正交试验优化滇重楼核苷提取工艺

Optimization of extraction of nucleosides from Paris polyphylla var. yunnanensis by orthogonal test

【目的】优化滇重楼核苷的提取工艺条件,为滇重楼资源的开发和利用提供技术支持。【方法】以滇重楼根茎为试验材料,6种核苷类物质(胞苷、尿苷、鸟苷、胸苷、腺苷和2'-脱氧腺苷)总提取量为考察指标,采用高效液相色谱法测定其提取量,先分析超声波提取法和热水浸提法对滇重楼核苷提取效果的影响,然后在单因素试验的基础上,以甲醇体积分数、料液比、提取时间和提取次数为考察因素,采用L_9(3~4)正交试验对滇重楼核苷类物质提取工艺进行优化,确定最佳工艺条件。【结果】超声波提取法得到的滇重楼根茎中6种核苷类物质总提取量高于热水浸提法;4个因素对滇重楼核苷总提取量的影响排序为甲醇体积分数>提取时间>提取次数>料液比,最佳提取工艺条件为:以纯净水(甲醇体积分数为0)为提取溶剂,在料液比1∶10(g/mL)、30℃条件下超声波提取60 min,提取2次,得到滇重楼根茎中胞苷、尿苷、鸟苷、胸苷、腺苷和2'-脱氧腺苷的总提取量为1.4216±0.0198 mg/g,高于理论值1.3434 mg/g。【结论】正交试验优化得到的滇重楼核苷超声波提取工艺操作简便、稳定可靠,较热水浸透法的提取效率高,适合用于滇重楼核苷的提取。

【Objective】The technical conditions of extracting nucleosides from Paris polyphylla var. yunnanensis were optimized, which provided technical support for the exploitation and utilization of P. polyphylla var. yunnanensis. 【Method】With rhizome of P. polyphylla var. yunnanensis as experimental material and total extracted amount of six nucleosides（cytidine,uridine,guanosine,thymidine,adenosine and 2＇-deoxyadenosine） as examining indexes,high performance liquid chromatography（HPLC） method was adopted to determine the extraction amount. First, the effects of ultrasonic extraction and hot water extraction on the extracted amount of nucleosides from P. polyphylla var. yunnanensis were analyzed, then based on single factor experiments, methanol volume fraction, material liquid ratio, extraction time and extraction frequency were selected as experimental factors. Extraction of nucleosides from P. polyphylla var. yunnanensis was optimized by using L9（34） orthogonal experiment to select the optimal technical conditions. 【Result】The total extracted amount of six nucleosides by ultrasonic extraction was higher than that by hot water extraction. The effects of four experimental factors on total extracted amount of six nucleosides from P. polyphylla var. yunnanensis were ranked as follows： methanol volume fraction〉extraction time〉extraction frequency〉material-liquid ratio. The optimal extraction conditions were as follows： pure water（methanol volume fraction was 0） was used as extraction solvent, material-liquid ratio 1∶10（g/mL）, extraction time 60 min at 30 ℃ and extracting twice. Under these conditions, the total extracted amount of cytidine, uridine, guanosine, thymidine, adenosine and 2＇-deoxyadenosine in rhizome of P. polyphylla var. yunnanensis was 1.4216±0.0198 mg/g, which was higher than theoretical value 1.3434 mg/g. 【Conclusion】The ultrasonic extraction method optimized by orthogonal test is simple and reliable, the extraction rate of ultrasonic extraction is higher than hot water extraction. It is suitable for the extraction of nucleosides from P. polyphylla var. yunnanensis.