目的 研究苦丁茶冬青叶多糖含量的测定方法。方法 以水提醇沉的方法从苦丁茶冬青叶中提取得到苦丁茶冬青叶多糖并以正交实验优化显色条件，采用苯酚-硫酸法测定其多糖含量。结果 测定波长为482nm，最佳显色条件为6％苯酚用量1．0ml，浓硫酸用量7．0ml，反应时间40min。标准曲线回归方程为A=0．0546C-0．0033（n=10），r=0．9998，并在2．51-45．11mg·L^-1范围内呈良好线性关系。平均回收率为98．55％，RSD=1．86％（n=5）。测得多糖平均含量为31．90％，RSD=1．75％（n=6）。结论 该方法简单、准确、重现性好，可为其继续研究开发及应用提供参考依据。
Objective To study the determination method for polysaccharides in the leaves of llex kudincha C. J. Tseng. Methods The leaves of llex kudincha C. J. Tseng were defatted with 95% EtOH, extracted with water and precipitated with 80% EtOH, and then was deproteined three times by Scrag method, and from which, a crude polysaccharide noted as KPSⅡ was obtained. The traditional colofimetric method applied for the determination of polysaccharides from the leaves of Ilex kudincha C. J. Tseng was developed by the use of phenylhydrate - sulfuric acid, and was optimized by orthogonal experiment. Results The results showed that the wavelength for measurement was 482 nm. The best colorimetric conditions were the time of reaction of 40 rain and the dosage of 6% phenylhydrate and H2SO4 of 1.0 and 7.0 ml, respectively. Linear range was 2.51 - 45.11 mg·L^-1 （A =0.054 6C -0. 003 3 ,r =0.999 8 ）. The average recovery was 98.55% and RSD was 1.86% （ n = 5 ）. The pelysaccharide content of KPS Ⅱ was 31.90% and RSD was 1.75% （n =6）. Conclusion The method used in this paper is simple, accurate and stable. The result obtained may act as a base for the further study and application of it.
Lishizhen Medicine and Materia Medica Research
Polysaccharides from the leaves of Ilex kudincha C. J. Tseng
Phenylhydrate -sulfuric acid method