Shinomiya Kazufusa, Ito Yoichiro
College of Pharmacy, Nihon University, 7-7-1, Narashinodai, Funabashi-shi, Chiba 274-8555, Japan.
J Liq Chromatogr Relat Technol. 2009 Jan 1;32(8):1096-1106. doi: 10.1080/10826070902841547.
Countercurrent chromatographic performance of the locular multilayer coil separation column newly designed in our laboratory was evaluated in terms of theoretical plate number, peak resolution and retention of the stationary phase in protein separation with an aqueous polymer phase system using the small-scale cross-axis coil planet centrifuge (X-axis CPC) fabricated in our laboratory. The locular column was made from 1.0 mm I.D., 2.0 mm O.D. or 1.5 mm I.D., 2.5 mm O.D. PTFE tubing compressed with a pair of hemostat at 2 or 4 cm intervals. The protein separation was performed using a set of stable proteins including cytochrome C, myoglobin and lysozyme with the 12.5% (w/w) polyethylene glycol 1000 and 12.5% (w/w) dibasic potassium phosphate system under 1000 rpm of column revolution. The 1.5 mm I.D., 2.5 mm O.D. locular tubing compressed at 2 cm intervals yielded better partition efficiencies than the non-clamped tubing using both lower and upper mobile phases with satisfactory retention of the stationary phase. The overall results suggest that the newly designed locular multilayer coil is useful to the preparative separation of proteins with aqueous-aqueous polymer phase system using our small-scale X-axis CPC.
使用我们实验室制造的小型横轴盘管行星离心机（X轴CPC），在蛋白质分离中，以理论塔板数、峰分辨率和固定相保留率为指标，评估了我们实验室新设计的腔室多层盘管分离柱的逆流色谱性能。腔室柱由内径1.0毫米、外径2.0毫米或内径1.5毫米、外径2.5毫米的聚四氟乙烯管制成，用一对止血钳以2或4厘米的间隔压缩。蛋白质分离使用一组稳定的蛋白质，包括细胞色素C、肌红蛋白和溶菌酶，在12.5%（w/w）聚乙二醇1000和12.5%（w/w）磷酸氢二钾体系中，柱转速为1000转/分钟。内径1.5毫米、外径2.5毫米、间隔2厘米压缩的腔室管，在使用上下流动相时，比未夹紧的管具有更好的分配效率，且固定相保留令人满意。总体结果表明，新设计的腔室多层盘管对于使用我们的小型X轴CPC在水-水聚合物相体系中进行蛋白质的制备分离是有用的。
