6520 accurate mass q tof mass spectrometer

Plant fractions were subjected to LC-HRMS. Identification of metabolites from an active fraction of water and ethyl-acetate extract of A. catechu, ethyl acetate fraction of S. robusta, ethyl acetate fraction of M. melabathrium, and hexane fraction of F. religiosa was carried out at Sophisticated Analytical Instrument Facility (SAIF), CSIR-Central Drug Research Institute, Lucknow. Samples were analyzed on an Agilent 6520, Accurate-Mass Q-TOF Mass Spectrometer equipped with a G1311A quaternary pump, G1329A autosampler, and G1315D diode array detector (DAD). The solvent system consists of acetonitrile (ACN) and 5mM acetate buffer and water at the flow rate of 0.5 ml/min. The initial condition started from 5% ACN for 0.1 min to 30% ACN for 6 min, 80% ACN for 20 min, and back to its initial conditions. During the whole process, column temperature was maintained at 30°C. After passing through the DAD flow cell, the column elute was directed to Q-TOF HRMS fitted with an electrospray interface (ESI). The MS analysis was carried out using an ESI-positive ionization mode with mass ranges from 100–3000 Da. Raw data obtained from the LC/HRMS system were converted to mzML format using the ProteoWizard tool MSConvertGUI [71 (link)]. MZmine 2 was used for peak detection, peak alignment, and identification (target compound annotation) using centroid data [71 (link)–73 (link)].

Structural determination of the antimicrobial compound was performed by spectroscopic techniques and literature comparison. The antimicrobial compound was analyzed by thin-layer chromatography on a silica 60 plates (Merck, Germany) with chloroform-acetic acid-ethanol at 95:5:2.5 (v/v) as the mobile phase, followed by spraying of an iron reagent (0.1 M FeCl₃ in 0.1 M HCl). LC-MS was performed with an Agilent 1290 Infinity LC system coupled to Agilent 6520 Accurate-Mass Q-TOF mass spectrometer (dual ESI source) equipped with an Agilent Zorbax Eclipse XDB-C18 column. The ultraviolet/visible absorption spectrum was recorded with the photodiode array detector equipped with the above-mentioned HPLC. The mobile phase was composed of water (A, 0.5% formic acid) and acetonitrile (B, 0.5% formic acid), the gradient program of which was 0–12.00 min 90% A and 10% B and 12.00–15.00 min 100% B. The follow rate of the mobile phase was 0.3 ml/min, and the column temperature was set to 25°C. The injection volume was 10 μl.

The LC-MS was used to identify the chemical contents of the extracts. The phytochemical compounds of the methanol extracts were performed using LC-MS followed by [31 (link)]. LC-MS analysis was performed using Agilent spectrometry equipped with a binary pump. The LC-MS was interfaced with Agilent 1290 Infinity LC system coupled to Agilent 6520 accurate-mass Q-TOF mass spectrometer with a dual ESI source. Full-scan mode from m/z 50 to 500 was performed with a source temperature of 125°C.
The column of Agilent zorbax eclipse XDB-C18, narrow-bore 2.1x150 mm, 3.5 microns (P/N: 930990–902) was used with the temperature 30°C for the analysis. A- 0.1% formic acid in water and B -0.1% formic acid in methanol were used as solvents. Isocratic elution was used to supply solvents at a total flow rate of 0.1 mL minutes^-1. MS spectra were collected in both positive and negative ion modes. The drying gas was 300°C, with a 10L min-1 gas flow rate and a 45-psi nebulizing pressure. Before analysis, 1 ml of concentration. sample extracts were diluted with methanol and filtered through a 0.22 m nylon filter. The extracts were injected into the analytical column in 1 μl volume for analysis. The mass fragmentations were discovered using an Agilent mass hunter qualitative analysis B.07.00 (Metabolom-ics-2019.m) tool and a spectrum database for organic chemicals.

MAA-Boc-lysine, MAA-lysine, and MAA-6ACA were characterized with an Agilent Technologies liquid chromatograph-mass spectrometer system consisting of a series 1200 HPLC and 6520 Accurate Mass Q-TOF mass spectrometer (Santa Clara, CA). Products were injected on a Waters Acquity UPLC CSH Fluoro-phenyl column, 2.1 mm x 100 mm, 1.7 μm particle size. Mobile phase was delivered isocratically at 0.2 mL/min using 70% water containing 0.1% formic acid and 30% acetonitrile. Solvent flow was diverted to waste for the first 1.2 min of the analysis. Mass spectrometer parameters were set to the following values: positive ionization mode, capillary voltage of 3500 V, nebulizing gas pressure of 40 psi, drying gas temperature of 300°C, drying gas flow of 12 L/min, and fragmentor voltage of 150 V. Scans from m/z 100 to m/z 1700 were acquired at a rate of 1 scan/s in the high-resolution, low-mass instrument mode. Reference masses used for real-time mass axis adjustment were purine, m/z 121.050873 and HP-0921, m/z 922.009798.