ABSTRACT
Liquid chromatography-mass spectrometry (LC-MS) analysis of methanol extracts of Oenanthe crocata roots revealed that oenanthotoxin co-eluted with another major polyalkyne, 2,3-dihydro-oenanthotoxin, using the existing high performance liquid chromatography (HPLC) method (isocratic elution from C18 with aqueous methanol) for investigating Oenanthe poisoning. Positive ES or APCI gave [(M+H)-H(2)O](+) and its methanol adduct as major ion species for oenanthotoxin, whereas 2,3-dihydro-oenanthotoxin formed [M+H](+) and its methanol adduct. The two polyalkynes could be chromatographically resolved on C18 by gradient elution with aqueous acetonitrile. This provides superior analysis for oenanthotoxin using HPLC with photodiode array (PDA) detection alone, but for LC-MS/MS aqueous acetonitrile was less suitable due to poor ionisation and, with APCI, an increase in the relative abundance of a [M-1](+) species, which could confuse compound assignment. HPLC-PDA and LC-MS/MS methods using an aqueous acetonitrile or aqueous methanol mobile phase, respectively, were successful when applied to the analysis of the stomach contents of a pony suspected to have eaten O. crocata. Relevant product ion spectra, by ion trap MS/MS, accurate mass data and complete sets of (1)H and (13)C NMR spectral assignments are given for the two compounds.
Subject(s)
Alkynes/analysis , Chromatography, High Pressure Liquid/methods , Fatty Alcohols/analysis , Horse Diseases/diagnosis , Oenanthe/chemistry , Spectrometry, Mass, Electrospray Ionization/methods , Alkynes/chemistry , Alkynes/poisoning , Animals , Enediynes , Fatty Alcohols/chemistry , Fatty Alcohols/poisoning , Hazardous Substances/analysis , Hazardous Substances/poisoning , Horse Diseases/chemically induced , Horses , Magnetic Resonance Spectroscopy , Methanol/chemistry , Molecular Structure , Plant Roots/chemistry , Reproducibility of Results , Sensitivity and Specificity , StereoisomerismABSTRACT
Résumé- Les commémoratifs, les symptômes, l'histologie et la réponse au traitement d'un cheval de 15 ans atteint de lymphome cutané sont décrits. La jument présente de multiples nodules cutanés depuis 11 ans. Un diagnostic de lymphome cutané est poséà l'examen histologique de biopsies réalisées 2 fois à 4 ans d'intervalle. Des marquages immunihistochimiques identifient ces cellules comme appartenant à la lignée T. Ceci est la première description du marquage immunophénotypique d'un lymphome cutané chez le cheval. La jument a été traitée par des glucorticoïdes intralésionnels et des progestagènes oraux ce qui a entrainé une régression complète des nodules. Resumen- Describimos la historia clinica y los hallazgos clinicos e histológicos de un caballo de 15 años con un linfoma cutáneo. La yegua tenia una historia clinica de multiples nédules cutáneos en los últimos 11 años. Se diagnosticó un linfoma cutáneo mediante estudio histológico de dos biosias tomadas con unos cuatro años de separación. Tinciones inmunohistoquimicas ulteriores identificaron la linea celular de ünfocitos T. Este es el primer estudio que caracteriza el linfoma equino a nivel inmunofenotipico. La yegua fue tratada con glucocorticoides intralesionales y progestágenos orales, lo que resultó en curación total de los nódulos. Abstract- The history, clinical and histological features and response to treatment of a 15-year-old horse with cutaneous lymphoma are described. The mare had an 11 year history of multiple cutaneous nodules. A diagnosis of cutaneous lymphoma was made on histological examination of biopsy material taken on two separate occasions some 4 years apart. Subsequent immunohistochemical staining identified the cell lineage as T-lymphocytes. This is the first report of immunophenotypic characterisation of cutaneous lymphoma in the horse. The horse was treated with intralesional glucocorticoids and oral progestins, and complete resolution of the nodules resulted.
