Studies on a new cross-axis coil planet centrifuge for performing counter-current chromatography. I. Design of the apparatus, retention of the stationary phase, and efficiency in the separation of proteins with polymer phase systems.
An improved model of the cross-axis synchronous flow-through coil planet centrifuge has been designed in light of previous studies. The apparatus has a versatile feature in that both analytical and preparative columns can be accommodated in both off-center and central positions. Each has merit in separations. Retention of stationary phase was examined with various two-phase solvent systems used for the separation of biopolymers. Both analytical and preparative columns showed satisfactory retention of the stationary phase under optimum conditions. The apparatus was evaluated in separation of a set of protein samples using a polyethylene glycol-potassium phosphate biphasic system. In both types of columns all proteins were resolved with partition efficiencies of 260 to 670 theoretical plates. Further studies indicated that the relatively low partition efficiency of proteins is mainly attributed to their high molecular mass or molecular heterogeneity within each species rather than due to the high viscosity of the polymer phase system.
Shinomiya
K
K
Laboratory of Biophysical Chemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
Menet
J M
JM
Fales
H M
HM
Ito
Y
Y
D002498:Centrifugation / Q000295:instrumentation*; D002845:Chromatography / Q000295:instrumentation*; D007202:Indicators and Reagents; D011092:Polyethylene Glycols; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D012997:Solvents
D007202:Indicators and Reagents; D011506:Proteins; D012997:Solvents; D011092:Polyethylene Glycols