Ito Yoichiro, Clary Robert, Sharpnak Frank, Metger Howard, Powell Jimmie
Center for Biochemistry and Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 50, Room 3334, 50 South Drive, Bethesda, MD 20892-8014, USA.
J Chromatogr A. 2007 Nov 23;1172(2):151-9. doi: 10.1016/j.chroma.2007.09.078. Epub 2007 Oct 10.
A novel system for performing high-speed counter-current chromatography has been developed for separation of biopolymers using polymer phase systems. The spiral disk assembly consisting of eight units, each equipped with over 300 mixer-settler sets, was constructed and performance evaluated in terms of retention of the stationary phase and separation efficiency. A series of experiments was performed with a polymer phase system composed of polyethylene glycol 1000 (12.5%, w/w) and dibasic potassium phosphate (12.5%, w/w) using two stable protein samples of myoglobin and lysozyme at various experimental conditions of flow rates and revolution speeds. The best results were obtained with revolution speeds of 800-1000rpm at flow rates of 0.25-0.5ml/min where the partition efficiency of several 100 theoretical plates was achieved with over 50% stationary phase retention.
已开发出一种用于执行高速逆流色谱的新型系统，用于使用聚合物相系统分离生物聚合物。构建了由八个单元组成的螺旋盘组件，每个单元配备300多个混合沉降器组，并根据固定相保留率和分离效率对性能进行了评估。使用聚乙二醇1000（12.5%，w/w）和磷酸氢二钾（12.5%，w/w）组成的聚合物相系统，在流速和转速的各种实验条件下，对肌红蛋白和溶菌酶这两种稳定的蛋白质样品进行了一系列实验。在流速为0.25 - 0.5ml/min、转速为800 - 1000rpm时获得了最佳结果，此时实现了几百个理论塔板的分配效率，固定相保留率超过50%。
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