Separation and Determination of Uridine Diphosphoglucose (UDPGlu) and Uridine Diphosphogalactose (UDPGal) in the Cell Free Extracts of Corynebacterium glutamicium by High Performance Liquid Chromatography

A novel high performance liquid chromatographic method for the separation and determination of UDPGlu and UDPGal in the cell free extracts of Corynebacterium glutamicium CCTCC M201005 has been developed. Effects of different columns and mobile phase compositions on the retention behaviors and resolution of UDPGlu and UDPGal were investigated. Experimental results show that UDPGlu and UDPGal are baseline resolved within 22 min under the following optimum conditions: Zorbax NH2 column (250 mm×46 mm id, 5 μm) with acetonitrile0125 mol/L potassium phosphate buffer (pH 36) (60∶40, v/v) as the mobile phase at the flow rate of 10 mL/min and the column temperature of 30 ℃. Good linearity was found for UDPGlu and UDPGal over the range from 10 to 200 mg/L with the correlation coefficients of 0999?6 and 0999?3 respectively. The corresponding detection limits were 32 and 35 ng with average recoveries of 9826% and 9902%, and RSDs of 271% and 354%, respectively, for UDPGlu and UDPGal. 