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The mass selective detector was operated in the selected ion monitoring mode (with electron impact) and set at m/z [M+] 214, 242 and 277 and m/z 376 and 439 for the detection of diclofenac and 4-hydroxydiclofenac, respectively.
The mass spectrometric detection was done on a triple quadrupole instrument (Waters Quattro Premier XE) in multiple reactions monitoring mode with positive electrospray ionization for optimal selectivity and sensitivity.
For quantification purposes, data were collected in the multiple reaction monitoring mode with positive-ion detection for chloroquine and negative-ion detection for fosmidomycin, tracking the transition of the parent and product ions specific for each compound: 320.3/141.9 and 320.3/246.7 for chloroquine and 182.0/79.0 and 182.0/136.0 for fosmidomycin.
Data were acquired in the selected ion monitoring mode with ions at m/z = 117 and 148 for isoflurane and m/z = 117 and 198 for halothane.
The MS detection was performed in the multiple reaction monitoring mode with the positive ion mode of electrospray ionization.
Acquisition took place in multiple-reaction monitoring mode, with two transitions per compound (one for quantification and one for confirmation) in positive ionization mode.
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To further verify the formation of specific metabolites, samples from 30 min incubations were analyzed on an Agilent 7890A gas chromatograph with a 5975 C mass selective detector in both total and selective ion monitoring modes with an HP-5 MS column (30 m × 0.32 mm I.D., 0.25 μm film thickness; Agilent) following a published method.
Triple quadrupole scans were acquired in the multiple reaction monitoring mode (MRM) with Q1 and Q3 set at "unit" resolution.
The specific ion transitions for cotinine and cotinine-d3 were monitored in a multiple reaction monitoring mode (MRM) with a dwell time of 0.3 s.
Amino acid isotopologues were analyzed by an ISQ GC/MS instrument (Thermo Scientific, Waltham, MA) operating in selected ion monitoring mode (SIM) with a DB-5 column (30 × 0.25 mm i.d., Restek, Bellefonte, PA) using a ramp profile of 100 °C for 4 min, then 100 200 °C at 4 °C/min, and then 200 300 °C at 10 °C/min with a helium carrier gas flow rate of 1.2 mL/min.
Caffeine, CAF-d3, PAX, TOL, and TOL-d9 were detected by positive electrospray ionization (ESI+) in the positive multiple reaction monitoring mode (MRM+) with the following ion transitions [ m/z]: 195.1 → 138.0 for CAF, 198.0 → 138.0 for CAF-d3, 181.1 → 124.0 for PAX, 271.3 → 91.1 for TOL, and 280.2 → 90.8 for TOL-d9.
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