High performance liquid chromatography associated with resonance Rayleigh scattering for synchronous determination of three antihistamines and mechanism study.

A highly sensitive and selective method of high performance liquid chromatography (HPLC) combined with resonance Rayleigh scattering (RRS) spectra was developed for the detection of three antihistamine drugs, including pyrilamine (PY), carbinoxamine (CAR) and tripelennamine (TRI). The three antihistamines were separated by a C18 column. The mobile phase contained 25% acetonitrile (ACN) and 75% phosphate buffered solution (pH 3.2) with the flow rate of 0.4 ml min . In medium of Britton-Robinson (BR) buffer solution (pH 4.6), the PY, CAR and TRI separated by HPLC and then reacted with Erythrosine B (EryB), forming 1:1 ion-association complexes, which led to significant signal enhancement of RRS spectra. The RRS spectra was detected at the wavelength λ =λ = 370 nm. The calibration curves of PY, CAR and TRI were linear in the range from 0.02 to 25 μg ml , and the detection limit [signal-to-noise ratio (S/N) = 3] were 3.38, 4.48 and 5.50 ng ml , respectively. In addition, under the optimum experiment condition, the reaction mechanism and the reasons for RRS enhancement were investigated in this work. The developed method was applied to the simultaneous detection of three antihistamines in water samples with satisfying results.