Analyses were carried out using a Shimadzu LC-20AD Prominence chromatograph (Shimadzu, Tokyo, Japan) equipped with a binary gradient pump, cooled autosampler SIL-20AC and column oven. A column packed with a reversed-phase sorbent ProntoSil 120-AQC18 (2 × 75 mm, 5 µm, EcoNova, Novosibirsk, Russia) was used for chromatographic separations. Mobile phase was water (eluent A) and MeOH (eluent B). The following gradient was used: 0 min—10% B; 1 min—90% B; 4.6 min—90%; 4.7 min—100% B; 6.0 min—100% B, followed by the equilibration of the column. Flow rate was 330 µL/min; injection volume was 10 µL. Mass spectrometric detection was performed on an ABSCIEX 6500 QTRAP mass spectrometer (AB SCIEX, Framingham, MA, USA) using negative electrospray ionization. The following parameters were set for the detection: scan mode—MRM, curtain gas (CUR) = 30 psi, collision-induced dissociation gas (CAD) = Medium, ion source voltage (IS) = 5500 V, gas drier temperature (TEM) = 250 °C, sprayer gas (GS1) = 15 psi, drier gas (GS2) = 20 psi, entrance potential (EP) = 10 V and dwell time = 80 msec. Detection parameters for agents 4c and 5d are shown in Table S1 (Supporting Information) . The instruments were controlled, and the data were collected using Analyst 1.6.3 software (AB SCIEX); data processing was performed using MultiQuant 2.1 software (AB SCIEX).
Absciex 6500 qtrap mass spectrometer
Manufactured by AB Sciex
Sourced in United States, United Kingdom, Japan
The ABSCIEX 6500 QTRAP mass spectrometer is a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) system designed for qualitative and quantitative analysis of a wide range of analytes. It features a triple quadrupole mass analyzer that enables sensitive and selective detection and quantification of target compounds in complex matrices.
Automatically generated - may contain errors
Lab products found in correlation
Analyst 1, by AB Sciex (2 mentions)
Multiquant 2, by AB Sciex (1 mentions)
Lc 20ad prominence chromatograph, by Shimadzu (1 mentions)
Omni tip, by Omni International (1 mentions)
Column oven, by Shimadzu (1 mentions)
Exionlc ad, by Shimadzu (1 mentions)
Acquity uplc hss t3 column, by Waters Corporation (1 mentions)
Vacutainer tube, by BD (1 mentions)
5 protocols using absciex 6500 qtrap mass spectrometer
1
LC-MS/MS Quantification of Compounds 4c and 5d
2
Quantifying Tryptophan Metabolites
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Tryptophan, kynurenine, and kynurenine metabolites were measured by liquid chromatography–tandem mass spectrometry, using a CTC PAL HTS-XT, Agilent 1260 Infinity liquid chromatograph and an AB SCIEX 6500 QTRAP mass spectrometer (AB Sciex UK, Warrington, United Kingdom). All results were calculated in Analyst 1.6.
3
Retinoid Quantification in Liver Slices
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Liver slices were preweighed and homogenized under yellow‐lights in 720 μL saline using a handheld homogenizer and disposable soft‐tissue OmniTip plastic homogenizing probe (Omni International). Homogenates were transferred to glass borosilicate tubes on ice, spiked with internal standard mixture, and retinol, retinyl esters, and retinoic acid were extracted using a two‐round liquid‐liquid extraction protocol, as previously described.30 Culture media was protein precipitated with acetonitrile (1:3), centrifuged at 16,000 × g for 15 min and supernatants collected for analysis. LC‐MS/MS analysis of retinoids was performed as previously described using an AB Sciex 6500 QTRAP mass spectrometer (AB Sciex LLC) coupled with Shimadzu UFLC XR DGU‐20A5 (Shimadzu Corp.).30 Analytes were monitored using the multiple reaction monitoring (MRM) transitions: m/z 269 > 93,95 (retinol and retinyl palmitate); m/z 277 > 98, 102 (retinol‐d8); m/z 275 > 98,102 (retinol‐d6 and retinyl palmitate‐d6); m/z 273 > 94,98 (retinyl palmitate‐d4); m/z 269 > 93 (retinyl acetate); 301 > 205 (atRA); m/z 307 > 211 (atRA‐d6); m/z 327 > 177 (Acitretin). Analyte peaks were integrated against a standard curve using Multiquant.
4
LC-MS/MS Analysis of Chemical Compounds
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LC and MS conditions LC-MS/MS analysis was conducted on ExionLC AD consisting of binary pumps, an on-line degassing unit, an autosampler, and a column oven (Shimadzu Corporation, Kyoto, Japan), which is coupled with an AB Sciex 6500 + QTRAP mass spectrometer consisting of an electrospray ionization (ESI) source (AB SCIEX, Framingham, MA, USA). Chromatographic separation was achieved on Waters Acquity UPLC HSS T3 Column, 100Å, 1.8 μm, 2.1 mm X 100 mm maintained at 40 °C, at a flow rate of 0.3 mL/min. The mobile phases consisted of Solution A (0.1% Formic acid in water) and Solution B (100% acetonitrile). The following gradient was used: 0–1 min, 98% A; 1–5 min, 98%-45% A; 5–8 min, 45%-0% A; 8–13 min, 100% A; 13-13.1 min, 100%-2% A; 13.1–18 min, 98% A, with a total run time of 18 min. The ion source was operated in mix mode: curtain gas, 35 psi; nebulizer gas 50 psi; auxiliary gas 50 psi; ion spray voltage, 5500 V/-4500 V (positive/negative); and temperature 500 °C. Multiple reaction monitoring (MRM) transitions were identified for all analyses and isotope-labeled standard. Data acquisition and analysis were all performed with Analyst 1.6.3 software (AB SCIEX) and OS software (AB SCIEX).
5
Plasma Lipid Mediator Profiling after Blast
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For acute LPA measurements in the plasma, the rats were euthanized at 1 and 4 h post-blast by drawing blood through cardiac puncture under isoflurane anesthesia. Blood was collected in vacutainer tubes (Becton, Dickinson and Company, Franklin Lakes, NJ) containing ethylenediaminetetraacetic acid as the anticoagulant and the plasma separated by centrifugation was used for LPA measurements. The plasma samples were stored at −80°C until analysis. LPA measurements were carried out as described earlier (43 (link)). Briefly, lipids were extracted from plasma using acidified organic solvents. Different LPA species were measured by Ultra High Performance Liquid Chromatography (UHPLC) coupled electrospray ionization tandem mass spectrometry using an AB Sciex 6500 Q-Trap mass spectrometer operated in multiple reaction monitoring mode to identify the molecules as based on their specific precursor and product ion pairs. 17:0 LPA was used as an internal standard. Thus, plasma LPA values were reported as μmoles per liter.
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