Journal of Lipid Research, Vol 21, 1137-1141, Copyright © 1980 by Lipid Research, Inc.
Enhancement of the Yu and Ledeen gas-liquid chromatographic method for sialic acid estimation: use of methane chemical ionization mass fragmentography
J Ashraf, DA Butterfield, J Jarnefelt and RA Laine
The sialic acid present in erythrocyte ghosts was estimated by methane
chemical ionization mass fragmentography of the trimethylsilyl methyl
glycosides. The internal standard was 3,4,6-tris-trimethylsilyl-alpha-
phenyl-2-deoxy-2-acetamide-D-glucosaminide, as proposed by Yu and Ledeen
(J. Lipid Res. 1970. 11: 506-516). The [MH-16]+ions (m/e 610 for the
TMS-methylglycoside of N-acetylneuraminic acid and m/e 498 for the internal
standard) were used for quantifying nanogram levels of sialic acid in the
presence of other contaminating substances. This effectively raises the
signal-to-noise ratio for the Yu and Ledeen method by at least two orders
of magnitude. The sensitivity and linearity of the method, without use of
isotopic carriers, were tested using known quantities of N-acetylneuraminic
acid. The limit of detection was below 0.4 nanograms (approximately one
picomole). The useful range of detection was 10 ng-1 microgram, showing a
large dynamic range.
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