Chromatographic conditions
A 2 mL sample extract aliquot was injected into a Thermo LC-MS
Surveyor system (Rodano, Italy) equipped with a 2.1 100 mm
Symmetry C18 3.5 mm column and a 2.1 10 mm Symmetry C18
3.5 mm guard column (Waters, Saint-Quentin En Yvelines, France).
Separation was achieved at 35 C using a gradient elution of a
formic acid (0.1%) in water (A) and methanol (B) mixture, at a flow
rate of 200 mL/min. After an initial time of 3 min at 60% A, the
percentage of B was linearly increased from 40% to 50% between 3
and 10 min, then to 98% between 10 and 18 min, followed by a
holding-time of 6 min at 98%. After a washing step using 98% of
acetonitrile (C) and 2% of A at a flow rate 250 mL/min for 6 min, the
mobile phase compositionwas restored to the initial conditions for
10 min prior to the next injection.
Chromatographic conditions
A 2 mL sample extract aliquot was injected into a Thermo LC-MS
Surveyor system (Rodano, Italy) equipped with a 2.1 100 mm
Symmetry C18 3.5 mm column and a 2.1 10 mm Symmetry C18
3.5 mm guard column (Waters, Saint-Quentin En Yvelines, France).
Separation was achieved at 35 C using a gradient elution of a
formic acid (0.1%) in water (A) and methanol (B) mixture, at a flow
rate of 200 mL/min. After an initial time of 3 min at 60% A, the
percentage of B was linearly increased from 40% to 50% between 3
and 10 min, then to 98% between 10 and 18 min, followed by a
holding-time of 6 min at 98%. After a washing step using 98% of
acetonitrile (C) and 2% of A at a flow rate 250 mL/min for 6 min, the
mobile phase compositionwas restored to the initial conditions for
10 min prior to the next injection.
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