Quantitative Determination of Glycosphingolipids in Human Plasma

Abstract

A method is described for the quantitative isolation and estimation of the four glycosphingolipid fractions from 10 ml of human plasma. The procedure consists of acetylation of the total lipids after extraction, separation of acetylated glycosphingolipids from non-glycolipids on a Florisil column, deacetylation, dialysis in water, separation of the single fractions by thin-layer chromatography on silica gel, and quantitative analysis of each glycosphingolipid by sulfuric acid-orcinol reagent, carried out in the presence of silica gel. The concentrations of the four glycosphingolipids in human plasma derived from a group of 23 healthy subjects are presented. The procedure described is sufficiently sensitive for clinical investigation. Quantitative Bestimmung der Glycosphingolipide im menschlichen BlutplasmaZusammenfassung: Über eine Methode zur quantitativen Isolierung und Bestimmung der vier neutralen Glycosphingolipidfraktionen aus 10 ml menschlichem Blutplasma wird berichtet. Das Verfahren besteht aus: Acetylierung der extrahierten Gesamtlipide, Abtrennung der acetylierten Glycosphingolipide durch Säulenchromatographie an Florisil, Deacetylierung, Dialyse gegen Wasser, Auftrennung der einzelnen Fraktionen durch Dünnschichtchromatographie auf Kieselgelplatten und quantitative Bestimmung der verschiedenen Glycosphingolipide im Schwefelsäure-Orcinol-Test in Gegenwart von Kieselgel. Die ermittelten Konzentrationen der vier Glycosphingolipide in einem Kollektiv von 23 gesunden Personen werden mitgeteilt. Das beschriebene Verfahren ist empfindlich und geeignet für klinische Untersuchungen.

High-Performance Liquid Chromatography Analysis of Ganglioside Carbohydrates at the Picomole Level after Ceramide Glycanase Digestion and Fluorescent Labeling with 2-Aminobenzamide

“…An advantage of this method was that only 10 -40 l of plasma was required, whereas typically 1 ml or more is used for TLC methods based on resorcinol staining (30). In agreement with previous reports (30,31), GM3, GM2, GD3, and GD1a were the main gangliosides detected (Fig. 6).…”

Lipid Investigation of Central and Peripheral Nervous System in Connatal Pelizaeus‐Merzbacher's Disease

“…Determinations of phosphorus were according to Bartlett (1959, modified by Debuch et al, 1968; plasmalogen (Feulgen et al, 1951, varied by Klenk andDebuch, 1963), cholesterol (Zak et al, 1954), and hexoses by two-dimensional T L C (Svennerholm, 1956, modified by Atzpodien andKremer, 1977).…”