Profiling of polyphenols from samples was achieved by an already published method [20 (link)] by using HPLC system (Agilent 1200 series) coupled to Agilent 6520 Accurate-Mass Q-TOF LC/MS (Agilent Technologies, Santa Clara, CA, USA) with an electrospray ionisation source (ESI). Separation of compounds was performed with RP-80A Column (250 mm × 4.6 nm, 4 µm). Mobile phases (A) and (B) were composed of water/acetic acid solution (98:2 v/v) and water/acetonitrile/acetic acid (50:49.5:0.5, v/v/v) respectively and elution program was set as: 0–20 min linear gradient from 90% to 75% eluent A; 20–30 min linear gradient from 75% to 65% eluent A; 30–40 min linear gradient from 65% to 60% eluent A; 40–70 min linear gradient from 60% to 45% eluent A; 70–75 min linear gradient from 45% to 20% eluent A; 75–77 min linear gradient from 20% to 0% eluent A; 77–79 min linear gradient from 0% eluent A; 82–85 min linear gradient from 0% to 90% eluent A. The flow rate was maintained at 0.8 mL/min. Sample injection volume was 6 µL. Peaks were identified in negative and positive ionisation modes. Mass spectra were obtained in the m/z range of 50–1300. Data were analyzed on an Agilent LC/MS/QTOF Mass Hunter Data Acquisition Software (Version B.03.01).
Agilent 6520 accurate mass q tof lc ms
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 6520 Accurate Mass Q-TOF LC/MS is a high-performance liquid chromatography-mass spectrometry system that provides accurate mass measurement for qualitative and quantitative analysis. It features a quadrupole time-of-flight (Q-TOF) mass analyzer that delivers high mass resolution and mass accuracy.
Automatically generated - may contain errors
Lab products found in correlation
Pgc nano chip, by Agilent Technologies (3 mentions)
Agilent 1200 series, by Agilent Technologies (2 mentions)
Masshunter qualitative analysis b08, by Agilent Technologies (2 mentions)
Orbitrap fusion lumos, by Thermo Fisher Scientific (1 mentions)
Ultimate wps 3000rs nanolc system, by Thermo Fisher Scientific (1 mentions)
Byonic software, by Protein Metrics (1 mentions)
Agilent 1200 series hplc, by Agilent Technologies (1 mentions)
Masshunter workstation software, by Agilent Technologies (1 mentions)
Combiflash rf 200i, by Teledyne (1 mentions)
Avance 500 mhz, by Bruker (1 mentions)
6 nbdg, by Thermo Fisher Scientific (1 mentions)
Multiskan spectrometer, by Thermo Fisher Scientific (1 mentions)
Streptozotocin (stz), by Sisco Research Laboratories (1 mentions)
Metformin, by Merck Group (1 mentions)
α glucosidase, by Sisco Research Laboratories (1 mentions)
High performance liquid chromatography (hplc), by Shimadzu (1 mentions)
Lc 20ad, by Shimadzu (1 mentions)
Agilent 1260 hplc, by Agilent Technologies (1 mentions)
10 protocols using agilent 6520 accurate mass q tof lc ms
1
Polyphenol Profiling by HPLC-QTOF-MS
2
Glycoproteomic Analysis of N-Glycans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The detailed procedures were reported previously.15 (link) Briefly, the N-glycans were released from cell membrane using PNGase F after overnight incubation at 37 °C. The released N-glycans were purified using porous graphitic carbon (PGC) SPE plates, the glycan samples were analyzed with an Agilent 6520 Accurate Mass Q-TOF LC/MS equipped with a PGC nano-chip (Agilent, CA), and the results were extracted with the MassHunter Qualitative Analysis B08 software (Agilent, CA). For glycoproteomic analysis, glycopeptides after trypsin digestion were enriched by solid-phase extraction using iSPE®-HILIC cartridges (Nest Group, MA). The enriched glycopeptides were characterized using a UltiMate™ WPS-3000RS nanoLC system coupled with an Orbitrap Fusion Lumos (ThermoFisher Scientific), and an in-house human N-glycan database was applied to the raw results using the Byonic software (Protein Metrics, CA).
