U.S. Department of Agriculture: Animal and Plant Health Inspection Service


Date of this Version

January 2006


Published in Journal of Chromatographic Science 44(January 2006), pp. 1-5.


A reversed-phase ion-pair liquid chromatogaphic analysis combined with a solid-phase extraction clean-up method is used to assess the quantity of diphacinone residue found in invertebrates. Three invertebrate species are exposed to commercially available diphacinone-fortified bait used for rat control. The invertebrate samples are collected, frozen, and shipped to the laboratory. The samples are homogenized after cryogenic freezing. A portion of the homogenized samples are extracted with acidified chloroform-acetone, followed by cleanup with a silica solid-phase extraction column. Diphacinone is detected by UV absorption at 325 nm after separation by the chromatographic system. The method limit of detection (MLOD) for snail and slug samples averaged 0.055 and 0.066 mg/kg, respectively. Diphacinone residues in snail tissue ranges from 0.83 to 2.5 mg/kg for Oxychilus spp. The mean recoveries from snails at 0.20 and 2.0 are 97 ± 21 % and 84 ± 6%. Diphacinone residues in slug tissue ranges from 1.3 to 4.0 mg/kg for Deroceras laeve and < MLOD to 1.8 mg/kg for Limax maximus, respectively. The mean recoveries from slugs at 0.20 and 2.0 mg/kg are 91% ± 15% and 86% ± 5%.