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大尺寸弱阴离子交换晶胶层析一步法分离ATP的实验研究
  • ISSN号:1003-9015
  • 期刊名称:Gao Xiao Hua Xue Gong Cheng Xue Bao/Journal of Che
  • 时间:0
  • 页码:768-773
  • 语言:英文
  • 分类:TQ425.232[化学工程] TQ028.8
  • 作者机构:[1]浙江工业大学化学工程与材料学院绿色合成技术国家重点实验室培育基地,浙江杭州310032
  • 相关基金: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.
  • 相关项目:复合晶胶介质的成孔机理和孔内接枝及对pDNA载体的层析特性
中文摘要:

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。

英文摘要:

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.

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期刊信息
  • 《高校化学工程学报》
  • 北大核心期刊(2011版)
  • 主管单位:国家教育部
  • 主办单位:浙江大学
  • 主编:陈纪忠
  • 地址:杭州浙大路38号浙江大学玉泉校区化工系
  • 邮编:310027
  • 邮箱:gxhgxb@zju.edu.cn
  • 电话:0571-87951235
  • 国际标准刊号:ISSN:1003-9015
  • 国内统一刊号:ISSN:33-1141/TQ
  • 邮发代号:
  • 获奖情况:
  • 2000年获浙江省科技期刊编辑学会颁发的“1999-200...,中国期刊方阵“双效”期刊
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  • 俄罗斯文摘杂志,美国化学文摘(网络版),荷兰文摘与引文数据库,美国工程索引,美国剑桥科学文摘,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:14205