Multiresidue assay for benzimidazole anthelmintics by liquid chromatography and confirmation by gas chromatography/selected-ion monitoring electron impact mass spectrometry.
A multiresidue method utilizing all-disposable labware has been developed for 8 benzimidazole anthelmintics from ovine, bovine, and swine muscle and liver tissues. After an initial extraction with ethyl acetate and subsequent evaporation, a 3-component extraction using hexane, ethanol, and 0.2N HCl was used for final cleanup. Clean extracts were produced for separation and determination by reverse-phase liquid chromatography at 298 nm, using methanol and aqueous buffer as mobile phase. A synthesized internal standard, 2-(n-butylmercapto)benzimidazole, was used for quantitation of all drugs. Results are included along with statistical information verifying the performance of the method. Spiked control tissues and incurred drug tissues were used for an intralaboratory study with a concentration range of 50-1470 ppb. A series of standard curves at 0, 50, 100, and 200 ppb were analyzed. Overall recovery at the 100 ppb level averaged 92% (CV 8%) in liver tissues, across all 3 species and 88% (CV 5%) in muscle tissues across all 3 species. Results were confirmed by gas chromatography/mass spectrometry with acid hydrolysis of the remaining extract in 2N HCl followed by re-extraction of the amine and derivatization to the tert-butyldimethylsilyl derivative. The anthelmintics were identified by gas chromatography/selected ion monitoring electron-impact mass spectrometry. Ion ratio measurements were taken and compared to standard material. CVs averaged 10% or less for all drugs tested.
Wilson
R T
RT
U.S. Department of Agriculture, Food Safety and Inspection Service, Midwestern Laboratory, St. Louis, MO 63115.
Groneck
J M
JM
Henry
A C
AC
Rowe
L D
LD
D000818:Animals; D000871:Anthelmintics / Q000032:analysis*; D001562:Benzimidazoles / Q000032:analysis*; D002417:Cattle; D002853:Chromatography, Liquid; D004350:Drug Residues / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry; D007202:Indicators and Reagents; D008099:Liver / Q000737:chemistry; D009132:Muscles / Q000737:chemistry; D012756:Sheep; D013552:Swine
D000871:Anthelmintics; D001562:Benzimidazoles; D007202:Indicators and Reagents