Fluorescence line narrowing-nonphotochemical hole burning spectrometry: femtomole detection and high selectivity for intact DNA-PAH adducts.
A new fluorescence line narrowing (FLN) apparatus is described and evaluated through experiments on intact DNA-PAH (polycyclic aromatic hydrocarbon) and globin-PAH adducts, as well as polar PAH metabolites. A detection limit of approximately 3 modified bases in 10(8) for a DNA adduct formed with a diol-epoxide of benzo[a]pyrene (BPDE-DNA) is reported for 20 micrograms of DNA at a spectral resolution of approximately 8 cm-1. The methodology employed avoids or minimizes spectral degradation and loss of sensitivity due to photooxidation and nonphotochemical hole burning (NPHB). A new double selection technique that employs both FLN and NPHB is described and found to lead to a significant improvement in selectivity over that obtained with conventional FLN.
Jankowiak
R
R
Ames Laboratory--USDOE and Department of Chemistry, Iowa State University, Ames 50011.
Cooper
R S
RS
Zamzow
D
D
Small
G J
GJ
Doskocil
G
G
Jeffrey
A M
AM
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000032:analysis*; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D018736:DNA Adducts; D008832:Microchemistry; D011083:Polycyclic Compounds / Q000032:analysis / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods
D018736:DNA Adducts; D011083:Polycyclic Compounds; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D004247:DNA