6230 ms tof mass spectrometer

Qualitative analysis was carried out using a 1260 Infinity liquid chromatograph (Agilent, Santa Clara, CA, USA) coupled to a 1290 Infinity photodiode array detector and a 6230 MS-TOF mass spectrometer. Separation of compounds was achieved using mobile phases such as ultrapure water (A) and acetonitrile (B), acidified with 2% (v/v) formic acid. A 5 μL sample was injected onto a pre-column guarded Zorbax SB-C18 column (250 × 4.6, 5 µm) and eluted using a gradient program (0–23 min, 0–20% B; 23–27 min, 20–22.5% B; 27–28 min, at 24.4% B; 28–30 min, 24.4–26.2% B; 30–50 min, 26.2–30% B; 50–60 min, 30–100% B; 6 min recondition). Solvent flow was 1.2 mL/min. The separation took place at 40 °C. The mass spectrometric conditions were as follows: an electrospray ionization (ESI) source was used in the negative ion mode with drying and sheath gas flow set to 11 L/min and temperature to 350 °C. A nebulizer pressure of 60 psi was used, capillary voltages of 2500 and 1000 V were applied, and the fragmentor was set to 60–320 V. The data were collected in the 120–3000 m/z range and analyzed in MassHunter Qualitative 10.0. analysis software. The compounds were characterized based on UV-Vis and MS spectra and the retention time of the standards.

The assessment of chemical composition of each extract (RE, LBE) was carried out on a 1260 Infinity chromatograph (Agilent, Santa Clara, CA, USA) consisting of binary pump, a column oven, and photo-diode array (PDA) detector over 55 min period. The separation was performed using a Kinetex XB C18 column (150 × 3 mm, 2.6 µm) (Phenomenex, Torrance, CA, USA). The mobile phase was 0.1% (v/v) FA in water (A) and 0.1% FA in acetonitrile (B). The separation was achieved by a gradient of 0–2 min 1% B; 2–20 min 1–20% B; 20–40 min 20–75% B; 40–45 min 75–95% B; 45–48 min 95% B; 48–49 min 95–1% B; 49–55 min 1% B. The flow rate was 0.5 mL/min and the column temperature was maintained at 25 ± 0.8 °C. The UV–Vis spectra was recorded from 190 to 540 nm with selective wavelength monitoring at 280 nm. Mass spectrometry (MS) detection was carried out on a 6230 MS/TOF mass spectrometer (Agilent, Santa Clara, CA, USA) equipped with an electrospray ionization source with Agilent Jet Stream thermal focusing. The parameters used for ionization source were set as follows: drying and sheath gas flow: 12 L/min; nebulizer: 35 psi; source temperature 350 °C; ion spray voltage 4500 V for the positive mode analysis. The data were collected in 115–1900 m/z range and processing was performed using Mass Hunter qualitative analysis software.