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Issue 86, 2014
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Systematic investigation of quinoxaline derivatization of sialic acids and their quantitation applicability using high performance liquid chromatography

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Abstract

Quinoxaline derivatization has been the most commonly used approach for sialic acid quantitation of biological samples and glycoproteins by either HPLC or LC-MS/MS. However, the reported methods still suffer from many disadvantages, such as instable/expensive reagents, instable derivatives, and/or multiple products, which prevent their practical applications for SA-focused clinical screening and biopharmaceutical analysis. In this study, we investigated the quinoxaline derivatization of SAs with different commercially available phenyldiamines, and found a stable and inexpensive 4,5-dimethylbenzene-1,2-diamine (DMBA), that selectively reacted with SA to form a stable quinoxaline derivative efficiently and with strong UV and fluorescence absorption. The optimized DMBA derivatization conditions were successfully applied to quantitation of free and total SAs (N-acetyl-D-neuraminic acid (Neu5Ac) and N-glycolyl-D-neuraminic acid (Neu5Gc)) in fetal bovine serum (FBS) and glycoprotein fetuin using HPLC with fluorescence detection. The limit of detection (LOD) for Neu5Ac and Neu5Gc were 6.00 and 8.80 pg, and the limits of quantitation (LOQ) for Neu5Ac and Neu5Gc were 18.0 and 29.0 pg on column, respectively. This method was also applied to determine total SA in human plasma samples. This DMBA derivatization method provides a robust, easy-handling, sensitive and cost effective approach for quantitation of SAs in glycoconjugates and biomatrices.

Graphical abstract: Systematic investigation of quinoxaline derivatization of sialic acids and their quantitation applicability using high performance liquid chromatography

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Publication details

The article was received on 20 Aug 2014, accepted on 11 Sep 2014 and first published on 12 Sep 2014


Article type: Paper
DOI: 10.1039/C4RA08930H
Author version available: Download Author version (PDF)
Citation: RSC Adv., 2014,4, 45797-45803
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    Systematic investigation of quinoxaline derivatization of sialic acids and their quantitation applicability using high performance liquid chromatography

    L. Wang, D. Wang, X. Zhou, L. Wu and X. Sun, RSC Adv., 2014, 4, 45797
    DOI: 10.1039/C4RA08930H

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