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In this system, feed and mobile phase streams are subsequently delivered through parallel columns to mimic their countercurrent movement with respect to the fluid flow.
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Chromatography, technique for separating the components, or solutes, of a mixture on the basis of the relative amounts of each solute distributed between a moving fluid stream, called the mobile phase, and a contiguous stationary phase.
A further optimization was applied to sample treatment by evaporation of solvent under a nitrogen stream and redissolution in the mobile phase to achieve high sensitivity of the assay.
flow rate of mobile phase (mL/min).
The collected samples were extracted with 0.2 ml dichloromethane, evaporated by nitrogen stream, and reconstituted in 100 μl mobile phase.
The methanolic extract was evaporated to dryness under nitrogen stream and reconstituted in 2 mL mobile phase.
The supernatants were dried under a stream of nitrogen and reconstituted in mobile phase (100 μl).
The mobile phase was removed from the plate by drying with a stream of cool air from a heating gun.
The organic phase was evaporated under a stream of nitrogen and resuspended in HPLC mobile phase (methanol : water 1 : 1 v v−1) (Bugianesi et al, 2000).
In all experiments, the plates were dried under air stream before they were developed with a new mobile phase.
These sample extracts were evaporated to dryness at room temperature under a stream of nitrogen, reconstituted with 200 μL of mobile phase, vortexed for 1 min, filtered through a 0.22 μm pore size filter (Millipore), and then 5 μL of the solution was injected into the LC--MS/MS system.
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