3
Comprehensive Metabolite Profiling via LC-ESI-QTOF/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LC-ESI-QTOF/MS analysis was performed on an Agilent 1200 series HPLC (Agilent Technologies, CA, USA) equipped with an Agilent 6520 Accurate-Mass Q-TOF LC/MS (Agilent Technologies, CA, USA). Peak identification was performed in both positive and negative modes. Nitrogen gas nebulization was set at 45 psi with a flow rate of 5 L/min at 300 °C and the sheath gas was set at 11 L/min at 250 °C. The capillary and nozzle voltage were set at 3.5 kV and 500 V respectively. A complete mass scan ranging from m/z 50 to 1300 was used. Instrument control, data acquisition and processing were performed using MassHunter workstation software (Qualitative Analysis, version B.03.01, Agilent). The same column and conditions described in HPLC-PDA analysis maintained except for sample injection volume of 6 µL. The LC-ESI-QTOF/MS identified compounds with more than 80 library identification score were further selected for characterisation and m/z verification.
4
ESI-HRMS Analysis Protocol
5
Glycan Analysis by Q-TOF LC/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The glycan samples were reconstituted in nanopure water, and 5 μL of the sample was injected and analyzed with an Agilent 6520 Accurate Mass Q-TOF LC/MS (Agilent, CA) equipped with a PGC micro-fluidic chip, which incorporates an enrichment column, an analytical column, and a nano-electrospray tip in a single assembly. A binary gradient using solvent A with 3% (v/v) ACN and 0.1% (v/v) FA in water and solvent B with 90% (v/v) ACN and 1% (v/v) FA in water was applied to separate samples at a flow rate of 300 nL min–1. MS spectra were collected with a mass range of m/z 600–2000 at a rate of 1.5 s per spectrum in positive ionization mode. Six of the most abundant precursor ions in each MS spectrum were subjected to fragmentation through collision-induced dissociation (CID) with nitrogen gas.
6
N-Glycan Analysis by Mass Spectrometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell membrane fractions were resuspended with 100 μL of 5 mM DTT in 100 mM ammonium bicarbonate. The mixture was heated in boiling water for 3 minutes. The cleavage of N-glycans was performed by adding 2 μL of PNGase F followed by incubation in a 37 °C water bath overnight. The released N-glycans were separated using centrifugation at 200 000×g for 30 minutes, and the supernatant was purified using porous graphitic carbon (PGC) on an SPE plate. The glycan samples were dried and reconstituted in 30 μL nanopure water. 5 μL of the sample was injected and analyzed with an Agilent 6520 Accurate Mass Q-TOF LC/MS equipped with a PGC nano-chip (Agilent, CA). A binary gradient using solvent A with 3% (v/v) ACN and 0.1% (v/v) formic acid in water and solvent B with 90% (v/v) ACN and 1% (v/v) formic acid in water was applied to separate N-glycans at a flow rate of 300 nL min−1. The resulting chromatographs of glycans were extracted with the MassHunter Qualitative Analysis B08 software (Agilent, CA). N-glycan compounds were identified with an in-house library that contains the accurate mass and formula of human N-glycans, and the N-glycan structures were confirmed through tandem MS fragmentation.
