Des (Sciex, MA, supply IonDriveTM Turbo Vin optimistic (ESI)unfavorable ion
Des (Sciex, MA, source IonDriveTM Turbo Vin positive (ESI)negative ion modes (Sciex, MA, USA). The ESI USA). operation parameters were as follows: source temperature, 450 C; ion spray 450 ; ion The ESI supply operation parameters have been as follows: supply temperature, voltage, 5500 and voltage, 5500 and 500 V; supply gas gas 1, ion supply gas two and 25 psi, respectively; spray500 V; ion source gas 1, ion ion source 2 and curtain gas, 50, 50 and curtain gas, 50, 50 collision gas (CAD), medium. The MS/MS system was operated within the various reaction and 25 psi, respectively; collision gas (CAD), medium. The MS/MS program was operated monitoring (MRM) mode using the optimized collision. The ionization energy, MRM in the several reaction monitoring (MRM) mode with the optimized collision. The ionitransition ions (Tasisulam site molecular and product ions), collision energy (CE), declustering possible zation power, MRM transition ions (molecular and product ions), collision energy (CE), (DP), entrance prospective (EP) and collision cell exit potential (CXP) have been optimized by aMolecules 2021, 26,four ofSciex Analyst software package (Table S1, Supplementary material). Analytical information were processed applying Analyst 1.6.3 software platform (Sciex, MA, USA). 2.4. Analysis of Carotenoids Carotenoids in tea had been extracted following the process previously described by Dautermann and Lohr (2017) [17]. Roughly 150 mg of each and every ground tea sample had been weighed into a two mL Eppendorf tube. Two stainless steel beads (four.8 mm in diameter) and 250 of acetone had been added. The mixture was homogenized for ten min at 20 Hz employing the TissueLyzer II, then centrifuged for five min at 16,000g. The supernatant was transferred into an LC vial prior injection into an ultra-performance liquid chromatograph (UPLC). All the sample preparation measures were performed under yellow light to guard carotenoids from UV degradation. Carotenoids in tea samples were analyzed making use of an Agilent 1290 Infinity II UPLC program coupled with a 1290 Infinity II diode array detector (DAD). The technique was equipped with an Acclaim C30 reverse-phase column (150 mm 2.1 mm; 3 particle size). The mobile phases consisted of (A) acetonitrile/methanol (2:1, v/v) and (B) methanol/ethyl acetate (1:1, v/v). The gradient conditions had been as follows: 0 min, one hundred A; eight min, 95 A and 5 B; 104 min, 85 A and 15 B; 14.five min, 95 A and five B; 20 min, 100 A. The flow rate was 0.3 mL/min and column temperature was set at 30 C. Detection was set at 440 and 450 nm. The injection volumes for carotenoid requirements and tea extracts had been two and 1 , GLPG-3221 Membrane Transporter/Ion Channel respectively. The processing of chromatographic information was performed utilizing OpenLab ChemStation application platform (Agilent Technologies, CA, USA). two.5. Evaluation of Saccharides Extraction of saccharides in ground tea samples was conducted in the exact same way as that of phenolics. The supernatant was evaporated till dry using a SpeedVac vacuum concentrator (ThermoFisher Scientific, CA, USA). The pellet was resuspended in 200 of Milli-Q water. The mixture was shaken for five min and centrifuged at 21,120g for five min. The soluble saccharides were analyzed utilizing a 1260 Infinity ELSD (evaporative light scattering detector) coupled with an Agilent 1290 Infinity II UPLC program equipped with a Shodex Sugar SP0810 (300 mm 8.0 mm ID) column. A supply of oil-free clean nitrogen was utilised to operate the detector. The mobile phase was ten acetonitrile. Flow rate was set at 0.four mL/min and column temperature was 80 C. Total ru.