E. Negroni et al., EVALUATION OF THE EFFICIENCY OF AN ASSAY PROCEDURE FOR GANGLIOSIDES IN HUMAN SERUM, Glycoconjugate journal, 13(3), 1996, pp. 347-352
An efficiency assessment of a ganglioside assay procedure was carried
out on human serum gangliosides from healthy subjects of different sex
and age. The analysis of the gangliosides, extracted with chloroform/
methanol and purified by lipid partitioning, ion exchange column chrom
atographic separation and desalting procedures as described by Senn et
al. (1989) Eur J Biochem 181: 657-62, was performed by HPTLC followed
by densitometric quantification. The yield of the procedure, expresse
d as radioactivity recovery, was determined by adding GM3 ganglioside,
tritium labelled at the sialic acid acetyl group and at the C3 positi
on of sphingosine, to the lyophilized serum or by associating it with
the serum lipoproteins. Tn spite of the fact that the extraction and p
urification procedures were performed exactly as described we found th
e radioactivity recovery to be variable (25-50%) and much lower than t
hat proposed. Much of the radioactivity was found in the organic phase
after lipid partitioning, whilst all the ganglioside purification ste
ps led to some further loss. After the introduction of some modificati
ons to the procedure the recovery improved, reaching 67-79%. The analy
ses on 33 samples of 5 ml showed a human serum ganglioside content of
about 10 nmol ml(-1) (as corrected for the recovery), and confined tha
t GM3 ganglioside is the main component of the total serum ganglioside
mixture.