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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
Journal of Chromatography B
  • Dan Wang, Cleveland State University
  • Xiang Zhou, Cleveland State University
  • Lin Wang, China Medical University
  • Sihe Wang, Cleveland Clinic
  • Xue-Long Sun, Cleveland State University
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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 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.

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Creative Commons Attribution-Noncommercial-No Derivative Works 4.0
Citation Information
Wang, D.; Zhou, X.; Wang, L.; Wang, S.; Sun, X. Quantification of free sialic acid in human plasma through a robust quinoxalinone derivatization and LC–MS/MS using isotope-labeled standard calibration. Journal of Chromatography B 2014, 944, 75-81.