Sequential GC/MS analysis of sialic acids, monosaccharides, and amino acids of glycoproteins on a single sample as heptafluorobutyrate derivatives

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Pons, A (author)

Sequential GC/MS analysis of sialic acids, monosaccharides, and amino acids of glycoproteins on a single sample as heptafluorobutyrate derivatives

Article/chapterEnglish2003

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2003-06-18
American Chemical Society (ACS),2003

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LIBRIS-ID:oai:lup.lub.lu.se:f7a02a4a-dc11-4c5c-bf47-91f40dd25981
https://lup.lub.lu.se/record/306878URI
https://doi.org/10.1021/bi034250eDOI

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Language:English
Summary in:English

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A GUMS procedure was developed for the analysis of all major constituents of glycoproteins. The rationale for this approach is that by using GC/MS analysis of the constituents as heptafluorobutyrate derivatives, it was possible to quantitatively determine the sialic acid, monosaccharide, fatty acids (when present), and the amino acid composition with the sample remaining in the same reaction vessel during the entire procedure. A mild acid hydrolysis was used to liberate sialic acids and was followed by formation of methyl-esters of heptafluorobutyrate (HFB) derivatives. After GC/MS analysis of sialic acids, the remaining material was submitted to acid-catalyzed methanolysis followed by the formation of HFB derivatives. After GUMS analysis of the monosaccharides, the sample was supplemented with norleucine (as internal standard) and hydrolyzed with 6 M HCl followed by the formation of isoamyl-esters of HFB derivatives and GC/MS analysis. His and Trp residues were modified during the step of acid-catalyzed methanolysis, but the resulting derivatives were stable during acid hydrolysis and quantitatively recovered by GUMS analysis. As a result, all constituents of glycoproteins (sialic acids, monosaccharides (or di- and trisaccharides) and amino acids) are identified in the electron impact mode of ionization and quantified using three GUMS analysis in the same chromatographic conditions and using a limited number of reagents, a considerable advantage over previous techniques. This method is very sensitive, all data (qualitative and quantitative) being obtained at the sub-nanomolar level of initial material.

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Richet, C (author)
Robbe, C (author)
Herrmann, AnnkatrinLund University,Lunds universitet,Institutionen för experimentell medicinsk vetenskap,Medicinska fakulteten,Department of Experimental Medical Science,Faculty of Medicine(Swepub:lu)medk-ahe (author)
Timmerman, P (author)
Huet, G (author)
Leroy, Y (author)
Carlstedt, IngemarLund University,Lunds universitet,Institutionen för experimentell medicinsk vetenskap,Medicinska fakulteten,Department of Experimental Medical Science,Faculty of Medicine(Swepub:lu)medk-ica (author)
Capon, C (author)
Zanetta, JP (author)
Institutionen för experimentell medicinsk vetenskapMedicinska fakulteten (creator_code:org_t)

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In:Biochemistry: American Chemical Society (ACS)42:27, s. 8342-83530006-29601520-4995

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By the author/editor: Pons, A; Richet, C; Robbe, C; Herrmann, Annkat ...; Timmerman, P; Huet, G; show more...; Leroy, Y; Carlstedt, Ingem ...; Capon, C; Zanetta, JP; show less...

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NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

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By the university: Lund University

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