Exion uplc qtrap 6500 plus

Manufactured by AB Sciex

Sourced in China

The Exion UPLC-QTRAP 6500 PLUS is a liquid chromatography-mass spectrometry system that combines an ultra-high-performance liquid chromatography (UPLC) module with a QTRAP 6500 PLUS tandem mass spectrometer. The system is designed for rapid, sensitive, and selective analysis of a wide range of analytes in complex samples.

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Lab products found in correlation

6890n gas chromatograph, by Agilent Technologies (1 mentions) J w db wax capillary column, by Agilent Technologies (1 mentions) Luna 3 m silica column, by Phenomenex (1 mentions)

Spelling variants of this product

QTRAP 6500 (337 citations) QTRAP 6500 Plus (29 citations) UPLC-QTRAP 6500 Plus (2 citations)

5 protocols using exion uplc qtrap 6500 plus

Analytical Workflow for Polar Lipid Profiling

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The polar lipids were separated by NP-HPLC using Phenomenex Luna 3 μm micron silica column (150 × 2.0 mm; Phenomenex Inc.) treated with mobile phase A chloroform: methanol: ammonia water (89.5:10:0.5) and mobile phase B chloroform: ethanol: ammonia water: water (55:39:0.5:5.5) at a flow rate of 0.25 mL/min. Gradient elution was performed with 5% B for 5 min, a step to 40% B until 12 min and maintained for 4 min, a linear decrease to 30% B and maintained for 15 min, and re-equilibration from 33 min to 38 min with 5% B. Mass spectrometry multiple reaction monitoring (MRM) mode was established for comparative analysis of various polar lipids.
All experiments were carried out by Exion UPLC-QTRAP 6500 PLUS (Sciex Inc.) liquid chromatography-mass spectrometry with the analysis carried out in the electrospray ionization (ESI) mode. The conditions were as follows: curtain gas = 20 psi, ion spray voltage = 5500 V, turbo spray source temperature = 400 °C, ion source gas 1 = 35 psi, ion source gas 2 = 35 psi [19 (link)].

Zhao Y.C., Zhou M.M., Zhang L.Y., Cong P.X., Xu J., Xue C.H., Yanagita T., Chi N., Zhang T.T., Liu F.H, & Wang Y.M. (2020). Recovery of brain DHA-containing phosphatidylserine and ethanolamine plasmalogen after dietary DHA-enriched phosphatidylcholine and phosphatidylserine in SAMP8 mice fed with high-fat diet. Lipids in Health and Disease, 19, 104.

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Pharmacokinetics of 25-Hydroxycholesterol

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Peripheral blood and plasma samples were collected following standard protocols, and the complete blood count and biochemical tests were performed with Sysmex automatic modular animal blood and body fluid analyzer XN-1000 V and Hitachi Automatic Aralyzer 3100 by Guangdong Laboratory Animals Monitoring Institute.
To study the preliminary pharmacokinetics of 25-hydroxycholesterol, lipids were extracted from plasma samples using an improved Bligh/Dyer extraction method (two extractions). The lipids were dried in a vacuum rotary thickener and then remelted in an ethanol and isotope mixture containing 1% (W/V) butylated hydroxytoluene (BHT). The lipids were incubated at 37 °C for 1 h, deionized water and ethanol were added, and the samples were incubated for another 15 min. Subsequently, the upper organic phase was obtained by centrifugation, and pyridine acid derivatization was carried out. After derivatization, a liquid-mass spectrometer (Exion UPLC-QTRAP 6500 PLUS, Sciex) was used for analysis, and electrospray ionization (ESI) mode was used for quantitative analysis. The isotopic internal standard is d₆-25-hydroxycholesterol.

Wu C., Zhao J., Li R., Feng F., He Y., Li Y., Huang R., Li G., Yang H., Cheng G., Chen L., Ma F., Li P, & Sun C. (2021). Modulation of Antiviral Immunity and Therapeutic Efficacy by 25-Hydroxycholesterol in Chronically SIV-Infected, ART-Treated Rhesus Macaques. Virologica Sinica, 36(5), 1197-1209.

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Lipid Extraction and Analysis Protocol

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Lipid extraction was conducted according to a modified Bligh/Dyer extraction method (Song et al., 2020 (link)). Samples were redissolved in isotopic mixed standards and then analyzed via Exion UPLC-QTRAP 6500 Plus (Sciex) with the electrospray ionization mode under the following conditions: curtain gas = 20; ion spray voltage = 5,500 V; temperature = 400°C; ion source gas 1 = 35; and ion source gas 2 = 35.

