Fluorimetric Microdetermination of Carbohydrates.

Abstract

excess sodium carbonate from the fusion acted as a spectrographic buffer, and in conjunction with the boiler cap electrode, resulted in a 100% increase in sensitivity. "Moving plate" studies, Figure 2, showed that the volatilization of silicon from the sodium carbonate buffer with boiler cap elec-(6) Ko, R., Appl. Spectr. 13, 10 (1959). ( 7) Snell, F. D., Snell, C. T., "Colorimetric Methods of Analyses," 3rd ed.

Fluorescently labelled glycans and their applications

“…Early work in this area included fluorogenic labels that react with reducing sugars under strongly acidic conditions, such as 5-hydroxytetralone [16], phenylenediamine [17], and N-(1-naphthyl)ethylenediamine dihydrochloride [18], however, these conditions can lead to cleavages of glycosidic linkages that might be present in the reducing sugar sample. Fluorogenic reagents that react with carbohydrates under mildly basic or neutral conditions were also reported.…”

Fluorometric Analysis

“…Ketone bodies in urine can be determined (after isolation by steam distillation) by their effect on the fluorescence of 2-naphthol (219). A microdetermination of alphaketo acids, based on the formation of fluorescent quinoxalines by reaction w7ith o-phenylenediamine, is described by Spikner and Tow7ne (460), who use a similar scheme for carbohydrates as well (489). A fluorometric method for blood glucose reported earlier has been adapted to pediatric work, where only small amounts of blood are available (62).…”

2-Pyridylfuran: A New Fluorescent Tag for the Analysis of Carbohydrates

“…The labeling step serves the dual purpose of installing a hydrophobic group on the polar sugar to improve chromatographic properties and enabling sensitive detection due to the fluorophore. The typical such method is reductive amination with an aromatic amine such as 2-aminobenzoic acid (2-AA; anthranilic acid), 2-aminobenzamide (2-AB), or 2-aminopyridine (2-AP), which react with the oligosaccharide reducing end under mild conditions with in situ reduction of the intermediate Schiff base with sodium cyanoborohydride. − A second common technique is derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) to afford products containing two chromophores per product molecule and deliver a detection limit in the high femtomole range with UV detection. , Many of these fluorescent/UV labeling methods result in intense HPLC peaks due to the excess reagent required, which can complicate the analysis by obscuring peaks of interest or necessitate sample cleanup steps that extend the analysis time and result in sample loss. , Several fluorogenic reagents for reducing sugar analysis have also been reported in the literature dating as far back as the 1960s, including 1,2-diarylethylenediamines, aromatic amidines, o -phenylenediamine, 2-cyanoacetamide, resorcinol, and ethylenediamine sulfate, although the majority of such methods relied only on total fluorescence measurements in a spectrometer rather than separation of various sugars, and these protocols do not appear to have become part of the standard carbohydrate analysis toolkit. Fluorogenic reagents specific for sialic acids are also known, such as thiobarbituric acid and acetoacetanilide/ammonia …”

“…5,6 Many of these fluorescent/UV labeling methods result in intense HPLC peaks due to the excess reagent required, which can complicate the analysis by obscuring peaks of interest or necessitate sample cleanup steps that extend the analysis time and result in sample loss. 2,3 Several fluorogenic reagents for reducing sugar analysis have also been reported in the literature dating as far back as the 1960s, including 1,2-diarylethylenediamines, 7 aromatic amidines, 8 o-phenylenediamine, 9 2-cyanoacetamide, 10 resorcinol, 11 and ethylenediamine sulfate, 12 although the majority of such methods relied only on total fluorescence measurements in a spectrometer rather than separation of various sugars, and these protocols do not appear to have become part of the standard carbohydrate analysis toolkit. Fluorogenic reagents specific for sialic acids are also known, such as thiobarbituric acid 13 and acetoacetanilide/ ammonia.…”