色谱柱及标准曲线的选择对HPGPC法测定右旋糖酐分子量及分子量分布的影响

目的： 比较不同色谱柱及标准曲线的选择对高效凝胶渗透色谱（HPGPC）法测定右旋糖酐分子量及分子量分布的影响。方法： 分别采用TSK G4000 PWXL、SHODEX OHPAK SB-803 HQ和Agilent PL AQUAGEL-OH MIXED-M 3种色谱柱对样品进行测定，选择已知分子量的右旋糖酐分子量标准（套）D1~D8系列对照品进行标准曲线的组合，采用GPC软件对结果进行分析，计算分子量及分子量的分布。结果： 3种色谱柱对右旋糖酐分子量与分子量分布的测定结果有显著差异，重均分子量（M_w）相对偏差达到3%，数均分子量（M_n）相对偏差达到12%，峰位相对分子质量（M_p）相对偏差达到25%，多分散性（D）相对偏差达到16%；标准曲线的选择结果比较中，相对偏差达到10%。结论： 不同色谱柱及标准曲线的选择对右旋糖酐分子量与分子量分布的测定结果有一定的影响。

Objective: To study the influence of different chromatograph columns and standard curves on molecular mass and molecular mass distribution of dextran by high performance gel permeation chromatography(HPGPC). Methods: Molecular mass and molecular mass distribution of dextran were determined by HPGPC with TSK G4000 PWXL,SHODEX OHPAK SB-803 HQ and Agilent PL AQUAGEL-OH MIXED-M columns,respectively. Selected dextran standard (set) D1-D8 series of reference materials with known molecular weight for the combination of standard curves,and then compare the results calculated by different serious of standard curves that handled with the WATERS GPC softwares. Results: There were obvious differences between results of different chromatograph columns. The deviation of weight average molecular mass(M_w) was up to 3%,the number average molecular mass(M_n) and the peak molecular mass(M_p) were up to 12% and 25%,respectively,and the molecular mass distribution width(D) between three columns was up to 16%. The deviation of different serious of standard curves was up to 10%. Conclusion: There are differences in the determination of molecular mass and molecular mass distribution by different chromatograph columns and standard curves.

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