不同产地刺五加中多糖含量测定、聚类分析及其超声提取工艺优化

Determination of Polysaccharide Content,Cluster Analysis in Acanthopanax senticosus from Different Producing Areas and Optimization of Its Ultrasonic Extraction Technology

目的:建立测定刺五加中多糖含量的方法,并对其进行聚类分析及超声提取工艺优化。方法:采用硫酸-苯酚法测定刺五加中多糖的含量。采用SPSS 23.0软件对17个不同产地刺五加进行聚类分析。采用L9(34)正交试验设计以提取温度、料液比、提取时间为考察因素,多糖含量为考察指标对其超声提取工艺进行优化并验证。结果:葡萄糖质量浓度线性范围为0.007 75~0.151mg/mL(r=0.999 1);定量限、检测限分别为2.854、0.856μg/mL;精密度、稳定性、重复性试验的RSD均小于2%;加样回收率为98.41%~101.58%(RSD=1.23%,n=6)。聚类分析结果显示,17批药材样品可聚为3类,S1、S6、S10、S12、S13聚为一类,S3~S5、S7聚为一类,其余聚为一类。最优超声提取工艺为提取温度55℃、料液比1∶10(g/mL)、提取时间35 min;3次验证试验结果显示,最优工艺所得多糖平均含量为4.36%(RSD=0.92%,n=3)。结论:所建含量测定方法操作简便、重复性较好,可用于测定刺五加中多糖含量;优化后的超声提取工艺稳定、可行。

OBJECTIVE:To establish a determination method for content of polysaccharide in Acanthopanax senticosus,and to conduct cluster analysis and ultrasonic extraction technology optimization. METHODS: The content of polysaccharide in A.senticosus was determined by phenol-sulfuric acid method. Cluster analysis was conducted by using SPSS 23.0 software of the polysaccharide in A. senticosus from 17 habitats. The ultrasonic extraction technology was optimized with L9(34)orthogonal test using extraction temperature,the ratio of material to liquid,extraction time as factors,the content of polysaccharides as index,and then validated. RESULTS:The linear range of glucose ranged from 0.007 75 to 0.151 mg/mL(r=0.999 1);the limits of quantification and detection were 2.854,0.856 μ g/mL;RSDs of precision,stability,and repeatability tests were less than 2%;recovery rates of the sample were 98.41%-101.58%(RSD=1.23%,n=6). Cluster analysis results showed that 17 batches of samples could be clustered into three classes;S1,S6,S10,S12 and S13 were clustered into one class;S3-S5 and S7 were clustered into one class;and the rest samples were clustered into one class. The optimal ultrasonic extraction technology was extraction temperature 55 ℃,ratio of material to liquid 1 ∶ 10(g/mL),extraction time 35 min. Results of validation tests showed the content of the polysaccharide under the best process condition was 4.36%(RSD=0.920%,n=3). CONCLUSIONS:Established method is simple,reproducible and suitable for determination of polysaccharide in A. senticosus;the optimized ultrasonic extraction technology is stable and feasible.