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Both of these samples were diluted to give 10−5 M stock solutions and then mixed with a 10−5 M MB stock solution to give two sets of mixtures of increasing MB concentrations.
Physiologically based pharmacokinetic (PBPK) models represent a potentially useful framework for predicting the consequences of interactions in mixtures of increasing complexity.
Sub-fraction C (8.0 g) obtained with Hexane-CHCl3 4:6 was subjected to CC (silica gel 60, 50 g) and eluted with Hex-CHCl3 mixtures of increasing polarity to give 6 new sub-fractions (C1-C6).
The EtOAc fraction was further chromatographed on a silica gel column (120 g) eluting with n-hexane-CH2Cl2 (100 0 to 0 100), then CH2Cl2-MeOH (100 0 to 0 100) mixtures of increasing polarity to yield 11fractionss.
The column was initially eluted with dichloromethane (DCM, 200 ml), and then eluted with ethyl acetate:dichloromethane (EtOAc: DCM) mixtures of increasing polarity (EtOAc: DCM 1 9, 1 4, 3 7, 1 1, 4 1 and 1 0).
Flash chromatography of HBFEC (12.0 g) using n-hexane and EtOAc as binary mixtures of increasing polarity afforded 30 fractions, which were pooled in 9 fractions after thin layer chromatography (TLC) analysis.
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The mobile phase consisted of a mixture of increasing proportions of CH3OH in water with trifluoroacetic acid 0.01% (TFA).
For instance, a mixture of increasing the dose of recombinant sulfamidase and modifying the chemical structure to remove mannose-6-phosphate glycans has been attempted.
Part of FPR (200 g) was submitted to silica gel 60 (0.04-0.063 mm, 200 g) vacuum flash chromatography using as eluent, hexane (Hex), hexane-ethyl acetate (Hex-EtOAc) mixture of increasing polarity, and methanol (MeOH).
Fraction E (10.0 g) obtained with CHCl3 was subjected to CC (silica gel 60, 50 g) and eluted with CHCl3-MeOH mixture of increasing polarity to give 7 sub-fractions (E1-E7).
The resulting yellowish residue was extracted with diethyl ether (3 x 150 mL), dried, concentrated and purified by column chromatography eluting with cyclohexane/ethyl acetate mixture of increasing polarity to give the corresponding alkaloids (8a–' o').
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