Journal of Lipid Research, Vol 20, 1028-1035, Copyright © 1979 by Lipid Research, Inc.
Quantitative analysis of monosialogangliosides by high-performance liquid chromatography of their perbenzoyl derivatives
EG Bremer, SK Gross and RH McCluer
A quantitative high-performance liquid chromatographic method for the
analysis of monosialogangliosides as their perbenzoyl derivatives has been
devised. Samples containing as little as 3 nmol were converted to their
perbenzoyl derivatives by reaction with 0.1 ml of 10% benzoyl chloride in
pyridine at 60 degrees C for 1 hr. The products were purified by silicic
acid chromatography and analyzed by high- performance liquid chromatography
(HPLC). The HPLC analysis was performed with a 50 cm X 2.1 mm LiChrosphere
SI 4000 column and a linear gradient of 7-23% dioxane in hexane in 18 min.
Detection was at 230 nm. The detector response was found to be proportional
to the amount of monosialoganglioside analyzed. As little as 50 pmol of
injected material could be conveniently quantitated. The overall yield from
derivatization and chromatography, as determined with radiolabeled GM1, was
found to be 86%. To take advantage of the high sensitivity of the HPLC, a
small-scale isolation method for gangliosides was devised. This method
coupled with HPLC isotope dilution analysis was used to analyze the GM3
content of 1 ml of human plasma.
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