Agilent 6520 q tof mass spectrometer

Manufactured by Agilent Technologies

Sourced in United States, Germany, Canada

The Agilent 6520 Q-TOF mass spectrometer is a high-resolution, accurate-mass instrument designed for analytical applications. It features a quadrupole time-of-flight (Q-TOF) configuration, enabling precise mass measurement and identification of unknown compounds.

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6520 Q-TOF mass spectrometer (67 citations) Agilent 6520 Q-TOF (16 citations) Agilent 6520 (16 citations) Spectrometers (10 citations) Agilent 6520 Accurate-Mass Q-TOF mass spectrometer (10 citations) Agilent 6520 Q-TOF tandem mass spectrometer (3 citations) Agilent 6520 HPLC-Q-TOF mass spectrometer (2 citations) Agilent 6520 Accurate-Mass Q-TOF LC/MS spectrometer (2 citations) Agilent 6520 Q-TOF LC-MS mass spectrometer (2 citations) Agilent 6520 Accurate-Mass Q-TOF ESI mass spectrometer (2 citations)

29 protocols using agilent 6520 q tof mass spectrometer

Synthesis of Lipid-Based Nanoparticles

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All starting materials for synthesis were purchased from Sigma-Aldrich, TCI or Alfa Aesar and were used as received unless stated otherwise. Labrafac WL 1349 (medium chain triglycerides, MCT) was obtained from Gattefossé (Saint-Priest, France), Vitamin E Acetate (VEA) was purchased from Tokyo Chemical Industry (Tokyo, Japan), Kolliphor ELP was from BASF (Ludwigshafen, Germany). NMR spectra were recorded in deuterated chloroform (CDCl₃) at a concentration of 15 mg.mL⁻¹ on a Bruker Avance III 400 MHz spectrometer (Rheinstetten, Germany). Mass spectra were obtained using an Agilent Q-TOF 6520 mass spectrometer (Agilent Technologies, Santa Clara, CA, USA); the samples were submitted in dichloromethane at a concentration of 0.1 mg.mL⁻¹. The protocol for synthesis of all new compounds as well as NMR and mass spectra can be found in the Supporting Information.

Wang X., Bou S., Klymchenko A.S., Anton N, & Collot M. (2021). Ultrabright Green-Emitting Nanoemulsions Based on Natural Lipids-BODIPY Conjugates. Nanomaterials, 11(3), 826.

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HPLC-MS Analysis of Catalytic Products

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The catalytic products were analyzed using an Agilent 1200 HPLC system coupled with an Agilent Q-TOF 6520 mass spectrometer (Agilent, Santa Clara, CA, USA) equipped with an electrospray ionization (ESI) device. Gradient elution was performed on an Agilent XDB-C18 column (150 mm × 2.1 mm, 3.5 μm) at room temperature with a flow rate of 0.3 mL/min using a linear gradient with water containing 0.1% formic acid (A) and acetonitrile (B) as the mobile phases as follows: 0–3 min, 5% B; 3–9 min, 25% B; 9–11 min, 25%–55% B; 11–14 min, 55%–95% B; 14–27 min, 95% B; 27–30 min, 95%–5% B. The injection volume was 20 μL.

Yin Y., Li Y., Jiang D., Zhang X., Gao W, & Liu C. (2019). De novo biosynthesis of liquiritin in Saccharomyces cerevisiae. Acta Pharmaceutica Sinica. B, 10(4), 711-721.

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Molecular Mass Determination of SCF-Lα-GCSF and GCSF-Lα-SCF

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The molecular mass of SCF-Lα-GCSF and GCSF-Lα-SCF was determined by the integrated method of high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS). The protein samples were diluted with an aqueous solution containing 1% formic acid (FA) and 2% acetonitrile to the concentration of 0.1 µg/µL. The samples of 10 μL were loaded onto a C₈ reverse-phase column (Poroshell 300SB-C8, 2.1 × 75 mm; Agilent Technologies, Santa Clara, CA, USA). Chromatographic separation of proteins was performed in an acetonitrile gradient (mobile phase A—1% FA in water, mobile phase B—1% FA in acetonitrile) at a flow rate 0.4 mL/min on an Agilent 1290 Infinity HPLC system coupled to an Agilent Q-TOF 6520 mass spectrometer (Agilent Technologies, Santa Clara, CA, USA), as follows: (1) initial equilibration at 2% B, (2) a 5-min gradient to 98% B, (3) a 1-min isocratic elution at 98% B, (4) a 2 min gradient to 2% B, and a final (5) a 1-min isocratic elution at 2% B. The temperature of the column was maintained at 30 °C. The mass analyzer was set to 100–3,200 m/z range in a positive ionization mode. The data were analyzed with Agilent MassHunter Workstation Software.

Mickiene G., Dalgėdienė I., Zvirblis G., Dapkunas Z., Plikusiene I., Buzavaite-Verteliene E., Balevičius Z., Rukšėnaitė A, & Pleckaityte M. (2020). Human granulocyte-colony stimulating factor (G-CSF)/stem cell factor (SCF) fusion proteins: design, characterization and activity. PeerJ, 8, e9788.

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Quantitative Analysis of Metabolites

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All analyses were carried out on an Agilent Q-TOF 6520 mass spectrometer (Agilent, Santa Clara, CA, USA) equipped with a DAD detector, autosampler and column compartment. Chromatographic separations were performed on a Agilent Proshell 120 SB-C18 column (3.0 mm × 150 mm, 2.7 μm) by gradient elution using a mobile phase consisting of HPLC-grade acetonitrile (solvent A) and ultrapure water (solvent B). The gradient elution was carried out as follows: 0–7.0 min, 25–55% A; 7.0–7.1 min, 55–100% A and 100% of solvent A holding for 2.9 min. The flow rate was kept at 0.5 mL/min. The column temperature was set to 45 °C and UV measurements were obtained at 267 nm. An aliquot of 2 μL of each sample block solution was injected onto the UPLC. Samples were filtered with 0.22 nm filter membrane prior to chromatographic analysis.

Yuan X.Y., Wang M., Lei S., Yang Q.X, & Liu Y.Q. (2017). Rapid Screening of Active Components with an Osteoclastic Inhibitory Effect in Herba epimedii Using Quantitative Pattern–Activity Relationships Based on Joint-Action Models. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(10), 1767.

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SILAC-PP Protocol for Quantitative Proteomics

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The SILAC-PP samples were analyzed on a Agilent 6520 Q-TOF mass spectrometer equipped with a Chip Cube interface (Agilent Technologies, Inc). Solvent A consisted of H₂O with 0.1 % formic acid and Solvent B consisted of acetonitrile with 0.1 % formic acid. The solvent gradient increased linearly from 3 to 5 % B over 2 min, 5 to 15 % B over 2 min, 15 to 60 % B over 18 min, 60 to 90 % B over 3 min, 90 to 100% B over 0.1 min, 100 to 5 % B over 1.9 min, and then isocratic at 5% B for 3 min. The flow rate was 0.4 μL/min and the inclusion window for precursor ions was 4 m/z. An HPLC chip with a 40 nL trapping column and 75 μm × 43 mm column with 300 Å Zorbax C18 packing (5 μM) was employed. The collision induced dissociation energy was achieved using the equation 3.50 V/100 m/z with an off-set of −4.80 V. The drying gas was set to 6 L/min at 350 °C, the capillary voltage ranged from 1800–1850, the skimmer was set to 65 V, and the fragmentor was set to 175 V. Four precursor ions were selected for fragmentation in each cycle. The mass spectrometry data from the SILAC-PP experiment have been deposited to the ProteomeXchange Consortium (http://proteomcentral.proteomeexchange.org) via the PRIDE partner repository^{30 (link)} with the dataset identifier to be released at the time of publication.

Geer Wallace M.A., Kwon D.Y., Weitzel D.H., Lee C.T., Stephenson T.N., Chi J.T., Mook R.A., Dewhirst M.W., Hong J, & Fitzgerald M.C. (2016). Discovery of Manassantin A Protein Targets using Large-Scale Protein Folding and Stability Measurements. Journal of proteome research, 15(8), 2688-2696.

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Column Chromatography and Spectroscopic Analysis

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Column chromatography (CC): silica gel H (10–40 μm) and silica gel (200–300 mesh) (Marine Chemical Factory, Qingdao, P. R. China). MCI gel CHP-20P: (Daiso, Co., Japan) and RP-C18 gel (40–63 μm; Daiso, Co., Japan) were used for column chromatography. Sephadex LH-20 (Pharmacia Fine Chemicals, Piscataway, NJ, USA); TLC: silica gel plates, visualization by spraying with 10% H₂SO₄ in EtOH and Dragendorff's reagent. Semi-preparative HPLC: Agilent 1260 series (Agilent Technologies, US) with a Zorbax SB-C₁₈ (5 μM, 9.4 mm × 25 cm) column. NMR spectra: Bruker Avance III-500 spectrometer (Bruker, Switzerland). MS: Agilent MSD-Trap-XCT (for ESI) and Agilent-6520 Q-TOF mass spectrometer (for HR-ESI). IR: Thermo Scientific Nicolet 6700 (Thermo Scientific, US). UV spectra: Agilent 1260 series DAD detector (Agilent Technologies, US). CD spectrum: Brighttime Chirascan (Applied Photophysics Ltd, UK). Optical rotation: Rudolph Autopo V (Rudolph Research Analytical, Hackettstown, NJ).

Wu Z.L., Li J.Y., Huang P.L., Sun Z.S., Li H.L, & Zhang W.D. (2022). New ent-kaurane diterpenoid acids from Nouelia insignis Franch and their anti-inflammatory activity. RSC Advances, 12(18), 11155-11163.

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Comprehensive Analytical Characterization of Natural Products

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IR spectra were obtained with a Thermo Scientific Nicolet iS50 FT-IR spectrometer (Thermo Scientific, Waltham, MA, USA). ¹H- and ¹³C-NMR spectra were obtained with a Bruker AVANCE III 500 spectrometer (Bruker BioSpin, Fällanden, Switzerland) using tetramethylsilane as an internal standard. HR-ESI-MS were obtained with an Agilent 6520 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). Sephadex LH-20 (25–100 μm, Pharmacia Fine Chemical Co., Ltd., Uppsala, Sweden), Lichroprep RP-18 gel (40–63 μm, Merck, Darmstade, Germany), MCI gel CHP-20P (75–150 μm, Mitsubishi Chemical Co., Tokyo, Japan), Silica gel (200–300 mesh and 300–400 mesh), and Silica gel H (Qingdao Oceanic Chemical Co., Qingdao, China) were used for column chromatography. Thin-layer chromatography (TLC) was performed on glass-backed plates coated with 0.25 mm layers of Silica gel H (Qingdao Oceanic Chemical Co., Qingdao, China). Fractions were monitored by TLC and spots were visualized by heating silica gel plates sprayed with 5% H₂SO₄ in EtOH. All solvents and chemicals used were of analytical reagent grade (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China), and water was doubly distilled before use.

Ni J.C., Shi J.T., Tan Q.W, & Chen Q.J. (2017). Phenylpropionamides, Piperidine, and Phenolic Derivatives from the Fruit of Ailanthus altissima. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(12), 2107.

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UHPLC-Q-TOF/MS Analysis of Compounds

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UHPLC-Q-TOF/MS system was composed of Agilent 1290 UHPLC instrument (Agilent Technologies, Waldbronn, Germany) and Agilent 6520 Q-TOF mass spectrometer (Agilent Corporation, Santa Clara, CA, USA). The mass spectra data was acquired in the negative electrospray ion (ESI) mode. The chromatographic peaks were separated on an ACQUITY UPLC BEH C₁₈ Column (2.1 × 150 mm, 1.7 µm, Waters) at a flow rate of 0.3 ml/min. The temperature of column was at room temperature (28°C). Mobile phase consisted of 0.1% formic acid–water (A) and acetonitrile (B). The gradient elution program was set as: 0–2.5 min, 5%–10% B; 2.5–7 min, 10%–13% B; 7–10 min, 13%–15% B; 10–11 min, 15%–18% B; 11–15 min, 18%–30% B; 15–17 min, 30%–45% B; 17–22 min, 45%–95% B; 22–27 min, 95%–5% B. The post run time was 5 min. The injection was 5 µl. The related Q-TOF/MS parameters were listed as follows: drying gas, N₂; gas flow rate, 11 L/min; drying gas temperature, 330^◦C; nebulizer gas pressure, 40 psig; capillary voltage, 3500 V; fragmentor voltage, 120 V; skimmer voltage, 65 V; octopole RF, 750 V; collision energy (CE), 20 and 30 V. The scan range of mass spectra was m/z 100 – 1,500.

