PMID- 9860490
OWN - NLM
STAT- MEDLINE
DCOM- 19990223
LR  - 20131121
IS  - 0893-228X (Print)
IS  - 0893-228X (Linking)
VI  - 11
IP  - 12
DP  - 1998 Dec
TI  - DNA adducts of 2,3-epoxy-4-hydroxynonanal: detection of 7-(1', 
      2'-dihydroxyheptyl)-3H-imidazo[2,1-i]purine and 1,N6-ethenoadenine by gas 
      chromatography/negative ion chemical ionization/mass spectrometry.
PG  - 1474-80
AB  - 2,3-Epoxy-4-hydroxynonanal (EH) is a bifunctional aldehyde formed by epoxidation 
      of trans-4-hydroxy-2-nonenal, a peroxidation product of omega-6 polyunsaturated 
      fatty acids. EH is mutagenic and tumorigenic and capable of modifying DNA bases 
      forming etheno adducts in vitro. Recent studies showed that etheno adducts are 
      present in tissue DNA of humans and untreated rodents, suggesting a potential 
      endogenous role of EH in their formation. A sensitive assay is needed so we can 
      determine whether EH is involved in etheno adduct formation in vivo and study the 
      biological significance of the etheno adducts in DNA. In this study, we developed 
      a gas chromatography/negative ion chemical ionization/mass spectrometry assay for 
      the analysis of 1, N6-ethenoadenine (epsilonAde) and 7-(1', 
      2'-dihydroxyheptyl)-3H-imidazo[2,1-i]purine (DHH-epsilonAde) in DNA; both are 
      products from the reaction of adenine with EH. The assay entails the following 
      sequence of steps: (1) addition of [15N5]epsilonAde and [15N5]DHH-epsilonAde to 
      DNA as internal standards, (2) acid hydrolysis of DNA, (3) adduct enrichment by 
      C18 solid phase extraction (SPE), (4) derivatization by pentafluorobenzylation 
      (PFB), (5) separation of PFB-epsilonAde and PFB-DHH-epsilonAde on a Si SPE 
      column, (6) acetonide (ACT) formation of PFB-DHH-epsilonAde, and (7) GC/MS 
      analysis with selective ion monitoring (SIM). The limit of detection by on-column 
      injection for PFB-epsilonAde monitoring of the (M - PFB)- ion at m/z 158 was 30 
      amol and for ACT-PFB-DHH-epsilonAde monitoring of the (M - PFB)- ion at m/z 328 
      was 0.4 fmol; the detection limits for the entire assay were 6.3 fmol for 
      epsilonAde and 36 fmol for DHH-epsilonAde. In calf thymus DNA modified with EH at 
      37 degreesC for 50 h, both epsilonAde and DHH-epsilonAde were detected at high 
      levels by this method, 4.5 +/- 0.7 and 90.8 +/- 8.7 adducts/10(3) adenine, 
      respectively. These levels were also verified by HPLC fluorescence analysis, 
      indicating that EH extensively reacts with adenine in DNA, forming etheno 
      adducts. The high sensitivity of the assay suggests that it may be used in the 
      analysis of ethenoadenine adducts in vivo.
FAU - Chen, H J
AU  - Chen HJ
AD  - American Health Foundation, Valhalla, New York 10595, USA. chehjc@ccu.edu.tw
FAU - Zhang, L
AU  - Zhang L
FAU - Cox, J
AU  - Cox J
FAU - Cunningham, J A
AU  - Cunningham JA
FAU - Chung, F L
AU  - Chung FL
LA  - eng
GR  - CA 43159/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Chem Res Toxicol
JT  - Chemical research in toxicology
JID - 8807448
RN  - 0 (7-(1',2'-dihydroxyheptyl)-3h-imidazo(2,1-i)purine)
RN  - 0 (Aldehydes)
RN  - 0 (DNA Adducts)
RN  - 0 (Epoxy Compounds)
RN  - 0 (Imidazoles)
RN  - 0 (Mutagens)
RN  - 0 (Purines)
RN  - 133398-57-9 (2,3-epoxy-4-hydroxynonanal)
RN  - 13875-63-3 (1,N(6)-ethenoadenine)
RN  - JAC85A2161 (Adenine)
SB  - IM
MH  - Adenine/analogs & derivatives/chemistry
MH  - Aldehydes/*chemistry
MH  - Animals
MH  - Calibration
MH  - Chromatography, High Pressure Liquid
MH  - DNA Adducts/*drug effects
MH  - Epoxy Compounds/*chemistry
MH  - Gas Chromatography-Mass Spectrometry
MH  - Imidazoles/*analysis
MH  - Liver/chemistry/drug effects
MH  - Magnetic Resonance Spectroscopy
MH  - Mutagens/chemistry
MH  - Purines/*analysis
MH  - Rats
MH  - Spectrometry, Fluorescence
EDAT- 1998/12/22 00:00
MHDA- 1998/12/22 00:01
CRDT- 1998/12/22 00:00
PHST- 1998/12/22 00:00 [pubmed]
PHST- 1998/12/22 00:01 [medline]
PHST- 1998/12/22 00:00 [entrez]
AID - tx980107o [pii]
AID - 10.1021/tx980107o [doi]
PST - ppublish
SO  - Chem Res Toxicol. 1998 Dec;11(12):1474-80. doi: 10.1021/tx980107o.