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EnglishA 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 α-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.
A hydrolysis method using microwaves: determination of N-acetyl-and N-glicolylneuraminicacids in biological systems by fluorometric high-performance liquid chromatography / Lagana', Aldo; Marino, Aldo; Fago, Giovanna; B., Pardo Martinez. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - ELETTRONICO. - 215(1993), pp. 266-272. [10.1006/abio.1993.1585]
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| Titolo: | A hydrolysis method using microwaves: determination of N-acetyl-and N-glicolylneuraminicacids in biological systems by fluorometric high-performance liquid chromatography | |
| Autori: | ||
| Data di pubblicazione: | 1993 | |
| Rivista: | ||
| Citazione: | A hydrolysis method using microwaves: determination of N-acetyl-and N-glicolylneuraminicacids in biological systems by fluorometric high-performance liquid chromatography / Lagana', Aldo; Marino, Aldo; Fago, Giovanna; B., Pardo Martinez. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - ELETTRONICO. - 215(1993), pp. 266-272. [10.1006/abio.1993.1585] | |
| Handle: | http://hdl.handle.net/11573/452905 | |
| Appartiene alla tipologia: | 01a Articolo in rivista |
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