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目的:考察流动相对高效凝胶渗透色谱法(HPGPC)测定灰树花多糖(GFP)相对分子质量(Mr)的影响因素。方法:采用HPGPC法分别以不同的流动相、盐浓度、pH测定GFP的Mr,并对测定结果进行分析。结果:分别以纯水、0.02%NaN_3、0.1mol·L~(-1)NaNO_3和0.1 mol·L~(-1)NaCl溶液为流动相进行测定,重均相对分子质量(WAMr)分别为822410、177520、29112、25066道尔顿,不同的流动相GFP的Mr有明显不同,以0.02%NaN_3较为理想;在低盐浓度(0～0.025mol·L~(-1))下,Mr变化明显,在0.1～0.2mol·L~(-1)硫酸钠溶液中趋于平稳;在pH 3～6时,Mr随pH增加而急剧增加,在pH 6～9范围内变化不明显。结论:不同流动相、盐浓度、pH对GFP的Mr测定结果有明显影响。
OBJECTIVE: To investigate the influencing factors for the determination of the relative molecular mass (Mr) of Grifola frondosa by mobile phase high performance gel permeation chromatography (HPGPC). Methods: The HPGPC method was used to determine the Mr of GFP with different mobile phases, salt concentration and pH, respectively, and the results were analyzed. Results: The mobile phase consisted of pure water, 0.02% NaN 3, 0.1 mol·L -1 NaNO 3 and 0.1 mol·L -1 NaCl respectively. The average relative molecular weights (WAMs) were 822410 , 177520, 29122 and 25066 Daltons, the different mobile phase GFP Mr significantly different to 0.02% NaN_3 is more ideal; at low salt concentrations (0 ~ 0.025mol·L -1)), Mr significant changes , Which is stable in 0.1 ~ 0.2mol·L ~ (-1) sodium sulfate solution. At pH 3 ~ 6, Mr increases sharply with the increase of pH, but does not change obviously in the range of pH 6 ~ 9. Conclusion: Different mobile phase, salt concentration and pH have significant effect on the determination of Mr of GFP.