Caffeine and 11 downstream metabolites were separated by high-performance liquid chromatography (Gilson, Middleton, WI) using a Synergi Fusion RP column (100 × 3.0 mm, 2.5 μm particle, 100 Å; Phenomenex, Torrance, CA). Column temperature was set at 50°C. The high-performance liquid chromatography system was connected to TSQ Quantum Ultra AM mass spectrometer (Thermo Fisher Scientific; Waltham, MA). The target compounds were analyzed in selected reaction monitoring positive ionization mode.
Tsq quantum ultra am
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TSQ Quantum Ultra AM is a high-performance triple quadrupole mass spectrometer designed for quantitative and qualitative analysis. It features enhanced sensitivity, resolution, and mass accuracy for a wide range of applications in analytical chemistry, environmental monitoring, and life sciences research.
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Lab products found in correlation
7 protocols using tsq quantum ultra am
1
Caffeine Metabolite Separation and Analysis
2
Metabolite Structural Elucidation Using NMR and LC/MS
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The isolated metabolites were analyzed using a nuclear magnetic resonance (NMR) spectrometer Varian INOVA600 (Palo Alto, CA, USA) to obtain 1H-NMR spectra. When further NMR analysis was needed to estimate a chemical structure, Heteronuclear Multiple-Bond Coherence (HMBC), Heteronuclear Single Quantum Coherence (HSQC), Heteronuclear Single Quantum Coherence ADiabatic (HSQCAD), Nuclear Overhauser and Exchange SpectroscopY (NOESY), TOtally Correlated SpectroscopY (TOCSY), and decoupling experiments were conducted. In addition to NMR analysis, liquid chromatography-mass spectrometry (LC/MS) analysis was conducted using a Shiseido NANOSPACE SI-2 system coupled to a Thermo Fisher Scientific TSQ Quantum Ultra AM (Waltham, MA, USA) with an electrospray ionization probe. To compare mass spectra and retention times (RT) between isolated metabolites and authentic standards using LC/MS, chromatography was performed on a Develosil C30-UG-5 (4.6 mm I.D × 250 mm, 5 μm; Nomura Chemical, Seto, Japan) at 50 °C with a mixture of 0.05% formic acid aqueous solution and acetonitrile at a flow rate of 0.5 mL/min. The mass spectrometer was operated in the positive and negative ion mode with capillary temperature of 350 °C, sheath gas of nitrogen at 80 units, and auxiliary gas of nitrogen at 20 units.
3
Quantitative Analysis by LC-MS/MS
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Quantitative analysis was performed with a Dionex Ultimate 3000 chromatographic system (Thermo Scientific, MA, USA) coupled to a TSQ Quantum Ultra AM triple quadrupole mass spectrometer (Thermo Scientific, MA, USA) operated in positive electrospray ionization (ESI) through multiple reaction monitoring (MRM) mode. System control and data acquisitions were conducted by LCQUAN™ 2.9QF1 (Thermo Scientific, MA, USA).
4
Quantifying Cocaine Levels in Mouse Striatum
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Mice were administered cocaine (20 mg/kg, i.p) and 30 minutes later, were sacrificed by rapid decapitation under isoflurane anesthesia. The brain was quickly dissected, blocked, and the striatum was punched bilaterally with a 0.75 mm inner diameter punch. Tissue punches were placed into Eppendorf tubes on dry ice and stored at -80°C until processing.
Tissue was homogenized in 150 μL 100 mM sodium carbonate. Following centrifugation, samples were extracted using acetonitrile containing 50 nM cocaine-d3 as an internal standard. The resulting solution was dried under nitrogen and brought up in Mobile Phase A. Tissue cocaine content was quantified using LC-MS/MS on a Thermo TSQ Quantum ultra AM triple-quadrupole mass spectrometer in positive-ion mode using 0.1% HCOOH in Water (solvent A) and 0.1% HCOOH in Acetonitrile (solvent B). The major transition, m/z 304 to 182, was used to determine cocaine concentration.
Tissue was homogenized in 150 μL 100 mM sodium carbonate. Following centrifugation, samples were extracted using acetonitrile containing 50 nM cocaine-d3 as an internal standard. The resulting solution was dried under nitrogen and brought up in Mobile Phase A. Tissue cocaine content was quantified using LC-MS/MS on a Thermo TSQ Quantum ultra AM triple-quadrupole mass spectrometer in positive-ion mode using 0.1% HCOOH in Water (solvent A) and 0.1% HCOOH in Acetonitrile (solvent B). The major transition, m/z 304 to 182, was used to determine cocaine concentration.
5
Mass Spectrometry Analysis of Nitroaromatic Compounds
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Mass Spectrometry analysis was done using a Thermo Scientific TSQ Quantum Ultra AM triple stage quadrupole mass spectrometer equipped with a heated electrospray ionization (HESI) source in the positive ionization mode. The typical ion source parameters were as follows: spray voltage: 3000 V; sheath gas pressure (N2): 60 units; auxiliary gas pressure (N2): 20 units; ion transfer tube temperature: 275 °C; collision gas (Ar): 1.5 mTorr; Q1/Q3 peak resolution: 0.7 Da; and scan width: 0.002 Da. Samples were direct infusions (0.1 mM) of: NBT, TNBT, NBT + S-nitrosocysteine, or TNBT + S-nitrosocysteine, for 30 s and the data was collected in the mass range between 200 and 1000 m/z. Tandem mass spectrometry experiments were performed as above, but analyzing the 747 Da and 808 Da mass peaks with a collision energy of 40 eV. Experiments were performed in triplicate with no difference between the three samples when tested. All data was collected in centroid mode and were processed using Xcalibur 2.2 software (Thermo Fisher Scientific).
6
HPLC-MS Analysis of Migration Solutions
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The migration solutions (carbonated mineral water and 20% ethanol) were analysed by HPLC/MS. HPLC equipment: Thermo Finnigan Surveyor; mass spectrometer: Thermo Finnigan TSQ Quantum Ultra AM (Dreieich, Germany). Column: Phenomenex Synergi polar RP 150×3 mm, particle size 4 µm. Analyses were performed with a gradient mobile phase using 0.5% formic acid (aqueous) and methanol at a flow of 0.6 ml min−1. The gradient method started at 20% methanol and linearly increased to 100% methanol in 3 min and was hold for 4 min. Injection volume: 5 µl, column oven temperature: 45°C. Ion source: heated electrospray ionisation (HESI), positive mode (spray voltage: 3000; vaporiser temperature: 350°C; capillary temperature: 250°C; sheath gas pressure: 40; aux gas pressure: 5). Analyte was detected by single reaction-monitoring mode (SRM positive, parent ion 137 Dalton, daughter ion 120 Dalton.
7
Quantifying 2-Arachidonoylglycerol in DRG
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