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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,0.1 mol/L HCl、90℃酸解4 h,乙酸乙酯充分萃取酸解产物———苯并(a)芘-四醇。以CenturySIL C18-BDS色谱柱(150 mm×4.6 mm,5μm)分离;洗脱流动相:V(甲醇)/V(水)=55/45;流速:1.0 mL/min;进样量:20μL;激发波长:265 nm,发射波长:395 nm;荧光检测苯并(a)芘-四醇的含量。结果该方法的检测限为0.3 ng/mL,在0.5~100 ng/mL范围内呈良好的线性关系(r2=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 establish a HPLC method for the determination of (+) - trans-7,8-dihydroxy-9,10-epoxybenzo [a] pyrene (BPDE) -DNA adduct in bivalve aquatic products HPLC / FD). Methods Tissue genomic DNA extraction kit was used to extract the DNA of 6 kinds of bivalve aquatic tissues, 0.1 mol / L HCl and acid hydrolysis at 90 ℃ for 4 h. The acid hydrolyzate - benzo (a) Pyrene-tetrol. The separation was performed on a CenturySIL C18-BDS column (150 mm × 4.6 mm, 5 μm); the elution mobile phase was V (methanol) / V (water) = 55/45; the flow rate was 1.0 mL / Excitation wavelength: 265 nm, emission wavelength: 395 nm; Fluorescence detection of benzo (a) pyrene-tetraol content. Results The limit of detection (RSD) was 0.3 ng / mL and the linear range was 0.5-100 ng / mL (r2 = 0.9960). The relative standard deviations (RSDs) were 2.8% -4.2% (+) - anti-BPDE-DNA in bivalve aquatic products ranged from 13.44 to 152.7 μg / kg with a RSD of 3.0% to 6.5%. The recoveries of spiked recoveries ranged from 86.9% to 91.6% From 3.1% to 7.3%. Conclusion The method is simple, rapid and sensitive and can be used for the detection of (+) - anti-BPDE-DNA adduct in bivalve aquatic products.