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The GBE product was obtained after the eluant was concentrated and dried, then weighted.
The neutral lipid fraction was redissolved in n-hexane after removal of the C-I eluant and transferred to fresh Bond Elut columns to elute cholesteryl esters with n-hexane and finally to elute TAGs with n-hexane containing 1% diethylether and 10% dichloromethane (by vol).
Bis(glycinato)copper(II) [[Cu[([L.sub.2]).sub.2]].sub.2] (3.11 g, 0.01 M) column eluted with water gave blue eluant. Yield: 1.92 g, 63%; d.p.: 212[degrees]C; UV-Vis (nm): 296, 386, 628.
The diluted extracting solution was chromatographed with HZ816 macroreticular resin using a gradient eluant of [H.sub.2]O-ethanol ([H.sub.2]O, 10%, 30%, 60%, and 90% ethanol).
The reaction mixture was refluxed for 2.5 h, and then the solvent was evaporated and the residue purified by column chromatography using Si[O.sub.2] as adsorbent and petroleum ether purified as eluant and its purity was > 98% using the HPLC methods described in a later section.
The resin which had adsorbed Pb(II) was shaken with the eluant. After reaching equilibrium, the concentration of Pb(II) in the aqueous phase was determined and then the percentage of elution was obtained.
After drying with [Na.sub.2]S[O.sub.4] and concentrating under reduced pressure, the EtOAc phase (2 g) was chromatographed over Sephadex LH-20 (30 x 2 cm), using MeOH as the eluant. This procedure provided 42 fractions (15 mL each), which were pooled into four groups (I-IV), after TLC analysis.
The oil obtained after extraction was purified by flash chromatography using silica gel (60 g) as the stationary phase and hexane: diethyl ether (80:20) as the eluant to yield 7.8 g of colorless, catalyst-free conjugated soybean oil.
The eluant was an unbuffered mixture of ethanol and water at a flow rate of 0.5 ml/min (except in the case of Potassium Fluoride when it was set at 1.0 ml/min).
Therefore, 0.01 M NaOH was used as eluant for desorption studies.
The sample injected and eluant of acetonitrile: water 9:1 was pumped at the rate of 1 mL/min in column controlled at 55[degrees]C.
The solvent was removed under reduced pressure and the residue was purified by flash chromatography, using dichloromethane/hexane 6/4 as eluant, to give 4 as a dark yellow solid (66.1 mg; yield 65%).
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