The LC–MS/MS instrument consisted of a binary ultra‐high‐performance liquid chromatography (UHPLC) system, with two LC‐30 AD pumps, a SIL30‐ACmp autosampler, a CTO‐20 AC column oven and a DGU‐20A5R degasser, all from Shimadzu. A triple quadrupole mass spectrometer, AB SCIEX quadrupole linear ion trap (QTRAP®5500, Ontario, Canada), was coupled to the LC system with a Turbo V™ TurboIonSpray® source and was equipped with an inlet valve. The LC system and mass spectrometer were controlled and data were collected using Analyst® software version 1.6.2 (Sciex, Ontario, Canada). Quantitative data processing was done using the MultiQuant™ software version 3.0.1 (Sciex, Ontario, Canada).
Analyst software version 1
Manufactured by AB Sciex
Sourced in United States, Canada, Germany, Switzerland
Analyst software version 1.6.2 is a data processing and analysis software tool for analytical instruments. It provides functionality for data acquisition, processing, and reporting.
Automatically generated - may contain errors
Lab products found in correlation
Multiquant software version 3, by AB Sciex (4 mentions)
Qtrap 5500 mass spectrometer, by AB Sciex (4 mentions)
5500 qtrap triple quadrupole mass spectrometer, by AB Sciex (3 mentions)
4000 qtrap, by AB Sciex (3 mentions)
Multiquant 3, by AB Sciex (3 mentions)
Qtrap 6500 mass spectrometer, by AB Sciex (3 mentions)
Api 4000 qtrap, by AB Sciex (3 mentions)
Turbo 5 ion source, by AB Sciex (2 mentions)
Cto 20ac column oven, by Shimadzu (2 mentions)
Sil 20ac autosampler, by Shimadzu (2 mentions)
3200 qtrap mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Lc 20a pump, by Shimadzu (2 mentions)
C8 guard column, by Phenomenex (2 mentions)
Hplc system, by Thermo Fisher Scientific (2 mentions)
Sil 20ac ht autosampler, by Shimadzu (2 mentions)
Api 4000 qtrap mass spectrometer, by AB Sciex (2 mentions)
Acquity uplc system, by Waters Corporation (2 mentions)
C1129, by Merck Group (2 mentions)
L6250, by Merck Group (2 mentions)
T4009, by Merck Group (2 mentions)
69 protocols using analyst software version 1
1
LC-MS/MS Quantification Method Development
2
Validated Hair Quantification of Drugs
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Basic drugs and their metabolites (AMP, MAMP, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), morphine, 6-acetyl-morphine, codeine, dihydrocodeine, cocaine, benzoylecgonine, cocaethylene, methadone/EDDP, MPH) were determined in hair by a validated achiral LC-MS/MS method. This method comprises chromatographic separation of the analytes on a Kinetex 2.6 μm F5 column (150 x 2.1 mm, Phenomenex, Torrance, CA, USA) using gradient elution with water/acetonitrile containing 0.1% formic acid, followed by tandem-mass spectrometric detection with a 5500 QTRAP® hybrid triple quadrupole/linear ion trap mass spectrometer and a Turbo V ion source (SCIEX, Brugg, Switzerland) operated in positive ESI and SRM mode. Data were acquired and analyzed with Analyst software version 1.6.2 (SCIEX, Brugg, Switzerland). For all analytes that were quantitatively determined, linearity ranges are from 100–5000 pg/mg and the LLOQ is 100 pg/mg. This method has been validated and is used in the accredited laboratory. Further details of this method can be obtained from the authors on request.
