<jats:title>ABSTRACT</jats:title><jats:p>The pharmacokinetics of diniconazole enantiomers in quail (<jats:italic>Coturnix coturnix japonica</jats:italic>) were investigated by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Quails were exposed to racemic diniconazole in capsule by oral at dose of 10 mg/kg (body weight). The maximal concentrations observed in blood, heart, liver, and kidney were 3.18, 11.35, 12.32, 15.03 µg/g for S‐diniconazole, and 1.13, 3.70, 6.00, 2.60 µg/g for R‐diniconazole. The elimination of enantiomers all met the one‐compartment model in blood, heart, liver, and kidney well. The elimination half‐lives (T<jats:sub>1/2</jats:sub>) of S‐diniconazole were 2.87, 3.85, 5.29, and 4.42 h in blood, heart, liver, and kidney, respectively; the T<jats:sub>1/2</jats:sub> of R‐diniconazole were 2.44, 3.42, 146.23, and 74.02 h in blood, heart, liver, and kidney, respectively. The enantiomer fractions (EFs) steadily increased from 0.50 to 0.92 in blood samples and 0.91 in heart samples. Meanwhile, the values increased to 0.70 and 0.80 in liver and kidney initially, and then decreased to 0.33 and 0.44 at the end of the experiment. Metabolism was examined as well and it was found that diniconazole was metabolized to 1, 2, 4‐triazole, (E)‐3‐(1H‐1, 2, 4‐triazol‐1‐yl) acrylaldehyde, (E, S)‐(R, S)‐4‐(2, 4‐dichlorophenyl)‐2, 2‐dimethyl‐5‐(1H‐1, 2, 4‐triazol‐1‐yl) pent‐4‐ene‐1, 3‐diol, (E)‐4‐(2, 4‐dichlorophenyl)‐3‐hydroxy‐2, 2‐dimethyl‐5‐(1H‐1, 2, 4‐triazol‐1‐yl) pent‐4‐enoic acid, and 1, 3‐dichlorobenzen in all samples of quail. <jats:italic>Chirality 25:910–916, 2013</jats:italic>. © 2013 Wiley Periodicals, Inc.</jats:p>