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Purification was performed under isocratic elution using two mobile phases at a flow rate of 3 mL/min: first with 100% water for 10 min then switching to 40% MeCN in water.
In this work, 10μL samples of a 50 mM probe solution were injected into C18-bonded columns using a series of five buffered mobile phases at SWpH between 2 and 12.
The results indicated that these separations could be tuned by variation of the temperature with fully aqueous mobile phases at specific ionic strength and pH values, without the need to use an organic solvent as a component in the mobile phase.
Sample loading (8 µL/sample) from the microtiter plate into the enrichment column was performed at a flow rate set to 4 µL/min with the mix of the two following mobile phases at a ratio 98∶2 (mobile phase A: 0.2% formic acid in H2O; mobile phase B: 100% ACN).
HPLC separation was performed at 30°C on a XBridge Shield RP18 column (2.1 × 50 mm, 3.5 μm, Waters) using (A) 1 mM formic acid and (B) acetonitrile as mobile phases at a flow rate of 0.4 ml/min.
Similar(55)
Chloroform was used as the mobile phase at a flow rate of 1.0 ml min−1.
Acetonitrile was used as a mobile phase at a flow rate of 1.0 mL/min.
Tetrahydrofuran was used as the mobile phase, at a flow rate of 1 mL/min.
Figure 5 Plot of HETP of glycine and ammonium chloride versus the interstitial velocity of mobile phase at 50°C. Figure 6 Plot of HETP of glycine and ammonium chloride versus the interstitial velocity of mobile phase at 60°C.
Separation was performed at 45 °C with 5 mM H2SO4 as mobile phase at a flow rate of 0.6 mL/min.
Separation was performed at 80°C, with H2O as mobile phase at a flow rate of 0.6 mL/min.
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