目的：建立云南部分兰科植物多糖含量测定的方法，并对其提取工艺进行研究。方法：采用传统水提取方法提取多糖，以葡萄糖为标准品，用苯酚-硫酸法测定多糖的含量，并以其为考察指标，采用提取条件的优化筛选最佳提取工艺。结果：葡萄糖在0.000~0.020 mg/mL范围内线性关系良好，线性回归方程为A=55.55c（mg/mL）+0.001（ r=0.9998），平均加标回收率为99.7%，RSD=3.13%（n=6）；最佳显色条件：苯酚用量1.0 mL、浓硫酸用量7.0 mL、显色温度80℃、显色时间30 min；提取兰科植物多糖的最佳工艺条件为：提取温度70℃，料液比1：40，提取时间2 h。结论：该方法简便、准确，优选出的提取工艺重复性好。%Objective: To establish the method for determining the contents of polysaccharides and to explore the extraction technique for polysaccharides from Yunnan orchidaceae plants. Methods:The polysaccharide was extracted with the traditional water extraction, using glucose as the standard, and the contents of polysaccharides was measured by the phenol-sulfuric acid method. With the content of polysaccharides as an index, the univariate test were performed to find the optimal extraction technique. Results:A good linear relationship with a regression equation of A=55.55c(mg/mL)+0.001(r=0.999 8)was achieved when the concentration of glucose ranged between 0.000 and 0.020 mg/mL. The average recovery rate was 99.7%, and the relative standard deviation was 3.13%(n=6). The best conditions of color are as follows, the dosage of phenol and sulphuric acid was 1.0 mL and 7.0 mL respectively, temperature of color was at 80℃, and developing time was 30 minutes. The optimal extraction of polysaccharide was performed by 2 hours soaking using 1:40 volumes of solvent at 70 ℃. Conclusion: The established method is simple and accurate and the optimal extraction technique shows good reproducibility.
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