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Measurement of Alkenyl/Epoxy DNA Adducts by GC-MS.

Bouve College of Pharmacy and Health Sciences, and Barnett Institute Northeastern University, Boston, Massachusetts 1994 Dec:5 pages
A final performance report on the development of the gas chromatography/electron capture/mass spectrometry (GC/EC/MS) measurement of DNA adducts from occupational exposure to ethylene (74851), ethylene-oxide (75218), butadiene (106990), styrene (100425), and propene (115071) was presented. Adduct measurement involved the isolation of DNA from a biological sample, separation of the DNA adducts, derivatization of the adducts with an electrophoric agent, and detection of products by GC/EC/MS. A procedure was initially developed to measure endogenous levels of the N7-guanine (N7-G)/DNA adduct of ethylene-oxide. This procedure has been applied to the detection of styrene, butadiene, benzene N7- G/DNA, and N7-methyl-guanine adducts. Several refinements in the procedures were reported. Use of all glass vials for derivatization eliminated trace contamination from Teflon liner and plastic caps. The use of two injectors for high performance liquid chromatography cleanup of adduct derivatives was found to alleviate carryover problems. Detection of the O6-guanine ethylene-oxide adduct required conversion of the exocyclic amino to fluoro. The t-butyl derivative of the aliphatic hydroxyl was determined to be optimal for GC/EC/MS detection, leading to the general guideline of polar footprint derivative minimalization. Interferences were overcome by the use of a new electrophoric reagent to shift the retention and mass of an analyte. Sample loss during solid phase extraction was found to be associated with the masking of analyte by solvent impurities. Conditions were developed for lowering the detection limit of the analytes to the zeptomole level as well as guidelines for operation of a GC/EC/MS at the attomole level. The author concludes that this research, in addition to the development of an analytical procedure capable of detecting endogenous levels of DNA adducts, also led to solutions for loss, contamination and interference problems associated with GC/EC/MS trace organic analysis.
NIOSH-Grant; Grants-other; DNA-adducts; Unsaturated-compounds; Mass-spectrometry; Analytical-methods;
Medicinal Chemistry Northeastern University 360 Huntington Avenue Boston, MA 02115
74-85-1; 75-21-8; 106-99-0; 100-42-5; 115-07-1;
Publication Date
Document Type
Final Grant Report;
Funding Amount
Funding Type
Fiscal Year
NTIS Accession No.
NTIS Price
Identifying No.
NIOSH Division
Priority Area
Other Occupational Concerns; Grants-other;
Source Name
Bouve College of Pharmacy and Health Sciences, and Barnett Institute Northeastern University, Boston, Massachusetts
Performing Organization
Northeastern University, Boston, Massachusetts