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目的:建立一种基于苯酚-硫酸法结合校正因子法测定含半乳糖醛酸(GalA)的多糖中糖含量的方法。方法:优化苯酚-硫酸法的测定条件,以不同质量比的GalA-葡萄糖(Glc)混合对照品溶液绘制系列标准曲线并测定含GalA的党参多糖CPP1b样品中的糖含量,在此基础上,根据GalA-Glc质量比为0-100%时的回归方程,以党参多糖CPP1b为参照多糖计算校正因子,对样品中的糖含量测定结果进行校正,并以与党参多糖CPP1b组成相同的混合单糖对照品对此方法合理性进行验证。结果:经计算,党参多糖CPP1b对Glc的校正因子为3.33,验证试验中与党参多糖CPP1b组成相同的混合单糖对照品的糖含量测定结果为103.47%。精密度、稳定性试验的RSD<1%;加样回收率为93.52%~107.35%(RSD=5.09%,n=6)。结论:该研究建立的方法可准确测定类似于党参多糖CPP1b的含GalA的多糖中的糖含量。
OBJECTIVE: To establish a method for the content determination of saccharide in polysaccharides containing galacturonic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS: The determination condition of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%, the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide, and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b. RESULTS: The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33; in validation test, the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS: The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.
多糖半乳糖醛酸党参多糖CPP1b苯酚-硫酸法校正因子法糖含量测定
PolysaccharidesGalacturonic acidCodonopsis pilosula polysaccharides CPP1bPhenol-sulfuric acid methodCorrection factor methodSaccharide contentDetermination
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