Deferoxamine → Fluorescein Desferrioxamine

Participants

Reaction Steps

An aliquot of 120 mg of desferrioxamine and 233.64 mg of fluorescein isothiocyanate (FITC) are mixed in 50 mL of 0.1 M sodium carbonate pH 9.5. A molar ratio of 3:1 of FITC: desferrioxamine is used as described in Hentz et al. (1997) Anal. Chem. 69:4994-5000. The reaction mixture is wrapped in aluminum foil to avoid light exposure and allowed to react for at least 24 hours at room temperature. To separate fluorescently labeled desferrioxamine from free fluorescein and free desferrioxamine, the mixture is separated using a P-2 column (Bio-Rad) equilibrated in saline and samples are eluted in 2 mL fractions. The fluorescein-desferrioxamine is eluted in the void volume of the column. The first peak is collected and used for preparing fluorescein-desferrioxamine, desferrioxamine, and rhodamine liposomes.

Provenance & Evidence