该文研究了高速逆流色谱技术(high-speed counter-current chromatography,HSCCC)分离麦芽糖基β-环糊精的可行性及分离条件。以醇盐溶液作为溶剂体系,通过测定麦芽糖基-β-环糊精、β-环糊精和麦芽糖在上下相的分配系数,发现磷酸二氢钠作为麦芽糖基β-环糊精分离的盐,丙酮作为改善两相分离时间的改性剂时,麦芽糖基-β-环糊精、β-环糊精和麦芽糖三者的分配系数差异最大,且相分离时间小于30 s;采取体积比为6∶2∶1.5的磷酸二氢钠(300 g/L)-乙醇-丙酮溶液为溶剂体系,以上相为固定相,下相为流动相,主机转速为800 r/min,流速为2 mL/min,每隔15 min收集流出液,经HPLC检测,30~35 min内收集的组分为麦芽糖基β-环糊精,纯度为93.2%,固定相保留率为60%。以上结果表明,HSCCC在麦芽糖基-β-环糊精的分离纯化中具有潜在应用价值。
High speed countercurrent chromatography (HSCCC) is a rapid and efficient separation method based on liquid-liquid separation principle. In this article, HSCCC was investigated to separate maltosyl-β-cyclodextrin, maltose and β-cyclodextrin simultaneously. The partitioning of sugars was investigated in ethanol-salt aqueous two-phase systems addressing the effects of phase-forming salt type (NaH2PO4/K2HPO4/(NH4)2SO4) and concentration, organic reagents type and concentration. By measuring the partition coefficients of sugars in the upper and lower phases, it was found that the partition coefficients were the most different among maltosyl-β-cyclodextrin, β-cyclodextrin and maltose when NaH2PO4 and acetone were used, and the phase separation time was less than 30 s. V(300 g/L NaH2PO4)∶V(ethanol)∶V(acetone)=6∶2∶1.5 was optimized finally for the solvent system, and the above phase was the stationary phase and another was the mobile phase. The operating conditions were determined to be 800 r/min and with 2 mL/min of the flow rate. The effluent was collected every 15 minutes. In 30-35 min, the target compound was collected and analyzed by HPLC, the result was shown that the purity was up to 93.2% which increased by 61%, and the retention rate of the stationary phase was 60%. Therefore, HSCCC is a fast and efficient separation method, and it has broad application scenarios in the separation and purification of maltosyl-β-cyclodextrin.
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