苯并(a)芘(B(a)P)被认为是高活性致癌剂,在体内可转化为二氢二醇环氧苯并(a)芘(BPDE),具有直接致癌性。采用HPLC-荧光检测技术建立BPDE纯品及细胞裂解液中BPDE的测定。以HypersilC18(250mm×46mm,10μm)为色谱柱配C。8保护柱;以甲醇和水(体积分数为90:10)为流动相,流速为1.0mL/min;柱温为室温;检测波长:激发波长为349nm,发射波长为387nm;进样量为20μL;跑样时间为10min。结果:BPDE浓度在0.01~1.0μg/mL范围内线性关系良好,其线形回归方程为Y=424298X+24598,相关系数为0.9921,最低检测限为0.005μg/mL,日内和日间精密度分别为3.2%~4.6%和5.9%~7.4%。HPLC荧光检测方法由于具有较高的灵敏度,使得BPDE标准品及其和细胞裂解液的混合溶液中的检测限都比紫外检测方法有所提高,方法操作简便,结果准确可靠。
Objective BPDE in DMSO solution cell lysate was detected by using high performance liquid chroma- tography with fluorescence detector (HPLC-FD). Method HPLC was operated with HypersilC18 column (250 min×46 ram,10 μm) at room temperature as the fixed phase,mixture of methanol and water (90 : 10) as the mobile phase. The flow rate of mobile phase was controlled at 1.0 mL/min. The wave lengths of excitation and emission were set at 349 nm and 387 nm, respectively. The result is that a good linear correlation between the concentrations of BPDE in Cell lysate and its peak areas was obtained at the range of 0.01--1.00 μg/mL. The equation of linear regression of BPDE in cell lysate is y=424298x+24598, and the correlation coefficient is 0. 9921. The lowest detection limit is 0. 005μg/mL. The intra-assay and inter assay relative standard deviations were 3.2--4.60% and 5.9--7.4%, respectively. It is concluded that HPLC FD detection is an effective way to the analysis of BPDE in the metabolism of BaP.