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wuzhirong

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Quantitative analysis of sugars was carried out using a post-column labelling system. Isocratic separation of sugars was carried out on a DEAE-Si100 column (250*4.6mm, 5mm particle size, Serva,C42308) by elution with acetonitrile-water (82:12 v/v;HPLC-grade acetonitrile; Sigma) at a flow rate of 0.7ml min-1.The column effluent was mixed with tetrazolium reagent in a T-connector to label sugars. The alkaline tetrazolium reagent consisted of 2 g l-1tetrazolium blue (Sigma) in 0.18M NaOH and was introduced at a flow rate of 0.2ml min-1 with a second HPLC pump. The T-connector effluent was lead into a Teflon reaction coil of 3m length that was mounted in a water bath and kept at a temperature of 85¡æ. The resulting reaction time was 80 s. The labelled sugars were detected at 487 nm using a Pharmacia LKB2141 variable wavelength monitor.
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