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Quantitation of lower levels of the DNA adduct of thymidylyl(3′‐5′)thymidine methyl phosphotriester by liquid chromatography/negative atmospheric pressure chemical ionization tandem mass spectrometry
Author(s) -
Zhang Fagen,
Bartels Michael J.,
Pottenger Lynn H.,
Gollapudi B. Bhaskar,
Schisler Melissa R.
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2933
Subject(s) - chemistry , atmospheric pressure chemical ionization , chromatography , adduct , mass spectrometry , tandem mass spectrometry , detection limit , chemical ionization , hydrolysate , liquid chromatography–mass spectrometry , metabolite , ionization , biochemistry , hydrolysis , organic chemistry , ion
A sensitive method has been developed for the direct quantitation of the methyl phosphotriester DNA adduct of thymidyl(3′‐5′)thymidine (dTp(Me)dT) from enzymatic DNA hydrolysates prepared from cultured cells treated with low levels of N ‐methyl‐ N ‐nitrosourea (MNU) and methyl methane sulfonate (MMS), by rapid and selective liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry (LC/APCI‐MS/MS). The lower limit of quantitation was 0.1 ng/mL (6.4 adducts per 10 8 nucleotides). Linearity of the calibration curve was greater than 0.999 from 0.1 to 50 ng/mL. Intra‐day precision for three levels of quality control samples ranged from 4.27 to 15.62%. Interday precision ranged from 2.46 to 11.95%. Using this method, the levels of dTp(Me)dT in DNA enzymatic hydrolysates obtained from a series of incubations of mouse lymphoma cells with low doses of MNU (50 µM) were quantified. Copyright © 2007 John Wiley & Sons, Ltd.