able to measure full-length INSL3 (6.1 kDa) down to 0.2 ng/ml with acceptable accuracy and precision. Conclusion: The final assay was applied to support an exploratory pharmacokinetic study to evaluate steady-state concentrations of dosed INSL3 in rat plasma.

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

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