王莹, 许玮仪, 李丽潇, 金红宇, 倪健, 马双成. 注射用黄芪多糖相对分子质量测定方法的比较及研究J. 药学学报, 2019,54(2): 348-353. doi: 10.16438/j.0513-4870.2018-0612
引用本文: 王莹, 许玮仪, 李丽潇, 金红宇, 倪健, 马双成. 注射用黄芪多糖相对分子质量测定方法的比较及研究J. 药学学报, 2019,54(2): 348-353. doi: 10.16438/j.0513-4870.2018-0612
WANG Ying, XU Wei-yi, LI Li-xiao, JIN Hong-yu, NI Jian, MA Shuang-cheng. Relative molecular weight determination of astragalus polysaccharides for injectionJ. Acta Pharmaceutica Sinica, 2019,54(2): 348-353. doi: 10.16438/j.0513-4870.2018-0612
Citation: WANG Ying, XU Wei-yi, LI Li-xiao, JIN Hong-yu, NI Jian, MA Shuang-cheng. Relative molecular weight determination of astragalus polysaccharides for injectionJ. Acta Pharmaceutica Sinica, 2019,54(2): 348-353. doi: 10.16438/j.0513-4870.2018-0612

注射用黄芪多糖相对分子质量测定方法的比较及研究

Relative molecular weight determination of astragalus polysaccharides for injection

  • 摘要: 采用不同的方法对注射用黄芪多糖相对分子质量进行测定与比较。以SHODEX GS-620为色谱分离柱,右旋糖酐葡聚糖作为对照品,采用高效凝胶渗透色谱-示差检测器(GPC-RI)测定注射用黄芪多糖的重均分子量(Mw)。同时采用多角度激光光散射法(MALLS)及基质辅助激光解吸/电离飞行时间质谱法(MALDI-TOF-MS)对其相对分子质量进行测定。GPC-RI法测得注射用黄芪多糖依次呈现4个色谱峰,其Mw分别为:138万,23.1万,1.8万,480;MALLS法测得3个信号较强的色谱峰,其Mw分别为:114.8万,18万,4.3万;MALDI-TOF-MS法测得在15.5万和1.8万有较强的信号,并获得黄芪多糖的糖残基单元为己糖。通过3种方法的测定与相互验证,初步确定了注射用黄芪多糖各分离峰的Mw,同时获取了部分黄芪多糖单元结构信息。最终经过普适校正后的GPC-RI法可应用于黄芪多糖的Mw测定。在研究中3种方法各有其优势,采用相互比较和验证的方式测定中药多糖可获得更为准确的结果。

     

    Abstract: To determine relative molecular weight of astragalus polysaccharides (APs), we used Shodex GS620 gel permeation chromatographic column and differential refraction detector (GPC-RI) with dextran as a reference. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and GPC combined with multi-angle laser light scattering detection (GPC-MALLS) were also used.GPC-RI measure showed four peaks of APs, with the Mw of 1 380 000, 231 000, 18 000, and 480, respectively. Three main peaks were found by GPC-RI-MALLS with the Mw as 1 148 000, 180 000, and 43 000, respectively. Strong signals in 155 000 and 18 000 were detected by MALDI-TOF-MS, which also indicated the sugar moieties of the APs as hexoses. From our study, we found that the GPC-RI method with universal correction is most suitable for Mw determination of the APs. Nevertheless, the three methods should be combined and contrasted with each other to obtain accurate information in research of polysaccharides from Chinese medicine.

     

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