两色金鸡菊提取物大剂量给药致小鼠死亡化学成分的研究

Induction of mice death by toxicity components from large-dose Coreopsis tinctoria extract

目的比较两色金鸡菊各种提取物对小鼠的急性毒性作用,分析对大剂量给药致小鼠死亡贡献最大的成分,为后期研究提供安全性数据并优化提取工艺。方法分别测定各提取物对小鼠的最大给药量或最大耐受量（MTD）,用Bliss法测定半数致死量（LD50）,记录小鼠死亡情况和体质量变化情况;高效液相色谱（HPLC）法测定各提取物指纹图谱,根据各物质吸收峰,结合多元线性回归,分析对大剂量给药致小鼠死亡贡献最大的成分。结果两色金鸡菊喷雾干燥水提物（SD）、醋酸乙酯萃余组分（AR）最大给药量均为36 g/kg,真空干燥水提物（VD）MTD为26 g/kg,乙醇提取物（ETE）LD50（95%可信限）为19.565（17.558~21.734）g/kg,醋酸乙酯萃取组分（AC）LD50（95%可信限）为16.414（13.987~34.725）g/kg;3,5-二咖啡酰奎宁酸可能是对大剂量给药致小鼠死亡贡献最大的成分。结论 ETE和AC大剂量给药会致小鼠死亡,3,5-二咖啡酰奎宁酸可能是对两色金鸡菊提取物大剂量给药致小鼠死亡贡献最大的成分。

objective By comparing the acute toxicity of different extracts from Coreopsis tinctoria on mice, combined with the HPLC fingerprint and multiple linear regression to analyze the element which plays the most important role in causing the death of mice, and to provide the safety data for improving the extraction technology. Methods To measure the maximum dose and maximal tolerance dose（MTD） of all the extracts, to measure the median lethal dose（LD50） by Bliss, and to record the death and weight changes; To measure the fingerprints of the extracts by HPLC, and to determine the element which mostly induced the death of mice by analyzing the absorption peak of the extracts by HPLC fingerprint with multiple linear regression. Results The extracts include aqueous extract by spray drying（SD）, aqueous extract by vacuum drying（VD） process, ethanol extract（ETE）, ethyl acetate extracted component（AC）, and the ethyl acetate extracted residuum（AR）. Among those extracts, the maximum dose of SD and AR is 36 g/kg, the MTD of the VD is 26 g/kg, the LD50（95% confidence limits） of ETE and AC are 19.565（17.558—21.734） g/kg and 16.414（13.987—34.725） g/kg, respectively; Under the high dose situation, 3,5-dicaffeoylquinic acid properly is the component which mostly contributes to the death of mice. Conclusion Under the high dose situation, the ETE and AC will lead the death, and 3,5-dicaffeoylquinic acid properly is the component which mostly contributes to the death of mice.