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After switching to the elution position the analyte was eluted with the mobile phase (gradient from 90% acidic solution of ammonium acetate and 10% acetonitrile to 100% acetonitrile in 5 seconds, flow rate 1.5 ml /min) and transferred to the mass spectrometric detector (split approximately 1 10).
The program was set as follows: 95% solvent A for 1 minute 30 seconds (flow rate 0.4 ml/minute), followed by 6 minutes in which solvent B increased till 98% (0.2 ml/min) which continued for 2 minutes 30 seconds at the same flow rate, followed by 1 minute 30 seconds at an increased flow rate (0.4 ml/min), subsequently returning to 95% solvent A for 1 minute until the end of the run.
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Second, flow rates recorded from the driving ventilator were scaled by the ratio between the tidal volume measured by the support ventilator and that reported by the driving ventilator.
At first the flow rate increases as the density increases, since you simply get more stuff through in the same period of time.
To prevent loss of analytes during the online analyses, the first-dimension flow rate did not exceed the 0.2 μL min−1.
Modulation between the first- and second-dimension, which may be achieved by focussing and desalting on trap columns before analyses on the D column [ 36], may allow reducing the D column diameter and, thus, the second-dimension flow rate.
The UV detector was equipped with either a 3-nL Z-shaped detector cell for the 1D capillary measurements (typically operated at 3 5 μL min−1) or a 180-nL Z-shaped detector cell for the 2D measurements (typical second-dimension flow rate (F) of 50 200 μL min−1).
The evaluation is conducted through simulations and practical essays with two different plants: a first order water flow rate problem and a second order under-damped electronic circuit.
A 1 mL injection in 10 seconds (360 mL/h flow rate) generated a pressure of 24.0 ± 3.4 kPa, whereas 10 mL delivered in 10 minutes (60 mL/h flow rate) generated a pressure of 7.4 ± 7.8 kPa.
Kinetic experiments were carried out with a contact time of 90 seconds at a flow rate of 30 µl/minute at 25°C.
Association of IFN- γ-SC with receptors was adjusted to 90 seconds at a flow rate of 100 μL/min and dissociation occurred in PBST running buffer for 10 min at the same flow rate.
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