手性农药甲基异柳磷对映体的细胞毒性
毕业论文关键词：手性农药；甲基异柳磷；细胞毒性；对映体选择性
The enantioselective cytotoxicity of chiral pesticides isofenphos-methyl
Abstract：In this study, the enantiomer separation of isofenphos-methy was achieved by HPLC coupled with a Lux Cellulose-3 chiral column. The absolute configuration of first eluted peak was cofirmed as S and the second was cofirmed as R by comparing the experimental and predicted electron circular dichroism spectroscopy. And then the cytotoxicity of isofenphos-methyl on the Hep G2 cell was studied. The results showed that both enantiomers were highly toxic, there was no significant difference between enantiomers, at the high concentration group. At low concentration, the toxicity of R-isofenphos-methy was significantly higher than that of S-isofenphos-methy (pharmacy concentration of 39.5mg/L, the toxicity of R was 3.2 times the S). Combined the influence of isofenphos-methyl on the SOD, CAT, LDH activity and the content of MDA,The activity of SOD and CAT decreased gradually with the increase of the pharmacy concentration. The activity of extracellular LDH and the content of MDA in the cells increased gradually with the pharmacy concentration. the mechnism of the enantioselective cytotoxicity was explored which provided a scientific and comprehensive theoretical basis for the corresponding risk assessment.
Key words: chiral pesticide;isofenphos-methyl;cytotoxicity; enantioselectivity
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 1
1实验材料 3
1.1细胞株 3
1.2实验药剂 3
1.3主要仪器和试剂 3
2实验方法 3
2.1甲基异柳磷手性拆分仪器条件的确定 3
2.2相关用液的配制 5
2.3 细胞的培养及处理 5
2.4细胞活性测定（CCK-8法） 5
2.5乳酸脱氢酶（LDH）的释放量的测定 5
2.6 细胞氧化应激反应 6
3结果分析 8
3.1 甲基异柳磷对Hep G2细胞毒性对映体选择性影响8
3.2 甲基异柳磷对Hep G2细胞LDH释放量的对映体选择性影响 9
3.3 甲基异柳磷对Hep G2细胞产生氧化应激的对映体选择性9
4讨论 11
致谢 12
参考文献 13
手性农药甲基异柳磷对映体的细胞毒性
引言
据报道，全世界范围内每年因农药中毒死亡的人数多达30万人[1]，化学农药的大量使用是造成土壤、 地表水污染乃至食品安全问题的主要因素之一[2]。当前世界范围内生产和使用的农药品种中有30% 具有手性，在手性农药中仅有20%的农药为富含高效对映体，并且这一比例会随着农药研发技术的提高及新化合物结构不断出现而呈递增趋势。手性对映体具有相同的理化性质，然而当这些手性化合物进入到手性环境中时，往往会在生理，生化等方面表现出显著地立体选择性[3,4]。大量实验研究数据表明，手性农药在进入自然界后，其在环境媒介中的降解代谢过程往往都是具有立体选择性的[5]。研究手性农药在对映体水平的毒性、活性以及在环境中的降解代谢对手性农药的安全评估具有重大的意义。但通过传统的非手性分析检测手段对手性农药评估得出的毒理学数据与实际生态毒理学效应有较大差异，导致手性农药的风险评估不准确，给人类健康和环境安全带来了潜在威胁。因此开展手性农药对映体水平的安全评价是对传统农药残留研究的补充，是深层次的农药安全评价研究[6]。