HyunYoung Chae (Animal and Plant Quarantine Agency, Kyungpook National University)

JeongWoo Kang (Animal and Plant Quarantine Agency, Kyungpook National University)

HyunYoung Chae (Animal and Plant Quarantine Agency, Kyungpook National University)

Jae-Won Byun (Animal and Plant Quarantine Agency)

Bok-Kyung Ku (Animal and Plant Quarantine Agency)

JeongWoo Kang (Animal and Plant Quarantine Agency, Kyungpook National University)

Antifreeze poisoning among companion animals, particularly cats, has seen a concerning rise in the Republic of Korea recently. Ethylene glycol (EG), a principal component of antifreeze, is dangerous due to its colorless, odorless nature and sweet taste that disguises its toxicity. The increasing number of such poisoning cases highlights the need for quick and accurate diagnostic methods to detect EG and its toxic metabolites in biological samples. Traditional techniques often require lengthy and complex derivatization processes. This study proposes an innovative approach that efficiently identifies EG and glycolic acid (GA), addressing these challenges. We utilize gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), simplifying sample preparation and speeding up detection in feline stomach tissue. The GC-MS method for EG showed high specificity and sensitivity without significant matrix interference, demonstrating a linear response across a 5 to 200 μg/mL concentration range and a detection limit of 25 ng/mL. Recovery rates for EG were between 83.05% and 91.73%. For GA, the LC-MS/MS method provided efficient identification with minimal matrix effects, achieving a detection limit of 5 ng/mL and a quantification limit of 20 ng/mL. Recovery rates for GA ranged from 80.92~ 95.30%, with excellent precision. Practical application of these methods was confirmed by analyzing samples from five suspected antifreeze poisoning cases, revealing elevated levels of EG and GA, supported by pathological evidence such as calcium oxalate crystals in renal tissues. The methodologies developed in this study for detecting EG and GA offer a rapid and precise analytical solution to the rising cases of antifreeze poisoning among companion animals in South Korea. By eliminating the derivatization step, these techniques reduce analysis time, minimize experimental errors, and ensure high accuracy. This advancement is expected to significantly improve responses to intentional poisoning incidents, greatly contributing to animal welfare and veterinary forensic science.

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