Compound discoverer software

Liquid chromatography-mass spectrometry analysis was carried on LC-MS (Thermo, Ultimate 3000LC, Q Exactive) platform. The parameter for both ESI+ and ESI− ion mode is listed below Heater Temp 300°C; Sheath Gas Flow rate, 45arb; Aux Gas Flow Rate, 15 arbs; Sweep Gas Flow Rate, 1arb; spray voltage, 3.0KV; Capillary Temp, 350°C; S-Lens RF Level, 30%. The data was performed with feature extraction and preprocessed with Compound Discoverer software (Thermo). Two thousand fifteen features at (ESI+) ion mode and 1,601 features at (ESI−) ion mode in this experiment, the data after editing were performed Multivariate Analysis (MVA) using SIMCA-P software (Umetrics AB, Umea, Sweden).

The samples were thawed at room temperature, 100 μL of them was then transferred into Centrifuge Tubes (1.5 mL) by pipette. All samples were extracted with 300 μL of methanol, and 10 μL of internal standard (3 mg/mL, DL-o-Chlorophenylalanine) was added. The samples were then ultra-sonicated at 4 K Hz on ice bath for 30 min. The samples were vortexes for 30 s, and centrifuged at 12,000 rpm and 4°C for 15 min. Two hundred microliter of supernatant was transferred to vial for LC-MS analysis. Analysis platform: LC-MS (Thermo, Ultimate 3000LC, Orbitrap Elite) Column: Waters ACQUITYUPLC HSS T3column (2.1 mm × 100 mm, 1.8 μm) Chromatographic separation conditions: Column temperature: 40°C; Flow rate: 0.3 mL/min; Mobile phase A: water + 0.1% formic acid; Mobile phase B: acetonitrile + 0.1% formic acid; Injection volume: 4 μL; Automatic injector temperature: 4°C. The data was performed feature extraction and preprocessed with Compound Discoverer software (Thermo), and then normalized and edited into two-dimensional data matrix by excel 2010 software, including Retention time(RT), Compound Molecular Weight (comp MW), Observations (samples) and peak intensity.

The bioactive molecule profile of cinnamon extract before and after digestion and the methodology utilised for the analyses were reported in a previous paper [9 (link)]. Briefly, a Thermo Vanquish UHPLC system coupled with a Thermo Orbitrap Exploris 120 mass spectrometer and a Vanquish Diode Array Detector (Thermo Scientific, Rodano, Italy) was used to identify the target bioactive molecules. The chromatographic separation was carried out on a Luna Omega Polar C18 (150 mm × 2.1 mm, 3 µm) (Phenomenex, Castelmaggiore, Italy). Phenolic compounds were identified based on the corresponding spectral characteristics (UV and MS/MS spectra), accurate molecular mass, characteristic MS fragmentation pattern, and library comparison in a semi-automatic way through Compound Discoverer Software (2.1, Thermo Scientific, Rodano, Italy). A semi-quantification was carried out to reveal the changes in the compound composition of the cinnamon extract before and after digestion. All compounds were quantified in duplicate at 280 nm by external calibration lines, dividing them into two major groups according to their structural similarity: trans-cinnamic acid or catechin. The data are reported in Pagliari et al., 2023 [9 (link)].