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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 

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A hydrolysis method using microwaves: determination of N-acetyl- and N-glycolylneuraminic acids in biological systems by fluorometric high-performance liquid chromatography.

A simple, rapid, and extremely sensitive method for determining N-acetyl- and N-glycolylneuraminic acids in serum and in submandibular, sublingual, and parotid glands using high-performance liquid chromatography with a fluorometric detector is described. The neuraminic acids contained in the samples are released in the presence of 2 M CH3COOH by means of microwave hydrolysis (only 10 min required) and are subsequently converted using 1,2-diamino-4,5-methylenedioxybenzene, a fluorogenic reagent for alpha-ketoacids, into highly fluorescent derivatives. In order to optimize the release of sialic acids and to minimize the effect of destruction, the following analytical variables were investigated: temperature, time, and concentration of the acid. Within 12 min after derivatization, the compounds were separated on a reversed-phase column by means of isocratic elution using a mobile phase of water (pH 3 with H3PO4)-methanol-acetonitrile (86:6:8, v/v). Fluorescence detection was performed at an excitation wavelength of 373 nm and an emission wavelength of 448 nm.[1]

References

  1. A hydrolysis method using microwaves: determination of N-acetyl- and N-glycolylneuraminic acids in biological systems by fluorometric high-performance liquid chromatography. Laganà, A., Marino, A., Fago, G., Martinez, B.P. Anal. Biochem. (1993) [Pubmed]
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