Mass spectrometric detection and measurement of N2-(2'-deoxyguanosin-8-yl)PhIP adducts in DNA.
Capillary column gas chromatography-electron-capture mass spectrometry (GC-MS) and microbore liquid chromatography-positive ion electrospray mass spectrometry (LC-MS) have been used to measure the carcinogenic, food-derived, heterocyclic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) adducted at C-8 of deoxyguanosine in DNA. For GC-MS analysis, PhIP was released from adducted DNA by alkaline hydrolysis and analysed as the di(3,5-bistrifluoromethylbenzyl) derivative, while for LC-MS analysis, the nucleoside N2-(2'-deoxyguanosin-8-yl)PhIP was generated by enzymic digestion of DNA and analysed intact. A deuterated analogue of N2-(2'-deoxyguanosin-8-yl)PhIP was used as an internal standard in both assays, which each had a limit of quantification of 200 pg/500 microg DNA. The two methods were used to analyse DNA extracted from h1A2v2 cells and HCT116 cells that had been exposed to PhIP.
Crosbie
S J
SJ
Division of Medicine, Imperial College School of Medicine, Hammersmith Hospital, London, UK.
Murray
S
S
Boobis
A R
AR
Gooderham
N J
NJ
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D018736:DNA Adducts / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006801:Humans; D006868:Hydrolysis; D007093:Imidazoles / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods*; D008862:Microsomes, Liver / Q000378:metabolism; D013950:Thymus Gland / Q000737:chemistry; D014407:Tumor Cells, Cultured
D018736:DNA Adducts; D007093:Imidazoles; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine