Agilent 6530 q tof mass spectrometer

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The Agilent 6530 Q-TOF mass spectrometer is a high-resolution, quadrupole time-of-flight (Q-TOF) mass analyzer designed for accurate mass measurements and structural elucidation. It provides precise mass data and can be used for a variety of analytical applications.

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6530 Q-TOF (34 citations) Q-TOF 6530 mass spectrometer (19 citations) Spectrometers (10 citations) Agilent 6530 Accurate-Mass Q-TOF LC-MS spectrometer (5 citations) Agilent 6530 Accurate-mass Q-TOF mass spectrometer (5 citations) Agilent 6530 Accurate-Mass Quadrupole Time-of-Flight (Q-TOF) mass spectrometer (5 citations) Agilent 6530 Q-TOF spectrometer (3 citations) Agilent 6530 Accurate-Mass Q-TOF-LC-MS mass spectrometer (2 citations)

24 protocols using agilent 6530 q tof mass spectrometer

Serum Stability Evaluation of Linkers

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Example 1

The following procedure was used for testing the blood serum stability of linkers in mouse, rat, or human serum

5 μL of text compound (0.5 mM in DMSO) was transferred separately to individual tubes containing 120 μl of 1× phosphate buffered saline, mouse, rat or human serum. The samples were incubated at 37° C. for 0, 1, 2, 4, and 24 hours. After each of the time points, an aliquot of 20 μl was taken from the samples and quenched with 60 μl of 75:25:0.1 MeOH:acetonitrile:formic acid. After quenching, all samples were held at −20° C. for 1 hour and further centrifuged at 14000 rpm for 15 mins. The supernatant was transferred to a fresh vial and stored at −20° C. until analysis.

Samples were analyzed using LC-MS/MS on Agilent 6530 Q-TOF mass spectrometer connected to an Agilent 1290 UPLC. 3 μL of samples were injected onto a Waters BEH C18 Column (2.1×50 mm, 1.7 μm) maintained at 60° C. The compounds were eluted from column at a flow rate of 0.4 mL/min, using a gradient of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. The total run time is 9.5 min.

US10898578B2. Modified self-immolating moieties for use in prodrugs and conjugates and methods of using and making (2021-01-26). BRISTOL-MYERS SQUIBB COMPANY [US]. Inventors: Yam B. Poudel [US], Sanjeev Gangwar [US].

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LC-Q/TOF-MS for Metabolite Profiling

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The assay was performed on LC-Q/TOF-MS using Agilent 1290 Infinity LC coupled with an electrospray ionization (ESI) source and Agilent 6530Q-TOF mass spectrometer (Agilent Technologies, Palo Alto, CA, USA). The LC separation was carried out on a ZORBAX RRHD SB-C₁₈ column (2.1 × 100 mm, 1.8 μm, Agilent) with the column temperature maintained at 40°C. The mobile phases consisted of ultrapure water (A) and acetonitrile (B) both containing 0.1% (v/v) formic acid; the gradient program was shown in Table 1. The injected volume of sample was 4 μL and temperature of autosampler was 4°C.
Nitrogen was used as both nebuliser gas and cone gas. Detection mode of flight tube was type V. The mass spectrometric data was collected in both positive and negative modes with the following parameters: capillary voltage 4 kV (positive mode) and 3.5 kV (negative mode), sampling cone voltage 35 kV (positive mode) and 50 kV (negative mode), source temperature 100°C, desolvation temperature 350°C (positive mode) and 300°C (negative mode), cone gas flow rate 50 L/h, desolvation gas flow rate 600 L/h (positive mode) and 700 L/h (negative mode), extraction cone voltage 4 V, and full scan mode scanning from m/z 50–1000 with a scan time of 0.03 s and an interscan time of 0.02 s. Leucine enkephalin was used as the lock mass ([M+H]⁺ = 556.2771 in the positive mode and [M−H]⁻ = 554.2615 in the negative mode).

Zhao L., Wan L., Qiu X., Li R., Liu S, & Wang D. (2014). A Metabonomics Profiling Study on Phlegm Syndrome and Blood-Stasis Syndrome in Coronary Heart Disease Patients Using Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry. Evidence-based Complementary and Alternative Medicine : eCAM, 2014, 385102.

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HPLC-QTOF Analysis of Compounds

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HPLC analysis was performed on an Agilent 1260 series system (Agilent, Santa Clara, CA, USA) with an auto-sampler, binary pump, degasser, and diode array detector. The sample of 10 μL infection volume was separated on Shiseido CapCell PAK C18 column (5μm, 4.6 mm I.D. × 150 mm) with flow rate of 0.6 mL/min. The mobile phase for separation of sample consisted of water (solvent A) and acetonitrile (solvent B) containing 0.1% formic acid each. The gradient elution: 10% of B at 0–3 min, 10–25% of B at 3–25 min, 25–95% of B at 25–30 min. Mass spectrometer was performed on Agilent 6530 Q-TOF mass spectrometer (Agilent, Santa Clara, CA, USA). All acquisition parameters are customized with MassHunter Workstation software LC/MS Data Acquisition. The electrospray ionization (ESI) interface was adjusted positive and negative mode, and mass detection range was m/z 50–1700.
The quantitative analysis was performed on Waters 2695 (Waters, Milford, MA, USA) linked with Waters 996 Photodiode Array Detector (PAD). The condition of analysis is same as above HPLC system.

Pan J.H., Lim Y., Kim J.H., Heo W., Lee K.Y., Shin H.J., Kim J.K., Lee J.H, & Kim Y.J. (2017). Root bark of Ulmus davidiana var. japonica restrains acute alcohol-induced hepatic steatosis onset in mice by inhibiting ROS accumulation. PLoS ONE, 12(11), e0188381.

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Metabolomic Analysis of Plasma and Urine

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The frozen samples of plasma and urine were briefly diluted with an acetonitrile:methanol:water (3:3:4) mixture (HPLC-grade, Millipore, Bedford, MA, USA) and 10% methanol, respectively. Each sample (5 μL) was loaded onto an Acquity UPLC BEH C18 (1.7 μm, 2.1 mm × 100 mm; Waters Corp., Milford, MA, USA) column and analyzed using an Agilent 6530 QTOF mass spectrometer (Agilent Technologies). The experimental procedure is described in detail in our previous studies (12 (link), 15 (link)). All samples were examined simultaneously in a single run to avoid the batch effect and the overall quality of the analysis procedure was monitored using repeat extracts of a pooled plasma sample.

Kim D.J., Oh J.Y., Rhee C.K., Park S.J., Shim J.J, & Cho J.Y. (2021). Metabolic Fingerprinting Uncovers the Distinction Between the Phenotypes of Tuberculosis Associated COPD and Smoking-Induced COPD. Frontiers in Medicine, 8, 619077.

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Chromatographic Analysis of Formulations

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Chromatographic analysis was performed on an Agilent 1290 Series (Agilent Corp., Santa Clara, CA, USA,) UPLC system equipped with a binary pump, micro degasser, an auto sampler and a thermostatically controlled column compartment. Chromatographic separation was carried out at 25 ^oC on a Zorbax RRHD Eclipse Plus C₁₈ column (2.1 × 50 mm, 1.8 μm). The mobile phase consisted of 0.1% formic acid solution (A) and ACN (B) using a gradient elution of 0–5% B at 0–6 min, 5–8% B at 6–15 min, 8–15% B at 15–20 min, 15–20% B at 20–30 min, 20–30% at 30–35 min, 30–35% at 35–45 min, 35–40% at 45–60 min. The flow rate was kept at 0.2 mL/min, and the sample volume injected was set at 5 μL. Detections were carried out by Agilent 6530 Q/TOF mass spectrometer (Agilent Corp., Santa Clara, CA, USA) equipped with an ESI interface. The parameters of operation were as follows: drying gas N₂ flow rate, 10.0 L/min; temperature, 330 ^oC; nebulizer, 35 psig; capillary, 3000 V; skimmer, 60 V; OCT RFV, 250 V. Each sample was analyzed in both the positive and negative modes due to the selective sensitivities to different components of the formulation-providing better information for molecular formulae and structural identification. Mass spectra were recorded across the range m/z 100–1000 with accurate mass measurements.

