正交设计优化双斛胶囊中总生物碱的醇提取工艺

Optimization of the Ethanol Extraction Technology of Total Alkaloids from Shuanghu Capsules by Orthogonal Design

目的:优化双斛胶囊总生物碱的醇提取工艺。方法:以石斛碱为对照品,采用酸性染料比色法测定双斛胶囊中主药金钗石斛和铁皮石斛的总生物碱含量。以干膏得率和总生物碱含量计算的综合评分作为评价指标,采用单因素试验考察浸泡时间、乙醇体积分数、提取时间、料液比和提取次数对提取效果的影响;根据单因素试验结果采用L9(34)正交试验进一步对乙醇体积分数、提取时间、料液比和提取次数进行优化,并对优化工艺进行验证试验。结果:石斛碱质量浓度在4.16~14.56μg/mL范围内与吸光度呈良好的线性关系(r=0.999 2);重复性试验、精密度试验的RSD均小于5%;平均加样回收率为93.01%(RSD=1.97%,n=6)。优化后的醇提取工艺为浸泡12 h后,按乙醇体积分数为70%、料液比为1∶12(g/mL)提取28 min,共提取3次;验证试验结果显示,优化工艺所得干膏的平均得率为12.80%(RSD=4.39%,n=3),平均生物碱含量为0.359 0 mg/g(RSD=0.66%,n=3)。结论:所建立的酸性染料比色法操作简便,精密度和准确度均良好,可用于总生物碱的含量测定;优化的醇提取工艺稳定、可行,可用于双斛胶囊中总生物碱的提取。

OBJECTIVE: To optimize the ethanol extraction technology of total alkaloids from Shuanghu capsules.METHODS:Using dendrobine as control,the contents of total alkaloids from Dendrobium nobile and Dendrobium officinale in Shuanghu capsules were determined by acidic dyes colorimetry. Using comprehensive scores calculated by the yield of the extract and the contents of total alkaloids as evaluation indexes,the effects of soaking time,ethanol volume fraction,extraction time,solid-liquid ratio and extraction times were investigated with single factor tests. L9(34)orthogonal test was used to optimize ethanol volume fraction,extraction time,solid-liquid ratio and extraction times according to the results of single factor test. The optimized technology was validated. RESULTS:The linear range of dendrobine were 4.16-14.56 μg/mL(r=0.999 2). RSDs of repeatability and precision tests were all lower than 5%. Average recovery tests were 93.01%(RSD=1.97%,n=6). The optimal ethanol extraction technology included soaking for 12 h,ethanol volume fraction of 70%,solid-liquid ratio of 1 ∶12(g/mL),extracting for28 min,extracting 3 times. Results of validation test showed that the average yield of extract was 12.80%(RSD=4.39%,n=3),and the content of alkaloids was 0.359 0 mg/g(RSD=0.66%,n=3). CONCLUSIONS:Established acidic dyes colorimetry is simple,precise and accurate,which can be used for the content determination of total alkaloids. The optimized ethanol extraction technology is stable and feasible,and can be used for the extraction of total alkaloids from Shuanghu capsules.