A dual-wavelength overlapping resonance Rayleigh scattering method for the determination of chondroitin sulfate with nile blue sulfate.

A dual-wavelength overlapping resonance Rayleigh scattering (DWO-RRS) method was developed to detect chondroitin sulfate (CS) with nile blue sulfate (NBS). At pH 3.0-4.0 Britton-Robinson (BR) buffer medium, CS interacted with NBS to form an ion-association complex. As a result, the new spectra of resonance Rayleigh scattering (RRS), second order scattering (SOS) and frequence doubling scattering (FDS) appeared and their intensities were enhanced greatly. Their maximum wavelengths were located at 303 nm (RRS), 362 nm (RRS), 588 nm (SOS) and 350 nm (FDS), respectively. The scattering intensities of the three methods were proportional to the concentration of CS in certain ranges. The methods had high sensitivity and the detection limits were between 1.5 and 7.1 ng mL(-1). The DWO-RRS method had the highest sensitivity with the detection limit being 1.5 ng mL(-1). The characteristics of the spectra and optimal reaction conditions of RRS method were investigated. The effects of coexistent substances on the determination of CS were evaluated. Owing to the high sensitivity, RRS method had been applied to the determination of CS in eye drops with satisfactory results. The recovery range was between 99.4% and 104.6% and the relative standard deviation (RSD) was between 0.4% and 0.8%. In addition, the reasons for RRS enhancement were discussed and the shape of ion-association complex was characterized by atomic force microscopy (AFM).