植物灭螺成分的提取工艺及作用机理研究

发布时间：2018-04-21 06:49

  本文选题：钉螺 + 灭螺植物　； 参考：《华中师范大学》2009年硕士论文

【摘要】： 控制钉螺是防治血吸虫病的关键技术之一。研发高效、持久、安全及价廉(WHO标准)的新型植物灭螺剂是灭螺药物研究的热点。由于缺乏对植物灭螺成分单体水平的构效关系、环境安全及经济性的同步研究,以致尚未研究出一种符合WHO标准的植物灭螺药物。本文研究灭螺植物有效成分的提取分离工艺及对Nic抑螺增效作用、钉螺转氨酶的动力学特征及对钉螺药物的敏感性,为新型灭螺药物研发提供设计依据和物质基础。 1.RC灭螺皂甙的提取分离研究 RC原粉经MeOH索氏提取和正丁醇萃取、TLC及CCL分离纯化,得到皂甙结晶RCI,其纯度为83.53%。采用WHO试验法测定,5.0 mg·L~(-1)的RCI浸杀钉螺72h,其死亡率为80.0%。结果说明RCI具有较高的灭螺活性。 2.SX生物碱的提取工艺研究 采用L_16(4~4)正交试验与极、方差分析,以SX果实灭螺生物碱SXⅡ和SXⅢ的质量分数为指标,研究确定其最佳提取条件。SXⅡ的最佳提取条件为:浸提温度40℃、浸提时间15h、水与原料配比10g·mL~(-1)、浸提剂pH3.0;SXⅢ的最佳提取条件为:浸提温度10℃、浸提剂pH3.0、浸提时间15h、水与原料配比10g·mL(-1)。结论为SX生物碱生产应用提供实验基础。3.SX生物碱对Nic抑螺增效研究 采用WHO试验法测定,SXⅠ、Ⅱ和Ⅲ浸泡钉螺24h,其死亡率达100%的浓度分别为100、8和200 mg·L~(-1)。用Nic抑螺无效浓度0.25、0.5及1.0 mg·L~(-1),分别与SXⅠ、Ⅱ及Ⅲ抑螺无效浓度50、2.7及67 mg·L~(-1)联用时,Nic抑螺率分别提高100、60及100%。结果说明SXⅡ具有高效灭螺和增加Nic抑螺效应的作用。 4.钉螺GPT和GOT反应动力学特征及对灭螺药物的敏感性研究 采用L_25(5~6)正交试验与极、方差分析,以钉螺软体60-90%饱和度盐析蛋白为材料,研究确定了钉螺GPT的反应最适条件为:酶浓度0.1 mg·mL~(-1)、α-酮戊二酸5μmol·L~(-1)、反应体系pH6.5、反应温度30℃、反应时间30min;GOT反应最适条件为酶浓度0.2mg·mL~(-1)、L-天门冬氨酸2.5μmol·L~(-1)、反应体系pH6.0、反应温度40℃、反应时间30min。 在4mg·L~(-1)的CuSO_4、Nic及SXⅡ药液中,GPT活性抑制率分别为81.2、-32.2及16.2%,GOT活性抑制率分别为89.7、-18.8及-11.4%。结论说明离体钉螺GPT和GOT对CuSO_4敏感性较高,对Nic和SXⅡ的敏感性较低。
[Abstract]:Snail control is one of the key techniques for schistosomiasis control. Research and development of new plant molluscicides with high efficiency, durability, safety and low cost is a hot spot in the research of snail control drugs. Due to the lack of the structure-activity relationship at the level of plant molluscicidal components, and the simultaneous study of environmental safety and economic efficiency, a plant snail killing drug conforming to the WHO standard has not been developed. In this paper, the extraction and separation of effective components from snail control plants, the synergistic effect of Nic on snail inhibition, the kinetic characteristics of snail aminotransferase and its sensitivity to snail drugs were studied, which provided the design basis and material basis for the research and development of new snail killing drugs. Study on extraction and Separation of 1.RC saponins The RC powder was extracted by MeOH Soxhlet and purified by TLC and CCL extraction with n-butanol, and the purity of the saponins was 83.53. The method of WHO test was used to determine the RCI dipping of Oncomelania hupensis for 72 h, and the mortality rate was 80.0%. The results showed that RCI had high snail killing activity. Study on extraction process of 2.SX Alkaloids The optimum extraction conditions of SX 鈪，

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