Analysis of the malondialdehyde-2'-deoxyguanosine adduct pyrimidopurinone in human leukocyte DNA by gas chromatography/electron capture/negative chemical ionization/mass spectrometry.
A method is described for the assay of the major malondialdehyde-deoxyguanosine adduct (M1G) based on immunoaffinity purification and gas chromatography/electron capture/negative chemical ionization/mass spectrometry. A stable isotope of M1G-deoxyribose ([2H2]M1G-dR) was used as an internal standard. Recovery of internal standard throughout the entire assay procedure was approximately 40%. The assay showed a linear response over a range of 10-1000 pg of M1G-dR and was verified by analysis of a synthetic. M1G-containing oligomer. The limit of detection in biological samples was 100 fmol/sample, corresponding to 3 adducts/10(8) bases for 1 mg of DNA. DNA was isolated from the blood of 10 healthy human donors, and M1G levels were measured. A mean value of 6.2 +/- 1.2 adducts/10(8) bases was obtained, with no obvious differences bases on age or cigarette smoking. A small, but statistically significant difference was observed between the levels in females (5.1 +/- 0.4 adducts/10(8) bases) and males 6.7 +/- 1.1 adducts/10(8) bases). The presence of M1G in leukocyte DNA was further verified by analysis using liquid chromatography/electrospray ionization mass spectrometry.
Rouzer
C A
CA
A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Chaudhary
A K
AK
Nokubo
M
M
Ferguson
D M
DM
Reddy
G R
GR
Blair
I A
IA
Marnett
L J
LJ
D000328:Adult; D002138:Calibration; D004247:DNA / Q000097:blood*; D018736:DNA Adducts / Q000097:blood*; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000097:blood* / Q000302:isolation & purification; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000592:standards; D006801:Humans; D007962:Leukocytes / Q000737:chemistry*; D008297:Male; D008315:Malondialdehyde / Q000097:blood*; D008875:Middle Aged; D009841:Oligonucleotides / Q000737:chemistry
D018736:DNA Adducts; D009841:Oligonucleotides; D008315:Malondialdehyde; C089188:3-(2'-deoxyribofuranosyl)pyrimido(1,2-a)purin-10(3H)-one; D004247:DNA; D003849:Deoxyguanosine