Vanquish flex uhplc

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Vanquish Flex UHPLC is a high-performance liquid chromatography system designed for reliable and efficient separation of complex samples. It features advanced technologies for precise flow control, temperature regulation, and enhanced detection capabilities. The Vanquish Flex UHPLC is a versatile instrument capable of handling a wide range of analytical applications.

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Lab products found in correlation

Q exactive orbitrap mass, by Thermo Fisher Scientific (1 mentions) Q exactive, by Thermo Fisher Scientific (1 mentions) Acquity uplc t3 column, by Waters Corporation (1 mentions) Q exactive focus mass spectrometer, by Thermo Fisher Scientific (1 mentions) Q exactive hf x, by Thermo Fisher Scientific (1 mentions) Acquity uplc beh c18 column, by Waters Corporation (1 mentions) Absolute idq p400 hr kit, by Biocrates (1 mentions)

Close spelling variants of this product

Vanquish UHPLC (149 citations) Vanquish Flex UHPLC system (32 citations) Vanquish Flex (26 citations) Vanquish Flex Binary UHPLC system (5 citations) Vanquish Flex Quaternary UHPLC system (2 citations) Vanquish Flex Binary UHPLC (2 citations)

8 protocols using vanquish flex uhplc

Proteome Profiling by LC-MS/MS

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Reverse-phase chromatography coupled to electrospray ionization tandem mass spectrometry (LC–MS/MS) was performed on an Orbitrap Q Exactive mass spectrometer connected to a Vanquish™ Flex UHPLC (Thermo Scientific). The protein digest was separated on a C18 column (ACQUITY UPLC Peptide BEH, 300 Å, 1.7 μm, 2.1 mm × 150 mm). The mobile phase was 0.1% formic acid in ddH₂O (double-distilled water, mobile phase A) and 0.1% formic acid in acetonitrile (mobile phase B) with a 90-min gradient with mobile phase B ranging 1 to 90%. The flow rate was 0.3 mL/min, and 20 μL of the digests was injected. Online desalting was achieved with a divert valve that switched LC from waste to MS detection at 1.5 min.
Mass spectrometry (MS) data acquisition was performed using a high-energy collision dissociation (HCD) method. MS1 scans were performed at a resolution of 70,000 over m/z 200–2000, and the automatic gain control (AGC) was set as 3e6, with the maximum injection time of 200 ms. Data-dependent HCD tandem mass spectra were acquired at a resolution of 17,500 with normalized collision energies (NCE) of 20, 30, and 40%.

Huang J., Hou S., An J, & Zhou C. (2023). In-depth characterization of protein N-glycosylation for a COVID-19 variant-design vaccine spike protein. Analytical and Bioanalytical Chemistry, 415(8), 1455-1464.

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HILIC-LC-MS/MS for Glycan Analysis

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Hydrophilic interaction chromatography (HILIC) coupled to electrospray ionization tandem mass spectrometry (LC–MS/MS) was performed on an Orbitrap Q Exactive mass spectrometer connected to a Vanquish™ Flex UHPLC (Thermo Scientific). The labeled glycans were separated on an ACQUITY BEH Amide column (130 Å, 1.7 μm, 2.1 mm × 150 mm). The mobile phase was 50 mM ammonium formate in ddH₂O (pH 4.4, mobile phase A) and acetonitrile (mobile phase B) with the following gradient: 75–54% B (0–51.9 min); 0% B (53.4–56 min); 0–75% B (56–59.9 min); 75% B (63.8–71.6 min); and 0% B (76.8 min). The flow rate was 0.4 mL/min except for a change to a lower rate of 0.2 mL/min in the time interval from 53.4 to 59.9 min, and 20 μL of the labeled glycans was injected.
Mass spectrometry data acquisition was performed using a high-energy collision dissociation (HCD) method. MS1 scans were performed at a resolution of 70,000 over m/z 380–4000, and the automatic gain control (AGC) was set as 3e6, with the maximum injection time of 200 ms. Data-dependent HCD tandem mass spectra were acquired with a resolution of 17,500 with normalized collision energies (NCE) of 20%, 30%, and 40%.

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UPLC-MS/MS Analysis of CR Composition

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Analysis of CR chemical composition was performed using UPLC-MS/MS. UPLC conditions: Chromatographic separation was achieved using Vanquish Flex UHPLC (Thermo Fisher Scientific, Bremen, Germany) (Wang et al., 2021 (link)), and the chromatographic column used was ACQUITY UPLC T3 column (100 mm*2.1 mm, 1.7 µm, Waters, Milford, United States). The mobile phase consisted of solvent (A) 0.1% formic acid and (B) Acetonitrile. The gradient elution was as follows: 0–0.8 min, 2% B; 0.8–2.8 min, 2%–70% B; 2.8–5.6 min, 70%–90% B; 5.6–6.4 min, 90%–100% B; 6.4–8 min, 100% B; 8–8.1 min, 100%–2% B; 8.1–10 min, 2% B. MS conditions: Mass spectra were obtained on Q-Exactive (Thermo Scientific). Both positive and negative ion modes were performed on Q-Exactive (Wang et al., 2021 (link)).

Li J., Huang H.Y., Lin Y.C., Zuo H., Tang Y, & Huang H.D. (2023). Cinnamomi ramulus inhibits cancer cells growth by inducing G2/M arrest. Frontiers in Pharmacology, 14, 1121799.

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Quantifying Empty/Full Ratio in AAV

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The full and empty ratio of produced AAV samples was assessed via High-Performance Liquid Chromatography (Vanquish Flex UHPLC, ThermoFisher, Waltham, MA, USA). The AAV samples, SPE purified, were injected into an HPLC system equipped with a size-exclusion chromatography (SEC) column, chosen for its ability to separate AAV particles based on size. Calibration standards with known empty/full ratios were injected to generate a standard curve. Detection was carried out using UV absorbance (Variable Wavelength Detector F, VF-D40-A, ThermoFisher, Waltham, MA, USA). Empty and full capsids eluted at the same time on this column. The A260:A230 ratio was used to measure empty/full capsid (Chromeleon 7.3 Chromatography Data System Software).

Chen Y., Hu S., Lee W., Walsh N., Iozza K., Huang N., Preston G., Drouin L.M., Jia N., Deng J., Hebben M, & Liao J. (2024). A Comprehensive Study of the Effects by Sequence Truncation within Inverted Terminal Repeats (ITRs) on the Productivity, Genome Packaging, and Potency of AAV Vectors. Microorganisms, 12(2), 310.

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LC-MS for Vaccine Protein Analysis

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The LC-MS system is configured with a Thermo Scientific™ Vanquish™ Flex UHPLC (Waltham, MA, USA), and a Thermo Scientific™ Q Exactive™ Focus mass spectrometer (Waltham, MA, USA) equipped with a heated electrospray ionization (HESI) interface. This setup was used for both peptide mapping and bioanalysis of S and N proteins in vaccines. Mobile phases were 0.1% FA in water (A) and 0.1% FA in acetonitrile (B). 10 µL of samples were injected onto Bio C18 column (2.1 × 150 mm, 3 µm) with a column oven temperature set at 35 °C. LC flow rates and gradient conditions were outlined in Table 1. For all sample runs, a diverter valve was used to stream the effluent to waste for the first 2 min before switching back to MS for the remainder of runs. Data acquisition was performed with Xcalibur™ 4.4 software and data analysis and relative quantification was performed with Proteome Discoverer™ 2.5 software.Table 1

Flow rate and mobile phase gradient for peptide mapping and bioanalysis.

	Peptide mapping		Bioanalysis
Flow rate	0.2 mL/min		0.3 mL/min
Gradient	Time(min)	B%	Time(min)	B%
	0	3	0	5
	3	3	8	40
	85	32	8.1	90
	90	90	10	90
	95	90	10.1	5
	95.1	3	15	5
	100	3

Long Z., Wei C., Dong X., Li X., Yang H., Deng H., Ma X., Yin S., Qi Y, & Bo T. (2021). Simultaneous quantification of spike and nucleocapsid protein in inactivated COVID-19 vaccine bulk by liquid chromatography-tandem mass spectrometry. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 1181, 122884.

