Tata Alessandra, Pallante Ivana, Zacometti Carmela, Moressa Alessandra, Bragolusi Marco, Negro Alessandro, Massaro Andrea, Binato Giovanni, Gallocchio Federica, Angeletti Roberto, Pozzato Nicola, Piro Roberto
Istituto Zooprofilattico Sperimentale delle Venezie, Laboratorio di Chimica Sperimentale, Vicenza, Italy.
Istituto Zooprofilattico Sperimentale delle Venezie, Laboratorio di Medicina Forense Veterinaria, Vicenza, Italy.
Front Chem. 2022 Aug 25;10:982377. doi: 10.3389/fchem.2022.982377. eCollection 2022.
Animal poisoning and dissemination of baits in the environment have public health and ethological implications, which can be followed by criminal sanctions for those responsible. The reference methods for the analysis of suspect baits and autopsy specimens are founded on chromatographic-based techniques. They are extremely robust and sensitive, but also very expensive and laborious. For this reason, we developed an ambient mass spectrometry (AMS) method able to screen for 40 toxicants including carbamates, organophosphate and chlorinated pesticides, coumarins, metaldehyde, and strychnine. Spiked samples were firstly purified and extracted by dispersive solid phase extraction (QuEChERS) and then analyzed by direct analysis in real time high-resolution mass spectrometry (DART-HRMS). To verify the performance of this new approach, 115 authentic baits ( = 59) and necropsy specimens (gastrointestinal content and liver, = 56) were assessed by the official reference methods and combined QuEChERS-DART-HRMS. The agreement between the results allowed evaluation of the performances of the new screening method for a variety of analytes and calculation of the resultant statistical indicators (the new method had overall accuracy 89.57%, sensitivity of 88.24%, and a specificity of 91.49%). Taking into account only the baits, 96.61% of overall accuracy was achieved with 57/59 samples correctly identified (statistical sensitivity 97.50%, statistical specificity 94.74%). Successful identification of the bitter compound, denatonium benzoate, in all the samples that contained rodenticides (28/28) was also achieved. We believe initial screening of suspect poison baits could guide the choice of reference confirmatory methods, reduce the load in official laboratories, and help the early stages of investigations into cases of animal poisoning.
动物中毒以及诱饵在环境中的散播具有公共卫生和行为学方面的影响,对此,相关责任人可能会受到刑事制裁。分析可疑诱饵和尸检标本的参考方法基于色谱技术。这些方法极其可靠且灵敏,但成本高昂且操作繁琐。因此,我们开发了一种常压质谱(AMS)方法,能够筛查40种有毒物质,包括氨基甲酸盐、有机磷和氯化农药、香豆素、聚乙醛和士的宁。加标样品首先通过分散固相萃取(QuEChERS)进行纯化和提取,然后通过实时直接分析高分辨率质谱(DART-HRMS)进行分析。为验证这种新方法的性能,采用官方参考方法和QuEChERS-DART-HRMS联用,对115份真实诱饵(n = 59)和尸检标本(胃肠道内容物和肝脏,n = 56)进行了评估。结果之间的一致性使得能够评估新筛查方法对各种分析物的性能,并计算所得的统计指标(新方法的总体准确率为89.57%,灵敏度为88.24%,特异性为91.49%)。仅考虑诱饵时,57/59个样品被正确识别,总体准确率达到96.61%(统计灵敏度为97.50%,统计特异性为94.74%)。在所有含有灭鼠剂的样品(28/28)中,还成功鉴定出了苦味化合物苯甲地那铵。我们认为,对可疑毒饵进行初步筛查可以指导参考确证方法的选择,减轻官方实验室的负担,并有助于动物中毒案件调查的早期阶段。