DEVELOPMENT AND VALIDATION OF HPLC-METHOD FOR QUANTITATIVE DETERMINATION OF PROPOXUR IN COLLARS AND SPRAYS

Abstract

Propoxur is a carbamate non-systemic insecticide, an acetylcholinesterase blocker of invertebrates, leading to their paralysis and death. Propoxur is used in insect acaricidal sprays and collars to kill fleas and ticks on pets. The aim of the work was to develop a method of identification and quantification of propoxur in spray for external use and collars with different matrices. The method was developed and validated by indicators of selectivity, linearity, suitability parameters of the chromatographic system and repeatability. Spray and collars with different polymer composition were used as a sample-object for the development of the technique. The standard sample and the test sample of the spray were dissolved in acetonitrile to a propoxur concentration of 50 μg/ml. The collar was dissolved in tetrahydrofuran and acetonitrile was added to precipitate the matrix. The samples were separated on a Dionex Ultimate 3000 chromatograph equipped with chromatographic column Acclaim C18 150×4.6, 5 μm. The mobile phase was a mixture of acetonitrile and water in a volume ratio of 80:20. Propoxur was detected at an absorption wavelength of 270 nm. Under the above conditions, it was possible to completely separate propoxur (retention time of the chromatographic peak – 2.2 min) and other components of the studied drugs. At the same time, the suitability parameters of the chromatographic system did not exceed the limits specified in the recommendations of the USA Food and Drug Association. For propoxur, the efficiency of the chromatographic system was 10,300–10,540 theoretical plates. The relative standard deviation (RSD) for the peak areas of the active substance was ± 0.37%, and the peak separation ratio (RS) of propoxur and other components of the drug was 3.1–5.7. The symmetry coefficient of the propoxur peak was 1.49. The calibration curve was linear in the recommended DFU 2.0 range (80–120% of the nominal concentration of the active substance). The coefficient of linearity (R2) for propoxur was 0.9985.