Feng C., Pan L., Tang S., He L., Wang X., Tao Y., Xie Y., Lai Z., Tang Z., Wang Q, & Li T. (2021). Integrative Transcriptomic, Lipidomic, and Metabolomic Analysis Reveals Potential Biomarkers of Basal and Luminal Muscle Invasive Bladder Cancer Subtypes. Frontiers in Genetics, 12, 695662.

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Lipid Profiling of Mouse Cortex

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After eight weeks of intervention, the mice were anesthetized by an intraperitoneal injection of sodium chloride solution containing 1％ pentobarbital sodium, and then all the mice were decapitated. Cortex was quickly separated from the brain. Cortex was crushed in phosphate buffer using tissue homogenizer and then was extracted with chloroform:methanol (2:1, v/v) . NP-HPLC was used to separate the polar lipids from total lipids using Phenomenex Luna 3 μm micron silica column (150×2.0 mm) . Mobile phase A was chloroform:methanol:ammonia water at the ratio of 89.5:10:0.5, and mobile phase B was chloroform: ethanol:ammonia water:water at the ratio of 55:39:0.5:5.5. The flow rate was 0.25 mL/min. The gradient of mobile phase A was maintained at 95％ for 5 min, then decreased linearly to 60％ within 7 min and kept for 4 min. The gradient of mobile phase A was further increased to 70％ and kept for 15 minutes. Finally, re-equilibration was performed from 33 min to 38 min with 95％ A. All experiments were carried out by Exion UPLC-QTRAP 6500 PLUS (Sciex) liquid chromatography-mass spectrometry, and the analyses were performed in the electrospray ionization (ESI) mode. The conditions were: curtain gas＝20 psi, ion spray voltage＝5500 V, temperature＝400℃, ion source gas＝35 psi, ion source gas 2＝35 psi 15) .

Zhang C., Zhou M.M., Zhang T.T., Cong P.X., Xu J., Xue C.H., Yanagita T., Wei Z.H, & Wang Y.M. (2021). Effects of Dietary Supplementation with EPA-enriched Phosphatidylcholine and Phosphatidylethanolamine on Glycerophospholipid Profile in Cerebral Cortex of SAMP8 Mice fed with High-fat Diet. Journal of oleo science, 70(2).

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Quantitative Analysis of Fatty Acids and Phosphatidic Acids

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FAs were extracted from several mg of lyophilized powder as described by Guo et al. (2019b (link)). FA methyl esters were analyzed with an Agilent GC–MS device (5973 inert mass spectrometer combined with 6890N gas chromatograph) equipped with an Agilent J&W DB-WAX capillary column (30 m × 0.25 mm × 0.25 µm) in a splitless mode. The temperature was set at 150°C for 3 min, then increased to 240°C at 10°C min⁻¹, and kept at 240°C for 5 min. For PA analysis, 5–10 mg of lyophilized powder was treated with 0.01% (w/v) butylated hydroxytoluene in isopropanol (100 µL mg⁻¹) at 75°C for 18 min to deactivate lipid-hydrolyzing enzymes, and extracted with chloroform:methanol:300 mM ammonium acetate (30:41.5:3.5; 300 µL mg⁻¹) at 4°C for 30 min. The samples were clarified by centrifugation at 12,000 rpm, 4°C for 5 min. After another round of extraction, the supernatants were combined and dried under N₂ gas. The extracted lipids were subject to normal phase analysis at LipidALL Technologies Company Ltd. (Changzhou, China) using an Exion UPLC-QTRAP 6500 Plus (Sciex) LC–MS/MS equipped with a Phenomenex Luna 3 µm silica column (internal diameter 150 × 2 mm) as reported previously (Liu et al., 2020 (link)). Individual PA species were quantified by reference to the spiked internal standards d7-PA33:1(15:0/18:1) and DMPA.

Lung S.C., Lai S.H., Wang H., Zhang X., Liu A., Guo Z.H., Lam H.M, & Chye M.L. (2021). Oxylipin signaling in salt-stressed soybean is modulated by ligand-dependent interaction of Class II acyl-CoA-binding proteins with lipoxygenase. The Plant Cell, 34(3), 1117-1143.

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