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Quantification of free sialic acid in human plasma through a robust quinoxalinone derivatization and LC-MS/MS using isotope-labeled standard calibration

  • Cleveland State University
  • China Medical University Shenyang
  • Cleveland Clinic Foundation

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

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 13C3-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 20ng/mL and a total running time of 4.2min, which is the most sensitive and quick measurement for free SA in biomatrices. © 2013 Elsevier B.V.
Original languageEnglish
Pages (from-to)75-81
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume944
DOIs
StatePublished - Jan 1 2014

Keywords

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

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