6490 triple quadrupole qqq mass spectrometer

Manufactured by Agilent Technologies

Sourced in United States

The 6490 triple quadrupole (QQQ) mass spectrometer from Agilent Technologies is a high-performance analytical instrument designed for sensitive and accurate detection and quantification of compounds in complex samples. It utilizes triple quadrupole technology to provide precise and selective mass spectrometric analysis.

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Lab products found in correlation

1290 uhplc, by Agilent Technologies (2 mentions) Zorbax eclipse plus c18 column, by Agilent Technologies (2 mentions) Uhplc, by Agilent Technologies (1 mentions) Mass hunter metabolite id software, by Agilent Technologies (1 mentions) Agilent 1290 series hplc system, by Agilent Technologies (1 mentions) Eclipse plus c18 column, by Agilent Technologies (1 mentions) 1290 series hplc system, by Agilent Technologies (1 mentions) Waters acquity uplc beh amide column, by Waters Corporation (1 mentions) Agilent 1290 uhplc, by Agilent Technologies (1 mentions) Agilent 1260 hplc system, by Agilent Technologies (1 mentions)

8 protocols using 6490 triple quadrupole qqq mass spectrometer

Comprehensive Metabolomics Analysis via UHPLC-MS/MS

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Metabolomics analysis was done using in-house protocols^{41 (link),42}. Briefly, samples were analyzed using an Agilent 1290 UHPLC and 6490 Triple Quadrupole (QqQ) Mass Spectrometer (LC-MS/MS). We employed two LC methods: reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC). The RPLC separates hydrophobic or non-polar metabolites, while the HILIC separates polar or ionic metabolites. This approach generated two data sets per batch resulting in relative quantification ~220 metabolites across the major metabolic pathways in central carbon metabolism, including glycolysis, the TCA cycle, pentose phosphate pathway, amino acid and nucleotide metabolism, among others⁴².

Ji L., Lee H.J., Wan G., Wang G.P., Zhang L., Sajjakulnukit P., Schacht J., Lyssiotis C.A, & Corfas G. (2019). Auditory metabolomics, an approach to identify acute molecular effects of noise trauma. Scientific Reports, 9, 9273.

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Lipid Profiling by HPLC-MS/MS

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Lipid extracts were analyzed using an Agilent 6490 triple quadrupole (QqQ) mass spectrometer coupled to an Agilent 1290 high performance liquid chromatography (HPLC) system as previously published^{45 (link)}. Briefly, we used a ZORBAX eclipse plus C18 column (2.1×100mm 1.8pm, Agilent) with thermostat set to 45°C. The final mass spectrometry analysis on each cell population was performed in positive mode with dynamic scheduled MRM. Solvents consisted of solvent A (50% H20, 30% acetonitrile, 20% isopropanol with 10 mM ammonium formate) and solvent B (1% H20, 9% acetonitrile, 90% isopropanol with 10 mM ammonium formate) and followed a modified 20-minute gradient as follows:

Time (min)	Solvent A (%)	Solvent B (%)	Flow (mL/min)
0.00	85.00	15.00	0.4
2.50	48.00	52.00	0.4
10.00	40.00	60.00	0.4
13.00	15.00	85.00	0.4
17.00	7.00	93.00	0.4
17.10	0.00	100.00	0.4
17.90	0.00	100.00	0.4
18.00	85.00	15.00	0.4
20.00	85.00	15.00	0.4

A wash vial comprising of 1:1 butanol:methanol was used after each sample injection. To further improve chromatic peak shape for many anionic/acidic lipid species (notably PS, PA, PIP, S1P), an additional pre-run passivation step was done with phosphoric acid to minimize interaction between the HPLC unit and these lipids.

Barrachina M.N., Pernes G., Becker I.C., Allaeys I., Hirsch T.I., Groeneveld D.J., Khan A.O., Freire D., Guo K., Carminita E., Morgan P.K., Collins T.J., Mellett N.A., Wei Z., Almazni I., Italiano J.E., Luyendyk J., Meikle P.J., Puder M., Morgan N.V., Boilard E., Murphy A.J, & Machlus K.R. (2023). Efficient megakaryopoiesis and platelet production require phospholipid remodeling and PUFA uptake through CD36. bioRxiv.

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Targeted Metabolite Analysis by QQQ-MS

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Samples were analyzed on a Agilent 6490 triple quadrupole (QQQ) mass spectrometer connected to an Agilent UHPLC. Metabolites were identified with isotopically labeled standards added to the tissue homogenates as well as using Agilent MassHunter Metabolite ID software.

Davis E.L., Salisbury E.A., Olmsted‐Davis E, & Davis A.R. (2015). Anaplerotic Accumulation of Tricarboxylic Acid Cycle Intermediates as Well as Changes in Other Key Metabolites During Heterotopic Ossification. Journal of Cellular Biochemistry, 117(4), 1044-1053.

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Lipidomic Analysis using LC-ESI-MS/MS

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Lipidomic analysis was performed using LC electrospray ionisation MS/MS (LC-ESI-MS/MS). An Agilent 6490 triple quadrupole (QQQ) mass spectrometer (Agilent 1290 series HPLC system and a ZORBAX eclipse plus C18 column [2.1 × 100 mm × 1.8 μm; Agilent, Santa Clara, CA, USA]) in positive ion mode was used (details of the method and chromatography gradient have been described previously [16 (link)]). The solvent system consisted of solvent A, 50% H₂O/30% acetonitrile/20% isopropanol (v/v/v) containing 10 mM ammonium formate, and solvent B, 1% H₂O/9% acetonitrile/90% isopropanol (v/v/v) containing 10 mM ammonium formate. We used a linear gradient with a 14-minute cycle time and a 1-μL sample injection per sample. The following mass spectrometer conditions were used: gas temperature, 150°C; gas flow rate, 17 L/min; nebuliser, 20 psi; sheath gas temperature, 200°C; capillary voltage, 3,500 V; and sheath gas flow, 10 L/min. Given the large sample size, samples were run across several batches, as described above.

