AI Article Synopsis
- A new method has been developed to separate and quantify gangliosides from 5 ml of human cerebrospinal fluid (CSF) using ultrafiltration, methanol/chloroform extraction, and high performance thin layer chromatography (HPTLC) for analysis.
- The recovery rate of this method is 78%, with no loss of gangliosides during the process, and the ganglioside patterns in normal CSF samples are similar to those found in brain tissue, particularly the cerebellum.
- The analysis identified specific percentages of various ganglioside types, with total ganglioside content ranging from 616-944 micrograms/l, and demonstrated high precision in assay results for both predominant and less prominent ganglioside fractions.
Article Abstract
A method for the separation and quantification of a complex ganglioside mixture from a clinically available amount (5 ml) of human cerebrospinal fluid (CSF) is described. After reduction of the CSF volume by ultrafiltration, gangliosides are extracted with methanol/chloroform, then separated and quantified by high performance thin layer chromatography (HPTLC) and direct densitometry. For purification of crude ganglioside extract, the method of choice was microdialysis against water. Recovery for the present method including all methodological steps was 78%. No delective loss of gangliosides was demonstrated. The CSF ganglioside pattern from 'normal' CSF samples resembles that of brain gangliosides, particularly cerebellum gangliosides. Based on chromatographic comparison with standards, the percentages of lipid-bound NeuAc-positive fractions were: GM1 = II3NeuAc-GgOse4Cer (3%), GD3 = II3NeuAc2-Lac-Cer (3%), GD1a = IV3NeuAc,II3NeuAc-GgOse4Cer (15%), X1 (3%), GD1b = II3(NeuAc)2-GgOse4Cer (16%), X2 (3%), GT1b = IV3NeuAc,II3NeuAc2-GgOse4-Cer (41%), and GQ1b = IV3NeuAc2-,II3NeuAc2-GgOse4-Cer (16%). The total ganglioside content varied between 616-944 micrograms/l. Within-run and between-run assay precision (relative standard deviation) for 'normal' pooled CSF ranged from 0.04 to 0.12 for the predominant CSF ganglioside fractions (GD1a, GD1b, GT1b, GQ1b), and from 0.13 to 0.23 for the less pronounced fractions (GM1, GD3).
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| http://dx.doi.org/10.1515/cclm.1990.28.11.863 | DOI Listing |
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