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Processing involved evaporating samples of known volume to dryness.
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The solvent volume was reduced to dryness and the residue was dissolved in n-hexane (200 μL).
One half of the total volume was evaporated to dryness and used for the BNI bioassay, while the other half was concentrated 100-fold (from 1.5 L to 15 mL) using rotary evaporators.
Total dissolved solids (TDS) were estimated by weighing the solid residue obtained by evaporation of a measured volume of water samples to dryness (Chopra and Kanwar 1980).
This volume was evaporated (45°C) to dryness in a vacuum concentrator and then reconstituted with 300 μL of mobile phase.
The volume of 1 mL was reduced to dryness and the pellet resuspended in 30 μL water.
After three times repeated protein precipitation by addition of an equal volume of acetonitrile followed by evaporation to dryness, the samples were re-dissolved in 100 μl acetonitrile/water (1 : 9, v/v) containing 0.2% v/v formic acid.
Supernatant in volumes of 500 μl was evaporated to dryness under vacuum in a CentriVap™ Concentrator (Labconco).
The volumes obtained (1200 mL) were concentrated to dryness under reduced pressure at 40°C to yield 26 g of n-hexane, 12.5 g of EtOAc, and 29.0 g of EtOH fractions.
A residual supernatant (2.5 L) was extracted twice with an equal volume of ethylacetate, and concentrated in vacuo to dryness to produce red brown materials (1.2 g).
After three repetitions of protein precipitation by the addition of an equal volume of acetonitrile followed always by evaporation to dryness, the samples were redissolved in 100 μL of acetonitrile/water (1/9 vol/vol) containing 0.2% vol/vol formic acid.
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