Savela K, King L, Gallagher J, Lewtas J
US EPA, Research Triangle Park, NC 27711, USA.
Carcinogenesis. 1995 Sep;16(9):2083-9. doi: 10.1093/carcin/16.9.2083.
Diesel exhaust extracts contain many carcinogenic compounds which have been shown to form polycyclic aromatic hydrocarbon (PAH)- and nitrated PAH-DNA adducts in rodent skin and lung. The aim of this study was to characterize by 32P-postlabeling, TLC and HPLC the primary postlabeled PAH-DNA adduct(s) formed in vitro and in vivo by diesel extracts. The diesel particle extracts had known concentrations of benzo[a]pyrene, benzo[b,j,k]-fluoranthenes (B[b,j,k]F) and chrysene. DNA adducts were analyzed in calf thymus DNA incubated in vitro with PAHs activated by S9 mix and in skin and lung DNA from topically treated mice. The main diesel-derived DNA adduct formed in vitro and in vivo did not co-migrate on HPLC and large TLC plates with (+/-)-r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti BPDE)-, B[b]F-,B[j]F-,B[k]F-or chrysene-DNA adduct standards. By co-chromatography DNA adducts formed by chrysene from both in vitro and in vivo samples were identified. Nissan diesel extract containing higher PAH concentrations than Volkswagen automobile extract formed skin DNA adducts that co-migrated with chrysene- and anti BPDE- DNA-derived adducts. We conclude that the use of a highly sensitive 32P-postlabeling method combined with HPLC improves the identification of PAH adducts formed by complex mixtures such as diesel exhaust extracts.
柴油废气提取物含有许多致癌化合物，这些化合物已被证明可在啮齿动物的皮肤和肺部形成多环芳烃（PAH）和硝化PAH-DNA加合物。本研究的目的是通过32P后标记、薄层色谱法（TLC）和高效液相色谱法（HPLC）来表征柴油提取物在体外和体内形成的主要后标记PAH-DNA加合物。柴油颗粒提取物含有已知浓度的苯并[a]芘、苯并[b,j,k]荧蒽（B[b,j,k]F）和屈。对在体外与经S9混合物激活的多环芳烃一起孵育的小牛胸腺DNA以及经局部处理的小鼠的皮肤和肺部DNA中的DNA加合物进行了分析。在体外和体内形成的主要柴油衍生DNA加合物在HPLC和大型TLC板上与（+/-）-r-7,t-8-二羟基-t-9,10-环氧-7,8,9,10-四氢苯并[a]芘（反式BPDE）、B[b]F、B[j]F、B[k]F或屈-DNA加合物标准品的迁移情况不同。通过共色谱法鉴定了体外和体内样品中由屈形成的DNA加合物。日产柴油提取物中多环芳烃浓度高于大众汽车提取物，其形成的皮肤DNA加合物与屈和反式BPDE衍生的DNA加合物迁移情况相同。我们得出结论，使用高灵敏度的32P后标记方法结合HPLC可提高对由柴油废气提取物等复杂混合物形成的PAH加合物的鉴定能力。
