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通过同步荧光分光光度法检测苯并(a)芘二环氧物-DNA加合物。

Benzo(a)pyrene diolepoxide-DNA adducts detected by synchronous fluorescence spectrophotometry.

作者信息

Vahakangas K, Trivers G, Rowe M, Harris C C

出版信息

Environ Health Perspect. 1985 Oct;62:101-4. doi: 10.1289/ehp.8562101.

Abstract

Using benzo(a)pyrene (BP) as a model carcinogen we are currently applying a fluorescence technique to detect the very low levels of carcinogen-DNA adducts in human populations due to environmental exposure. In synchronous fluorescence spectrophotometry for detection of BP-diol epoxide-DNA, excitation and emission wavelengths are scanned simultaneously with a fixed wavelength difference (delta lambda) of 34 nm. Compared to conventional fluorescence methods only one peak emerges because excitation and emission peaks have to match delta lambda to show. Because of the quenching effect of DNA, samples are hydrolyzed by acid. After this, BP-diol epoxide (BPDE)- -modified DNA gives a peak at the same wavelength and of the same fluorescence yield as BP-tetrols. When DNA from peripheral blood lymphocytes of 44 coke oven workers were analyzed, 10 had a sharp peak at 379. Among 36 coke oven workers from another factory, 4 had detectable levels of adducts. A much smaller percentage of samples was positive in a group of aluminum plant workers. We have also found BPDE-DNA adducts in DNA from pulmonary alveolar macrophages and peripheral blood lymphocytes from tobacco smokers and some of the nonsmokers.

摘要

以苯并(a)芘(BP)作为模型致癌物,我们目前正在应用一种荧光技术来检测由于环境暴露导致的人群中极低水平的致癌物 - DNA加合物。在用于检测BP - 二醇环氧化物 - DNA的同步荧光分光光度法中,激发波长和发射波长以34 nm的固定波长差(Δλ)同时扫描。与传统荧光方法相比,只出现一个峰,因为激发峰和发射峰必须匹配Δλ才能显示出来。由于DNA的猝灭效应,样品用酸水解。在此之后,BP - 二醇环氧化物(BPDE)修饰的DNA在与BP - 四醇相同的波长处给出一个峰,且荧光产率相同。当分析44名炼焦炉工人外周血淋巴细胞的DNA时,10人在379处有一个尖锐峰。在另一家工厂的36名炼焦炉工人中,4人有加合物可检测水平。在一组铝厂工人中,阳性样本的比例要小得多。我们还在吸烟者和一些不吸烟者的肺泡巨噬细胞和外周血淋巴细胞的DNA中发现了BPDE - DNA加合物。

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