Design and validation of analytical methods for quantitative determination of active ingredients in extemporal combined medicine in spray form

Abstract

The aim of the work was the development and study of the validation characteristics of the method of quantitative determination of phenylephrine hydrochloride, nitrofural and diphenhydramine hydrochloride when they are simultaneously present in an extemporaneous combined medicinal product in the form of a spray using liquid chromatography method.

Materials and methods. Agilent 1260 liquid chromatographs, equipped with a diode-matrix and UV detector from the company "Agilent technologies", USA. Chromatographic columns 250×4.6 mm filled with octadecylsilyl silica gel for chromatography (Zorbax StableBond SB-Aq, Agilent company), mobile phase - 0.1 % aqueous solution of trifluoroacetic acid R - methanol R, elution mode - gradient; mobile phase speed – 1.2 ml/min; the detection wavelength is 220 nm.

Results. The determined chromatographic conditions ensure proper separation of the peaks of the substances to be determined: phenylephrine hydrochloride, nitrofural and diphenhydramine hydrochloride in their joint presence, and do not have a negative effect on the quantitative assessment of their content. Validation tests were conducted to confirm the suitability of the analytical method for the performance of the task - control of the quantitative content of active substances in the combined medicinal product in the form of a spray. The determined validation characteristics indicate that the method is characterized by appropriate specificity, linearity, correctness and convergence in the range of application for phenylephrine hydrochloride (range 0.499-0.749 mg/ml, Δ_Z=0.44 ≤ max Δ_Z=3.20, d=0.22 ≤ max d=1.02, a=0.01  max a=5.1, r = 0.9997  min r= 0.9924), nitrofural (range 0.154-0.231 mg/ml, Δ_Z=0.44 ≤ max Δ_Z=3.20, d=0.62 ≤ max d=1.02, a=0.0006  max a=5.1, r = 0.9996  min r= 0.9924) and diphenhydramine hydrochloride (range 0.499-0.749 mg/ml, Δ_Z=0.50 ≤ max Δ_Z=3.20, d=0.05 ≤ max d=1.02, a=0.076  max a=5.1, r = 0.9999  min r= 0.9924).

Conclusions. An analytical technique for the quantitative determination of phenylephrine hydrochloride, nitrofural and diphenhydramine hydrochloride when simultaneously present in an extemporaneous combined medicinal product in the form of a spray by the method of high-performance liquid chromatography was developed. The determined validation parameters confirm the correctness of the methodology. The proposed HPLC technique was used to study the chemical stability of the spray for the treatment of allergic rhinitis