Method for monitoring alginate released in biological fluids by high-performance anion-exchange chromatography with pulsed amperometric detection.
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.
Desille
Mireille
M
INSERM U456-Détoxication et Réparation Tissulaire, Université de Rennes I, Rennes, France.
Allain
Nathalie
N
Anger
Jean Pierre
JP
Mahler
Stephan
S
Lognoné
Vincent
V
Mallédant
Yannick
Y
Clément
Bruno
B
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
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid