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Selective post-column fluorigenic reaction with benzamidine for trace level detection of reducing saccharides in liquid chromatography

Authors
Journal
Journal of Chromatography A
0021-9673
Publisher
Elsevier
Publication Date
Volume
553
Identifiers
DOI: 10.1016/s0021-9673(01)88496-x

Abstract

Abstract A liquid chromatographic post-column fluorigenic reaction with benzamidine for the trace level detection of reducing saccharides is described. The system consists of a Sugar Pak-1 gel column (300 mm × 6.5 mm I.D.) packed with a microparticulate resin in the calcium form and heated at 70°C. Elution is carried out with water containing 10 -4 M CaEDTA at 0.4 ml mm -1. The post-column reaction is carried out with a mixture of 30 m M benzamidine and 1 M potassium hydroxide solution at 1 ml min -1 using a 530-μl reaction coil thermostated at 100°C. Fluorescence was monitored at 470 nm (emission) with an excitation wavelength of 360 nm. The system had good reproducibility (relative standard deviation 1–3%) for an injected amount of 0.9 μ (5 nmol). Linear calibration graphs were obtained between 16 and 500 nmol ml -1 ( r0.999) and detection limits of 16–63 pmol at a signal-to-noise ratio of 3 were achieved. The detection of saccharides in plants is also described.

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