摘要
使用一种新型的大尺寸弱阴离子交换晶胶柱从酵母细胞发酵液中一步分离纯化广泛应用于临床治疗的高能化合物三磷酸腺苷(ATP)。该晶胶柱高150mm,直径140mm,柱体体积达到2300mL。实验测定了晶胶柱的孔隙率和等板高度等基本性能参数。层析过程以去离子水为冲洗液,分别用0.03、0.4和1mol·L-1NaCl溶液在2cm·min-1流速下进行洗脱。发现0.03mol·L-1NaCl溶液能把大部分杂质洗脱,0.4mol·L-1NaCl溶液洗脱后能获得纯度较高的ATP。洗脱下来的ATP样品通过高效液相色谱(HPLC)定量分析,回收的ATP纯度达95%～97%,总回收率49.1%。与其它已有方法相比,本法耗时少,工艺简单,降低了ATP变性的风险;在层析过程中以去离子水代替酸液作为冲洗剂和洗脱液溶剂,降低分离过程成本。
Adenosine triphosphate （ATP） is an important high-energy compound widely used in clinical treatment,and,a novel separation method of using large-size anion-exchange supermacroporous cryogel was developed in this paper to isolate the ATP directly from crude fermentation broth of Saccharomyces cerevisiae. The volume,height and diameter of the cryogel column used are 2300 mL,150 mm and 140 mm,respectively. The porosity and HETP （height equivalent to a theoretical plate） of the cryogel column used were measured. The deionised water was used as the running buffer and the elutions of 0.03,0.4 and 1 mol·L^-1 NaCl solution at flow velocity of 2 cm·min^-1 were carried out respectively during chromatographic separation. The majority of the impurities can be eluted by the 0.03 mol·L^-1 NaCl solution and higher purity ATP can be obtainted by elution of 0.4 mol·L^-1 NaCl solution. The purities of the obtained ATP were analyzed quantitatively by high performance liquid chromatography （HPLC） and the results show that the purity of ATP obtained by one-step separation method can be 95%～97% and the recover ratio is 49.1%. Compared with other bioparticles ion-exchange separation methods in existence,this method has the advantages of simpler process and less time-consuming which reduces the risk of ATP degeneration. In the chromatography process of this method,the deionised water was used as running buffer instead of hydrochloric acid which is commonly used in other bioparticles ion-exchange separation methods in existence; this substitution results in the reduction in cost of separation process.
出处
《高校化学工程学报》
EI
CAS
CSCD
北大核心
2009年第5期768-773,共6页
Journal of Chemical Engineering of Chinese Universities
基金
国家自然科学基金项目(20876145)
浙江省自然科学基金项目(Y4080329)
浙江工业大学东晖科技创新基金
