Simultaneous determination of amitraz and its metabolite in human serum by monolithic silica spin column extraction and liquid chromatography–mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2008-05, Vol.867 (1), p.99-104
• Main Authors: Saito, Takeshi, Yamamoto, Rie, Inoue, Shigeaki, Kishiyama, Izumi, Miyazaki, Shota, Nakamoto, Akihiro, Nishida, Manami, Namera, Akira, Inokuchi, Sadaki
• Format: Article
• Language: English
• Subjects: Adult; Amitraz; Amitraz metabolite; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Calibration; Chromatography, Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Humans; Insecticides - blood; LC–MS; Male; Mass Spectrometry - methods; Medical sciences; Monolithic silica spin column; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity; Serum; Silicon Dioxide; Toluidines - blood
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2008.03.018

A simple, rapid, sensitive, and specific liquid chromatography–mass spectrometry (LC–MS) method was developed and validated for the quantification of amitraz and its metabolite in human serum. Both the compounds were extracted using monolithic silica spin columns with acetonitrile. The chromatographic separation was performed on a reverse-phase C 18 column with a mobile phase of 10 mM ammonium formate–acetonitrile. The protonated analyte was quantitated in positive ionization by mass spectrometry. The method was validated over the concentration range of 25–1000 ng/ml for amitraz and its metabolite in human serum. For both compounds, the limit of detection was 5 ng/ml. The method was applied to serum samples taken from an attempted suicide patient, and only small volumes of serum were required for the simultaneous determination of these compounds.