Determination of vitamin E and its metabolites in equine urine using liquid chromatography–mass spectrometry

• DOI: 10.1002/dta.3006
• Publication Date: 2021-02-03T22:59:09Z
• Authors: Gianna Favro; Hadi Habib; Ingrid Gennity; Birgit Puschner; Erin N. Hales; Carrie J. Finno; Benjamin C. Moeller
• Keywords: Atmospheric-pressure chemical ionization

Equine neuroaxonal dystrophy/degenerative myeloencephalopathy (eNAD/EDM) is a hereditary, deteriorating central nervous disease in horses. Currently, the only way to confirm eNAD/EDM through postmortem histological evaluation of system. Vitamin E, specifically isoform alpha-tocopherol (α-TP), known protect susceptible horses from developing clinical phenotype. While vitamin E an essential nutrient diet horses, there are no diagnostic tests able quantitate and its metabolites urine. An ultra-performance liquid chromatography-atmospheric-pressure chemical ionization mass spectrometry (UPLC-APCI-MS/MS) method was developed validated following acidic hydrolysis solid phase extraction equine A blank control horse urine matrix used spiked with different concentrations analytes form standard curve using either alpha-tocopherol-d6 or chlorpropamide as internal standard. Inter-day intra-day statistics were performed evaluate for accuracy (90% 116%) precision (0.75% 14%). Matrix effects, percent recovery, stability also assessed. The successfully analyzed alpha-carboxyethyl hydroxychroman (α-CEHC), alpha-carboxymethylbutyl hydroxychromans (α-CMBHC), gamma-carboxyethyl γ-CEHC, α-TP determine baseline levels healthy can be allowing approach treatment eNAD/EDM.