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The separation was optimum under the mobile phase conditions described in the section " Reversed-phase HPLC of labeled N -glycans".
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The assay procedure involves chromatographic separation on a Zorbax-Eclipse C-18 column under isocratic conditions with the mobile phase consisting of acetonitrile and ammonium acetate buffer (pH 5.0, 10 mM) in different ratios depending upon the compound.
The plate was developed with a stationary phase of silica gel on aluminium paper and the extract was charged and moved under the influence of the mobile phase of chloroform and methanol in 9 1 ratio for 4 h.
Aliquots of 20 μL were injected (triplicate) and eluted with the mobile phase under the reported chromatographic conditions.
Twenty μL aliquots were injected (in triplicates) and eluted with the mobile phase under the reported chromatographic conditions.
Aliquots of 20 μL were injected (triplicate) and eluted with the mobile phase under the optimum chromatographic conditions.
20 μL aliquots were injected (triplicate) and eluted with the mobile phase under the reported chromatographic conditions.
Aliquots of 4.0 μL were injected (six injections) and eluted with the mobile phase under the reported chromatographic conditions.
Acetonitrile and water in 1 1 (v/v) ratio was chosen as the mobile phase under a column temperature of 25 C.
Acetonitrile 0.1% HCOOH/H2O was used as the mobile phase under a linear gradient elution mode (acetonitrile, 28 60%, 50 min (compound 1); acetonitrile, 35 60%, 50 min (compound 2)) at a flow rate of 1 mL/min.
Carotenoids were separated by RP-HPLC using a reverse phase RP-18 LiChroCART 125 4 (Merck) column with acetonitrile:methanol:isopropanol (85:10:5 v/v) as the mobile phase under isocratic conditions with a 1 ml/min flux.
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