Simultaneous determination of 2-keto-L-gulonic acid and 2-keto-D-gluconic acid in fermentation broth by HPLC

A simple and fastHPLCmethod for simultaneous determination of 2-keto- L-gulonic acid and 2-keto-D-gluconic acid in fermentation broth was described. Separation and quantitation were achieved on a shim-pack CLCNH 2 column (150mmi‚´6mm, 5i­m) using 0.015mol/Lammoniumdihydrogen phosphate solution (pH 4.1 adjusted by phosphoric acid) asmobile phase. Detection was by UV absorbance at a wavelength of 210nm, and the flow rate was 1mL/min. The elution of the analytes was achieved in less than 19.0min. The linearity, accuracy and precision of the method were found to be good and acceptable over the concentration ranges from 10 to 600i­giƒ—mL-1 for both 2-keto-L-Gulonic acid and 2-keto-D-Gluconic acid.