title: Method for monitoring alginate released in biological fluids by high-performance anion-exchange chromatography with pulsed amperometric detection.
abstract: The use of alginate-entrapped cells in cell therapy requires a method for monitoring possible released compound within biological fluids following either their implantation or inoculation in artificial organs. Oligomannuronic and oligoguluronic acids were prepared by enzymatic depolymerization with alginate lyase from Pseudomonas alginovora, characterized by high-performance anion-exchange chromatography with pulsed amperometric detection and quantitated in human, pig and rabbit blood, urine and tissue samples. The method was tested for linearity and detection limit, accuracy, intra- and inter-day precision. The limit of detection was 3 microgram/ml in both urine and plasma and 5 mg/g of tissues. The relative standard deviations (RSDs) of intra-day precision were 6.0-16.6% and 4.8-8.7% in plasma and urine, respectively; the RSDs of inter-day precision were 5.1-14.4% and 5.0-11.6% in plasma and urine, respectively. Thus, this method appears suitable for the measurement of released alginate from entrapped cells used in cell therapy.
authors.affiliation: INSERM U456-Détoxication et Réparation Tissulaire, Université de Rennes I, Rennes, France.
authors.forename: Mireille
authors.initials: M
authors.lastname: Desille
authors.forename: Nathalie
authors.initials: N
authors.lastname: Allain
authors.forename: Jean Pierre
authors.initials: JP
authors.lastname: Anger
authors.forename: Stephan
authors.initials: S
authors.lastname: Mahler
authors.forename: Vincent
authors.initials: V
authors.lastname: Lognoné
authors.forename: Yannick
authors.initials: Y
authors.lastname: Mallédant
authors.forename: Bruno
authors.initials: B
authors.lastname: Clément
mesh_terms: D000464:Alginates / Q000032:analysis* / Q000493:pharmacokinetics; D000818:Animals; D001682:Biological Availability; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004563:Electrochemistry / Q000379:methods*; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid