Method for the quantification of alginate in microcapsules.
Alginate is a key reagent in the preparation of microcapsules for cell transplantation. To address the question of the intracapsular alginate concentration, a sensitive assay has been developed to quantify the alginate content of microcapsules. The method is based on the metachromatic change induced by alginate binding to the dye, 1,9-dimethyl methylene blue (DMMB). The assay has a high sensitivity and precision. It covers a wide concentration range enabling the measurement of alginate in dilute supernatants as well as in microcapsules. For the latter, the membrane is initially dissolved by incubating the microcapsules in an alkaline medium. The effect of potentially interfering substances (poly-L-lysine (PLL), citrate, chloride, sodium) and of pH has been studied. Poly-L-lysine interfered with the assay at pH 6.5 but not at pH 13. Interference by sodium augmented with increasing sodium concentration and reached a plateau at 200 mM. This problem was overcome by routinely adjusting all samples to 500 mM sodium. The other substances tested had a negligible effect on the assay. The reliable measurement of alginate with this new assay will allow the optimization of the intracapsular alginate concentration.
Hallé
J P
JP
Centre de Recherche, Hôpital Maisonneuve-Rosemont, Montreal, Quebec, Canada.
Landry
D
D
Fournier
A
A
Beaudry
M
M
Leblond
F A
FA
D000464:Alginates / Q000032:analysis* / Q000592:standards; D002214:Capsules; D017690:Cell Transplantation; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000032:analysis; D008751:Methylene Blue / Q000031:analogs & derivatives; D011107:Polylysine; D012015:Reference Standards; D013053:Spectrophotometry / Q000592:standards / Q000706:statistics & numerical data
D000464:Alginates; D002214:Capsules; D006603:Hexuronic Acids; D007202:Indicators and Reagents; D011107:Polylysine; D020723:Glucuronic Acid; D008751:Methylene Blue; C016401:1,9-dimethylmethylene blue