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针对两种甲醛-DNA加合物,N~6-羟甲基腺嘌呤(N~6-HOMe-d A)和N2-羟甲基鸟嘌呤(N_2-HOMe-d G),本实验室建立了基于液相色谱串联质谱的分析方法,以便从DNA分子水平上探讨甲醛暴露对DNA的损伤作用机理,其中[~(15)N_5]N~6-Me-d A和[~(13)C_(10)~(15)N_5]N_2-Me-d G为内标;液相分离所用色谱柱为Thermo Polar Advantage II C_(18);方法的精密度较好(RSD5%)、灵敏度较高(检出限分别为0.014和0.0064 ng·m L~(-1))、回收率良好(94.9%~111%).后用小牛胸腺DNA进行体外染毒实验,设置甲醛染毒的浓度梯度(0.001,0.01,0.05,0.1,0.5和1 mmol·L~(-1))和染毒时间组(0、2、4、8、12、16、24 h),利用建立的LC-MS/MS方法检测了DNA中N~6-HOMe-d A和N_2-HOMe-d G的含量.结果表明,两种甲醛-DNA加合物N~6-HOMe-d A和N2-HOMe-d G的量与甲醛暴露存在剂量-反应和时间-反应关系. 相似文献
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The atmospheric polychlorinated dibenzo-p-dioxins (PCDDs) partition appreciably in the gas phase, where they undergo rapid oxidation. The atmospheric oxidation mechanisms of a few PCDDs, initiated by OH radical, are studied using density functional theory calculations. The oxidations start with OH-addition to the aromatic rings, dominantly at γ-sites, followed by the non-chlorinated β-sites; while additions to the α-sites or chlorinated sites are negligible. For PCDDs with all β-sites being chlorinated, formation of PCDD-γ-OH adducts become virtually the only reaction path. Under the atmospheric conditions, the PCDD-β/γ-OH adducts combine with O2 slowly at rates <1 s−1. Instead, the PCDD-β-OH adducts will react with O2 through hydrogen abstraction at rates <50 s−1, forming PCDD-β-ol, and the PCDD-γ-OH adducts will decompose to the substituted phenoxy radicals by fused-ring C-O bond cleavage at rates of 103 ∼ 105 s−1. The reaction mechanisms of PCDDs are drastically different from the peroxy mechanism for the atmospheric oxidations of benzene and dibenzofuran. 相似文献
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综述了环境中常见的7种醛类污染物与DNA形成加合物的结构研究现状,包括单核苷酸-醛试管反应、DBA-醛试管反应、细胞培养、动物体以及人体内所产生的加合物结构的研究方法与结论,并对加合物结构研究的难点和发展方向进行了探讨。 相似文献
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Adeline Tarantini Marie-Blanche Personnaz Jean-Luc Besombes Jean-Luc Jafrezzo 《毒物与环境化学》2013,95(5):941-954
Particulate organic matter (PM) present in the atmosphere is a complex mixture of chemicals like polycyclic aromatic hydrocarbons (PAH) that may exert adverse health effects including respiratory and cardiovascular disturbances and cancer. In this study, airborne samples from environmental or industrial areas exhibiting different physicochemical composition were compared for their capacities to induce DNA damage in human hepatocytes HepG2. DNA strand breaks and DNA adducts formed by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), the most reactive metabolite of the carcinogenic benzo[a]pyrene (B[a]P), were measured with the comet assay and by high-performance liquid chromatography (HPLC)/mass spectrometry, respectively. Cells were exposed to organic matter extracted from PM. Experiments were performed either at a constant concentration of B[a]P or at concentrations corresponding to fixed air volumes. Results show that industrial extracts tend to produce more benzo[a]pyrene diol epoxide-N 2-2′-deoxyguanosine (BPDE-N 2-dGuo) DNA adducts than strand breaks, whereas the opposite was observed with environmental extracts. The chemical composition of the extracts significantly impacts the nature and levels of DNA damage. The amount of B[a]P and interaction with other contaminants in the extracts need to be considered to explain the formation of DNA damage. These results emphasize the use of in vitro tests as promising and complementary tools to widely used toxic equivalent factor (TEF) approach in order to assess health hazards related to chemical exposure of the general population. 相似文献
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用~(32)P后标记方法,对一组22例接触苯乙烯的工人和8例未接触苯乙烯(空白)的工人进行测试,其静脉血中的淋巴细胞有4例(高浓度,大于40ppm)发现DNA-环氧苯乙烯加合物;而另外接触高浓度苯乙烯、低浓度苯乙烯和空白试样中,均未发现。在已测出的4例中,加合物形成水平为0.07—3.38加合物/10~7正常核酸,DNA的总损伤水平为5—9加合物/10~7正常核酸。 用小牛胸腺DNA、四种脱氧核苷3′-单磷酸分别与环氧苯乙烯进行体外反应,然后用~(32)P后标记法测定产物中的DNA加合物,初步确证有六种加合物及其衍生物存在,修饰位置是鸟苷碱基。并确证了其中三种加合物的化学结构,另外几种加合物的结构有待进一步鉴定。 ~(32)P后标记法测DNA加合物,灵敏度高达1加合物/10~(10)正常核酸,近于人体二倍体细胞基因水平(1.2×10~(10)碱基)。 相似文献
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^32P后标记法测DNA加合物 总被引:5,自引:2,他引:3
^32P后标记方法测DNA-致癌物的加合物过程是用小球菌核酸酶和脾核酸酶将DNA及加合物酶解成2′-脱氧核苷3′-单磷酸(3′-dNmP+3′-dxmP),以它为底物,用T4多核苷酸激酶催化,使γ^32PATP的放射^32P转移反应到底物上,形成2′-脱氧核苷3′5′.dNDP+3′5′*dxDP),用ODS-TLC和PEI-TLC结合将标记过的二磷酸分离分辨,用自显影制成加合物的指纹图,液闪计数 相似文献
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应用紫外及荧光光谱法研究三种醛类污染物对牛腺DNA的损伤作用 总被引:4,自引:0,他引:4
应用紫外光谱移动法和溴乙锭荧光法对三种醛类化合物与牛腺DNA的加合以及它们引起的DNA-DNA交联进行了检测。结果表明,甲醛和乙醛能够与DNA形成共价加合物,与四种脱氧核苷加合顺序分别为dG〉dC〉dA〉dT和dA〉dG〉dC〉dT,丙烯醛与DNA的加合不能用紫外光谱移动法测定。甲醛能够显著地引起DNA-DNA的交联;乙醛也有这种作用,但不显著;丙烯醛的交联作用未测出,可能是由于羰基碳正电性较弱, 相似文献
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Low levels (ng/g) of musk ketone (MK),used as a fragrance additive in the formulation of personal care products,are frequently detected in the water and other environment.Thus,aquatic organisms can be continuously exposed to MK.In this study,kinetics and dose-response assessments of 2-amino-MK (AMK) metabolite,bound to cysteine-hemoglobin (Hb) in rainbow trout,formed by enzymatic nitro-reduction of MK have been demonstrated.Trout were exposed to a single exposure of 0.010,0.030,0.10,and 0.30 mg MK/g fish... 相似文献