Beyene H.B., Olshansky G., T. Smith A.A., Giles C., Huynh K., Cinel M., Mellett N.A., Cadby G., Hung J., Hui J., Beilby J., Watts G.F., Shaw J.S., Moses E.K., Magliano D.J, & Meikle P.J. (2020). High-coverage plasma lipidomics reveals novel sex-specific lipidomic fingerprints of age and BMI: Evidence from two large population cohort studies. PLoS Biology, 18(9), e3000870.

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Lipidomics Analysis of SOD2 Mutant Muscles

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Lipidomics was performed on approximately 50 μg of soluble protein (homogenised and sonicated) from TA muscles taken from SOD2 WT + OIL, mCre + OIL, WT + TAM and mCre + TAM using LC electrospray ionisation MS/MS (LC-ESI-MS/MS) on an Agilent 6490 triple quadrupole (QQQ) mass spectrometer coupled with an Agilent 1290 series HPLC system and a ZORBAX eclipse plus C18 column as previously described [40 (link)].

Zhuang A., Yang C., Liu Y., Tan Y., Bond S.T., Walker S., Sikora T., Laskowski A., Sharma A., de Haan J.B., Meikle P.J., Shimizu T., Coughlan M.T., Calkin A.C, & Drew B.G. (2021). SOD2 in skeletal muscle: New insights from an inducible deletion model. Redox Biology, 47, 102135.

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Label-free Targeted Metabolomics Analysis

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Agilent 1290 UHPLC and 6490 Triple Quadrupole (QqQ) Mass Spectrometer (LC–MS) were used for label-free targeted metabolomics analysis, as described previously (Lee et al., 2019 (link)). Agilent MassHunter Optimizer and Workstation Software LC–MS Data Acquisition for 6400 Series Triple Quadrupole B.08.00 was used for standard optimization and data acquisition. Agilent MassHunter Workstation Software Quantitative Analysis Version B.0700 for QqQ was used for initial raw data extraction and analysis. For RPLC, a Waters Acquity UPLC BEH TSS C18 column (2.1 × 100 mm, 1.7 µm) was used in the positive ionization mode. For HILIC, a Waters Acquity UPLC BEH amide column (2.1 × 100 mm, 1.7 µm) was used in the negative ionization mode. Further details are found in our previous study (Lee et al., 2019 (link)). The unprocessed metabolomics data are presented in Supplementary file 5.

Boyer S., Lee H.J., Steele N., Zhang L., Sajjakulnukit P., Andren A., Ward M.H., Singh R., Basrur V., Zhang Y., Nesvizhskii A.I., Pasca di Magliano M., Halbrook C.J, & Lyssiotis C.A. (2022). Multiomic characterization of pancreatic cancer-associated macrophage polarization reveals deregulated metabolic programs driven by the GM-CSF–PI3K pathway. eLife, 11, e73796.

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Aconitum Alkaloids Quantification in Sera

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The concentration of Aconitum alkaloids in the sera was determined using Agilent 1290 UHPLC coupled with 6490 triple quadrupole (QQQ) mass spectrometer (Santa Clara, CA, United States). The detailed sample preparation, LC condition, parameters of mass spectrometry (Supplementary Table S1), and the chemical structures (Supplementary Figure S1) of detected compounds were shown in Supplementary Material.

Zhang Y., Bian X., Yang J., Wu H., Wu J.L, & Li N. (2019). Metabolomics of Clinical Poisoning by Aconitum Alkaloids Using Derivatization LC-MS. Frontiers in Pharmacology, 10, 275.

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Serum Metabolomics Analysis in Mice

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Metabolomics assessment was performed at the Johns Hopkins Medical Institution's Metabolomics Facility. Serum from mice at baseline and 6 weeks postinduction was processed according to the previously published protocol (8) (link). Briefly, following methanol extraction, the supernatant containing the metabolite fraction resuspended in 50% acetonitrile in mass-spectrometry grade water was measured using mass spectroscopy. The protein fraction was used to standardize metabolite intensities by protein concentration. Metabolomics data were obtained with an Agilent 6490 triplequadrupole (QQQ) mass spectrometer equipped with an Agilent 1260 HPLC system (Santa Clara, CA). Samples were maintained at a stable sample temperature of 4°C throughout the analysis.

Nidadavolu L.S., Cosarderelioglu C., Merino Gomez A., Wu Y., Bopp T., Zhang C., Nguyen T., Marx-Rattner R., Yang H., Antonescu C., Florea L., Talbot C.C., Smith B., Foster D.B., Fairman J.E., Yenokyan G., Chung T., Le A., Walston J.D, & Abadir P.M. (2023). Interleukin-6 Drives Mitochondrial Dysregulation and Accelerates Physical Decline: Insights From an Inducible Humanized IL-6 Knock-In Mouse Model. The journals of gerontology. Series A, Biological sciences and medical sciences, 78(10).

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