Alolga R.N., Fan Y., Zhang G., Li J., Zhao Y.J., Lelu Kakila J., Chen Y., Li P, & Qi L.W. (2015). Pharmacokinetics of a multicomponent herbal preparation in healthy Chinese and African volunteers. Scientific Reports, 5, 12961.

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HPLC-QTOF Analysis of JBT Components

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Characterization of the components of JBT was performed by a 1260 series HPLC instrument (Agilent, Waldbronn, Germany) connected to an Agilent 6530 Q-TOF mass spectrometer (Agilent Corp., USA) equipped with Dual Agilent Jet Stream Electrospray Ionization (Dual AJS ESI). The operating parameters were optimized in both positive and negative modes, as follows: capillary voltage, 3,500 V for ESI mode and 4000 V for ESI+ mode; nozzle voltage, 500 V; fragmentor, 110 V; nebulizer, 45 psi; drying gas temperature, 300°C; drying gas flow rate, 6 L/min; sheath gas temperature, 320°C; and sheath gas flow rate, 12 L/min. Separation of compounds in JBT was carried out on an ACE Excel 3 Super C18 column (100 mm × 2.1 mm; Advanced Chromatography Technologies Ltd., Aberdeen, Scotland). The mobile phase was composed of solvent A (water containing 0.1% formic acid) and solvent B (acetonitrile containing 0.1% formic acid), with a flow rate of 0.35 ml/min. Gradient elution was performed as follows: 0–30 min, 95%–23% A; 30–40 min, 23%–15% A; 40–45 min, 15%–10% A; 45–50 min: 10%–5% A; 50–55 min, 5% A; 55–56 min: 5%–95% A; and 56–66 min, 95% A. The column temperature was maintained at 40°C. System operations and data analysis were conducted on Masshunter Workstation software (Agilent Technologies, USA).

Wang T., Jia Q., Chen T., Yin H., Tian X., Lin X., Liu Y., Zhao Y., Wang Y., Shi Q., Huang C., Xu H, & Liang Q. (2020). Alleviation of Synovial Inflammation of Juanbi-Tang on Collagen-Induced Arthritis and TNF-Tg Mice Model. Frontiers in Pharmacology, 11, 45.

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Analysis of Hydroxycinnamic Acid Derivatives

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Analysis of the purified hydroxycinnamic acid derivatives was carried out using an Agilent 1200 series HPLC system coupled to an Agilent 6530 Q/TOF mass spectrometer (Agilent Technologies, USA) equipped with a dual spray ESI source operating in negative mode. Followed by an isocratic separation with reversed phase HPLC, elemental formulae of compounds were determined with the aid of MassHunter software based on high-resolution (>20,000 FWHM) accurate mass (<5 ppm) data completed with evaluation of isotope abundance matching and isotope spacing. Data was obtained from full-scan TOF-only peak spectra acquired in the mass range of m/z 50–1100. Next, manually selected ion peaks of the TOF-only spectra were subjected to Q/TOFMS analysis and elemental formulae of fragments were determined based on accurate mass (<20 ppm) data obtained from product ion peak spectra.

Villa-Rodriguez J.A., Kerimi A., Abranko L., Tumova S., Ford L., Blackburn R.S., Rayner C, & Williamson G. (2018). Acute metabolic actions of the major polyphenols in chamomile: an in vitro mechanistic study on their potential to attenuate postprandial hyperglycaemia. Scientific Reports, 8, 5471.

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HPLC-QToF Analysis of Serum Metabolites

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All analyses were carried out on the Agilent 1200 HPLC instrument (Agilent, Germany). The chromatographic separation was performed on an ACE Excel C18 column (100 mm × 2.1 mm, 3.0 mm) at 40°C. The mobile phases were composed of (A) 0.1% (v/v) formic acid aqueous solution and (B) acetonitrile containing 0.1% formic acid with a flow rate of 0.4 ml/min. The gradient elution program for serum samples was carried out as follows: 0–2 min, 5% B; 2–7 min, 5–65% B; 7–20 min, 65–95% B; 20–22 min, 95% B; 22–23 min, 95–5% B; and 23–28 min, 5% B. The sample injection volume was set to 3 μL.
Mass spectrometry analysis was conducted to confirm the peak identities using the Agilent 6530 Q-TOF mass spectrometer (Agilent Corp, United States) coupled with an electrospray ionization (ESI) source. MassHunter Workstation software (Agilent Technologies, United States) was employed for the system operation. The operation conditions of the mass spectrometer were as follows: capillary voltage of 4.0 kV for the positive ion mode and 3.5 kV for the negative ion mode; nebulizer pressure of 35 psig; gas temperature of 320°C; gas flow of 12 L/min; collision energy of 35 eV; drying gas temperature of 300°C; and drying gas flow rate of 6 L/min. The mass range was set from 50 to 1100 Da with the full scan mode.

