title: Determination of the active transport of fucoidan derived from Okinawa Mozuku across the human intestinal Caco-2 cells as assessed by size-exclusion chromatography.
abstract: BACKGROUND
  In order to clarify the mechanism of fucoidan transport, we developed the chromatographic determination method.
  
  
  METHODS
  A size-exclusion chromatography (SEC) method for the determination of Okinawa-fucoidan using Develosil 300 Diol-5 (60×8.0mm I.D., 30nm pore-diameter) with the eluent containing 1% non-ionic detergent is developed. Determination range (UV at 210nm) is from 0 to 100ng of fucoidan with the linear calibration line inserting to zero.
  
  
  RESULTS
  A transport activity of fucoidan is demonstrated by using Caco-2 cells (model of gut transport system); i.e., the initial transport velocity 12nmol/h/mg of protein (25-fold slower rate as compared to a bacterial l-alanine active-transport activity 300nmol/h/mg of protein) is found to occur. Since this fucoidan transport is inhibited by 10mM sodium azide (respiration inhibitor) and 0.05mM FCCP (uncoupler), this transport by Caco-2 cells is found to be an active one requiring energy-source. On the other hand, colchicine (inhibitor of phagocytosis/pinocytosis) and mannitol (putative competitive-inhibitor of tight-junction transport) cannot inhibit the fucoidan transport at all.
  
  
  CONCLUSIONS
  We firstly report that the active transport occurs for such a high molecular-weight sulphated-polyfucose of fucoidan in vitro using Caco-2 cells.
authors.affiliation: Graduate School of Health Sciences, Gunma University 3-39-15 Showa-machi, Maebashi, Gunma 371-8514, Japan.
authors.forename: Takeaki
authors.initials: T
authors.lastname: Nagamine
authors.affiliation: Graduate School of Health Sciences, Gunma University 3-39-15 Showa-machi, Maebashi, Gunma 371-8514, Japan.
authors.forename: Kou
authors.initials: K
authors.lastname: Hayakawa
authors.affiliation: Graduate School of Health Sciences, Gunma University 3-39-15 Showa-machi, Maebashi, Gunma 371-8514, Japan.
authors.forename: Kyoumi
authors.initials: K
authors.lastname: Nakazato
authors.affiliation: South Product Co., Ltd., 12-75 Suzaki, Uruma, Okinawa 904-2234, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Iha
mesh_terms: D000293:Adolescent; D000368:Aged; D001693:Biological Transport, Active; D018938:Caco-2 Cells; D002850:Chromatography, Gel / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009369:Neoplasms / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics* / Q000652:urine
chemical_list: D011134:Polysaccharides; C007789:fucoidan
keywords: Active transport; Azide; Caco-2; Fucoidan; SEC; Uncoupler