RESUMO
Traditionally, creatinine determination is made by a spectrophotometric method; however, some compounds present in biological samples can interfere with creatinine determination, decreasing the sensitivity of the method in urine samples. Consequently, we report the development of a new molecularly imprinted polymer as a sorbent phase for disposable pipette extraction to determine creatinine in urine samples by high-performance liquid chromatography with UV detection. The synthesized polymer showed a high superficial area and presented a first-order kinetic reaction and a high selectivity for creatinine extraction compared to the non-molecularly imprinted polymer. The main disposable pipette extraction variables evaluated included the number of draw/eject cycles, the pH of the solution and desorption solvent type. The developed method showed an inter and intra-day precision from 1.3% to 2.0% and 0.8-1.6% respectively, accuracy values ranging from 82.3% to 102.1% respectively and recovery values ranging between 96.5% and 101.3%, with a limit of quantification of 0.302 g L-1. The application of the developed method in real urine samples showed creatinine concentrations ranging from 0.55 to 6.61 g L-1. Thus, the developed method was revealed to be an efficient strategy for creatinine determination, reducing analysis time (3 min) and solvent use, and increasing selectivity compared with DPX commercial sorbents.
Assuntos
Impressão Molecular , Polímeros Molecularmente Impressos , Cromatografia Líquida de Alta Pressão/métodos , Creatinina , Humanos , Impressão Molecular/métodos , Polímeros/química , Extração em Fase Sólida/métodos , Solventes/químicaRESUMO
BACKGROUND: Metaldehyde is a toxic pesticide used mainly as a molluscicide, responsible for intoxication and deaths in both humans and animals. Accidental exposure to metaldehyde in dogs is considered rare, but severe. Data concerning clinical and veterinary forensic toxicology are largely incomplete, especially regarding case reports in dogs. The present work reports a complete and detailed description of a case from the history, clinical evolution, pathological exams and toxicological diagnosis in an accidental case of metaldehyde poisoning in dog. CASE PRESENTATION: An eleven-month-old, 3.0 kg, male German Spitz was presented for emergency care with acute vomiting and seizures 3 hours after suspected accidental ingestion of commercial molluscicide containing 3% metaldehyde (Lesmax®). The animal was in lateral recumbency and showed stuporous mentation, salivation, tonic-clonic status epilepticus, systemic tremors, bilateral miosis, absent palpebral, corneal, oculovestibular and gag reflexes, severely depressed spinal reflexes, dyspnea and tachycardia. Despite treatment, the patient progressed to comatose mentation and died. Necropsy examination revealed discrete lesions in the liver and central nervous system, while stomach examination revealed content of feed, activated charcoal and blue-green granules, compatible to the commercial formula of metaldehyde. Histology examination revealed extensive hemorrhage and severe centrolobular necrosis of the liver and tumefaction of Kupfer cells. Brain samples showed discrete hemorrhage and hyperemia. In order to confirm the diagnosis, samples from feces, stomach content, spleen, liver, heart, kidneys and brain were submitted gas chromatography analysis. Results confirmed the presence of metaldehyde in all samples. We describe clinicopathological abnormalities of a fatal case of metaldehyde poisoning in a dog, as well as postmortem diagnosis using gas chromatography. CONCLUSION: Metaldehyde poisoning is rarely reported, since the diagnosis is often difficult and the notifications scarce. To our knowledge, this is the first report describing clinical signs, pathological findings and chromatographic diagnosis. This report aims to contribute to the understanding of the pathogenesis of metaldehyde intoxication, to further explore veterinary forensic toxicology diagnosis.