Gas chromatography/mass spectrometry identification and quantification of isazophos in a famphur pour-on and in bovine tissues after a toxic exposure.

A sample identified as "Warbex pour-on," expected to contain 13.2% famphur, and bovine tissue samples from 2 heifers that died after exhibiting signs of organophosphate intoxication were analyzed by gas chromatography/mass spectrometry (GC/MS). A product formulation problem was suspected because brain cholinesterase activities were depressed in both animals. Electron impact (EI) GC/MS of the pour-on revealed 9.7% famphur and an unidentified peak with approximately 76% of the peak area of the famphur. The unidentified peak showed a molecular ion at m/z 313, with a single Cl isotope cluster. Methane chemical ionization (MeCI) MS confirmed the molecular weight at 313 (1 Cl). A search on the molecular formula C9H17N3O3PSCl yielded a single match, isazophos. EI and MeCI GC/MS of reference isazophos confirmed the identity of the suspect peak. The concentration of isazophos in the pour-on was determined to be 6.0%. Famphur and isazophos were identified by their EI spectra and GC retention times in extracts of liver and brain from the 2 deceased animals. A GC/MS procedure utilizing selected ion monitoring (SIM) was developed for quantification of isazophos in liver, kidney, muscle, and fat of additional affected animals sacrificed at various times after exposure. Isazophos remained in animal tissues for as long as 94 days after topical exposure. Isazophos was present in fetal liver 70 days after exposure of the dam. High levels (6-3,500 ppm) of isazophos and famphur remained on the skin at 39 days postexposure.