A new type-J CCC planet centrifuge with two spiral tube columns (2×15 mL, β value 0.3–0.7) was developed and evaluated for its retention ability of different solvent systems including less polar, moderately polar and polar organic-aqueous solvent systems, and aqueous two-phase systems in different operation conditions. The results suggested that the spiral tube column can produce good retention of stationary phase for less polar to moderately polar solvent systems, such as heptane-methanol (1:1, v/v) and hexane-ethyl acetate-methanol-water (1:1:1:1, v/v) (1mL/min, Sf=70%) by eluting lower mobile phase from head (H) to tail (T), or upper mobile phase from tail (T) to head (H), regardless of whether the inlet is located at the inner (I) or outer (O) terminals. It can also provide satisfactory retention for polar to aqueous two-phase systems such as n-butanol–acetic acid–water (4:1:5, v/v/v) (0.5 mL/min, Sf=60%) and PEG1000-K2HPO4-water (12.5:12.5:75, w/w) (0.5 mL/min, Sf=50%) by eluting lower mobile phase from inner terminal (I) to outer terminal (O), or upper mobile phase from outer terminal (O) to inner terminal (I), except for U-O-T mode. The results also clearly indicated that the spiral tube column could significantly increase the retention of four typical solvent systems compared with a multilayer coil column with similar parameters, the Sf for the former two solvents can be increased by about 10%, and for the latter two by 30–40%. The preliminary applications of this spiral tube column on the separation of small molecular compounds such as moderately polar theaflavins, polar anthocyanins and dipeptides were successful. Acceptable resolution can be obtained between cytochrome c and myoglobin, lysozyme and myoglobin when it was applied on protein separation, however it still needs to be improved regarding to its low column efficiency.
展开▼
