The MDMS-SL assay [26 (link),58 (link),59 (link)] was performed to determine differences in lipid composition of EVs isolated from young and old WT mice using the ExoQuick protocol described above. Quantitative analysis was performed on a triple-quadruple mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) equipped with an automated nanospray apparatus NanoMate and Xcalibur system. Internal standards for quantification of individual molecular species of the major lipid classes were added to each sample prior to extraction. Lipid extraction was performed via the utilization of the methyl-tert-butyl ether (MTBE) method with resuspension in a chloroform/methanol (1:1 v/v) solution with nitrogen flush. The identification and quantification of all reported lipid molecular species scans from the mass spectrometer was automatically performed with a customized sequence subroutine operated under Xcalibur software (v4.3, ThermoFisher) [58 (link),59 (link)]. The resulting MDMS-SL data were normalized to total protein content, which was assessed according to the Pierce BCA Protein Assay Kit (ThermoFisher). Abbreviations used for the classes assessed include: FA: fatty acyl chains in TAG, TAG: triacylglycerol; PC: phosphatidylcholine; PE: phosphatidylethanolamine; CAR: carnitine and acetyl carnitine; SM: sphingomyelin; CE: ceramide.
Triple quadruple mass spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The triple-quadrupole mass spectrometer is an analytical instrument used for the detection and quantification of chemical compounds. It consists of three consecutive quadrupole mass filters that allow for the selection and fragmentation of target molecules, followed by the analysis of the resulting fragment ions. This device is commonly utilized for applications such as targeted quantitative analysis, bioanalysis, and environmental monitoring.
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Lab products found in correlation
3 protocols using triple quadruple mass spectrometer
1
Lipidomic Analysis of Extracellular Vesicles
2
UPLC-MS/MS Quantification of Opioids
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Concentrations were quantified using a Transcend LX2 Multiplexed UPLC system coupled with a SCIEX (API4500 for morphine and API6500 for fentanyl and tramadol) triple quadruple mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). The study samples and triplicate standard curves spiked in control matrix (30 μL of 2% formic acid per 100 μL of monkey plasma) were prepared for analysis using a protein precipitation extraction method. The chromatographic separation was performed using an Acquity UPLC HSS T3 (50 × 2.1 × 1.8 μm) column. The flow rate was 0.750 mL/min, and the liquid chromatography (LC) gradient method was started with 80% water with 0.1% formic acid and ramped to 98% acetonitrile with 0.1% formic acid for morphine, 100% water with 0.1% formic acid and ramped to 98% acetonitrile with 0.1% formic acid for fentanyl, and 95% water with 0.1% formic acid and ramped to 98% acetonitrile with 0.1% formic acid for tramadol. The concentration of L-005346293-001D005 in the samples was determined using MultiQuant 3.0.1 based on triplicate standard curves ranging from 5 to 10,000 nM for morphine, from 0.1 to 1,000 nM for fentanyl, and from 2 to 10,000 nM for tramadol.
3
LC-MS Characterization of H. cordata
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The LC–MS characterization of the H. cordata was performed by injecting 10 µl of sample in a LC–MS Thermo Scientific Plus with Dionex Ultimate 3000 HPLC with column Hypersil Gold (C18) Diameter 150 × 2.1 mm, Particle Size 1.9 μ at room temperature. The mobile phases used were A (acetonitrile) and B (0.2% aqueous acetic acid, v/v). The run time was set for 20 min followed by flow rate of 0.6 mL/min. DAD detector was set at 280 nm for acquiring chromatograms. The mass spectrometer used was a Triple quadruple Mass Spectrometer (Thermo Scientific) equipped with ion sources ESI with Mass range for full scans m/z 50–6000. All raw data acquired were processed by METLIN database [32 (link)].
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