Nexera uhplc system

Flow injection combined with high-resolution accurate mass spectrometry (HRMS) was conducted using a Shimadzu Nexera UHPLC system connected to an AB SCIEX TripleTOF^® 5600 (Concord, Ontario, Canada) mass spectrometer equipped with a Turbo V ionization source operated in positive and negative electrospray ion mode. For negative ion mode acquisition, the following parameter settings were used to operate the mass spectrometer: spray voltage −4200 V; source temperature 550 °C, and a period cycle time of 150 ms was used. For positive ion mode acquisitions, the instrument settings were the same as those used in the negative ion mode except that the spray voltage was set to 4500 V. The mass spectrometer was equipped with a calibrant delivery system.
For the flow-injection analysis, the flow rate was set at 0.2 mL/min utilizing aqueous methanol (20% v/v). A 3 µL injection volume was used. The total run time per sample was 3 min. The acquired data were aligned, deconvoluted, and normalized using Progenesis QI^TM V2.4 (Nonlinear Dynamics, Waters Corporation, Milford, MA, USA). This deconvolution step assembles isotopologues and adducts from the same molecular species into one molecular feature [17 (link)]. For creating a GNPS network for CAW and derived fractions, MS/MS data were acquired in data-dependent acquisition mode as previously described [30 (link)].

The Shimadzu Nexera UHPLC system equipped with the 6600 QTOF mass spectrometer (AB Sciex) was applied for untargeted metabolomics analysis. 1 uL aliquots of sample was injected into a Waters Acquity UPLC T3 column (100mm×2.1mm, 1.7 μm) maintained at 45°C by gradient elution. Mobile phase A was deionized water with 5 mM ammonium acetate (NH₄AC) and 0.02% formic acid, and phase B was acetonitrile: water (95:5, v/v) with 5 mM NH₄AC and 0.02% formic acid. The flow rate was 0.3 mL/min. The ion source parameters were set as follows: scan range, 50-1200 Da; source temperature, 300°C; curtain gas, 35; collision gas, medium; two ion source gases, 60; ion spray voltage, 5500/-4500V; DP, 60; CE, 35 ± 15.

A Shimadzu Nexera UHPLC system was coupled to an AB Sciex 5,600⁺ Triple TOF mass spectrometer. Conditions for chromatographic separation was the same as the program used in UHPLC‐MS/MS.
The data station operating software was Analyst 1.7. The Q‐TOF/MS instrument was operated in ESI+ mode using scheduled information‐dependent acquisition (IDA) mode. The TOF‐MS method was tuned for the 100–1000 mass range under the following conditions: accumulation time, 0.1 s; ion source gas 1 (N₂), 55 μl/min; ion source gas 2 (N₂), 55 μl/min; curtain gas (N₂), 35 μl/min; source temperature, 550°C; ion spray voltage floating, 5,500 V; declustering potential, 80 V; and collision energy, 10 V. The product ion (+) IDA method was the same as above except that the accumulation time was 0.05 s, the collision energy was 35 V and the collision energy spread was 15 V.