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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 30, Issue 3, April 2006,
pp. 178-186
Quantitation of Cotinine in
Nonsmoker Saliva Using Chip-Based Nanoelectrospray Tandem Mass Spectrometry
Bruce A. Tomkins[1,2], Gary J. Van Berkel[1,2], Roger
A. Jenkins[2], and Richard W. Counts[3]
[1]Organic and Biological Mass Spectrometry Group,
[2]Chemical Sciences Division,
[3]Statistics and Data Sciences Group, Computer Science and Mathematics Division,
Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831-6120
A new analytical procedure was developed for the quantitation
of nonsmoker salivary cotinine. Small volumes of saliva were diluted with water,
fortified with cotinine-d3 (internal standard), then passed through small extraction
columns. The analyte and internal standard were eluted with 0.1% (v/v) acetic
acid/acetonitrile. Aliquots of each extract were analyzed directly, without
chromatographic separation, using chip-based (NanoMate™) nanospray tandem
mass spectrometry. The calculated detection limit was 0.49 ng cotinine/mL saliva.
This method was used to quantify salivary cotinine collected from nonsmoking
human subjects living in one of three environmental tobacco smoke (ETS) exposure
categories or “cells”: 1. smoking home/smoking workplace; 2. smoking
home/nonsmoking workplace; and 3. nonsmoking home/smoking workplace. Samples
were collected during five sequential days, including Saturday, as part of a
larger study to evaluate potential variability in exposure to ETS. Salivary
cotinine measurements were made for the purpose of excluding misclassified smokers
and for comparison with known levels of exposure to airborne nicotine in each
exposure category. The concentrations observed were consistent with those reported
from other large studies reported elsewhere. A non-parametric statistical test
was applied to the data within each cell. No statistically significant differences
were found between the mean cotinine concentrations collected on a weekday as
compared to those collected on a weekend day. When the non-parametric test was
applied to the three cells, a statistically significant difference was observed
between cell 1 compared to cells 2 and 3. The salivary cotinine concentrations
were thus statistically invariant over a five-day exposure period, and they
were greatest under the conditions of smoking home and smoking workplace.
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