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A sensitive method using ion-pair extraction was developed by liquid chromatography tandem mass spectrometry (LC-MS/MS) for measurement of 4-methylimidazole (4-MI) in NMRI mice plasma and cerebrospinal fluid (CSF). Detection was done by electrospray positive ionization mass spectrometry in the multiple-reaction monitoring (MRM) mode. The validation method was applied to quantification of 4-MI in plasma and CSF samples using oral doses of 100, 200, and 300 mg/kg in NMRI mice. The efficiency of the method was evaluated in terms of linearity (R 2 &gt; 0.99), recovery (98-107%, 3 levels) and precision (8-10%, 3 levels, n = 6). Limit of detection (LOD) and limit of quantification (LOQ) were 25 ng/mL and 50 ng/mL, respectively. The results obtained showed that the exposure to oral doses of 4-MI in mice makes different concentrations in plasma and CSF and causes significant changes in mice. This study was the first report for determination of 4-MI in plasma and CSF samples in mice. Our results suggest that LC-MS/MS-based on ion-pair extraction is a robust method with high detection ability for measurement of 4-MI in plasma and CSF samples. Therefore, the developed method can be useful for evaluation and monitoring of imidazole derivatives in biological samples.

Quantification of 4-Methylimidazol in NMRI Mice Plasma and Cerebrospinal Fluid (CSF) by Using Liquid Chromatography Tandem Mass Spectrometry.