期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1346, 期 -, 页码 88-96出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2014.04.051
关键词
Acetylneuraminic acid; Glycolylneuraminic acid; Methyl ester acetylneuraminic acid; Serum; LC-MS/MS; On-line SPE
资金
- Spanish Ministerio de Economia y Competitividad (MINECO) [CTQ2012-37428]
- European FEDER program [CTQ2012-37428]
- Spanish Ministerio de Ciencia e Innovacion (MICINN) [RYC-2009-03921]
- FPU scholarship [AP2009-0499]
N-acetylneuraminic acid (Neu5Ac) and N-acetylglycolylneuraminic acid (Neu5Gc), two acylated derivatives of 9-C carboxylated monosaccharides, are involved in a number of biological processes as modulators of glycoconjugates. A partially automated method is here presented for determination of these sialic acids in the two most important biofluids for clinical analysis: serum and urine. For this purpose, a solid-phase extraction (SPE) workstation was on-line connected to an LC-MS/MS triple quadrupole mass detector. Hydrolysis to release sialic acids bound to glycoconjugates and derivatization were the two steps implemented as sample preparation prior to SPE-LC-MS/MS analysis. Following thorough optimization of the SPE and LC-MS/MS conditions, the analytical method was validated using the standard addition approach to assess the presence of matrix effects. The proposed method affords detection limits of 0.03 ng/mL and 0.04 ng/mL for Neu5Ac and Neu5Gc, respectively. The precision (expressed as relative standard deviation) was 1.7 and 4.6% for within-day variability, and 4.8 and 7.2% for between-days variability. Accuracy, estimated using spiked (between 1 and 50 ng/mL) and non-spiked samples of both biofluids, ranged from 95.2 to 99.6%. The method was applied to human serum and urine of healthy volunteers, thus showing its suitability for application in both clinical and research laboratories. (C) 2014 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据