高效凝胶渗透色谱法测定多糖纯度及分子量

用已知平均分子量(w)的T系列葡聚糖为标准,分别用两种不同的流动相,在Bio-Gel TSK柱上测定多糖的纯度及分子量。用线性回归法得出校正曲线。分子量w的平均偏差不超过5%。

DETERMINATION OF PURITY AND MOLECULAR WEIGHT OF POLYSACCHARIDES BY HIGH PERFORMANCE GEL PERMEATION CHROMATOGRAPHY

Polysaccharides, PIB-I, PIB-II-A, PIB-II-Band PIA-I, were prepared in our laboratory. Their purity and molecular weight (Mw) were determined by high performance gel permeation chromatography (HPGPC). The HPGPC system consisted of a two-column set, Bio-Gel TSK-50 and TSK-40 columns. 0.2 mol/L phosphate pH 6.0 buffer and 0.1 mol/L Tris-HCl pH 8.0 buffer were used as mobile phase, respectively. The void volume and total volume were measured using dextran T-2000 and glucose. The Mw of samples were determined from the standard curves which were made by standard dextran T-series. In two different mobile phases, the deviations of Mw were less than 6.0%, which showed that the method is highly sensitive, accurate, reproducible and rapid.