SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

M45
DETECTION OF COTININE IN EXHALED BREATH BY GAS CHROMATOGRAPHY-MASS
SPECTROMETRY: A PRELIMINARY STUDY
3
Michele L. Merves J *, Chris W. Chronister l , Bruce A. Goldberger l • , Richard J. Melker and Mark S. Gold 3 • 4 :
University of Florida College of Medicine, IDepartment of Pathology, Immunology and Laboratory Medicine;
2Department of Anesthesiology; JDepartment of Psychiatry, Division of Addiction Medicine; 4Departrnents of
Neuroscience, Community Health and Family Medicine; 'Gainesville, FL; 4800 S.W. 35 th Street, Gainesville,
FL 32608.
Exhaled breath analysis is a very useful diagnostic tool in the fields of medicine and forensic science. Exhaled
breath is routinely used to detect markers of disease, as well as correlate ethanol concentrations with effects on
human performance and behavior (DUI). Compared to other biological specimens, the collection of exhaled
breath is simple and non-invasive, with minimal exposure to infectious material. Because exhaled breath is a
physiological waste product, the quantity available exceeds those of other biological matrices such as blood and
urine. In addition, little sample preparation is required. Finally, exhaled breath accounts for all routes of drug
exposure including oral ingestion, injection, inhalation, and dermal absorption.
This is a preliminary study to determine if cotinine, an oxidative metabolite of nicotine, is detectable in exhaled
breath of humans. Cotinine was chosen as a metabolic marker of nicotine use because of its long half-life
compared to nicotine. Exhaled breath specimens were collected onto commercially-available XAD-4 traps
(Supelco) and analyzed by gas chromatography-mass spectrometry (GC-MS). The resin, XAD-4, is a polymeric
adsorbent available as beads within a small glass tube (70 mm x 6 mm). Upon collection of the exhaled breath,
the resin was removed, fortified with drcotinine (Cerilliant Corporation), and sonicated in methanol for 30
minutes. The organic layer was separated from the resin and dried under a gentle stream of nitrogen at 40°C. The
residue was reconstituted in phosphate buffer (PH 6). The cotinine was isolated from the buffer utilizing
CleanScreen® solid-phase· extraction (SPE) cartridges (ZSDAU020) manufactured by United Chemical
Technologies, Inc. Following elution from the SPE cartridges using a solution ofmethylene chloride, isopropanol,
and ammonium hydroxide, the extracts were dried under a gentle stream of nitrogen at 40°C and reconstituted in
methanol. The extracts were subjected to GC-MS analysis using an Agilent Technologies 6890N series gas
chromatograph equipped with a Agilent Technologies 5973 network mass spectrometer operated in electron
ionization and selected ion monitoring (SIM) modes. Helium was used as the carrier gas at a constant flow rate of
1.0 mLimin. Automated injections (2-3 ).iL) were made onto a capillary GC column. The ions (mlz) monitored
for cotinine were 98.1, 175.1 and 176.1, and the ions (mlz) monitored for d 3 -cotinine were lOLl and 179.1.
Initial studies have demonstrated cotinine in the exhaled breath of cigarette smokers, while cotinine was not
detected in control (non-smokers) exhaled breath. These data will serve as a model for detection of other
compounds (drugs and their metabolites) present in exhaled breath which may be beneficial for many other
medical, forensic, and toxicological applications.
Keywords: Cotinine, Exhaled Breath, GC-MS
Page 328