Open AccessSimple and Sensitive Spectrofluorometric Method for the Determination of Protein Using an Europium-thenoyltrifluoroacetone Probe
Author(s) -
Jianghong Tang,
Ning Lian
Publication year - 2009
Publication title -
analytical sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.392
H-Index - 73
eISSN - 1348-2246
pISSN - 0910-6340
DOI - 10.2116/analsci.25.1237
Subject(s) - thenoyltrifluoroacetone , chemistry , europium , human serum albumin , fluorescence , detection limit , fluorescence spectroscopy , analytical chemistry (journal) , chromatography , fluorescence spectrometry , ion , extraction (chemistry) , organic chemistry , physics , quantum mechanics , solvent extraction
The fluorescence intensity of the europium (Eu3+)-thenoyltrifluoroacetone (TTA) complex can be remarkably enhanced by human serum albumin (HSA) in a Britton-Robinson buffer solution. Based on this fact, a simple, rapid, and sensitive method has been developed for the determination of proteins at the nanogram level by fluorescence spectroscopy. Under the optimum conditions, the enhanced fluorescence intensity is proportional to the concentration of HSA. The linear ranges for HSA are 0-5.1 and 5.1-44.4 microg ml(-1) and the limit of detection is 20.7 ng ml(-1). Moreover, there is very little interference from common inorganic ions and other coexisting compounds. The binding site number and the binding constant of Eu(3+)-TTA used as a fluorescence probe to HSA were calculated by using the Rosenthal graphic method. This method has been applied to the determination of total protein in human serum samples. The results are good agreement with data obtained by clinical physicians.