Mixer-settler counter-current chromatography with multiple spiral disk assembly.
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.
Ito
Yoichiro
Y
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. itoy2@mail.nih.gov
Clary
Robert
R
Sharpnak
Frank
F
Metger
Howard
H
Powell
Jimmie
J
D000465:Algorithms; D002498:Centrifugation; D002853:Chromatography, Liquid / Q000379:methods; D003377:Countercurrent Distribution / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D009113:Muramidase / Q000032:analysis; D009211:Myoglobin / Q000032:analysis; D011092:Polyethylene Glycols / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D014867:Water
D009211:Myoglobin; D011108:Polymers; D012997:Solvents; D014867:Water; D011092:Polyethylene Glycols; D009113:Muramidase