Automated Solid Phase Extraction and Polarity-Switching
Tandem Mass Spectrometry Technique for High Throughput Analysis of
Urine Biomarkers for 14 Tobacco-related Compounds
posted on 2021-11-12, 18:42authored bySujeewa C. Piyankarage, Ernest McGahee, June Feng, Benjamin C. Blount, Lanqing Wang
Tobacco
use is the leading preventable cause of premature disease
and death in the United States. Approximately, 34 million U.S. adults
currently smoke cigarettes. We developed a method for automated sample
preparation and liquid chromatography-tandem mass spectrometry quantitation
of 14 tobacco-related analytes: nicotine (NICF), cotinine (COTF), trans-3′-hydroxycotinine (HCTF), menthol glucuronide
(MEG), anabasine (ANBF), anatabine (ANTF), isonicoteine (ISNT), myosmine
(MYOS), beta-nicotyrine (BNTR), bupropion (BUPR), cytisine (CYTI),
varenicline (VARE), arecaidine (ARD), and arecoline (ARL). The method
includes automated solid-phase extraction using customized positive-pressure
functions. The preparation scheme has the capacity to process a batch
of 96 samples within 4 h with greater than 88% recovery for all analytes.
The 14 analytes, separated within 4.15 min using reversed-phase liquid
chromatography, were determined using a triple-quadrupole mass spectrometer
with atmospheric-pressure chemical ionization and multiple reaction
monitoring in negative and positive ionization modes. Wide quantitation
ranges, within 1.2–72,000 ng/mL, were established especially
for COTF, HCTF, MEG, and NICF to quantify the broad range of biomarker
concentrations found in the U.S. population. The method accuracy is
above 90% while the overall imprecision is below 7%. Finally, we tested
urine samples from 90 smokers and observed detection rates of over
98% for six analytes with urinary HCTF and MEG concentrations ranging
from 200–14,100 and 60–57,100 ng/mL, respectively. This
high throughput analytical process can prepare and analyze a sample
in 9 min and along with the 14-compound analyte panel can be useful
for tobacco-exposure studies, in smoking-cessation programs, and for
detecting changes in exposure related to tobacco products and their
use.