GC/MS identification and quantification of constituents of bacterial lipids and glycoconjugates obtained after methanolysis as heptafluorobutyrate derivatives.
In previous articles [Anal. Biochem. 284 (2000) 201; J. Lipid Res. 43 (2002) 794], we reported that the GC/MS identification and quantification of nearly all constituents of glycolipids could be obtained on the same sample in a single GC/MS analysis as heptafluorobutyrate derivatives of the products liberated using acid-catalyzed methanolysis. The same type of data could be obtained on glycoproteins and proteoglycans [Biochemistry 42 (2003) 8342]. These experiments were performed on material from higher organisms, and there was no evidence that bacteria-specific constituents could also be identified and quantified. The current article reports that the GC/MS analysis of compounds liberated by acid-catalyzed methanolysis as heptafluorobutyrate derivatives allows the simultaneous qualitative and quantitative determinations of pentoses, deoxyhexoses, hexoses, hexosamines, uronic acids, Kdo, Mur, heptose, Kdn, and neuraminic acid as well as of most fatty acids (including hydroxylated fatty acids). This approach provides a way of obtaining fingerprints of bacterial constituents and quantification of the overall effect of gene inactivation or of culture conditions.
Bohin
Anne
A
CNRS Unité Mixte de Recherche 8576, Unité de Glycobiologie Structurale et Fonctionnelle, Université des Sciences et Technologies de Lille Bâtiment C9, 59655 Villeneuve d'Ascq Cedex, France.
Bouchart
Franck
F
Richet
Colette
C
Kol
Osarath
O
Leroy
Yves
Y
Timmerman
Philippe
P
Huet
Guillemette
G
Bohin
Jean-Pierre
JP
Zanetta
Jean-Pierre
JP
D001419:Bacteria / Q000737:chemistry*; D005227:Fatty Acids / Q000302:isolation & purification; D005466:Fluorocarbons / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry; D006001:Glycoconjugates / Q000302:isolation & purification*; D006880:Hydroxy Acids / Q000032:analysis; D008055:Lipids / Q000032:analysis / Q000302:isolation & purification*; D000432:Methanol / Q000737:chemistry; D009005:Monosaccharides / Q000302:isolation & purification; D009112:Muramic Acids / Q000032:analysis; D011073:Polyamines / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D005227:Fatty Acids; D005466:Fluorocarbons; D006001:Glycoconjugates; D006880:Hydroxy Acids; D008055:Lipids; D009005:Monosaccharides; D009112:Muramic Acids; D011073:Polyamines; C007924:heptafluorobutyric anhydride; D000432:Methanol