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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 To compare the acute toxic effects of various extracts of two species of Coreopsis chinense on mice and to analyze the components that contribute most to the death of mice caused by high dose administration and to provide safety data and optimize the extraction process for later studies. Methods The maximal dose and maximum tolerated dose (MTD) of each extract were determined in mice. The half-lethal dose (LD50) was measured by Bliss method and the death and body weight were recorded. High performance liquid chromatography HPLC method was used to determine the fingerprint of each extract. According to the absorption peak of each substance, combined with multiple linear regression analysis, the components that contributed the most to the death of mice caused by high dose administration were analyzed. Results The maximum dosages of spray dried water extract (SD) and ethyl acetate extractable fraction (AR) were 36 g / kg and VDD of vacuum dried water extract were 26 g / kg, The LD50 (95% confidence interval) of ethanol extract (ETE) was 19.565 (17.558-21.734) g / kg and the ethyl acetate extractable fraction (LD50) with a 95% confidence limit of 16.414 (13.987-34.725) g / kg; 3,5-dicaffeoylquinic acid may be the largest contribution to the death of mice caused by high doses of ingredients. Conclusion ETE and AC high-dose administration can cause death in mice. 3,5-Dicaffeoylquinic acid may be the most contributor to the death of mice induced by high dose of two-color Coreopsis extract.