正交试验优化灵芝多糖含量测定方法的研究

Study on Anthrone-Sulfuric Acid Method Optimized by Orthogonal Experiment for Quantitative Determination of Polysaccharide from Ganoderma Lucidum

以野生黑灵芝的粗多糖含量为指标,采用L18(37)正交试验设计建立优化的蒽酮-硫酸法。样品用料液比为30m L/g,在100℃条件下超声提取1h并重复3次,浓缩液5m L用70m L无水乙醇于4℃沉淀4h,用4000r/min离心20min分离沉淀,取一定量沉淀溶解后的溶液用0.2%蒽酮试剂在100℃显色15min,在620nm波长下测定吸收值。采用最佳实验条件测定灵芝的多糖含量并与药典方法对比,校准曲线回归方程为y=5.0724x-0.0033,相关系数R2=0.9990,6种灵芝的多糖含量为云芝1.639%、平盖灵芝4.350%、野生黑灵芝6.178%、桦褐孔菌2.041%、桦树桑黄菌4.751%、松针桑黄菌4.221%,RSD为0.38%~3.14%。与药典方法对比,结果准确性高、精密度高、稳定性好,操作更加快速简便。

The anthrone-sulfuric acid method was established and optimized by Lls（37） orthogonal experiment while detecting the polysaccharide content from wild ganoderma atrum was used as the index. The best parameters of method were following. The polysaccharide was ultrasonic extracted from sample by soaked in water at 100℃ for lh for three times repeatedly. While condensing the extract into 5mL, the polysaccharide settled down at 4℃ for 4h and separated with 4000r/rain for 20min, and dissolving slag into solution which was determined by 620 nm after color reaction at 100℃ for 15min used 0.2% anthrone-sulfuric acid. The equation of calibration curve and relative standard deviation were y=5.0724x- 0.0033, and 0.9990,respectively, and RSD was 0.38%-3.14% , the polysacharide content of coriolus versicolor, ganoderma applanatum, wild ganoderma atrum, chaga, birch phellinusigniarius and Pine needle phellinusigniarius were 1.639%, 4.350%, 6.178%, 2.041%, 4.751%, 4.221%, respectively. The method was accurate, precision and fast compared with pharmacopoeia of China.