大鼠BPDE-DNA加合物生成及其高效液相色谱测定  

The Formation of BPDE-DNA Adduct in Rat and Its Determination by High Performance Liquid Chromatography

作　　者：杨柳桦[1] 刘渠[2] 孙成均[1] 郭淑妍[2] 林少杰[2] 

机构地区：[1]四川大学华西公共卫生学院卫生检验教研室,成都610041 [2]深圳市龙岗区疾病预防控制中心

出　　处：《四川大学学报（医学版）》2008年第2期302-304,共3页Journal of Sichuan University(Medical Sciences)

基　　金：深圳市科技和信息局立项课题(JH200507120937A)资助

摘　　要：目的研究苯并(a)芘(BaP)诱导大鼠产生(7R,8S)-二羟基-(9S,10R)-环氧-7,8,9,10-四氢苯并(a)芘(BPDE)-DNA加合物的生成。建立以血液为样品的检测染毒大鼠BPDE-DNA加合物的高效液相色谱法。方法选用清洁级SD大鼠,一次性腹腔注射苯并(a)芘二甲基亚砜溶液(以1%羧甲基纤维素钠为助溶剂)100 mg/kg,5 h后取股静脉血。提取抗凝全血中的DNA,采用琼脂糖凝胶电泳确认DNA的提取效果。将提取的DNA在0.1 nmol/L HCl、90℃恒温水浴箱中酸水解4 h,乙酸乙酯提取酸水解产物——四醇-苯并(a)芘,高效液相色谱法检测,最后用高效液相色谱-质谱法确认。结果BaP染毒组与溶剂对照组和阴性对照组比较,高效液相色谱检测有新的色谱峰产生,质谱确定其相对分子质量与四醇-苯并(a)芘一致。结论本实验的染毒方法能使大鼠产生BPDE-DNA加合物;建立的高效液相色谱法可以血液为样品检测BPDE-DNA加合物。Objective To study the benzo （a） pyrene （BaP） induced DNA adduct in rat and establish a method to measure the DNA adduct in blood by high performance liquid chromatography （HPLC）. Methods The SD rats were treated with 100 mg/kg of BaP-DMSO （cosolvent： 1% sodium carboxymethyl cellulose） once by i.p., and the blood of femoral vein was collected 5 hours later. The blood DNA was extracted by kit and confirmed by agarose gel electrophoresis. After extraction, the DNA adducts were hydrolyzed in 0. 1 nmol/L HCl at 90 ℃ for 4 hours. The acid-hydrolysis products （BP-tetrols） of DNA adducts were extracted by ethyl acetate and measured by HPLC, and finally confirmed by HPLC-MS. Results In chromatogram there were new peaks to occur for rats treated by BaP, with compared to control. By HPLC-MS, one of the new peaks was confirmed to be BP-tetrol. Conclusion The rats ingesting BaP in the way described in this experiment can form DNA adducts, and the adducts in blood can be detected by HPLC.

关 键 词：(7R 8S)-二羟基-(9S 10R)-环氧-7 8 9 10-四氢苯并(a)芘 苯并(A)芘 DNA加合物 四醇-苯 并(a)芘 高效液相色谱法(HPLC) 液相色谱-质谱(HPLC/MS) 

分 类 号：R115[医药卫生—公共卫生与预防医学]