4500 q trap

The sample extracts were analyzed using an UPLC-ESI-MS/MS system (UPLC, SHIMADZU Nexera X2-Shimadzu-Kyoto, Japan, https://www.shimadzu.com.cn/, accessed on 1 October 2024); MS, Applied Biosystems 4500 Q TRAP, https://www.thermofisher.cn/cn/zh/home/brands/applied-biosystems.html, accessed on 1 October 2024). The analytical conditions were as follows: UPLC employed a column, Agilent SB-C18 (1.8 µm, 2.1 mm × 100 mm). The mobile phase consisted of solvent A, pure water with 0.1% formic acid, and solvent B, acetonitrile with 0.1% formic acid. Sample measurements were performed with a gradient program that employed the starting conditions of 95% A and 5% B. Within 9 min, a linear gradient to 5% A and 95% B was programmed, and a composition of 5% A and 95% B was kept for 1 min. Subsequently, a composition of 95% A and 5.0% B was adjusted over 1.1 min and kept for 2.9 min. The flow velocity was set to 0.35 mL per minute. The column oven was set to 40 °C. The injection volume was 4 μL. The effluent was alternatively connected to an electrospray ionization triple quadrupole linear ion trap mass spectrometer (ESI-Q-TRAP-MS).

The non-targeted metabolomics techniques were used to uncover the chemical profiles of tea samples with different PF durations, following the method of Qian et al. [20 (link)], and set up three biological replicates. Firstly, sample extraction was carried out in the following procedure: The vacuum freeze-dried tea samples were ground into powder, and 100 mg of powder was precisely weighed and dissolved in 0.6 mL of 70% methanol extract. Subsequently, the mixture was refrigerated at 4 °C overnight with six vortexing cycles. The next day, the extract was centrifuged (14,000 rpm, 10 min), and the resulting filtrate, passed through a nylon needle microporous filter membrane (SCAA-104, 0.22 μm pore size; ANPEL, Shanghai, China), was used for UPLC-MS/MS analysis.
The chromatograms were obtained using UPLC (Shim-pack UFLC SHIMADZU CBM30A, Shanghai, China) coupled with tandem mass spectrometry (MS/MS, Applied Biosystems 4500 QTRAP, Shanghai, China). The UPLC conditions were set as follows: The chromatographic column used was a Waters ACQUITY UPLC HSS T3 C18 (1.8 μm, 2.1 mm × 100 mm); The mobile phases A and B were ultrapure water and acetonitrile, respectively, both containing 0.04% acetic acid; The elution gradient started at 5% phase B, increased linearly to 95% over 10.00 min, held at 95% for 1 min, then decreased to 5% between 11.00 and 11.10 min, and remained at 5% until 14 min; The flow rate was set at 0.35 mL/minutes; The column temperature was maintained at 40 °C; The injection volume was 4 μL. The MS/MS conditions were as follows: electrospray ionization temperature set to 550 °C; mass spectrometry voltage at 5500 V; curtain gas at 30 psi; collision-activated dissociation parameter set to high; acquisition mode was the multi-reaction monitoring (MRM) mode in the data-dependent acquisition (DDA).

A previously described relative quantification method of widely targeted metabolites was used to analyze samples [16 (link)]. The freeze-dried peach flesh was crushed using a mixer mill (MM 400, Retsch) with zirconia beads for 1 min at 30 Hz. Sixty to eighty milligrams of powder was extracted overnight at 4 °C with 1 ml of 70% aqueous methanol. Following centrifugation at 12,000 rpm for 10 min at 4 °C, the extracts were absorbed (CNWBOND Carbon-GCB SPE Cartridge, 250 mg, 3 ml; ANPEL, Shanghai, China, www.anpel.com.cn) and filtrated (SCAA-104, 0.22 μm pore size; ANPEL, Shanghai, China, www.anpel.com.cn), and then analyzed using an LC-ESI-MS/MS system (HPLC, Shim-pack UFLC Shimadzu CBM30A system, www.shimadzu.com.cn; MS, Applied Biosystems 4500 QTRAP, www.appliedbiosystems.com.cn/). The analytical conditions were as follows: HPLC column, Waters ACQUITY UPLC HSS T3 C18 (1.8 μm, 2.1 mm × 100 mm); solvent system, water (0.04% acetic acid): acetonitrile (0.04% acetic acid); gradient program, 95:5 V/V at 0 min, 5:95 v/v at 11.0 min, 5:95 v/v at 12.0 min, 95:5 v/v at 12.1 min, 95:5 v/v at 15.0 min; flow rate, 0.35 ml/min; temperature, 40 °C; injection volume, 2 μl. The effluent was alternatively connected to an ESI-triple quadrupole-linear ion trap (QTRAP) MS.
Linear ion trap (LIT) and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap MS (QTRAP) using an API 4500 QTRAP LC/MS/MS System, which was equipped with an ESI Turbo Ionspray interface operated in positive ion mode and controlled by Analyst 1.6.2 software (ABSciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature, 550 °C; negative ion spray voltage (IS), 4500 V; ion source gas I (GSI), gas II (GSII), and curtain gas (CUR) were set at 55, 60, and 25 (35) psi, respectively; and the collision gas (CAD) was high (medium). Instrument tuning and mass calibration were performed with 10 and 100 mmol/l polypropylene glycol solutions in QQQ and LIT modes. The QQQ scans were acquired as multiple reaction monitoring (MRM) experiments with the collision gas (nitrogen) set to 5 psi. The declustering potential (DP) and collision energy (CE) for individual MRM transitions were performed with further DP and CE optimization. A specific set of MRM transitions was monitored for each period according to the metabolites that were eluted within this period.
Using the above method, a total of 70 representative samples (35 in 2015 and 35 in 2016 years) were selected and carried out metabolome library construction. The result showed that 2151 substances could be identified. After performing quality control, a total of 1858 metabolites were found to be stable. These 1858 metabolites were used as the references to identify metabolites in the 252 samples over 2 years.

The sample extracts were analyzed using an LC-ESI-MS/MS system (HPLC, Shim-pack UFLC SHIMADZU CBM30A system, www.shimadzu.com.cn/; MS, Applied Biosystems 4500 Q TRAP, www.appliedbiosystems.com.cn/). The analytical conditions were as follows, HPLC: column, Waters ACQUITY UPLC HSS T3 C18 (1.8 μm, 2.1 mm^*100 mm); solvent system, water (0.04% acetic acid): acetonitrile (0.04% acetic acid); gradient program, 100:0 V/V at 0 min, 5:95 V/V at 10.0 min, 5:95 V/V at 11.0 min, 95:5 V/V at 11.1 min, 95:5 V/V at 15.0 min; flow rate, 0.35 ml/min; temperature, 40°C; and injection volume: 5 μl. The effluent was alternatively connected to an ESI-triple quadrupole-linear ion trap (Q TRAP)-MS.
LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (Q TRAP) using an API 4500 Q TRAP LC/MS/MS System, which was equipped with an ESI Turbo Ion-Spray interface operated in a positive ion mode and controlled by Analyst 1.6.2 software (AB Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 550°C; ion spray voltage (IS) 5,500 V; ion source gas I (GSI), gas II (GSII), curtain gas (CUR) were set at 55, 60, and 25.0 psi, respectively; and the collision gas (CAD) was high. Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. The QQQ scans were acquired as MRM experiments with the collision gas (nitrogen) set to 5 psi. The DP and CE for individual MRM transitions were performed with further DP and CE optimization. A specific set of MRM transitions was monitored for each period according to the metabolites that were eluted within this period.