The separation of molecules was achieved using an Acquity UPLC I-class UHPLC (Waters, Milford, MA, US) with a PDA detector, coupled to a hybrid quadrupole-time of flight (Q-TOF) mass spectrometer TripleTOF 6600 (SCIEX Instruments, Concord, ON, Canada) with a DuoSpray Ion Source operating in negative and positive ion mode. Five µL of the samples in random order were injected in the UHPLC system and analyzed in a run time of 60 min. The separation was performed on a reverse-phase Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7-μm particle size) (Waters, Milford, MA, US). The solvents used were A) water + 0.1% FA and B) acetonitrile (ACN) + 0.1% FA, all LC-MS grade and the column was maintained at 50 °C during all the run time. The gradient was as follows: 0 min, 1% B; 4 min, 1% B; 16 min, 5% B; 35 min, 40% B; 45 min, 100% B; 50 min, 100% B; 54 min, 1% B; 60 min, 1% B, at 0.5 mL/min flow rate. UV-visible spectra were also acquired between 190 and 800 nm at a rate of 10 points/s.

The ESI parameters were set as follows: source temperature of 650 °C, ion spray voltage of −4500 V and 4500 V, for the negative and positive mode, respectively, curtain gas (nitrogen) of 30 psi, nebulizer gas (air) of 55 psi and turbine gas (air) of 50 psi. The declustering potential was set at −60 eV in negative mode and 60 eV in positive mode. The precursor charge state selection was set at 1. For information-dependent acquisition in high sensitivity mode, survey scans were acquired in 175 ms and the 10 most abundant product ion scans were collected during 200 ms if exceeding a threshold of 100 counts/s, the total cycle time being 2.225 s. A sweeping collision energy setting of 15 eV below and above 15 eV was applied to all precursor ions. The dynamic exclusion was set for 2 s after three occurrences before the precursor could be fragmented again. For MS1, full HR-MS spectra between 100 and 2000 mass-to-charge ratio (m/z) were recorded. MS2 scans were recorded between 50 and 2000 m/z, in profile mode.

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