Quantitative method for analysis of six anticoagulant rodenticides in faeces, applied in a case with repeated samples from a dog (original) (raw)
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Acta Veterinaria Scandinavica
Background: Exposure to anticoagulant rodenticides (ARs) in dogs is among the most common causes of poisoning in small animal practice, but information about toxicokinetic of these rodenticides in dogs is lacking. We analysed blood and faeces from five accidentally exposed dogs and 110 healthy dogs by reversed phase ultra-high performance liquid chromatography-tandem mass spectrometry. The aim of the study was to estimate elimination of brodifacoum, bromadiolone and difenacoum after acute exposure, calculate the half-lives of these rodenticides in dogs, estimate faecal elimination in a litter of puppies born, and further to identify the extent of AR exposure in a healthy dog population. Results: Three dogs were included after single ingestions of brodifacoum; two dogs ingested bromadiolone and one dog ingested difenacoum. Maximum concentrations in faeces were found after day 2-3 for all ARs. The distribution half-lives were 1-10 days for brodifacoum, 1-2 days for bromadiolone and 10 days for difenacoum. Brodifacoum and difenacoum had estimated terminal half-lives of 200-330 days and 190 days, respectively. In contrast, bromadiolone had an estimated terminal half-life of 30 days. No clinical signs of poisoning or coagulopathy were observed in terminal elimination period. In blood, the terminal half-life of brodifacoum was estimated to 8 days. Faeces from a litter of puppies born from one of the poisoned dogs were examined, and measurable concentrations of brodifacoum were detected in all samples for at least 28 days after parturition. A cross-sectional study of 110 healthy domestic dogs was performed to estimate ARs exposure in a dog population. Difenacoum was detected in faeces of one dog. Blood and faecal samples from the remaining dogs were negative for all ARs. Conclusions: Based on the limited pharmacokinetic data from these dogs, our results suggest that ARs have a biphasic elimination in faeces using a two-compartment elimination kinetics model. We have shown that faecal analysis is suitable and reliable for the assessment of ARs exposure in dogs and a tool for estimating the AR half-lives. Half-lives of ARs could be a valuable indicator in the exposed dogs and provides important information for veterinarians monitoring AR exposure and assessment of treatment length in dogs.
Anticoagulant rodenticide poisoning in animals of Apulia and Basilicata, Italy
This study evaluates the presence of anticoagulant rodenticides in animals with a diagnosis of suspected poisoning and in bait samples. The survey was carried out from 2010 to 2012, in 2 regions of South Italy (Puglia and Basilicata) on 300 organs of animals and 90 suspected bait samples. The qualitative and quantitative analyses were conducted using an analytical method based on high‑performance liquid chromatography (HPLC) with fluorimetric detection (FLD) for the simultaneous determination of 8 anticoagulant rodenticides (bromadiolone, brodifacoum, coumachlor, coumafuryl, coumatetralyl, difenacoum, flocoumafen, and warfarin). The presence of anticoagulant rodenticides was detected in 33 organs of animals (11% of the total) and 6 bait samples (7% of the total). The most commonly detected compound was coumachlor (47% of 39 positive samples) followed by bromadiolone (24%), and brodifacoum (11%). The species mostly involved in anticoagulant rodenticide poisoning were dogs and cats. This study emphasizes the relevance of the determinations of anticoagulant rodenticides in cases of suspected poisoning in veterinary practice.
Journal of Veterinary Diagnostic Investigation
Exposure of wildlife and domestic animals to anticoagulant rodenticides (ARs) is a worldwide concern, but few methods exist to determine residue levels in live animals. Traditional liver detection methods preclude determining exposure in live wildlife. To determine the value of assessing AR exposure by fecal analysis, we compared fecal and liver residues of ARs in the same animals. We collected liver and fecal samples from 40 apparently healthy red foxes ( Vulpes vulpes) potentially exposed to ARs, and quantified brodifacoum, bromadiolone, coumatetralyl, difenacoum, difethialone, and flocoumafen residues by liquid chromatography–tandem mass spectrometry. Residues of ARs were detected in 53% of the fecal samples and 83% of the liver samples. We found good concordance between AR residues in feces and liver for coumatetralyl, difenacoum, and difethialone. Bromadiolone occurred in significantly greater frequency in livers compared to feces, but no significant difference in concentration...
Journal of Veterinary Diagnostic Investigation, 2012
Poisoning of domestic animals is frequently caused by anticoagulant rodenticides. Validation and applications of a rapid and reliable method for the simultaneous determination of 8 anticoagulant rodenticides (bromadiolone, brodifacoum, coumachlor, coumafuryl, coumatetralyl, difenacoum, flocoumafen, and warfarin) in baits and animal livers using highperformance liquid chromatography with fluorescence detection are reported herein. The methodology was validated by an in-house validation model at 2.5 mg/kg, which is the level commonly found in the tissues of poisoned domestic animals. The 8 anticoagulants can be determined at the concentration range of 1.25-100 mg/kg with determination coefficients higher than 0.992. A recovery value from 70% to 109% was observed for all the studied molecules. The results of the validation process demonstrate suitability for application in official analysis and for monitoring purposes of animal poisoning by anticoagulant rodenticides.
