Determination of protein by hydroxypropyl-β-cyclodextrin sensitized fluorescence quenching method with erythrosine sodium as a fluorescence probe

Abstract

The fluorescence spectral behavior of interaction of erythrosine sodium (ES) and bovine serum albumin (BSA) was investigated in hydroxypropyl-β-cyclodextrin (HP-β-CD) medium at pH 5.8. The excitation and emission wavelengths were 527 nm and 549 nm, respectively. The fluorescence intensity of ES increased due to the formed inclusion complex of HP-β-CD and ES. But the fluorescence intensity of ES–HP-β-CD diminished when BSA was added, and there was a linear relationship between the fluorescence quenching value of the system (Δ F = F ES–HP-β-CD − F BSA–ES–HP-β-CD) and the concentration of BSA. Based on this, a novel fluorescence quenching method for the determination of protein with ES as a fluorescence probe has been developed. Under the optimal conditions, the linear range of calibration curve for the determination of BSA was 0.5–32.0 μg mL −1, and the detection limit was 49.2 ng mL −1. It has been applied to the determination of serum in serum samples with satisfactory results.