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The crude extract of S. dasyphyllum was subjected to solvent partitioning using n-hexane, dichloromethane ethylacetate and methanol (according to their polarity).
After confirmation of the anti-tumor activity, the 95% ethanolic extract was subjected to successive solvent partitioning to petroleum ether, dichloromethane, n-butanol, and water soluble fractions.
The presence of cells in the system at levels normally used in in vitro exposure systems did not significantly alter solvent partitioning.
The empirically calculated parameter LogPo/w, the log10 of the coefficient for solvent partitioning between 1-octanol and water, has been used to provide the key data for a unique non-covalent interaction force field called HINT (Hydropathic INTeractions).
However, addition of a few percent N,N-dimethyl formamide resulted in agglomeration of the dispersed montmorillonite layers and an overall mixed morphology, demonstrating solvent partitioning and the delicate enthalpic balance necessary to maintain layer dispersion.
It is interesting that the relatively crude HINT "electrostatics" [11], largely based on experimental solvent partitioning of small organic molecules, perform measurably better than the CNS partial charge-based Coulombic electrostatics.
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The total DHCA recovery was twice higher than that when the solvent partition was used.
A further purification of the SC-CO2 extract using a normal phase Silica-gel 60 resin column chromatography without two solvent partitions could yield a purified product containing 95 wt.% DHCA.
Countercurrent chromatography (CCC) using solely immiscible solvent systems allowed the fractionation of principal components and an enrichment of minor concentrated metabolites from a crude polar solvent partition of C. limetta peels for subsequent structural identification by LC ESI MS/MS analysis.
Reactions to determine the glycosyltransferase activity of MrSGT on fucosterol and gramisterol, were performed as follows; MrSGT dissolved in the above-mentioned reaction buffer was incubated with 0.6 mM each of fucosterol and gramisterol, respectively, along with 0.8 mM of UDP-Glc at 30 °C for 15 min, and then extracted by organic solvent partition.
As described in Material & Methods, gangliosides were extracted by several solvent partitions and their oligosaccharides were digested by an endoglycoceramidase.
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