This paper demonstrates a chemiluminescence-based method for the determination of proteins with hydrophobic sites. The measurement is based on the chemical excitation of a complex formed by the binding of a fluorescent molecule to the hydrophobic portion of the protein. The fluorophore is excited with an aqueous peroxyoxalate chemiluminescence reaction and the resulting chemiluminescence signal is enhanced by the hydrophobic environment of the protein. The major advantage is that chemical modification of the analyte is not required. Low nanogram per millilitre concentrations of human albumin have been detected using the method.