Simultaneous determination of glucose and 1-deoxyglucose in serum by anion-exchange chromatography with an immobilized pyranose oxidase reactor.

A liquid chromatographic system for the determination of glucose and 1-deoxyglucose in serum using an immobilized pyranose oxidase reactor and chemiluminescence detection is described. Separation was achieved on a TSK gel SAX column (2 cm x 4 mm I.D.) with an eluent of 0.1 M sodium hydroxide solution. Serum was diluted 30-fold with 0.1 M NaOH. Sample solution (50 microliters) was injected into the system. The hydrogen peroxide produced was detected by measuring the chemiluminescence emitted on admixing with luminol and potassium hexacyanoferrate (III). The calibration graphs were linear from 1 to 600 microM glucose and from 0.6 to 400 microM 1-deoxyglucose; the detection limits for glucose and 1-deoxyglucose were 0.5 and 0.3 microM, respectively. The sample throughput was 10/h. The immobilized enzyme reactor was stable for at least 2 months.