Bai J., Xiong T., Wang X., Cheng Y., Luo R., Yang X, & Fu C. (2022). Potential mechanisms of Lian-Zhi-Fan solution for TNBS-induced ulcerative colitis in rats via a metabolomics approach. Frontiers in Pharmacology, 13, 1014117.

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Metabolomic Profiling of Urine and Plasma

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Each sample (5 μL) was loaded onto an ACQUITY UPLC BEH C18 column (1.7 μm, 2.1 × 100 mm; Waters Corp., Milford, MA, USA), held at 40 °C, and eluted with 0.1% formic acid and 20 mM ammonium formate in water (solvent A) and 0.1% formic acid in methanol (solvent B) at a constant flow rate of 0.4 mL/min and the following gradient conditions: 0–0.1 min, 2% B; 0.1–13 min, 2–98% B; 13–15 min, 98% B; 15–15.1 min, 98–2% B; and 15.1–17 min, 2% B. Subsequently, the eluate was analysed using an Agilent 6530 QTOF mass spectrometer (Agilent Technologies). The overall quality of the analysis procedure was monitored using repeat extracts of a pooled plasma or urine sample (Fig. 2). Urine samples were analysed in a negative electrospray ionisation (ESI) mode, and plasma samples were analysed in a positive ESI mode. The intensity of each ion was normalised, scaled, z-transformed, and aligned according to the retention time using the Mass Hunter Profinder B.08.00 and Mass Profiler Professional (MPP) software package B.14.9 (Agilent Technologies) to generate a normalised data matrix consisting of the retention time, m/z value, and peak area. Subsequently, PCA was performed using the MPP software for both positive and negative ESI datasets to determine sample clustering and distinguishing ions (filtered by P < 0.05, adjusted for FDR between pre-dose and week 8).

Kim D.J., Yoon S., Ji S.C., Yang J., Kim Y.K., Lee S., Yu K.S., Jang I.J., Chung J.Y, & Cho J.Y. (2018). Ursodeoxycholic acid improves liver function via phenylalanine/tyrosine pathway and microbiome remodelling in patients with liver dysfunction. Scientific Reports, 8, 11874.

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HPLC-MS Analysis of Metabolites

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Reversed-phase chromatographic analysis was performed using an Agilent 1200 Series HPLC system (Agilent Technologies, Inc., Waldbronn, Germany) equipped with an Eclipse Plus C18 (ZORBAX, 3.5 µm, 2.1 × 150 mm, Agilent Technologies, Inc., Waldbronn, Germany) as described previously (Voigt et al. 2021 ). Briefly, a gradient of the eluent system acetonitrile/water, both acidified with formic acid, was applied.
An Agilent 6530 Q-ToF mass spectrometer (Agilent Technologies, Santa Clara, USA) equipped with a Jet-Stream Electrospray Ion Source (ESI) was coupled to the HPLC system and used in the positive ion mode. The fragmentation voltage was set to 125 V. For MS/MS spectra, the collision energy was set to 30 eV. The HPLC–MS was controlled using Mass-Hunter Workstation B.06.00 (Agilent Technologies, Santa Clara, USA).

Schmiemann D., Hohenschon L., Bartels I., Hermsen A., Bachmann F., Cordes A., Jäger M., Gutmann J.S, & Hoffmann-Jacobsen K. (2023). Enzymatic post-treatment of ozonation: laccase-mediated removal of the by-products of acetaminophen ozonation. Environmental Science and Pollution Research International, 30(18), 53128-53139.

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