1290 infinity uplc

Manufactured by Agilent Technologies

Sourced in France, United States

The 1290 Infinity UPLC is a high-performance liquid chromatography (HPLC) system designed for advanced analytical applications. It features a modular design and supports ultra-high-pressure liquid chromatography (UHPLC) techniques. The 1290 Infinity UPLC system is capable of delivering precise and reliable solvent flow and sample injection for efficient separation and analysis of complex samples.

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21 protocols using 1290 infinity uplc

Liver Metabolomics by UPLC-QTOF/MS

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Liver samples were extracted according to the method in the previously reported literature [25 ]. The extracts of liver tissue were analyzed using a UPLC-QTOF/MS system (Agilent 1290 Infinity UPLC equipped with Agilent 6530 MS). Assay conditions are detailed in the previously published literature [26 (link)]. SIMCA 15.0 software was used to perform multivariate statistical analysis. OPLS-DA was used to screen the liver metabolites with significant differences between the model and MP groups (VIP value > 1.0, p < 0.05). Finally, MetaboAnalyst 5.0. the online platform was used for pathway analysis.

Wu L., Zhou K., Yang Z., Li J., Chen G., Wu Q., Lv X., Hu W., Rao P., Ai L, & Ni L. (2022). Monascuspiloin from Monascus-Fermented Red Mold Rice Alleviates Alcoholic Liver Injury and Modulates Intestinal Microbiota. Foods, 11(19), 3048.

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Targeted LC-MS Analysis of CoA Thioesters

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3-hydroxypropionyl-Coa, acrylyl-CoA and propionyl-CoA were analyzed using an Agilent 6550 iFunnel Q-TOF LC-MS system equipped with an electrospray ionization source set to positive ionization mode through a 1290 Infinity UPLC (Agilent Technologies Inc. Santa Clara, CA, USA). Compounds were separated on a RP-18 column (50 mm x 2.1 mm, particle size 1.7 µm, Kinetex XB-C18, Phenomenex, Aschaffenburg, Germany) using a mobile phase system comprised of 50 mM ammonium formate pH 8.1 (A) and methanol (B). Chromatographic separation was carried out using the following gradient condition at a flow rate of 250 µl/min: 0 min 0% B; 1 min 0% B, 3 min 2.5% B; 9 min 23% B; 14 min 80 %B; 16 min 80%; 17 min 0 % B; 18 min 0 % B.
Capillary voltage was set at 3.5 kV and nitrogen gas was used as nebulizing (20 psig), drying (13 l/min, 225 °C) and sheath gas (12 l/min, 400°C). The TOF was calibrated using an ESI-L Low Concentration Tuning Mix (Agilent Technologies Inc. Santa Clara, CA, USA) before measurement (residuals less than 2 ppm for five reference ions). MS data were acquired with a scan range of 750-1200 m/z.
CoA-thioesters were additionally detected by UV absorbance at 260 nm using a diode array detector (1290 Infinity II, Agilent Technologies Inc. Santa Clara, CA, USA)
LC-MS data were analyzed using MassHunter Qualitative Analysis software (Agilent).

Bernhardsgrütter I., Vögeli B., Wagner T., Peter D.M., Cortina N.S., Bange G., Engilberge S., Girard E., Riobé F., Maury O., Shima S., Zarzycki J, & Erb T.J. (2018). The multi-catalytic compartment of propionyl-CoA synthase sequesters a toxic metabolite. Nature chemical biology, 14(12), 1127-1132.

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NMR and LC/MS Analysis Protocol

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¹HNMR spectra were recorded on a Bruker Avance 300 MHz spectrometer. High-resolution LC/MS analysis was performed on an Agilent system consisting of a 1290 Infinity UPLC coupled to a 6230 accurate-mass time-of-flight mass spectrometer with a Dual AJS ESI source. Mass spectral data were acquired in positive-ion mode over the range of 100–1700 m/z using a gas temperature of 350 °C, a nozzle potential of 1000 V, and a capillary potential of 3500 V.

Lewin A.H., Silinski P., Hayes J., Gilbert A., Mascarella S.W, & Seltzman H.H. (2017). Synthesis and Physicochemical Characterization of the One-Carbon Carrier 10-Formyltetrahydrofolate; A Reference Standard for Metabolomics. Metabolomics : Official journal of the Metabolomic Society, 13(10), 117.

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Integrated Proteomic and Metabolomic Analysis

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Proteomic analysis was performed on an ultra-high-resolution Orbitrap Fusion mass spectrometer equipped with a nanoflow Ultimate 3000 UPLC (both from Thermo Fisher, Germany) as described in Ref.^{91 (link)}. A survey of metabolomic compounds was conducted on a high-resolution G6550 Q-TOF mass spectrometer with a 1290 Infinity UPLC (both from Agilent, Germany). Details are given in Supplementary Appendix B (sections no. 3 and no. 7 for proteomic analysis, and sections no. 5 and no. 8 for metabolomic analysis). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE^{93 (link)} partner repository with the dataset identifier PXD035863 and 10.6019/PXD035863.

