Zhang Mei, Zhang Lina, Cheung Peter C K, Dong Jing
Department of Chemistry, Wuhan University, Wuhan 430072, China.
J Biochem Biophys Methods. 2003 Jun 30;56(1-3):281-9. doi: 10.1016/s0165-022x(03)00065-4.
A kind of regenerated cellulose gel (RCG) particles were treated by toluene to obtain particles with smaller mean pore size, then was mixed with the cellulose gel with pore size of 370 and 525 nm. A preparative size-exclusion chromatography (SEC) column (700 x 20 mm) packed with three gel particles was used to fractionate water-soluble polysaccharide (WEP) extracted from the sclerotia of Pleurotus tuber-regium by aqueous solution. The exclusion limit and fractionation range of the stationary phase of the preparative SEC were molecular mass 8 x 10(5) and 5 x 10(3) to 8 x 10(5), respectively. The calibration curve of the preparative SEC was represented as: log M=13.96-0.53 Ve. The WEP sample (weight-average molecular mass M(w)=2.2 x 10(4), polydispersity=2.4) was divided into three fractions with M(w) ranging from 1.4 x 10(4) to 3.4 x 10(4) by the preparative SEC column, and the fractions were characterized by gas chromatography GC, SEC combined with laser light scattering (SEC-LLS) and viscometry. The unfractionated WEP exhibited triple peaks due to different molecular mass, but each fraction exhibited single peak with the polydispersity of 1.1-1.8 in the SEC patterns. The results indicated that the preparative SEC was efficient for fractionation of polysaccharides having low molecular weight and for determination of their molecular mass.
一种再生纤维素凝胶（RCG）颗粒用甲苯处理以获得平均孔径较小的颗粒，然后与孔径为370和525 nm的纤维素凝胶混合。使用填充有三种凝胶颗粒的制备型尺寸排阻色谱（SEC）柱（700×20 mm）对通过水溶液从虎奶菇菌核中提取的水溶性多糖（WEP）进行分级分离。制备型SEC固定相的排阻极限和分级范围分别为分子量8×10⁵和5×10³至8×10⁵。制备型SEC的校准曲线表示为：log M = 13.96 - 0.53Ve。通过制备型SEC柱将WEP样品（重均分子量M(w)=2.2×10⁴，多分散性=2.4）分为三个级分，其M(w)范围为1.4×10⁴至3.4×10⁴，并且通过气相色谱GC、SEC结合激光散射（SEC - LLS）和粘度测定对这些级分进行表征。未分级的WEP由于分子量不同而呈现出三个峰，但在SEC图谱中每个级分均呈现单峰，多分散性为1.1 - 1.8。结果表明，制备型SEC对于低分子量多糖的分级分离及其分子量的测定是有效的。
