title: The analysis of DNA adducts: the transition from (32)P-postlabeling to mass spectrometry.
abstract: The technique of (32)P-postlabeling, which was introduced in 1982 for the analysis of DNA adducts, has long been the method of choice for in vivo studies because of its high sensitivity as it requires only <10μg DNA to achieve the detection of 1 adduct in 10(10) normal bases. (32)P-postlabeling has therefore been utilized in numerous human and animal studies of DNA adduct formation. Like all techniques (32)P-postlabeling does have several disadvantages including the use of radioactive phosphorus, lack of internal standards, and perhaps most significantly does not provide any structural information for positive identification of unknown adducts, a shortcoming that could significantly hamper progress in the field. Structural methods have since been developed to allow for positive identification of DNA adducts, but to this day, the same level of sensitivity and low sample requirements provided by (32)P-postlabeling have not been matched. In this mini review we will discuss the (32)P-postlabeling method and chronicle the transition to mass spectrometry via the hyphenation of gas chromatography, capillary electrophoresis, and ultimately liquid chromatography which, some 30years later, is only just starting to approach the sensitivity and low sample requirements of (32)P-postlabeling. This paper focuses on the detection of bulky carcinogen-DNA adducts, with no mention of oxidative damage or small alkylating agents. This is because the (32)P-postlabeling assay is most compatible with bulky DNA adducts. This will also allow a more comprehensive focus on a subject that has been our particular interest since 1990.
authors.affiliation: Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University, Boston, MA 02115, United States.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Klaene
authors.affiliation: Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University, Boston, MA 02115, United States.
authors.forename: Vaneet K
authors.initials: VK
authors.lastname: Sharma
authors.affiliation: Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University, Boston, MA 02115, United States.
authors.forename: James
authors.initials: J
authors.lastname: Glick
authors.affiliation: Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University, Boston, MA 02115, United States. Electronic address: p.vouros@neu.edu.
authors.forename: Paul
authors.initials: P
authors.lastname: Vouros
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D018736:DNA Adducts / Q000032:analysis* / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D010761:Phosphorus Radioisotopes; D012680:Sensitivity and Specificity
chemical_list: D018736:DNA Adducts; D010761:Phosphorus Radioisotopes