Journal: Chemical research in toxicology
Article Title: Development of a Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry Method for Analysis of Stable 4-Hydroxyequilenin-DNA Adducts in Human Breast Cancer Cells
Figure Lengend Snippet: Optimization of hydrolysis conditions for detecting stable 4-OHEN-DNA adducts. A) Method I, incubation with NP1 and ALP together in 25 mM ammonium acetate (pH 5.3) for 45 min at 37 °C. Method II, the DNA solutions were incubated with NP1 for 4 h in ammonium acetate at 55 °C, and then ALP and VPH were added to mixture for 4 h at 37 °C after pH adjustment to 9.8 by 0.1 M diethanolamine. B) Equal amounts of calf thymus DNA were hydrolyzed using Method I and Method II. The hydrolysates were analyzed by LC-MS/MS and compared by total relative intensity of each adduct peak.
Article Snippet: Alkaline phosphatase (ALP) and venom phosphodiesterase I (VPH) were purchased from Worthington (Lakewood, NJ).
Techniques: Incubation, ALP Assay, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry