Triple tof 5600 ms ms system

All reagents were purchased commercially. ¹H-NMR and ¹³C-NMR spectra were recorded on Bruker AC400 and Bruker AC600 NMR spectrometers, respectively (Billerica, MA, USA). Low-resolution mass spectra were recorded on a 6120 Quadrupole mass spectrometer (Agilent, Santa Clara, CA, USA) equipped with electrospray ionization (ESI). High-resolution mass spectra were determined on triple TOF 5600⁺ MS/MS system (AB Sciex, Concord, ON, Canada) in negative ESI mode. The purity of target compounds was determined by high-performance liquid chromatography (Agilent, Santa Clara, CA, USA, DIKMA Diamonsil Plus C18, 250 × 4.6 mm, 5 μm, 25 °C, UV 290 nM). All the biologically tested compounds achieved ≥95% purity.

Chromatographic analysis was carried out on a Prominence™ UHPLC system (Shimadzu, Japan) coupled with a triple TOF™ 5600+ MS/MS system (AB Sciex, Framingham, MA, USA). Separations were accomplished on a Welch C₁₈ column (2.1 mm × 100 mm, 1.8 μm, Shanghai, China) and 2 μL injected into UHPLC. The column oven was maintained at 40 °C. A linear gradient elution of eluents A (water containing 0.1% formic acid) and B (acetonitrile) was used for separation. The elution program was: 0.1–2 min, 1–5% B; 2.01–20 min, 8–37% B; 20–22 min, 37–50% B; 22–32 min, 50–90% B; 32.01–35 min, 95–95% B, 35.01–40 min, 1% B. The flow rate was 0.25 mL/min.
The mass spectrometer was operated both in negative and positive ion modes. The following parameter settings were used: ion spray voltage 4500 V; turbo spray temperature 600 °C; curtain gas 25 psi, nebulizer gas (GS 1) 50 psi, heater gas (GS 2) 50 psi, declustering potential 100 V. TOF MS and TOF MS/MS were scanned with the mass range of m/z 50–1250 and 50–1250, respectively. In the IDA-MS/MS experiment, collision energy was set at 35 eV and collision energy spread was (±) 10 eV. Accurate mass and composition for the precursor and fragment ions were analyzed using Peakview software (Version 1.2, AB Sciex) integrated with the instrument.

High-resolution magic-angle-spinning NMR (Bruker BioSpin, Germany) and solution NMR (Bruker BioSpin) were used for ¹H NMR-based metabolomic analysis of heart and serum samples, respectively. An Agilent 1290 Infinity LC and 6490 Triple Quadrupole MS system (Agilent Technologies, USA) was used to determine the concentration of SAAs. LC-ESI-MS/MS analysis for lipidomics was performed on a triple TOF™ 5600 MS/MS System (AB Sciex, Canada) combined with a UPLC system (Waters, USA). For further details, see Supplementary Information.