D'Ulivo Lucia, Feng Yong-Lai
Exposure and Biomonitoring Division, Environmental Health Science and Research Bureau, Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Canada.
PLoS One. 2016 Apr 7;11(4):e0153081. doi: 10.1371/journal.pone.0153081. eCollection 2016.
The interaction of chemicals with DNA may lead to genotoxicity, mutation or carcinogenicity. A simple open tubular capillary electrochromatographic method is proposed to rapidly assess the interaction affinity of three environmental contaminants (1,4-phenylenediamine, pyridine and 2,4-diaminotoluene) to DNA by measuring their retention in the capillaries coated with DNA probes. DNA oligonucleotide probes were immobilized on the inner wall of a fused silica capillary that was first derivatized with 3-(aminopropyl)-triethoxysilane (APTES). The difference in retention times and factors was considered as the difference in interaction affinity of the contaminants to the DNA probes. The interaction of the contaminants with both double-stranded (dsDNA) and single-stranded DNA (ssDNA) coatings was compared. Retention factors of 1,4-phenylenediamine, pyridine and 2,4-diaminotoluene in the capillary coated with ssDNA probe were 0.29, 0.42, and 0.44, respectively. A similar trend was observed in the capillary coated with dsDNA, indicating that 2,4-diaminotoluene has the highest affinity among the three contaminants. The relative standard deviation (RSD) for the retention factors was in the range of 0.05-0.69% (n = 3). The results demonstrated that the developed technique could be applied for preliminary screening purpose to provide DNA interaction affinity information of various environmental contaminants.
化学物质与DNA的相互作用可能导致遗传毒性、突变或致癌性。本文提出了一种简单的开管毛细管电色谱方法，通过测量三种环境污染物（1,4-苯二胺、吡啶和2,4-二氨基甲苯）在涂有DNA探针的毛细管中的保留情况，快速评估它们与DNA的相互作用亲和力。DNA寡核苷酸探针固定在经3-(氨丙基)-三乙氧基硅烷（APTES）衍生化的熔融石英毛细管内壁上。保留时间和因子的差异被视为污染物与DNA探针相互作用亲和力的差异。比较了污染物与双链（dsDNA）和单链DNA（ssDNA）涂层的相互作用。在涂有ssDNA探针的毛细管中，1,4-苯二胺、吡啶和2,4-二氨基甲苯的保留因子分别为0.29、0.42和0.44。在涂有dsDNA的毛细管中也观察到了类似的趋势，表明在这三种污染物中，2,4-二氨基甲苯具有最高的亲和力。保留因子的相对标准偏差（RSD）在0.05 - 0.69%范围内（n = 3）。结果表明，所开发的技术可用于初步筛选目的，以提供各种环境污染物的DNA相互作用亲和力信息。
