1290 series hplc system

Manufactured by Agilent Technologies

Sourced in Germany, United States

The 1290 series HPLC system is a high-performance liquid chromatography (HPLC) system designed and manufactured by Agilent Technologies. The system is capable of performing liquid chromatography analysis and separation of complex mixtures. It features advanced components and technology to deliver precise and reliable results.

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24 protocols using 1290 series hplc system

Quantitative Metabolite Analysis by LC-MS

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LC-MS analysis was performed using 6490 triple quadrupole mass spectrometer coupled to an Agilent 1290 series HPLC system using single reaction monitoring (SRM). An Agilent 1290 series HPLC system equipped with a degasser, binary pump, thermostatted auto sampler and column oven. This SRM based measurement of relative metabolite levels used normal phase chromatographic separation. 10 μl of suspended samples were injected and analyzed using source parameters as follows: gas temperature-250 °C; gas flow-14 l/min; nebulizer - 20psi; sheath gas temperature - 350 °C; sheath gas flow-12 l/min; capillary - 3000 V positive and 3000 V negative; nozzle voltage-1500 V positive and 1500 V negative. Approximately 8–11 data points were acquired per each detected metabolite.

Vantaku V., Donepudi S.R., Piyarathna D.W., Amara C.S., Ambati C.R., Tang W., Putluri V., Chandrashekar D.S., Varambally S., Terris M.K., Davies K., Ambs S., Bollag R., Apolo A.B., Sreekumar A, & Putluri N. (2019). Large-scale profiling of serum metabolites in African American and European American bladder cancer patients reveals metabolic pathways associated with patient survival. Cancer, 125(6), 921-932.

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Quantitative Plasma Lipid Profiling

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Lipid extraction was performed as previously described.⁸³ (link) 10 μL of plasma was mixed with 100 μL of butanol:methanol (1:1) with 10 mM ammonium formate which contained a mixture of internal standards. Samples were then vortexed and set in a sonicator bath for 1 h at RT. Samples were then centrifuged (14,000 x g, 10 min, 20°C) before transferring into the sample vials for analysis. Analysis of plasma extracts was performed on an Agilent 6495C QQQ mass spectrometer with an Agilent 1290 series HPLC system and a ZORBAX eclipse plus C18 column (2.1 × 100 mm 1.8 mm, Agilent) with the thermostat set at 45°C. Mass spectrometry analysis was performed with dynamic scheduled multiple reaction monitoring (dMRM). Chromatogram integration was performed using Agilent MassHunter v10.0 and quantification of lipid species was determined by comparison to the relevant internal standard.

Ryan F.J., Norton T.S., McCafferty C., Blake S.J., Stevens N.E., James J., Eden G.L., Tee Y.C., Benson S.C., Masavuli M.G., Yeow A.E., Abayasingam A., Agapiou D., Stevens H., Zecha J., Messina N.L., Curtis N., Ignjatovic V., Monagle P., Tran H., McFadyen J.D., Bull R.A., Grubor-Bauk B., Lynn M.A., Botten R., Barry S.E, & Lynn D.J. (2023). A systems immunology study comparing innate and adaptive immune responses in adults to COVID-19 mRNA and adenovirus vectored vaccines. Cell Reports Medicine, 4(3), 100971.

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Plasma Lipidomics Analysis by LC-MS/MS

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Analysis of plasma extracts was performed on an Agilent 6490 QQQ mass spectrometer with an Agilent 1290 series HPLC system and a ZORBAX eclipse plus C18 column (2.1×100 mm, 1.8μm, Agilent) with the thermostat set at 60°C [8 (link)]. Mass spectrometry analysis was performed in positive ion mode, with dynamic scheduled multiple reaction monitoring (MRM). Mass spectrometry settings and MRM transitions for each lipid class are shown in Supplementary Table 3. Detailed description of the method is presented in Huynh et al. [8 (link)].

