Measurement of free and total sialic acid by isotopic dilution liquid chromatography tandem mass spectrometry method

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• 作者：Abdellah Tebani ; Dimitri Schlemmer ; Apolline Imbard ; Odile Rigal ; Dominique Porquet ; Jean-Franç ; ois Benoist  ; ;   ; ^{jean-francois.benoist@rdb.aphp.fr}
• 关键词：Tandem mass spectrometry ; N-Acetylneuraminic Acid ; Sialic acid storage disease
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2011
• 出版时间：1 December 2011
• 年：2011
• 卷：879
• 期：31
• 页码：3694-3699
• 全文大小：521 K

文摘

The measurement of urine sialic acid (N Acetylneuraminic Acid: Neu5Ac) is useful for screening sialic acid storage disorders. We developed a new LC MS/MS method for the determination of a sialic acid. Urine samples were analyzed, after an HCl n-Butanol derivatization step, by a reverse phase based high-performance liquid chromatography method using 1,2,3-¹³C₃ N-Acetyl-d-neuraminic Acid (¹³C-Neu5Ac) as an internal standard. Selective detection was performed by tandem mass spectrometry using an electrospray source operating in positive ionization mode employing multiple reactions monitoring to monitor N-Acetylneuraminic Acid and the internal standard. The transitions m/z 366 ?330 and 369 ?333 for Neu5Ac and ¹³C-Neu5Ac were respectively monitored. The limit of the method quantification was 1.40 ¦ÌM of N-Acetylneuraminic Acid and the calibration curve showed a good linearity up to 1000 ¦ÌM. The within assay precision and accuracy of the method ranged from 3.22 to 5.95 % and 98.69 to 109.18 % , respectively and the between assay precision and accuracy ranged, respectively, from 5.15 to 7.65 % and 96.14 to 102.30 % . The method can be applied for the determination of N-Acetylneuraminic Acid concentrations in urine and other biological fluids (e.g., amniotic and peritoneal fluids).