Analysis of Trehalose in Transgenic Tobacco Extracts by High Performance Liquid Chromatography with Evaporative Light Scattering Detection
液相色谱/蒸发光散射测定转基因烟草提取液中的海藻糖

A high performance liquid chromatographic method with ethylenediamine dynamically modified silica column and evaporative light-scattering detection (ELSD) was developed for the direct determination of trehalose in transgenic tobacco extracts. The separation column (Zorbax Rx-SIL, 4.6 mm i.d. x 250 mm, 5 microns, Hewlett-Packard, USA) modified by ethylenediamine and a guard column (Zorbax Rx-SIL, 4.6 mm i.d. x 12.5 mm, 5 microns) were used. The mobile phase was a mixture of acetonitrile-water(2.6:1, V/V) containing 0.03% ethylenediamine. Regression equations revealed the linear relationships (r = 0.996-0.999) between the concentrations and peak areas of carbohydrates detected by ELSD. The detection limits of ELSD (S/N = 3:1) were 10 mg/L, 20 mg/L, 10 mg/L and 10 mg/L for fructose, glucose, sucrose and trehalose, respectively.