Kopylov A.T., Stepanov A.A., Butkova T.V., Malsagova K.A., Zakharova N.V., Kostyuk G.P., Elmuratov A.U, & Kaysheva A.L. (2023). Consolidation of metabolomic, proteomic, and GWAS data in connective model of schizophrenia. Scientific Reports, 13, 2139.

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UPLC Analysis of Phenolic Compounds

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The samples were dissolved in methanol (2 mL) and centrifuged at 18,000 rpm for three minutes, and the supernatant was analyzed by UPLC. Chromatographic analysis was performed using an Agilent Technologies 1290 Infinity UPLC apparatus equipped with a diode array detection (DAD), a binary solvent delivery pump, vacuum degasser, an autosampler (injection volume 2 µL) according to Grzegorczyk-Karolak et al. [16 (link)]. The follow rate was 0.3 mL/min. The detection wavelength was set at 328 nm. When a pure reference standard was not available, the phenolic compounds were quantified according to the calibration curve of another compound from the same phenolic group: flavonoids were quantified according to a apigenin-7-O-glucoside curve, phenylethanoids according to verbascoside, and phenolic acid according to caffeic acid. The total content of each different chemical group was calculated by summing the concentrations of the individual quantified compounds. The results were expressed in mg per g DW (dry weight).

Grzegorczyk-Karolak I, & Kiss A.K. (2018). Determination of the Phenolic Profile and Antioxidant Properties of Salvia viridis L. Shoots: A Comparison of Aqueous and Hydroethanolic Extracts. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(6), 1468.

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UPLC-MS Analysis of Metabolites

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Samples from either chemical hemi-synthesis or metabolic reactions were carried using a 1290 Infinity UPLC (Agilent Technologies, Courtaboeuf, France). The UPLC was coupled to an Esquire 3000 Plus ion trap mass spectrometer using an ESI source (Bruker-Daltonics, Billerica, MA, USA). 2 µL were injected into an Agilent SB-C18 column (1.8 µm, 2.1 × 100 mm). Samples were eluted with solvent A (H₂O 0.1% FA) and solvent B (acetonitrile 0.1% FA) by the following gradient program: 0–1.7 min, 10% B; 1.7–3.4 min, 10–20% B; 3.4–5.1 min, 20–30% B; 5.1–7.8 min, 30% B; 7.8–8.5 min, 30–35% B; 8.5–11.9 min, 35–60% B; 11.9–15.3 min, 60–100% B; 15.3–17 min, 100% B; 17–17.3 min, 100–10% B. The flow rate was set to 0.4 mL/min and the UV detector was set at the wavelength 320 nm. Total ion chromatograms were obtained using negative mode with a range of m/z 130–1400. The parameters were: capillary voltage, +4 kV; nebulizer pressure, 40 psi; dry gas, 10 L/min; dry temperature, 365 °C. Data analysis was performed with Bruker Data Analysis 3.2 (Bruker-Daltonics, Billerica, MA, USA). Metabolite concentrations were expressed as an equivalent of the ε-viniferin standard curve.

Courtois A., Jourdes M., Dupin A., Lapèze C., Renouf E., Biais B., Teissedre P.L., Mérillon J.M., Richard T, & Krisa S. (2017). In Vitro Glucuronidation and Sulfation of ε-Viniferin, a Resveratrol Dimer, in Humans and Rats. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(5), 733.

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UPLC-HRMS Quantification of Compounds

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200 mL of samples supplemented with 100 nM levofloxacin as external standard were transferred into glass vials and 10 μL of samples and standards were injected into the UPLC in triplicates. A UPLC/HRMS system Agilent 1290 Infinity UPLC coupled to an Agilent 6545 quadrupole time-of-flight mass spectrometer was used to quantify the concentrations of compounds. Samples were processed in positive ion mode. An Eclipse plus C18 Rapid Resolution HD column (1.8 μm, 2.1 × 50 mm, Agilent, USA) was used for separation. The flow rate was 0.6 mL/min using the following gradient: 0 to 1 min (0- 20% ACN), 1 to 4 min (20- 80% ACN), 4 to 5.1 min (80–100% ACN), 5.1 to 6.30 (100% ACN). Solvents contained 0.1% formic acid to promote positive ion formation in the electrospray. The MS parameters were as follows: ion-source gas temperature, 325 °C; capillary voltage, 4000 V; fragmentor voltage, 180 V; m/z range, 50–1100; data acquisition rate, 4 GHz; and 2 spectra were recorded per second. Data were analyzed using Agilent Masshunter workstation quantitative analysis v. B.10 software.

Leus I.V., Weeks J.W., Bonifay V., Shen Y., Yang L., Cooper C.J., Nath D., Duerfeldt A.S., Smith J.C., Parks J.M., Rybenkov V.V, & Zgurskaya H.I. (2022). Property space mapping of Pseudomonas aeruginosa permeability to small molecules. Scientific Reports, 12, 8220.

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Protein Identification Using UPLC-ESI/TOF

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Data was acquired as described before [26 (link)] using an Agilent 1290 Infinity UPLC in tandem with an Agilent 6230 electrospray ionization time-of-flight (UPLC-ESI/ TOF) mass spectrometer, operating in positive ion mode. Unconjugated protein was loaded onto a reverse phase (RP) PLRP-S column (Agilent) with dimension of 4.6 x 50 mm. Mobile phase A consisted of 0.05% trifluoroacetic acid (TFA) while mobile phase B consisted of 0.05% TFA and 80% acetonitrile (ACN), and was used for the gradient between 20–90% solvent B.

Shatz W., Hass P.E., Peer N., Paluch M.T., Blanchette C., Han G., Sandoval W., Morando A., Loyet K.M., Bantseev V., Booler H., Crowell S., Kamath A., Scheer J.M, & Kelley R.F. (2019). Identification and characterization of an octameric PEG-protein conjugate system for intravitreal long-acting delivery to the back of the eye. PLoS ONE, 14(6), e0218613.

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Quantification of Dopamine via UPLC-MS/MS

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The quantification of DA was performed using a 1290 Infinity UPLC (Agilent Technologies, Singapore) coupled with G6460 Electrospray Ionisation Triple Quad Mass Spectrometer (Agilent Technologies, Waldbronn, Germany). Chromatographic separation was done on a Zorbax SB-C18 RRHT 2.1 × 50 mm, 1.8 μm column with SB-C18 2.1 × 5 mm, 1.8 μm precolumn (Agilent Technologies, Santa Clara, CA, USA), thermostatted at 40 °C. The mobile phase A consisted of 0.2% (v/v) formic acid in water, while the mobile phase B was composed of 0.2% (v/v) formic acid in acetonitrile. An isocratic elution with a flow of 0.5 mL/min was applied, with the 30.0%-involvement of the organic mobile phase B, and the injection volume of 5 μL. For the sake of quality control, negative (blank) and positive (spiked) samples were analysed with every sample batch. Quantification was done using an external matrix-matched calibration curve ranging from 1 to 1500 µg/kg. The concentrations were corrected using the average recovery values obtained by validation.

Kvrgić K., Lešić T., Džafić N, & Pleadin J. (2022). Occurrence and Seasonal Monitoring of Domoic Acid in Three Shellfish Species from the Northern Adriatic Sea. Toxins, 14(1), 33.

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UPLC-MS/MS Phenolic Compound Analysis

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Phenolic compound analyses were carried out using a 1290 Infinity UPLC (Agilent Technologies, Les Ulis, France). The UPLC system was coupled to an Esquire 3000 plus mass spectrometer from Bruker Daltonics (Wissembourg, France). Five µL was injected into a column Zorbax SB-C18 (2.1 × 100 mm, 1.8 µm) (Agilent Technologies, Les Ulis, France). Two different solvents were used as a mobile phase: solvent A (water/formic acid 99.9:0.1, v/v) and solvent B (acetonitrile/formic acid 99.9:0.1, v/v), at a flow rate of 0.4 mL/min and a gradient as follows in solvent A: 0 min 1% B, 0.4 min 1% B, 2 min 10% B, 6 min 35% B, 7 min 50% B, 8.8 min 70% B, 10.8 min 92% B, 11 min 100% B, 12 min 100% B, 12.2 min 1% B, and 15.2 min 1% B. The MS/MS parameters were set as follows: negative mode; capillary tension +4000 V; nebulizer 40 psi; dry gas 10 L/min; dry temperature 365 °C; and scan range m/z 100 to 1400. Data were processed using HyStar 3.2 software (Bruker Daltonics, Wissembourg, France).

Wauquier F., Mevel E., Krisa S., Richard T., Valls J., Hornedo-Ortega R., Granel H., Boutin-Wittrant L., Urban N., Berger J., Descamps S., Guicheux J., Vinatier C.S., Beck L., Meunier N., Blot A, & Wittrant Y. (2019). Chondroprotective Properties of Human-Enriched Serum Following Polyphenol Extract Absorption: Results from an Exploratory Clinical Trial. Nutrients, 11(12), 3071.

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