目的考察高效凝胶渗透色谱法(HPGPC)测定注射用灰树花倍他葡聚糖(GFGI)相对分子质量(M_r)的影响因素。方法采用HPGPC分别以不同的流动相、色谱柱、上样浓度、上样体积、温度和辅料比例测定GFGI的M_r,并对测定结果进行比较。结果分别以纯水,质量浓度0.2g·L^-1NaN₃,0.1mol·L^-1NaNO₃和0.1mol·L^-1Na₂SO₄溶液为流动相进行测定,GFGI的M_r依次为822 410,177 520,29 112和25 066;以0.2g·L^-1NaN₃溶液为流动相时,采用TSK G4000PWxl和Shodex OHpak SB-804HQ色谱柱的Mr测定结果相差12.06%;在0.5～10g·L^-1的上样质量浓度内GFGI的M_r测定结果相差8.33%;在上样体积为5～30μL内相差8.65%;在温度为30～45℃范围内相差7.26%;但在不同的辅料比例下M_r的测定结果相近。结论流动相对GFGI的Mr测定结果有很大的影响,色谱柱、上样浓度、上样体积和测定温度对Mr测定结果有明显影响,辅料比例无明显影响。

OBJECTIVE To investigate the influence factors on relative molecular weight(M_r) determination of Grifola frondosa beta-Glucan for injection(GFGI) by high performance gel permeation chromatography (HPGPC).METHODS M_r of GFGI was determined by HPGPC using different mobile phases, chromatograph columns, concentration and volume of sample, temperature and ratios of excipient to Grifola frondosa beta-Glucan(GFG) separately.RESULTS The M_r of GFGI were 822 410,177 520,29 112 and 25 066 dalton determined by HPGPC using pure water,0.2 g·L^-1 NaN₃ solution,0.1 mol·L^-1 NaNO₃ solution and 0.1 mol·L^-1 Na₂ SO₄ solution as mobile phase respectively. The deviation of M_r of GFGI was 12.06% between Shodex OHpak SB-804HQ and TSK G4000PWxl columns determined by HPGPC using 0.2 g·L^-1 NaN₃ solution as mobile phase. The deviation of M_r of GFGI were 8.33%, 8.65%, 7.26% respectively determined by HPGPC in the concentration ranges of 0.5～10 g·L^-1,sample volume of 5～30 μL and temperature of 30～45 ℃.M_r of GFGI determined by HPGPC in different ratios of excipient to GFG were similar.CONCLUSION Mobile phases show great influence on M_r determination of GFGI.Chromatograph columns, concentration and volume of sample and temperature have obvious influence on M_r determination of GFGI, and ratios of excipient to GFG have little influence on M_r determination of GFGI.