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Determination of albumin in biological fluids by flow injection analysis using the peroxyoxalate chemiluminescent
L Gámiz-Gracia1, A M García-Campaña, F Alés Barrero
1Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Av. Fuentenueva s/n, 18071 Granada, Spain.
Abstract:
A new sensitive analytical procedure for the determination of albumin is presented, based on the participation of a fluorescent derivative of the protein in the peroxyoxalate chemiluminescent system using imidazole as a catalyst. The chemiluminescent emission is detected in a flow injection analysis (FIA) system, employing micellar medium as carrier, whereas the derivatisation reaction is carried out off-line using fluorescamine as a label. The optimisation of the operational and chemical variables of the method such as concentration of reagents, flow-rates, injection volume, etc., involves the use of experimental designs, including Draper-Lin small composite design, scarcely used in analytical chemistry. The performance characteristics were established and a detection limit of about 0.1 fmol of albumin was obtained. The method has been validated and satisfactorily applied to the determination of albumin in human serum and urine.
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