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Proteomic Analysis of Frozen Samples

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The method devised by Zhu et al. (2021) (link) was used. A frozen sample was taken and transferred to a centrifuge tube, and an appropriate amount of protein lysis buffer (8 mol/L urea + 1 % sodium dodecyl sulfate containing a protease inhibitor) was added. The peptides were redissolved in 2 % acetonitrile (pH 10) and were then fractionated by high-pH reversed-phase liquid chromatography (Vanquish Flex UHPLC, Thermo Fisher Scientific, Waltham, MA, USA) with an Acquity UPLC BEH C18 column (1.7 µm × 2.1 mm × 150 mm, Waters, Milford, MA, USA). The peptides were ionized using a nanospray ionization ion source and analyzed by tandem mass spectrometry (MS/MS) (Q-Exactive HF-X, Thermo Fisher Scientific, Waltham, MA, USA) with an ion source voltage of 2.4 kV. The MS scan range was 350–1300 m/z, and the peptides corresponding to the 20 strongest signals among the precursor ions were selected for secondary fragmentation. The data acquisition mode employed a data-dependent scanning program. The proteomics data were analyzed using MetaboAnalyst 5.0 and the free online platform of Majorbio Cloud Platform (https://www.majorbio.com).

Qian S., Liu K., Wang J., Bai F., Gao R., Zeng M., Wu J., Zhao Y, & Xu X. (2022). Capturing the impact of oral processing behavior and bolus formation on the dynamic sensory perception and composition of steamed sturgeon meat. Food Chemistry: X, 17, 100553.

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Targeted Metabolomic Profiling of Blood

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We used Vanquish Flex UHPLC (Thermo Fisher Scientific, Waltham, MA, USA) connected to Q ExactiveTM Hybrid Quadrupole-Orbitrap MS (Thermo Fisher Scientific) for targeted metabolic and lipid profiling. We employed the AbsoluteIDQ p400HR kit (Biocrates, Innsbruck, Austria) to analyze 408 metabolites and lipids in the blood, including 21 amino acids, 21 biogenic amines, 1 hexose, 172 phosphatidylcholines, 24 lysophosphatidylcholines, 31 sphingomyelins, 9 ceramides, 55 acylcarnitines, 14 cholesteryl esters, 18 diglycerides, and 42 triglycerides. The analysis was performed following the kit’s instructions. Quantities were calculated using software, including Quant Browser (Thermo Xcalibur 4.3.73.11) and MetIDQ (Oxygen-DB110-3023), based on obtained spectra from the instrument.

Kim Y.H., Chung J.S., Lee H.H., Park J.H, & Kim M.K. (2024). Influence of Dietary Polyunsaturated Fatty Acid Intake on Potential Lipid Metabolite Diagnostic Markers in Renal Cell Carcinoma: A Case-Control Study. Nutrients, 16(9), 1265.

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Metabolite Detection via HILIC-LCMS

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For LC chromatographic separation, 2 µL of reconstituted metabolite extracts, QC sample, and processing blank were run on a Millipore SeQuant ZIC-pHILIC (2.1 × 150 mm, 5 µm particle size, Millipore Sigma #150460) column with a ZIC-pHILIC guard column (20 × 2.1 mm, Millipore Sigma #150437) attached to a Thermo Vanquish Flex UHPLC. The mobile phase comprised Buffer A (20 mM (NH₄)₂CO₃, 0.1% NH₄OH (v/v)) and Buffer B (acetonitrile). The chromatographic gradient was run at a flow rate of 0.150 mL/min as follows: 0–21 min-linear gradient from 80 to 20% Buffer B; 20–20.5 min-linear gradient from 20 to 80% Buffer B; and 20.5–28 min-hold at 80% Buffer B. Data were acquired using a Thermo Q Exactive Classic MS operated in full-scan, polarity-switching mode with a spray voltage set to 3.0 kV, the heated capillary held at 275 °C, and the HESI probe held at 350 °C. The sheath gas flow was set to 40 units, the auxiliary gas flow was set to 15 units, and the sweep gas flow was set to 1 unit. MS data acquisition was performed in a range of m/z 70–1000, with the resolution set at 70,000 (out of maximum 120,000), the AGC target at 10⁶, and the maximum injection time at 200 ms. The QC sample was analyzed at the beginning and end of the LC–MS run and after every 10 samples.

Buchanan J.L., Tormes Vaquerano J, & Taylor E.B. (2022). Isolated Effects of Plasma Freezing versus Thawing on Metabolite Stability. Metabolites, 12(11), 1098.

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