[关键词]
[摘要]
目的 利用正交试验结合体外模型测定固公果根总三萜(TTROG)提取物抑制大鼠离体肠平滑肌收缩活性,优化其提取工艺。方法 以熊果酸为对照品,采用香草醛-冰醋酸显色,紫外分光光度法测定TTROG提取物中三萜含量,单因素考察提取溶剂、料液比、提取温度、提取时间、提取次数对TTROG提取率的影响;在单因素实验的基础上采用正交试验法考察提取溶剂、料液比、提取时间对TTROG提取率的影响;通过张力传感器与BL-420生物实验多通道生理信号采集处理系统记录不同提取物对体外肠平滑肌收缩的抑制作用,利用加权方法使生物活性与提取率相结合评价TTROG的提取工艺。结果 TTROG的最佳提取工艺条件为:提取溶剂80%乙醇,料液比1:10,提取时间2 h,提取温度80℃,提取次数3次;最优条件下TTROG的提取率可达42.12 mg/g;TTROG提取物对十二指肠、空肠、回肠、结肠4个肠段都具有显著的抑制离体肠平滑肌收缩的作用,呈剂量相关性,对空肠的作用最强,抑制率可达41.96%。结论 经优化后的TTROG提取工艺提取率高、稳定,且获取的TTROG提取物具有显著的抑制大鼠离体肠平滑肌收缩功能。
[Key word]
[Abstract]
Objective To optimize the extraction technology of total triterpenoid from root of Rose odorata var. gigantean (TTROG) by orthogonal test combined with the contraction effect of TTROG on the isolated intestinal smooth muscle of rats in vitro. Methods UV spectrophotometric method was used to determine the contents of total triterpenoids in the TTROG extractive at the wavelength of 550 nm by taking ursolic acid as standard substance, and vanillin acetic acid as chromogenic reagent. The extraction rate of total triterpenoids was used as index to evaluate the technology based on single factor test, in which three factors were considered as follows:the concentration of extraction solvent, ratio of material to liquid, extraction time, and their interaction on extraction were studied by orthogonal experimental design. The inhibition effect of different extracts obtained from the optimized extraction process on the contraction of intestinal smooth muscle were recorded by tension transducer to the BL-420 biological experimental multi-channel physiological signal acquisition and processing system. The extraction process of TTROG was evaluated by the combination of biological activity and extraction rate with weighting method. Results The optimal extraction conditions of TTROG were as follows:extraction solvent 80% ethanol, solid-liquid ratio 1:10, extraction time for 2 h, three times and extraction temperature of 80℃. The optimized extraction rate could reach 42.12 mg/g. TTROG obtained using the optimized method showed significantly contraction effect on rat intestinal smooth muscle with dose effect dependence, and the effect on jejunum was the strongest, and the inhibition rate was 41.96%. Conclusion The optimized extraction technology is stable and effective with high extraction rate. TTROG showed the significant inhibitory function on contraction of isolated rat intestinal smooth muscle.
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[基金项目]
国家自然科学基金项目(81673693)