Acquity ftn autosampler

ET fractions
were subjected to UPLC-ESI-MS/MS analysis using negative and positive
ion acquisition modes on a triple quadruple instrument (XEVO TQD,
Waters Corporation, Milford, MA) mass spectrometer; column: ACQUITY
UPLC-BEH C18 1.7 μm × 2.1 mm × 50 mm column. The samples
were injected automatically using a Waters ACQUITY FTN autosampler.
The mobile phase was filtered using a 0.2 μm filter membrane
disc and degassed by sonication before injection. Its flow rate was
0.2 mL/min using a gradient mobile phase (methanol and water acidified
with 0.1% formic acid that applied from 10% to 30% in 5 min, then
from 30% to 70% in 10 min, then from 70% to 90% in 5 min, then holds
the gradient for 3 min, then from 90% to 10% in 3 min). The instrument
was controlled by Masslynx 4.1 software.

Root and in
vitro induced callus extracts, 40 days old callus grown on
medium I (100 μg/mL), were dissolved in high-performance liquid
chromatography (HPLC) solvent grade and filtered using a 0.2 μm
membrane disc filter. Then, 10 μL of each of the prepared samples
was individually injected into the instrument equipped with a reversed-phase
C18 column (ACQUITY UPLC BEH C18 2.1 mm × 50 mm Column, 1.7 μm
particle size). The mobile phase was filtered using a 0.2 μm
filter membrane disc and degassed by sonication before injection.
The flow rate was adjusted to 0.2 mL/min using a gradient mobile phase
(methanol and water acidified with 0.1% formic acid that was applied
from 10 to 30% in 5 min, from 30 to 70% in 10 min, from 70 to 90%
in 5 min, then the gradient was held for 2 min, and finally from 90
to 10% in 2 min). The samples were injected automatically using a
Waters ACQUITY FTN autosampler. The instrument was controlled by Masslynx
4.1 software. The parameters for analysis were measured using negative
modes as follows: source temperature, 150 °C; cone voltage, 30
eV; capillary voltage, 3 kV; desolvation temperature, 450 °C;
cone gas flow, 50 L/h; and desolvation gas flow, 900 L/h. Mass spectra
were detected in the ESI negative ion and positive ion modes between m/z 50 and 1000.