7
Glycomic MS Analysis via PGC Nano-Chip
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Details of the glycomic MS analysis have been described previously ((Wu et al., 2010 (link); Wu et al., 2011 (link))). Glycan samples were reconstituted with 30 μl nanopure water and analyzed using an Agilent 6520 Accurate Mass Q-TOF LC/MS equipped with a PGC nano-chip (Agilent Technologies, CA, United States). The glycan separation was performed at a constant flow rate of 300 nl min −1, and a binary gradient was applied using (A) 0.1% (v/v) formic acid in 3% acetonitrile and (B) 1% (v/v) formic acid in 90% acetonitrile: 0–2 min, 0–0% (B); 2–20 min, 0–16% (B); 20–40 min, 16%–72% (B); 40–42 min, 72–100% (B); 42–52 min, 100–100% (B); 52–54 min, 100–0% (B); 54–65 min, 0–0% (B). MS spectra within the mass range of m/z 600–2000 were collected at a rate of 1.5 s per spectrum in positive ionization mode. The most abundant precursor ions in each MS1 spectrum were subjected to fragmentation through collision-induced dissociation (CID) based on the equation V collision = 1.8 × (m/z)/100–2.4 V.
8
Purification and Characterization of Bioactive Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Combiflash Rf 200i (Teledyne ISCO) fitted with glass column and silica gel 60 RP-18 (40–63 μm) were used for purification of fractions. Shimadzu HPLC, LC 20AD with PDA detector fitted with either an analytical column (Discovery RP Amide C-16 Supelco 5 μm 250 × 4.6 mm) or semi-preparative (Discovery Amide C-16 Supelco 5 μm 250 × 10 mm) were used for analysis and purification of compounds. Column chromatography was carried out with Merck silica gel (100–200 mesh size). Thin layer chromatography (TLC) was done on TLC plates pre-coated with silica gel 60F254 (0.25 mm normal phase Merck). α-Glucosidase (Maltase EC 3.2.1.20) and p-nitro phenyl-α-d -glucopyranoside were purchased from Sisco Research Laboratory (SRL). Streptozotocin was purchased from Sisco Research Laboratories, Mumbai and metformin from Sigma Aldrich USA. 6-NBDG was procured from Invitrogen. NMR spectra were recorded on a Bruker Avance 500 MHz instrument with TMS as internal standard. Chemical shifts are expressed in δ values. Agilent 6520 Accurate mass Q-TOF/LC-MS was used to determine molecular weight. Absorbance was measured by Thermo Scientific Multiskan spectrometer. All other chemicals and reagents were of analytical grade.
9
Untargeted Metabolomics of Chrysanthemum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Leaves (0.2 g) from the top of young cuttings of wild type and T0 chrysanthemum lines were used for nonvolatile analysis according to a protocol for untargeted metabolomics of plant tissues as previously described in detail (Yang et al., 2011 ). LC‐MS analysis was performed using an Agilent 1260 HPLC connected to an Agilent 6520 Accurate‐Mass Q‐TOF LC/MS (Agilent) operating in negative ionization. The column used was a ZORBAX Eclipse plus C18 (1.8 μm) LC column (2.1 × 100 mm). The gradient started at 5% B (acetonitrile:formic acid [1000:1, v/v]) and increased linearly to 35% in 50 min at 0.2 mL/min. The injection volume was 5 μL.
10
Polyphenol Characterization via HPLC-QTOF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polyphenols were characterised by our previously published method (Iqbal et al., 2021) on HPLC system (Agilent 1200 series) connected to Agilent 6520 Accurate-Mass Q-TOF LC/MS (Agilent Technologies, CA, USA) with an electrospray ionization (ESI) source. RP-80A Column (250 mm × 4.6 nm, 4 µm) was used for the analysis. Mobile phase A was composed of water/acetic acid solution (98:2 v/v) and mobile phase B was composed of water/acetonitrile/acetic acid (50:49.5:0.5, v/v/v). The gradient used was: 90% A, 0 min; 75% A, 20 min; 65% A, 30 min; 60% A, 40 min; 45% A, 70 min; 20% A, 75 min; 0% A, 77-79 min; 90% A, 82-85 min with a steady flow rate of 0.8 mL/min. Injection volume for each sample was set as 6 µL. Analysis of data was achieved on Agilent LC/MS/QTOF Mass Hunter Data Acquisition Software (Version B.03.01).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!