Journal of analytical toxicology, 2015
A rapid, highly sensitive and specific analytical method for the extraction, identification and quantification of nine rodenticides from whole blood has been developed and validated. Commercially available rodenticides in Australia include coumatetralyl, warfarin, brodifacoum, bromadiolone, difenacoum, flocoumafen, difethialone, diphacinone and chlorophacinone. A Waters ACQUITY UPLC TQD system operating in multiple reaction monitoring mode was used to conduct the analysis. Two different ionization techniques, ES+ and ES-, were examined to achieve optimal sensitivity and selectivity resulting in detection by MS-MS using electrospray ionization in positive mode for difenacoum and brodifacoum and in negative mode for all other analytes. All analytes were extracted from 200 µL of whole blood with ethylacetate and separated on a Waters ACQUITY UPLC BEH-C18 column using gradient elution. Ammonium acetate (10 mM, pH 7.5) and methanol were used as mobile phases with a total run time of 8 mi...
Evaluation of a point-of-care anticoagulant rodenticide test for dogs
Journal of Veterinary Emergency and Critical Care, 2014
Objective-To evaluate a point-of-care anticoagulant rodenticide lateral flow analyzer for the detection of various rodenticide compounds. Design-Prospective, laboratory study. Setting-University teaching hospital. Animals-The study utilized a serum sample from one healthy canine donor. Samples were centrifuged and serum samples were aliquoted and either used within 4 hours or frozen at −70 • C for further quantitative analysis. Interventions-Samples were spiked with clinically relevant concentrations of 1 of 6 rodenticide compounds (warfarin, pindone, chlorphacinone, brodifacoum, bromethalin, and its metabolite desmethylbromethalin). Seventy-five microliters of spiked serum (or unaltered serum) was introduced into the lateral flow test. Measurements and Main Results-Three readers who were blinded to the sample preparation interpreted the lateral flow test as either positive or negative for the presence of anticoagulant rodenticide. All readers were in agreement for the results of each serum sample. The point-of-care test kit was able to detect a single anticoagulant rodenticide (warfarin) at concentrations below the manufacturer's recommended limit of detection, but was unable to detect any other anticoagulant rodenticide. Conclusions-The results of this test and therapeutic interventions must be considered in light of history, physical examination, and other clinical data. Based on results from this study, the test kit only detects warfarin and not other more common second-generation anticoagulant rodenticides.
Multiresidue Analysis of Nine Anticoagulant Rodenticides in Serum
Journal of Agricultural and Food Chemistry, 1998
A rapid procedure was developed for the determination of nine anticoagulant rodenticides in animal serum by high-performance liquid chromatography (HPLC) with a fluorescence and photodiode array detector (DAD). The anticoagulants coumafuryl, pindone, warfarin, coumachlor, diphacinone, chlorphacinone, bromadialone, brodifacoum, and difethialone were extracted and quantitated simultaneously with this method. Anticoagulants were extracted at pH 5.5 with 5% (v/v) ethanol in ethyl acetate and 1% (w/v) trichloroacetic acid. Sample extracts were subjected to a cleanup on a Florisil solid-phase extraction (SPE) column. Separation was performed with a reversed phase amine-deactivated C 18 column with a gradient of phosphate buffer, acetonitrile, and methanol. The eluent was monitored with the fluorescence detection at an excitation wavelength of 310 nm and an emission wavelength of 390 nm, and the photodiode array detection was set to 325 nm. Recoveries from spiked serum samples at ranges of 0.004-1 ppm were between 73 and 105%. The precision (CV) data range was between 1.6 and 15.6%. Detection limits were between 0.005 and 0.002 ppm for the fluorescence detection and between 0.020 and 0.050 ppm for the photodiode array detection at 325 nm.
Toxicity of Two Anticoagulant Rodenticides to Rodent Species under Laboratory Conditions
A laboratory evaluated of 2 anticoagulant rodenticides, Kelerat super (Brodifacoum 0.005%) and Caid (Chlorophacinone 0.005%) against three rodent species, Rattus rattus frugivorus, Rattus rattus alexandrinus and Nile grass rat, A. niloticus fed on poison bait for 3 days, 4 days and 5 days. Results indicated that Kelerat was the most effective ones followed by Caid. The dead period for Caid was longer than in the case of Kelerat. It was found that there was a significant difference in the animal consumption of the tested rodenticide baits for rodent species. Also, significant difference in rodenticides consumed by males and females.