双壳类水产品中(+)-anti-BPDE-DNA加合物HPLC/FD检测  

Determination of(+)-anti-benzo[a]pyrene diol-epoxide-DNA adducts metabolited from benzo[a]pyrene in bivalves by high-performance liquid chromatography with fluorescence detection

作　　者：齐琳[1] 刘晓晨[1] 李健[2] 杨波[1] 孙文平[1] 于腾[1] 李发胜[1] 刘辉[1] 

机构地区：[1]大连医科大学检验医学院,辽宁大连116044 [2]大连医科大学附属第一医院检验科

出　　处：《中国公共卫生》2014年第3期371-373,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金资助项目(30770724)

摘　　要：目的建立检测双壳类水产品中（＋）-反-7，8-二羟基-9，10-环氧苯并（a）芘（BPDE）-DNA加合物的高效液相色谱荧光法（HPLC／FD）。方法用组织基因组DNA提取试剂盒提取6种双壳类水产品组织中的DNA，0．1mol／LHCl、90℃酸解4h，乙酸乙酯充分萃取酸解产物——苯并（a）芘一四醇。以CenturySILC18-BDS色谱柱（150mmX4．6mm，5Ixm）分离；洗脱流动相：V（甲醇）／V（水）=55／45；流速：1．0mL／min；进样量：20μL；激发波长：265nm，发射波长：395nm；荧光检测苯并（a）芘-四醇的含量。结果该方法的检测限为0．3ng／mL，在0．5～100ng／mL范围内呈良好的线性关系（，=0．9960）；日内相对标准偏差（RSD）为2．8％～4．2％，日间RSD为3．2％-5．8％；双壳类水产品中（＋）-anti—BPDE—DNA含量为13．44～152．7μg／kg，RSD为3．0％-6．5％．加标回收率为86．9％-91．6％，RSD为3．1％～7．3％。结论该法简便、快速、灵敏度高，可用于双壳类水产品中（＋）-anti—BPDE—DNA加合物的检测。Objective To develop a simple and highly sensitive high-performance liquid chromatography/fluores- cence method for the determination of benzo[ a] pyrene（BaP）-tetrols released after acid hydrolysis of（ ＋ ）-anti- benzo [ a] pyrene diol-epoxide （BPDE）-DNA adducts metabolited from benzo [ a] pyrene in bivalves. Methods The tissue DNA in bivalves were extracted by reagent kit and the（ ＋ ） -anti-BPDE-DNA adducts were hydrolyzed in 0.1 mol/L HCI at 90℃ for 4 hours. The acid-hydrolysis products（BaP-tetrols） of DNA adducts were extracted by ethylacetate and meas- ured by high-performance liquid chromatography/fluorescence detection. Samples were separated on CenturySIL C18- BDS column （ 150 mm x 4.6 mm, 5 μm） and the elution was isocratic using a 55 % methanol ： 45 % water （ v/v ） mobile phase at a flow rate of 1.0 mL/min; fluorescence detection was conducted at 265 nm（excitation） and 395 nm（ emis- sion）, with the injection volume of 20 μL. Results The operating linear range was at least 0. 5 - 100 ng/mL （ r2 = 0. 9960 ） and the detection limits（ S/N = 3 ） was 0. 3 ng mL ^- 1. Coefficients of variation for within-run and between- run assays were 2. 8% - 4. 2% and 3.2% - 5.8%, respectively. The content of（ ＋ ） - anti-BPDE-DNA adducts were 13.44 - 152. 7 μg/kg and the relative standard deviations （RSDs） were 3.0% - 6. 5%. The average recoveries were 86. 9% -91.6% and the RSDs were 3. 1% -7.3 %, respectively. Conclusion The method was proved to be sensitive, accurate, rapid and could be applied to the determination of（ ＋ ）-anti-BPDE-DNA adducts in bivalves.

关 键 词：高效液相色谱荧光法 (+)-反-二羟基-9 10-环氧苯并(a)芘-DNA加合物 苯并(a)芘-四醇 双壳类 

分 类 号：R115[医药卫生—公共卫生与预防医学]