Analytical HPLC Method

Samples were analyzed by the described ion-pair HPLC. After stopping an aliquot from the reaction mixture by heat (see Subheading 3.1.1., step 2) particulate matter must be removed from the sample by centrifugation at 12,000 rpm (18000^) for 10 min with Biofuge A. All solutions used in HPLC must be filtered with a 0.22-|m filter, and degassed using a vacuum pump to avoid particulate matter and air bubbles. The UDP-glucose concentration was read from a calibration chart (0.2-3 mM) of the compound vs peak area (absorption at 254 nm).

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