Department of Chemistry, Chemical and Biomedical Engineering and Center for Gene Regulation in Health and Disease (GRHD), Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, United States.
School of Pharmaceutical Science, China Medical University, 92 Bei-er Road, Shenyang, Liaoning 110001, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jan 1;944:75-81. doi: 10.1016/j.jchromb.2013.11.016. Epub 2013 Nov 14.
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)C3-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.
我们报道了一种使用 LC-MS/MS 方法结合同位素标记标准校准（ILSC）和稳健衍生化，准确量化人血浆中游离唾液酸（SA）的方法。具体来说，通过与稳定且廉价的 3,4-二氨基甲苯（DAT）衍生化 SA，可以得到具有高 MS 响应的稳定的 SA 产物，证明了游离 SA 的 LC-MS/MS 分析既方便又具有成本效益。此外，使用（13）C3-SA 作为校准标准，确保了测量的准确性。该测定法使用超高效液相色谱（UHPLC）分离天然/标记的 SA 和内标物，以避免基质干扰，并采用质谱在多重反应监测（MRM）模式下进行灵敏和选择性检测。我们实现了 20ng/mL 的定量下限（LLOQ）和总运行时间为 4.2min，这是生物基质中游离 SA 的最灵敏和快速的测量方法。
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