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
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摘要
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.
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.
引文
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[11]潘英,李宁,倪慧,等.金鸡菊属植物化学成分和药理活性研究进展[J].现代药物与临床,2012,27(5):512-518.
[12]李冬明.昆仑雪菊的药学研究进展[J].浙江中医志,2012,47(10):776-777.
[13]郑大成,阿布力孜木合布力,吐鲁洪阿依努尔.昆仑雪菊水溶性黄酮的制备及初步鉴定[J].亚太传统医药,2010,6(10):18-20.
[14]古扎丽努尔·艾尔肯.两色金鸡菊药材质量标准建立及黄酮提取物质量分析[D].乌鲁木齐:新疆医科大学,2013.
[15]张瑞,李新霞,毛新民,等.HPLC法测定两色金鸡菊提取物中flavanomarein和marein的含量[J].安徽农业科学2015,43(6):3-74.
[16]张瑞.两色金鸡菊黄酮提取物工艺及化学成分分析[D].乌鲁木齐:新疆医科大学,2015.
[17]谢秀琼.中药新制剂开发与应用[M].北京:人民卫生出版社,2006.
[2]刘伟新,邓继华,徐鸿.一种金鸡菊花的生药学研究[J].中国民族医药杂志,2009,15(1):24-25.
[3]木合布力·阿布力孜,张兰,张敏.昆仑雪菊中氨基酸的含量分析[J].医药导报,2011,30(4):431-432.
[4]张彦丽,阿布都热合曼·合力力,阿依吐仑·斯马义.苯酚-硫酸法测定维吾尔药昆仑雪菊多糖含量的研究[J].药物分析杂志,2010,30(11):2205-2207.
[5]Abdureyim A,Abliz M,Sultan A,et al.Phenolic compounds from the flowers of Coreopsis tinctoria[J].Chem Nat Compd,2013,48(6):1085-1086.
[6]钱宗耀,周晓龙,刘河疆,等.气相色谱-质谱联用技术分析两色金鸡菊中的脂肪酸[J].江苏农业科学,2012,40(7):293-294.
[7]赵军,孙玉华,徐芳,等.昆仑雪菊黄酮类成分研究[J].天然产物研究与开发,2013,25(1):50-52.
[8]张彦丽,韩艳春,阿依吐仑·斯马义.GC-MS对昆仑雪菊挥发油成分的研究[J].新疆医科大学学报,2010,33(11):1299-1300.
[9]木合布力·阿布力孜,张燕,景兆均,等.新疆昆仑雪菊化学成分的初步定性研究[J].新疆医科大学学报,2010,33(6):628-630.
[10]明婷,庞市宾,哈木拉提,等.金鸡菊提取物对微循环及抗凝血作用的实验研究[J].农垦医学,2012,34(1):17-19.
[11]潘英,李宁,倪慧,等.金鸡菊属植物化学成分和药理活性研究进展[J].现代药物与临床,2012,27(5):512-518.
[12]李冬明.昆仑雪菊的药学研究进展[J].浙江中医志,2012,47(10):776-777.
[13]郑大成,阿布力孜木合布力,吐鲁洪阿依努尔.昆仑雪菊水溶性黄酮的制备及初步鉴定[J].亚太传统医药,2010,6(10):18-20.
[14]古扎丽努尔·艾尔肯.两色金鸡菊药材质量标准建立及黄酮提取物质量分析[D].乌鲁木齐:新疆医科大学,2013.
[15]张瑞,李新霞,毛新民,等.HPLC法测定两色金鸡菊提取物中flavanomarein和marein的含量[J].安徽农业科学2015,43(6):3-74.
[16]张瑞.两色金鸡菊黄酮提取物工艺及化学成分分析[D].乌鲁木齐:新疆医科大学,2015.
[17]谢秀琼.中药新制剂开发与应用[M].北京:人民卫生出版社,2006.