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A high performance liquid chromatographic method for the separation of hexose monophosphates and UDP glucose from plant extracts and its use to determine specific activities in radiotracer experiments

, : A high performance liquid chromatographic method for the separation of hexose monophosphates and UDP glucose from plant extracts and its use to determine specific activities in radiotracer experiments. Phytochemical Analysis 5(1): 10-14

Anion exchange high performance liquid chromatography (HPLC) was used to separate UDP glucose (UDPGlc) and hexose monophosphates (HMPs) in crude ethanol extracts of potato tubers. Detection and quantification of UDPGlc and (U-14C)UDPGlc was achieved with UV (A-254) and radioactivity monitors connected in series. The 14C content in glucose 6-phosphate (Glc-6-P), fructose 6-phosphate (Fru-6-P) and glucose 1-phosphate (Glc-1-P) was obtained following their enzymatic conversion to gluconate 6-P and subsequent separation by HPLC. Individual HMPs were quantified by enzyme-coupled reaction systems using a microplate reader equipped with a 340 nm filter. This approach allowed the specific activities of UDPGlc, Glc-6-P, Fru-6-P and Glc-1-P to be quantified in radiolabeled potato extracts within 2 h.


DOI: 10.1002/pca.2800050104

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