PMID: 3753443May 1, 1986Paper

Determination of gangliosides as 2,4-dinitrophenylhydrazides by high-performance liquid chromatography

The Biochemical Journal
K MiyazakiY C Lee

Abstract

A specific, sensitive and easily performed method for the determination of gangliosides in tissue was developed. After removal of water-soluble compounds, total lipids were extracted from tissue and then treated with 2,4-dinitrophenylhydrazine hydrochloride and dicyclohexylcarbodi-imide in dimethylformamide at 0 degrees C to form ganglioside hydrazides. After removal of excess reagents by column chromatography on silicic acid, the ganglioside 2,4-dinitrophenylhydrazides were eluted from the column and analysed by h.p.l.c. with the use of a silica-gel normal-phase column eluted with an isocratic chloroform/methanol/water/acetic acid system. The addition of CaCl2 improved the separation of GM3 ganglioside containing N-acetylneuraminic acid from that containing N-glycollylneuraminic acid. 2,4-Dinitrophenylhydrazide peaks were measured by the absorbance at 342 nm. Quantification of GM3, GM2, GM1, GD1a, GD1b, GT1b and LM1 gangliosides was linear in a range 0.02-1.6 nmol. GM4, GD3, GT1a and GQ1b gangliosides also yielded distinct peaks, although the range of linearity was not examined. This method was applied to the analysis of the total lipids of rat brain and hepatocytes.

Citations

Mar 1, 1989·Veterinary Pathology·H S Singer, L C Cork
Oct 1, 1990·Clinical Genetics·J S O'BrienS L O'Brien
Nov 22, 2007·The FEBS Journal·E Martínez-AbundisC Zazueta
Jan 1, 1990·Journal of Pharmaceutical and Biomedical Analysis·C GandiniG Kirschner
Nov 6, 1987·Journal of Chromatography·S AndoK Kon
Nov 1, 1990·Analytical Biochemistry·S MorimotoY Kishimoto

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