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采用β-环糊精键合的带负电荷聚丙烯酰胺凝胶整体手性固定相通过毛细管电色谱法对阳离子和中性化合物进行对映体分离。

Enantiomeric separations of cationic and neutral compounds by capillary electrochromatography with monolithic chiral stationary phases of beta-cyclodextrin-bonded negatively charged polyacrylamide gels.

作者信息

Koide T, Ueno K

机构信息

Shionogi Research Laboratories, Shionogi & Co, Ltd, Osaka, Japan.

出版信息

J Chromatogr A. 2000 Sep 29;893(1):177-87. doi: 10.1016/s0021-9673(00)00699-3.

Abstract

Enantiomeric separation by capillary electrochromatography with beta-cyclodextrin-bonded negatively charged polyacrylamide gels was examined. The columns used are capillaries filled with a negatively charged polyacrylamide gel, a so-called monolithic stationary phase, to which allyl carbamoylated beta-CD (AC-beta-CD) derivatives covalently bind. The capillary wall is activated first with a bifunctional reagent to make the resulting gel bind covalently to the inner surface of the fused-silica tubing. Enantiomeric separations of 15 cationic compounds were achieved using the above-mentioned columns and mobile phases of 200 mmol l(-1) Tris-300 mmol I(-1) boric acid buffer (pH 7.0 or 9.0) or 200 mmol l(-1) Tris-300 mmol l(-1) boric acid buffer (pH 7.0) containing an achiral crown ether (18-crown-6). Enantiomeric separations of two neutral compounds were also achieved using 200 mmol l(-1) Tris-300 mmol l(-1) boric acid buffer (pH 9.0) as a mobile phase. High efficiencies of up to 150,000 plates m(-1) were obtained. Both the within- and between-run reproducibilities of retention time and separation factor were good. The reproducibilities of retention time and separation factor for three different columns prepared from a different batch of monomers were acceptable. The gel-filled capillaries were stable for at least 3 months with intermittent use, utilizing the mobile phase of 200 mmol I(-1) Tris-300 mmol I(-1) boric acid buffer (pH 9.0).

摘要

考察了采用β-环糊精键合的带负电荷聚丙烯酰胺凝胶的毛细管电色谱法进行对映体分离。所用的柱是填充有带负电荷聚丙烯酰胺凝胶的毛细管,即所谓的整体固定相,烯丙基氨甲酰化β-环糊精(AC-β-CD)衍生物与之共价结合。首先用双功能试剂活化毛细管内壁,使生成的凝胶与熔融石英管的内表面共价结合。使用上述柱以及200 mmol l(-1) Tris-300 mmol I(-1)硼酸缓冲液(pH 7.0或9.0)或含有非手性冠醚(18-冠-6)的200 mmol l(-1) Tris-300 mmol l(-1)硼酸缓冲液(pH 7.0)作为流动相,实现了15种阳离子化合物的对映体分离。使用200 mmol l(-1) Tris-300 mmol l(-1)硼酸缓冲液(pH 9.0)作为流动相,也实现了两种中性化合物的对映体分离。获得了高达150,000理论塔板数每米的高效能。保留时间和分离因子的批内和批间重现性均良好。由不同批次单体制备的三根不同柱子的保留时间和分离因子的重现性是可接受的。使用200 mmol I(-1) Tris-300 mmol I(-1)硼酸缓冲液(pH 9.0)作为流动相,间歇使用时,填充凝胶的毛细管至少稳定3个月。

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