Simultaneous determination of pyridostigmine bromide, N, N-diethyl- m-toluamide, permethrin, and their metabolites in rat plasma and urine by high-performance liquid chromatography

• Published in: Journal of chromatography. B, Biomedical sciences and applications Biomedical sciences and applications, 2000-12, Vol.749 (2), p.171-178
• Main Authors: Abu-Qare, Aqel W, Abou-Donia, Mohamed B
• Format: Article
• Language: English
• Subjects: Analysis; Animals; Biological and medical sciences; Calibration; Cholinesterase Inhibitors - blood; Cholinesterase Inhibitors - metabolism; Cholinesterase Inhibitors - urine; Chromatography, High Pressure Liquid - methods; DEET - blood; DEET - metabolism; DEET - urine; General pharmacology; Insect Repellents - blood; Insect Repellents - metabolism; Insect Repellents - urine; Insecticides - blood; Insecticides - metabolism; Insecticides - urine; Medical sciences; N, N-Diethyl- m-toluamide; Permethrin; Pharmacology. Drug treatments; Pyrethrins - blood; Pyrethrins - metabolism; Pyrethrins - urine; Pyridostigmine bromide; Pyridostigmine Bromide - blood; Pyridostigmine Bromide - metabolism; Pyridostigmine Bromide - urine; Quality Control; Rats; Rats, Sprague-Dawley; Reference Standards
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(00)00407-2

A rapid and simple method was developed for the separation and quantification of the anti nerve agent drug pyridostignmine bromide (PB; 3-dimethylaminocarbonyloxy- N-methyl pyridinium bromide) its metabolite N-methyl-3-hydroxypyridinium bromide, the insect repellent DEET ( N, N-diethyl- m-toluamide), its metabolites m-toluamide and m-toluic acid, the insecticide permethrin (3-(2,2-dichloro-ethenyl)-2,2-dimethylcyclopropanecarboxylic acid(3-phenoxyphenyl)methylester), and two of its metabolites m-phenoxybenzyl alcohol, and m-phenoxybenzoic acid in rat plasma and urine. The method is based on using C 18 Sep-Pak ® cartridges for solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) with reversed-phase C 18 column, and gradient UV detection ranging between 208 and 230 nm. The compounds were separated using gradient of 1 to 99% acetonitrile in water (pH 3.20) at a flow-rate ranging between 0.5 and 1.7 ml/min in a period of 17 min. The retention times ranged from 5.7 to 14.5 min. The limits of detection were ranged between 20 and 100 ng/ml, while limits of quantitation were 150–200 ng/ml. Average percentage recovery of five spiked plasma samples were 51.4±10.6, 71.1±11.0, 82.3±6.7, 60.4±11.8, 63.6±10.1, 69.3±8.5, 68.3±12.0, 82.6±8.1, and from urine 55.9±9.8, 60.3±7.4, 77.9±9.1, 61.7±13.5, 68.6±8.9, 62.0±9.5, 72.9±9.1, and 72.1±8.0, for pyridostigmine bromide, DEET, permethrin, N-methyl-3-hydroxypyridinium bromide, m-toluamide, m-toluic acid, m-phenoxybenzyl alcohol and m-phenoxybenzoic acid, respectively. The relationship between peak areas and concentration was linear over the range between 100 and 5000 ng/ml. This method was applied to analyze the above chemicals and metabolites following their administration in rats.