title: On-line identification of diastereomeric dibenzo[a,l]pyrene diol epoxide-derived deoxyadenosine adducts by capillary electrophoresis-fluorescence line-narrowing and non-line narrowing spectroscopy.
abstract: A capillary electrophoretic method for the separation and on-line identification of closely related analytes using low-temperature fluorescence spectroscopy is reported for the eight diastereomeric deoxyadenosine (dA) adducts derived from dibenzo[a,l]pyrene diol epoxide (DB[a,l]PDE). Electrophoretic separation of stereoisomers was accomplished by application of a mixed surfactant buffer [dioctyl sulfosuccinate (DOSS) and Brij-S], which was below the critical micelle concentration (CMC) due to the high concentration (approximately 25%) of organic solvent. Addition of multiple surfactant additives to the separation buffer provided electrophoretic resolution, which was unattainable under single surfactant conditions. It is shown that the CE-separated analyte zones could be identified on-line via low-temperature (4.2 K) fluorescence non-line narrowing and fluorescence line-narrowing (FLN) spectroscopy. In addition, it was determined that in CE buffer trans-syn-,cis-syn- and cis-anti-DB[a,l]PDE-14-N6dA diastereomeric adducts exist mostly with the -dA and DB[a,l]P moiety in an "open"-type conformation while the trans-anti-DB[a,l]PDE-14-N6dA adducts exist in two different conformations whose relative distribution depends on matrix composition. The above conformations have also been revealed by selective laser excitation. Thus, the low-temperature methodology not only provides fingerprint structure via vibrationally resolved 4.2 K fluorescence spectra for adduct identification, but also provides conformational information on the spatial relationship of the carcinogen and dA moiety. These results, taken together with those for DB[a,l]P-DNA adducts formed in standard glasses and mouse epidermis exposed to DB[a,l]P, support our earlier findings that DB[a,l]P-derived adducts exist in different conformations [Jankowiak et al., Chem. Res. Toxicol. 11 (1998) 674]. Therefore, the combination of the separation power of CE and spectral selectivity of low-temperature fluorescence spectroscopy at NLN and FLN conditions provides a powerful methodology which should prove useful for identification of closely related DNA adducts formed at low levels in biological systems.
authors.affiliation: Ames Laboratory-US Department of Energy, Iowa Sate University 50011, USA.
authors.forename: K P
authors.initials: KP
authors.lastname: Roberts
authors.forename: C H
authors.initials: CH
authors.lastname: Lin
authors.forename: R
authors.initials: R
authors.lastname: Jankowiak
authors.forename: G J
authors.initials: GJ
authors.lastname: Small
mesh_terms: D001580:Benzopyrenes / Q000032:analysis*; D002021:Buffers; D002273:Carcinogens / Q000032:analysis; D018736:DNA Adducts / Q000032:analysis*; D003839:Deoxyadenosines / Q000032:analysis* / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D004852:Epoxy Compounds / Q000032:analysis*; D009862:Online Systems; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D013696:Temperature
chemical_list: D001580:Benzopyrenes; D002021:Buffers; D002273:Carcinogens; D018736:DNA Adducts; D003839:Deoxyadenosines; D004852:Epoxy Compounds; C121930:dibenzo(a,l)pyrene diol epoxide