Abstract:

An analytical method for the determination of trehalose, maltose, and glucose in biotransformation samples was developed by using high performance anion exchange chromatography coupled with pulsed ampere detection (HPAEC-PAD). The analysis was performed on a CarboPac^TM10 column (250 mm×2 mm) with the gradient elution of NaOH-NaAc as the mobile phase. The column temperature was set at 30 ℃, the flow rate was 0.30 mL/min. The results showed that trehalose, maltose, and glucose in biotransformation system were completely separated and determined in 15 min. The linear ranges and the working curves were determined by using standard samples. The correlation coefficients of three kinds of carbohydrates were over 0.9998 . The detection limits (LODs) were 0.010-0.100 mg/L. Under the optimized separation conditions, the recoveries of saccharides in the transformation system at three different spiked levels ranged from 89.4% to 103.2%. In biotransformation system, 50 IU trehalose synthase were added into 200 g/L maltose for reaction of 8 h at 37 ℃, pH 8.0. Under the above conditions, the concentration of trehalose in biotransformation sample was 101.084 g/L, and the conversion rate of trehalose reached 50.5%. The method can be applied to determine the composition in the transformation system with the advantages of simplicity and convenience.

Key words: biotransformation sample, glucose, high performance anion exchange chromatography (HPAEC), maltose, trehalose