title: [Emulsion liquid membrane extraction with D2EHPA mobile carrier Hirudinaria manillensis hirudin in experimental study].
abstract: OBJECTIVE
  To study the extraction system of hirudin emulsion liquid membrane with the Poecilobdella manillensis as raw material, di-(2-ethylhexyl) phosphate (D2EHPA) as carrier, Span 80 as emulsifier, octane and D2EHPA mixed to constitute membrane solution, diluted HCl solutions as internal aqueous phase.
  
  
  METHODS
  Using the orthogonal experiment to optimize the extraction conditions of hirudin reference substance such as membrane phase, internal aqueous phase volume ratio (MIPVR), external aqueous phase pH, internal aqueous phase pH, mobile carrier concentration and so on, and then using hirudin crude extracts to do purifying experiment, and gaining experimental samples.
  
  
  RESULTS
  The optimal conditions of hirudin extraction were as follows: MIPVR 10: 3, internal aqueous phase pH 2.6, external aqueous phase pH 3.4, the mass fraction of carrier D2EHPA 2%. In the optimal extraction conditions, when the initial concentration of hirudin was one anti-thrombin activity units (ATU) x mL(-1), ATU recovery rate of the reference substance was 83.06%. In the purifying experiment of crude extracts, ATU recovery rate was 82.99%, and the specific activity of sample was 3 289.48 the ATU x mg(-1). Discontinuous polyacrylamide gel electrophoresis and spectral scanning, the results showed that the purity and reference substance were considerable.
  
  
  CONCLUSIONS
  The method of preparation hirudin was relatively simple, the purity of the experimental samples and ATU recovery were both high.
authors: Center of Modern Biochemistry, Guangdong Ocean University, Zhanjiang 524088, China. fang01638@163.com
authors: Fu-Yong
authors: FY
authors: Fang
authors: Yan-Li
authors: YL
authors: Miao
authors: Hui-Qin
authors: HQ
authors: Liu
authors: Yan-Jun
authors: YJ
authors: He
authors: Shao-Hong
authors: SH
authors: Chen
authors: Yan
authors: Y
authors: Liao
authors: Shao-Bin
authors: SB
authors: Liu
authors: Wen-Dong
authors: WD
authors: Song
authors: Yun-Tao
authors: YT
authors: Zhao
mesh_terms: D000818:Animals; D004655:Emulsions / Q000737:chemistry; D006629:Hirudins / Q000032:analysis / Q000302:isolation & purification*; D007865:Leeches / Q000737:chemistry*; D008567:Membranes, Artificial; D052616:Solid Phase Extraction / Q000295:instrumentation / Q000379:methods*
chemical_list: D004655:Emulsions; D006629:Hirudins; D008567:Membranes, Artificial