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6495c triple quadrupole mass spectrometer

Manufactured by Agilent Technologies
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The 6495C triple quadrupole mass spectrometer is a high-performance analytical instrument designed for precise and sensitive detection and quantification of analytes in complex samples. It features a triple quadrupole configuration that allows for multiple stages of mass selection and fragmentation, enabling accurate identification and measurement of target compounds.

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Lab products found in correlation

Zorbax eclipse plus c18 column, by Agilent Technologies (3 mentions) 1290 infinity, by Agilent Technologies (2 mentions) Infinitylab poroshell 120 ec c18 column, by Agilent Technologies (1 mentions) 1290 infinity 2 uhplc system, by Agilent Technologies (1 mentions) Kinetex f5, by Phenomenex (1 mentions) 1290 uhplc system, by Agilent Technologies (1 mentions) Zorbax rrhd eclipse plus c18 column, by Agilent Technologies (1 mentions) 1290 infinity uhplc system, by Agilent Technologies (1 mentions) Rapid resolution high definition, by Agilent Technologies (1 mentions)

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Spectrometers (10 citations)

7 protocols using 6495c triple quadrupole mass spectrometer

1

Glycopeptide Profiling in Serum

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Digested serum samples were injected into an Agilent6495C triple quadrupole mass spectrometer (Santa Clara, CA, USA), equipped with an Agilent 1290 Infinity ultra-high-pressure (UHP)-LC system and an Agilent ZORBAX Eclipse Plus C18 column (2.1 mm internal diameter × 150 mm length, 1.8 µm particle size). Separation of the peptides and glycopeptides was performed using a 70 min binary gradient. The aqueous mobile phase A was 3% acetonitrile and 0.1% formic acid in water (v/v), and the organic mobile phase B was 90% acetonitrile and 0.1% formic acid in water (v/v). The flow rate was set at 0.5 mL/min. Electrospray ionization was used as the ionization source and was operated in positive ion mode. The triple quadrupole MS was operated in dynamic multiple reaction monitoring (dMRM) mode. Samples were injected in a randomized fashion with regard to underlying phenotype. For quality control purposes, the ratios of glycopeptide abundance relative to their cognate nonglycosylated peptides were assessed in pooled serum replicates by run order. Five representative system suitability glycopeptide biomarkers from each abundance category were monitored, for a total of 15; 10 of these 15 glycopeptides had a coefficient of variation below 10%, while 14 were below 20%.
Pickering C., Zhou B., Xu G., Rice R., Ramachandran P., Huang H., Pham T.D., Schapiro J.M., Cong X., Chakraborty S., Edwards K., Reddy S.T., Guirgis F., Wang T.T., Serie D, & Lindpaintner K. (2022). Differential Peripheral Blood Glycoprotein Profiles in Symptomatic and Asymptomatic COVID-19. Viruses, 14(3), 553.
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2

Quantitative Proteomics Using UHPLC-MS/MS

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Tryptic digests were analyzed on a 6495C triple quadrupole mass spectrometer (Agilent) coupled to a 1290 Infinity II UHPLC system (Agilent). Prior to MS analysis, samples were chromatographically separated on an InfinityLab Poroshell 120 EC‐C18 column (2.1 × 50 mm, 1.9 μm, Agilent) heated to 45°C with a flow rate of 800 μl/min. The 6495C mass spectrometer was controlled by MassHunter Workstation software (LC–MS/MS Data Acquisition for 6,400 series Triple Quadrupole, Version 10.1 (Agilent)) and was operated in positive electrospray ionization mode. Samples were analyzed in dynamic multiple reaction monitoring (MRM) mode with both quadrupoles operating at unit resolution (Wang et al, 2022a (link)).
Wang Z., Tober‐Lau P., Farztdinov V., Lemke O., Schwecke T., Steinbrecher S., Muenzner J., Kriedemann H., Sander L.E., Hartl J., Mülleder M., Ralser M, & Kurth F. (2022). The human host response to monkeypox infection: a proteomic case series study. EMBO Molecular Medicine, 14(11), e16643.
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3

Targeted Quantitative Analysis via LCMS

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All samples were run on an Agilent 6495C triple quadrupole mass spectrometer with a 1290 Infinity II liquid chromatograph system. A 100 μL aliquot was injected onto a Phenomenex Kinetex F5 analytical column. A dynamic multiple reaction monitoring (DMRM) method was used to measure two transitions for all analytes (Table S2). See supplemental information for further details.
Weed R.A., Boatmand A.K, & Enders J.R. (2022). Recovery of Per- and Polyfluoroalkyl Substances After Solvent Evaporation. Environmental science. Processes & impacts, 24(12), 2263-2271.
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4

Lipidome Analysis by UHPLC-MS/MS

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Lipidome analysis was carried out on an Agilent 1290 UHPLC system coupled with an Agilent 6495C triple quadrupole mass spectrometer (Supplementary File S2). Samples were extracted with butanol:methanol, as in Lipid extraction. Concentrations for each lipid species were calculated based on area under the chromatographic curve relative to the labeled internal standard concentrations (23 (link)). For the UWAC samples, the chromatography was extended and retention time windows shifted appropriately, to include lower-abundance short chain fatty acid containing TG (SCFA-TG) with the lipidome.
George A.D., Paul S., Wang T., Huynh K., Giles C., Mellett N., Duong T., Nguyen A., Geddes D., Mansell T., Saffery R., Vuillermin P., Ponsonby A.L., Burgner D., Burugupalli S, & Meikle P.J. (2023). Defining the lipid profiles of human milk, infant formula, and animal milk: implications for infant feeding. Frontiers in Nutrition, 10, 1227340.
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5