Lim W.L., Huynh K., Chatterjee P., Martins I., Jayawardana K.S., Giles C., Mellett N.A., Laws S.M., Bush A.I., Rowe C.C., Villemagne V.L., Ames D., Drew B.G., Masters C.L., Meikle P.J, & Martins R.N. (2020). Relationships Between Plasma Lipids Species, Gender, Risk Factors, and Alzheimer’s Disease. Journal of Alzheimer's Disease, 76(1), 303-315.

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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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HPLC-MS/MS Analytical Platform

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The HPLC instrument was a 1290 series HPLC system (Agilent, Santa Clara, CA, USA), which comprised a pump (G4220A), an autosampler (G4226A), and a thermostatted column compartment (G1316C). An API 4000 triple quadrupole system (Sciex, Concord, ON, Canada) was employed as the mass spectrometric detector. The data were processed using the Analyst^® 1.6.2 software.

Sun L.N., Zhao Y., Qian Z.Y., Chen X.L., Ma H., Guo Y.J., Shen H, & Wang Y.Q. (2023). Simultaneous Determination of Orelabrutinib, Zanubrutinib, Ibrutinib and Its Active Metabolite in Human Plasma Using LC-MS/MS. Molecules, 28(3), 1205.

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Quantification of Fusarium Mycotoxins by HPLC and LC-MS/MS

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High performance liquid chromatography (HPLC) were used to quantify DON, T-2 and HT-2 from media. In addition, liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to quantify FUMS from media and Fusarium toxins from grains. HPLC used consisted of an Agilent 1200 Series system equipped with a UV diode array detector (DAD) set at 220.4 nm (Agilent Technologies, Palo Alto, CA, USA). The column used for the chromatographic separation was a Phenomenex^® Gemini C₁₈, 150 mm × 4.6 mm, 3 µm (Phenomenex, Macclesfield, UK) preceded by a Phenomenex^® Gemini 3 mm guard cartridge and the column temperature was set at 25 °C. LC-MS/MS used consisted of an QTrap 5500 LC-MS/MS System (Applied Biosystems, Foster City, CA, USA) equipped with a TurboIonSpray electrospray ionization (ESI) source and an 1290 Series HPLC System (Agilent, Waldbronn, Germany). Chromatographic separation was performed at 25 °C on a Gemini^® C₁₈-column, 150 × 4.6 mm i.d., 5 µm particle size, equipped with a C₁₈ 4 × 3 mm i.d. security guard cartridge (all from Phenomenex, Torrance, CA, USA).

Mylona K., Garcia-Cela E., Sulyok M., Medina A, & Magan N. (2019). Influence of Two Garlic-Derived Compounds, Propyl Propane Thiosulfonate (PTS) and Propyl Propane Thiosulfinate (PTSO), on Growth and Mycotoxin Production by Fusarium Species In Vitro and in Stored Cereals. Toxins, 11(9), 495.

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HPLC-MS/MS Analysis of Ribonucleosides

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Ribonucleosides were separated using a Synergy Fusion RP, 2.5 µm particle size, 100 Å pore size, 100 mm length, 2 mm inner diameter from Phenomenex (Torrance, CA), on an Agilent 1290 series HPLC system equipped with a diode array detector. Mobile phase A was 5 mM ammonium acetate adjusted to pH 5.3 with glacial acetic acid and mobile phase B was pure acetonitrile. Gradient elution started with 100% A for one minute, increase to 10% B after 10 minutes, 40% after 14 minutes and regeneration of starting conditions with 100% A for 3 additional minutes. The flow rate was 0.35 mL/minute and the column temperature 35°C. For mass spectrometric measurements an Agilent 6490 Triple Quadrupole mass spectrometer set to dynamic multiple reaction monitoring (MRM) mode was used. The MS was operated in positive ion mode with the following parameters: electro-spray ionization (ESI-MS, Agilent Jetstream), Fragmentor Voltage (set in tunefile to) 250 V, Cell Accelerator Voltage 2 V, N2-Gas temperature 150°C, N2-Gas flow 15 L/min, Nebulizer 30 psi, Sheath gas (N2) temperature 275 °C, Sheath gas flow 11 l/min, Capillary 2500 V and Nozzle Voltage 500 V. The mass transition for each modified nucleoside and its isotopomers are found in Table S2.

