Quantification of Free Sialic Acid in Human Plasma through a Robust Quinoxalinone Derivatization and LC–MS/MS Using Isotope-labeled Standard Calibration

Dan Wang, Xiang Zhou, Lin Wang, Sihe Wang, Xue-Long Sun

    Research output: Contribution to journalArticlepeer-review

    Abstract

    We report an accurate quantification of free sialic acid (SA) in human plasma using LC–MS/MS method with isotope-labeled standard calibration (ILSC) and robust derivatization. Specifically, derivatization of SA with a stable and inexpensive 3,4-diaminotoluene (DAT) provides a stable product of SA with high MS response, proving a convenient and cost-effective LC–MS/MS analysis of free SA. In addition, the use of 13 C 3 -SA as calibration standard ensured the accuracy for the measurement. This assay used ultra high performance liquid chromatography (UHPLC) for separation of native/labeled SA and IS from matrix interference, and employed mass spectrometry in multiple reaction monitoring (MRM) mode for sensitive and selective detection. We have achieved a lower limit of quantification (LLOQ) of 20 ng/mL and a total running time of 4.2 min, which is the most sensitive and quick measurement for free SA in biomatrices.

    Original languageAmerican English
    JournalJournal of Chromatography B
    Volume944
    DOIs
    StatePublished - Jan 1 2014

    Keywords

    • Free sialic acid; Human plasma; Isotope-labeled standard calibration; Liquid chromatography–tandem mass spectrometry; Quinoxaline derivatization

    Disciplines

    • Analytical Chemistry
    • Biochemistry
    • Chemistry

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