Guo J., Li J., Yang X., Wang H., He J., Liu E., Gao X, & Chang Y.X. (2020). A Metabolomics Coupled With Chemometrics Strategy to Filter Combinatorial Discriminatory Quality Markers of Crude and Salt-Fired Eucommiae Cortex. Frontiers in Pharmacology, 11, 838.

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Quantitative LC-QTOF-MS Metabolomic Analysis

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LC-QTOF-MS experiments were performed using an Agilent 1200 SL LC system coupled online with an Agilent 6520 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA). Each prepared sample (4 μL for positive ESI ionization, 8 μL for negative ESI ionization) was injected onto an Agilent Zorbax 300 SB-C8 column (2.1 × 50 mm, 1.8-micron), which was heated to 50 °C. The flow rate was 0.4 mL/min. Mobile phase A was 5 mM ammonium acetate and 0.1% formic acid in water, and mobile phase B was 5% water in ACN containing 5 mM ammonium acetate and 0.1% formic acid. The mobile phase composition was kept isocratic at 35% B for 1 min, and was increased to 95% B in 19 min; after another 10 min at 95% B, the mobile phase composition was returned to 35% B. The ESI voltage was 3.8 kV. The mass accuracy of our LC-MS system is generally less than 5 ppm; the Q-TOF MS spectrometer was calibrated prior to each batch run, and a mass accuracy of less than 1 ppm was often achieved using the standard tuning mixture (G1969-85000, Agilent Technologies, Santa Clara, CA). The mass scan range is 100–1600, and the acquisition rate was 1.5 spectra/s. The absolute intensity threshold for MS data collection was set to 100, and the relative threshold was 0.001%.

Buas M.F., Gu H., Djukovic D., Zhu J., Drescher C.W., Urban N., Raftery D, & Li C.I. (2015). Identification of novel candidate plasma metabolite biomarkers for distinguishing serous ovarian carcinoma and benign serous ovarian tumors. Gynecologic oncology, 140(1), 138-144.

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Qualitative Analysis of Ethyl Ferulate by UHPLC-QTOF-MS

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EFE was accurately weighed (50 mg) and dissolved in 10 mL of methanol with the aid of ultrasound. An Agilent 1290 liquid chromatography system (Agilent Technologies, Santa Clara, CA), connected with the Agilent 6520 Q-TOF mass spectrometer (Agilent Technologies, Seattle, WA) via an ESI interface was used for the qualitative analysis. The separation was carried out on an ACQUITY UHPLC BEH C18 column (2.1 mm Â100 mm, 1.7 lm), with a column temperature maintained at 35 C. The mobile phases A and B were acetonitrile and water, respectively. The flow rate was 0.5 mL/min, the injection volume was 5 lL and the gradient duration program was the following: 0-7 min, 10-20% A; 7-37 min, 20-50% A; 37-52 min, 50-60% A; 52-54 min, 60-65% A; 54-64 min, 65-71% A; 64-94 min, 71-100% A. The MS detection parameters were optimized as follows: drying gas temperature, 350 C; drying gas (N 2 ) flow rate, 10.0 sL/min; nebulizer gas pressure, 40 psig; fragmentor voltage, 150 V; capillary voltage, 4500 V; skimmer voltage, 65 V; octopole radio frequency voltage, 750 V. To gain more information on the structural identification, the sample was analyzed in negative ion mode. The collision energy was set at 15-40 V and the mass range was recorded from m/z 100 to 1000. Data acquisition and processing were controlled by Mass Hunter software (Agilent Technologies).

Gao J., Zhang J., Qu Z., Zhou H., Tong Y., Liu D., Yang H, & Gao W. (2016). Study on the mechanisms of the bronchodilator effects of Folium Eriobotryae and the selected active ingredient on isolated guinea pig tracheal strips. Pharmaceutical biology, 54(11).

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