Heiss M., Reichle V.F, & Kellner S. (2017). Observing the fate of tRNA and its modifications by nucleic acid isotope labeling mass spectrometry: NAIL-MS. RNA Biology, 14(9), 1260-1268.

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HPLC Analysis of Compounds

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HPLC was carried out
on an Agilent 1290
series HPLC system, equipped with an online-degasser, a binary pump,
and an autosampler. A fluorescence detector (model G1321B, Agilent,
U.S.A.) was adjusted at wavelengths of 265 and 360 nm for excitation
and emission. The analysis procedure was run with an isocratic mobile
phase consisting of ACN and 0.125 mol/L ammonium acetate (4:1, v/v)
at a flow rate of 1.3 mL/min. A Phenomenex Luna 5 μm C18 column
(250 mm × 4.6 mm; Torrance, CA, U.S.A.) was used. The injection
volume was 20 μL.

Dong J.X., Xu C., Wang H., Xiao Z.L., Gee S.J., Li Z.F., Wang F., Wu W.J., Shen Y.D., Yang J.Y., Sun Y.M, & Hammock B.D. (2014). Enhanced Sensitive Immunoassay: Noncompetitive Phage Anti-Immune Complex Assay for the Determination of Malachite Green and Leucomalachite Green. Journal of Agricultural and Food Chemistry, 62(34), 8752-8758.

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Liquid Chromatography Pigment Analysis

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Liquid chromatography ultraviolet detection (LC-UV) analysis was performed using an Agilent 1290 series HPLC system equipped with an Agilent G1315B DAD detector. The UV detector was set at 245 nm, and separation was performed on an Acquity UPLC BEH C18 column (2 × 100 mm, 1.7 µm; Waters, MA, USA). Mobile phases were (A) 0.1% formic acid and (B) methanol. LC conditions were identical to those described in the previous section. The extract of the pigment with 1% SDS was passed through an ultrafiltration membrane (nominal molecular weight limit of 3 kDa), and 10-fold diluted samples were used for LC analysis.

Fujiwara M., Kono N., Hirayama A., Malay A.D., Nakamura H., Ohtoshi R., Numata K., Tomita M, & Arakawa K. (2021). Xanthurenic Acid Is the Main Pigment of Trichonephila clavata Gold Dragline Silk. Biomolecules, 11(4), 563.

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HPLC-MS/MS Analysis of Ribonucleosides

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Ribonucleosides were separated using a Synergy Fusion RP, 2.5 μm particle size, 100 Å pore size, 100 mm length, 2 mm inner diameter from Phenomenex (Torrance, CA, USA), on an Agilent 1290 series HPLC system equipped with a diode array detector. Mobile phase A was 5 mM ammonium acetate adjusted to pH 5.3 with glacial acetic acid and mobile phase B was pure acetonitrile. Gradient elution started with 100% A for one minute, increase to 10% B after 5 min, 40% B after 7 min which was maintained for an additional minute. From minute 8 to 8.5 starting conditions are re-established and equilibrated for two additional minutes. The flow rate was 0.35 ml/min and the column temperature 35°C. The effluent from the column was directed through the DAD before entering the Agilent 6490 Triple Quadrupole mass spectrometer in dynamic multiple reaction monitoring (MRM) mode. The MS was operated in positive ion mode with the following parameters: electro-spray ionization (ESI-MS, Agilent Jetstream), Fragmentor Voltage (set in tunefile to) 250 V, Cell Accelerator Voltage 2 V, N₂-gas temperature 150°C, N₂-gas flow 15 l/min, Nebulizer 30 psi, sheath gas (N₂) temperature 275°C, sheath gas flow 11 l/min, capillary 2500 V and nozzle voltage 500 V. The mass transitions for each modified nucleoside are found in Supplementary Table S3.

Schäck M.A., Jablonski K.P., Gräf S., Klassen R., Schaffrath R., Kellner S, & Hammann C. (2020). Eukaryotic life without tQCUG: the role of Elongator-dependent tRNA modifications in Dictyostelium discoideum. Nucleic Acids Research, 48(14), 7899-7913.

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