UPLC-MS/MS Analysis of Compounds

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UPLC-MS/MS analysis was conducted using a 1290 Infinity Series high-performance LC (HPLC) system (Agilent), coupled to a 6495 C triple quadrupole mass spectrometer (Agilent) that was operated under the electrospray ionization (ESI) negative ion mode. A ZORBAX RRHD Eclipse Plus C18 column ( 2.1×50mm , 1.8μm ; Agilent) equipped with a ZORBAX Eclipse Plus C18 guard column ( 2.1×5mm , 1.8μm ; Agilent) maintained at 40°C. A ZORBAX Eclipse Plus C18 column ( 4.6×50mm , 5μm ; Agilent) was placed between the pump and the injector valve to remove interferences from the mobile phase. The injection volume was 5μL . The separation was achieved by gradient elution of mobile phase A (milli-Q water with 10 mM ammonium acetate) and mobile phase B (MeOH) at a flow rate of 0.4mL/min . A gradient program was used as follows: 0–0.75 min, 40% B; 2.75 min, 60% B; 6.75 min, 65% B; 9.70 min, 90% B; 13.70 min, 98% B with a final hold of 1.3 min (15.0 min). The mass spectrometer was operated in multiple reaction monitoring (MRM) modes. The temperature and flow of the sheath gas were set at 375°C and 15L/min , respectively. The nebulizer gas was 35 psi and the capillary voltage was −3,500V .
Zhan W., Qiu W., Ao Y., Zhou W., Sun Y., Zhao H, & Zhang J. (2023). Environmental Exposure to Emerging Alternatives of Per- and Polyfluoroalkyl Substances and Polycystic Ovarian Syndrome in Women Diagnosed with Infertility: A Mixture Analysis. Environmental Health Perspectives, 131(5), 057001.
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6

Targeted Proteomics of FXRN Protein

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Solutions (2 μL) containing the tryptic peptides were injected in triplicate on the UHPLC-MRM/MS system. This was equivalent to the injection of a trypsin digest from heart tissue spiked with 1.6 ng (105 fmol) of SILAC-hFXN-M protein internal standard. An Agilent 1290 Infinity UHPLC system equipped with a Zorbax Rapid Resolution High Definition (2.1 × 50 mm, 1.8 µm particle size) UHPLC column was used. It was coupled on-line to an Agilent 6495 C triple quadrupole mass spectrometer. Mobile solvent A was 0.1% formic acid in water and solvent B was 0.1% formic acid in ACN. The UHPLC column was maintained at 35 °C with a flow rate of 0.4 mL/min. The tryptic peptides were separated with the following linear gradient: 5% B at 0-min, 5% B at 1-min, 24% B at 2.75-min, 36% B at 3.50-min, 95% B at 5.0-min, 95% B at 6.50-min, 5% B at 7.0-min, 5% B at 8.5-min ready for the next injection. Mass spectrometer operating conditions were maintained as follows: nitrogen gas flow 13 L/min, gas temperature 230 °C, nebulizer gas 40 psi, sheath gas temperature 300 °C, sheath gas flow 10 L/min, capillary voltage 4500 V, and nozzle voltage 500 V. The MRM/MS transitions shown in Table 1 were used and care was taken to ensure that the retention times (ret time) were within 0.1 min of the times shown in Table 1.
Rojsajjakul T., Hordeaux J.J., Choudhury G.R., Hinderer C.J., Mesaros C., Wilson J.M, & Blair I.A. (2023). Quantification of human mature frataxin protein expression in nonhuman primate hearts after gene therapy. Communications Biology, 6, 1093.
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7

Glycopeptide Quantification by LC-MS/MS

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Digested serum samples were injected into an Agilent 6495C triple quadrupole mass spectrometer equipped with an Agilent 1290 Infinity ultra-high-pressure (UHP)-LC system and an Agilent ZORBAX Eclipse Plus C18 column (2.1 mm internal diameter x 150 mm length, 1.8 µm particle size). Separation of the peptides and glycopeptides was performed using a 70-min binary gradient. The aqueous mobile phase A was 3% acetonitrile, 0.1% formic acid in water (v/v), and the organic mobile phase B was 90% acetonitrile 0.1% formic acid in water (v/v). The flow rate was set at 0.5 mL/min. Electrospray ionization was used as the ionization source and was operated in positive ion mode. The triple quadrupole MS was operated in dynamic multiple reaction monitoring (dMRM) mode. Samples were injected in a randomized fashion with regard to underlying phenotype, and reference pooled serum digests were injected interspersed with study samples to allow for correction of within-run drift of baseline signal. For quality control purposes, the ratios of glycopeptide abundance relative to their cognate non-glycosylated peptides were assessed in pooled serum replicates by run order. Five representative system suitability glycopeptide biomarkers from each abundance category were monitored, for a total of 15; 10 of these 15 glycopeptides had a coefficient of variation below 10%, while 14 were below 20%.
Pickering C., Zhou B., Xu G., Rice R., Ramachandran P., Huang H.H., Pham T.D., Schapiro J.M., Xin C., Chakraborty S., Edwards K., Reddy S.T., Guirgis F.W., Wang T.T., Serie D., & Lindpaintner K. (2022). Differential Peripheral Blood Glycoprotein Profiles in Symptomatic and Asymptomatic COVID-19.
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