3
Quantitative LC-MS/MS Analysis of FIT039
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A LC-20 AD series unit (Shimadzu Corporation, Kyoto, Japan) with InertSustain C18 column (particle size: 3 µm, column size: 2.1 mm × 50 mm: GL sciences, Tokyo, Japan) was employed for liquid chromatography analysis. The autosampler was set at 10 °C, and the column oven was set at 40 °C. The mobile phase was a mixture of water and 1% formic acid (1000:1, v/v) (A) and acetonitrile (B). The gradient elution program started at a composition of 50% B for 0.10 min (initial condition) and another 1.50 min and then was ramped to 90% B at 1.50 min and held for another 2.00 min. The composition of mobile phase B was maintained at 50% from 2.01 min to 3.00 min, and the system was then returned to the initial condition at 3.00 min. An API 4000 triple quadrupole instrument (SCIEX, Framingham, MA, USA) was equipped with an electrospray ionization (ESI) source in the positive mode for mass spectrometric detection. The monitoring ions were FIT039; m/z 316.2 (Q1) → m/z 282.1 (Q3), and IS; m/z 350.1 (Q1) → m/z 239.3 (Q3). Internal standards (IS) for the analysis were deuterated forms of the analytes (GIF430, prepared in Kyoto University). Analyst software (version 1.6.2) (SCIEX, Framingham, MA, USA) was used for data processing.
4
Quantification of Oxygen Isotopes in Phosphate
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The concentration of 16O- and 18O- Pi following sequential P extraction was performed by direct infusion analysis on the 4000 QTRAP. The HPLC-MS/MS calibrations curves were produced in their respective sequential fractionation matrices from synthesized 16O- and 18O- Pi stocks following quantification on the SEAL segmented flow analyzer (AA3; Seal Analytical, Mequon, WI, USA). The optimized chromatographic and instrumental parameters for 16O- and 18O- Pi quantification on the HPLC-MS/MS are in S1 Table . The quality assurance (QA)/quality control (QC) for the method included: duplicates; spikes; and low, medium and high QC concentrations from the calibration curve in order to determine accuracy and any variation occurring intra- and inter-day. The concentration of the Pi in mg/L was determined by reporting the chromatographic peak areas of the samples versus standard solution concentrations using AB Sciex Analyst® Software version 1.6.2 (SCIEX. 2013. Analyst 1.6.2 Software Installation Guide. Framingham, MA, USA). The concentration of Pi was converted to mg/g dry soil by multiplying by the extraction volume and dividing by the mass of dry soil.
5
Quantitative Analysis of Efavirenz by LC-MS/MS
6
Quantitative LC-MS/MS Serum Screening
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For analysis of the serum samples, a routinely used LC–MS/MS screening method, currently covering 94 compounds with at least two ion transitions for each analyte and one ion transition for each internal standard in positive scheduled multiple reaction monitoring (+sMRM) was applied [26 ]. Briefly, a QTRAP™ 4000 triple quadrupole linear ion trap instrument (Sciex, Darmstadt, Germany) equipped with a TurbolonSpray® interface and coupled to a Shimadzu Prominence HPLC system consisting of two LC-20 AD SP isocratic pumps, SIL 20 AC autosampler, CTO-20A controller (Shimadzu, Duisburg, Germany) and Analyst® software version 1.6.2 were used (Sciex). Chromatographic separation was performed on a Kinetex® C18 column (2.6 µm, 100 Å, 100 × 2.1 mm; Phenomenex, Aschaffenburg, Germany) applying gradient elution as follows: starting concentration of 20% mobile phase B was held for 1 min, then linearly increased to 60% B for 1.5 min, further increased to 65% B for 1.5 min, held for 1.5 min, further increased to 90% B for 2.5 min, held for 2.0 min, decreased to starting conditions of 20% B for 0.1 min and held for 1.9 min for re-equilibration. The total flow rate was 0.5 mL/min. The autosampler and the column oven temperatures were set to 10 and 40 °C, respectively. The monitored ion transitions used for Cumyl-PINACA were m/z 350→215 and 350→232.
7
UPLC Analysis of Metabolite Samples
8
Quantitative Analysis of Active Metabolites
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Samples were analyzed using an Agilent® 1290 series liquid chromatograph (Agilent Technologies, Santa Clara, CA, USA), coupled with an ABSCIEX® API 5500 mass spectrometer (SCIEX, Framingham, MA, USA), and a SunfireTM C18 chromatographic column (Waters Corp., Milford, MA, USA) of 3.5 μm and 150 × 2.1 mm was used. The analytes were separated chromatographically using a mobile gradient of 0.1% formic acid in water (Phase A) and 0.1% formic acid in methanol (Phase B). The flow rate was adjusted to 0.2 mL/min, the injection volume was 20 µL, the duration was 25.423 min, and the column temperature was set at 35 °C ± 1 °C. The liquid chromatographic pump gradient is shown in Table 1 . The scans per peak are shown in Table S1 .
