Rapid detection of asperphenamate in a hay batch associated with constipation and deaths in dairy cattle. The application of DART-HRMS to veterinary forensic toxicology

Nicola Pozzato; Elena Piva; Ivana Pallante; Dino Bombana; Roberto Stella; Claudia Zanardello; Alessandra Tata; Roberto Piro

doi:10.1016/j.toxicon.2020.08.022

Rapid detection of asperphenamate in a hay batch associated with constipation and deaths in dairy cattle. The application of DART-HRMS to veterinary forensic toxicology

Toxicon. 2020 Nov:187:122-128. doi: 10.1016/j.toxicon.2020.08.022. Epub 2020 Sep 3.

Authors

Nicola Pozzato¹, Elena Piva², Ivana Pallante³, Dino Bombana⁴, Roberto Stella⁵, Claudia Zanardello⁶, Alessandra Tata⁷, Roberto Piro⁸

Affiliations

¹ Laboratorio di Diagnostica Clinica e Sierologia di Piano, SCT1 Verona e Vicenza, Istituto Zooprofilattico Sperimentale delle Venezie, Via San Giacomo, 5, Verona, 37135, Italy. Electronic address: [email protected].
² Laboratorio di Chimica Sperimentale, Istituto Zooprofilattico Sperimentale delle Venezie, Viale Fiume, 78, Vicenza, 36100, Italy. Electronic address: [email protected].
³ Laboratorio di Diagnostica Clinica e Sierologia di Piano, SCT1 Verona e Vicenza, Istituto Zooprofilattico Sperimentale delle Venezie, Via San Giacomo, 5, Verona, 37135, Italy. Electronic address: [email protected].
⁴ Veterinary Practitioner, Villafranca di Verona, 37069, Italy. Electronic address: [email protected].
⁵ Laboratorio residui e farmaci, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10, Legnaro, 35020, Italy. Electronic address: [email protected].
⁶ Laboratorio di Diagnostica Specialistica e istopatologia, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10, Legnaro, 35020, Italy. Electronic address: [email protected].
⁷ Laboratorio di Chimica Sperimentale, Istituto Zooprofilattico Sperimentale delle Venezie, Viale Fiume, 78, Vicenza, 36100, Italy. Electronic address: [email protected].
⁸ Laboratorio di Chimica Sperimentale, Istituto Zooprofilattico Sperimentale delle Venezie, Viale Fiume, 78, Vicenza, 36100, Italy. Electronic address: [email protected].

PMID: 32891666
DOI: 10.1016/j.toxicon.2020.08.022

Abstract

Direct analysis in real time (DART) coupled to high-resolution mass spectrometry (HRMS) was applied for the first time to veterinary forensic toxicology to investigate the presence of toxic compounds in hay after an episode of acute intoxication in a dairy cattle farm. In addition to gross field necropsy and histological examination, microbial cultures, and heavy metals analysis, the molecular fingerprinting of the suspected hay batch was investigated by DART-HRMS. DART-HRMS revealed a distinct signal of m/z 507.2289 in the hay batch thought to be associated with the digestive complications. A search on chemical structure databases matched the ion with asperphenamate, a toxin produced by Penicillium spp. and Aspergillus spp. Liquid Chromatography-HMRS analysis and electrospray-HRMS-MS/MS of the hay extracts further characterized the structure and confirmed the identification of the compound as asperphenamate. Asperphenamate is fungal metabolite which can have cytotoxic and antitumor activity in humans, and it is classified as acute toxicant and harmful if swallowed.

Keywords: Ambient mass Spectrometry; Asperphenamate; Cattle; DART-HRMS; Hay; Toxin.

MeSH terms

Animals
Aspergillus
Cattle
Cattle Diseases / diagnosis*
Constipation / complications
Constipation / diagnosis
Constipation / veterinary*
Forensic Toxicology
Phenylalanine / analogs & derivatives*
Phenylalanine / analysis
Tandem Mass Spectrometry

Substances

Phenylalanine
asperphenamate