The criteria to identify the different AMs and their active metabolites was the monitoring of the masses of the precursor and fragment ions (Table S2 ). In addition, different parameters were used for the operation of the mass detector (Table 2 ). The chromatographic integration of the samples was performed using Analyst® software version 1.6.2 (SCIEX, Framingham, MA, USA).
Samples were processed and analyzed at the Veterinary Pharmacology Laboratory (FARMAVET, as per initials in Spanish) of the Faculty of Veterinary and Animal Sciences at the University of Chile, which is accredited under ISO 17,025 standards.
The criteria to identify the different AMs and their active metabolites was the monitoring of the masses of the precursor and fragment ions (
Samples were processed and analyzed at the Veterinary Pharmacology Laboratory (FARMAVET, as per initials in Spanish) of the Faculty of Veterinary and Animal Sciences at the University of Chile, which is accredited under ISO 17,025 standards.
9
Quantification of Resolvin E4 by LC-MS/MS
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The liquid chromatography-tandem mass spectrometry (LC-MS/MS) consists of a 5500 QTRAP mass spectrometer (Sciex, Framingham, MA) equipped with a LC-20AD ultra-fast liquid chromatography (UFLC) system (Shimadzu, Japan). A Poroshell 120 EC-C18 (4.6x100 mm, 2.7 micron, Agilent Technologies, Santa Clara, CA) was kept in a column oven maintained at 50˚C. Resolvin E4 was eluted from this column with a gradient of LC-MS grade methanol (Thermo Fisher, Waltham, MA)/water (Thermo Fisher, Waltham, MA)/acetic acid (Sigma-Aldrich, St. Louis, MO) from 50/50/0.01% (vol/vol/vol) to 98/2/0.01% (vol/vol/vol) at a flow rate of 0.5 ml/min (19 (link)). Targeted multiple reaction monitoring (MRM) for m/z 333>115 and enhanced product ion (EPI) mass spectra in negative polarity were used with the following parameters to identify resolvin E4: Collision Energy= −22, Collision Cell Exit Potential= −13, Entrance Potential= −10V, Declustering Potential= −80V, Curtain Gas= 25, Collision Gas= Medium, Ion Spray Voltage= −4,000 V, Temperature= 500°C, Ion Source Gas1 = 60 pounds per square inch (psi), and Ion Source Gas2 = 60 psi. Data were acquired and analyzed with Analyst software version 1.6.2 (Sciex, Framingham, MA) as in (17 (link)). Data represented as screen captures from Analyst software.
10
Quantitative Analysis of Amphetamine Enantiomers
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A 5500 QTRAP hybrid triple quadrupole/linear ion trap mass spectrometer equipped with a Turbo V ion source (SCIEX, Brugg, Switzerland) was operated in selected reaction monitoring (SRM) mode for mass spectrometric detection. Electrospray ionization was performed in positive ion mode using the following settings: ion source voltage, 3500 V; source temperature, 600 °C; curtain gas, 40; collision gas, medium; gas 1, 40; and gas 2, 60 (arbitrary units for the gas settings). Optimal SRM parameters were determined by direct infusion of a 100 μg/L methanolic solution of (±)-AM and (±)-AM-d 5 , respectively, at a flow rate of 10 μL/min using a syringe pump. Quantifier and qualifier mass transitions were selected on the basis of signal intensity. Monitored transitions were m/z 136.1 → 91.0 (quantifier) and m/z 136.1 → 119.0 (qualifier) for AM enantiomers, and m/z 141.1 → 93.0 for AM-d 5 enantiomers. Collision energies were 26, 11 and 26 eV, respectively. Declustering potential, entrance potential, collision cell exit potential and dwell time were 45 V, 10 V, 8 V and 150 ms, respectively, for all transitions. Analyst software version 1.6.2 (SCIEX, Brugg, Switzerland) was used for data acquisition and processing. The HPLC system and the mass spectrometer were controlled separately. MS/MS data acquisition was started 2 min after sample injection and lasted for 6 min.
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