Rapid diagnosis of hypoglycin A intoxication in atypical myopathy of horses.
Abstract: Hypoglycin A (2-amino-3-(2-methylidenecyclopropyl)propanoic acid) is the plant toxin shown to cause atypical myopathy in horses. It is converted in vivo to methylenecyclopropyl acetic acid, which is transformed to a coenzyme A ester that subsequently blocks beta oxidation of fatty acids. Methylenecyclopropyl acetic acid is also conjugated with carnitine and glycine. Acute atypical myopathy may be diagnosed by quantifying the conjugates of methylenecyclopropyl acetic acid plus a selection of acyl conjugates in urine and serum. We describe a new mass spectrometric method for sample volumes of <0.5 mL. Samples were extracted with methanol containing 5 different internal standards. Extracts were analyzed by ultra-high-performance liquid chromatography-tandem mass spectrometry focusing on 11 metabolites. The total preparation time for a series of 20 samples was 100 min. Instrument run time was 14 min per sample. For the quantification of carnitine and glycine conjugates of methylenecyclopropyl acetic acid in urine, the coefficients of variation for intraday quantification were 2.9% and 3.0%, respectively. The respective values for interday were 9.3% and 8.0%. Methylenecyclopropyl acetyl carnitine was detected as high as 1.18 µmol/L in serum (median: 0.46 µmol/L) and 1.98 mmol/mol creatinine in urine (median: 0.79 mmol/mol creatinine) of diseased horses, while the glycine derivative accumulated up to 1.97 mmol/mol creatinine in urine but was undetectable in most serum samples. In serum samples from horses with atypical myopathy, the intraday coefficients of variation for C4-C8 carnitines and glycines were ≤4.5%. Measured concentrations exceeded those in healthy horses by ~10 to 1,400 times.
© 2015 The Author(s).
Publication Date: 2016-03-12 PubMed ID: 26965229DOI: 10.1177/1040638715624736Google Scholar: Lookup
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The research article focuses on a new method for the swift diagnosis of Hypoglycin A intoxication in horses suffering from atypical myopathy, using ultra-high-performance liquid chromatography-tandem mass spectrometry. This method requires less than 0.5 mL of urine or serum samples and is more efficient than previous methodologies.
Hypoglycin A and Atypical Myopathy
- The paper discusses Hypoglycin A, a plant toxin which causes atypical myopathy in horses.
- In the body, Hypoglycin A is converted into methylenecyclopropyl acetic acid, that turns into a coenzyme A ester and halts the beta oxidation of fatty acids.
- This acid also combines with glycine and carnitine, compounds which can be used as biomarkers for diagnosing acute atypical myopathy.
New Mass Spectrometric Method
- The researchers have developed a new mass spectrometric method for the diagnosis of Hypoglycin A intoxication in horses.
- By focusing on about 11 different metabolites, they were able to extract and analyze samples using ultra-high-performance liquid chromatography-tandem mass spectrometry.
- This method was found to be efficient, needing only 100 minutes to prepare 20 samples and 14 minutes per sample for running the tests.
Test Findings and Efficiency
- The test results for carnitine and glycine conjugates of methylenecyclopropyl acetic acid in urine showed intraday and interday variations of 2.9%, 3.0%, 9.3%, and 8.0% respectively.
- High levels of Methylenecyclopropyl acetyl carnitine were found in both the serum and urine samples of diseased horses, demonstrating that this test could effectively identify Hypoglycin A intoxication.
- Significantly higher concentrations of various detected compounds were found in the test horses compared to healthy ones, further showcasing the effectiveness of the new method.
Cite This Article
APA
Sander J, Cavalleri JM, Terhardt M, Bochnia M, Zeyner A, Zuraw A, Sander S, Peter M, Janzen N.
(2016).
Rapid diagnosis of hypoglycin A intoxication in atypical myopathy of horses.
J Vet Diagn Invest, 28(2), 98-104.
https://doi.org/10.1177/1040638715624736 Publication
Researcher Affiliations
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw) j.sander@metabscreen.de.
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
- Screening-Labor Hannover, Germany (J Sander, Terhardt, S Sander, Peter, Janzen)Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany (Cavalleri)Department of Clinical Chemistry, Hannover Medical School, Hannover, Germany (Janzen)Department of Animal Nutrition, Institute for Agricultural and Nutritional Sciences, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany (Bochnia, Zeyner)Department of Veterinary Pathology, College of Veterinary Medicine, Freie, Universität Berlin, Berlin, Germany (Zuraw).
MeSH Terms
- Animals
- Chromatography, High Pressure Liquid / veterinary
- Horse Diseases / blood
- Horse Diseases / chemically induced
- Horse Diseases / diagnosis
- Horse Diseases / urine
- Horses
- Hypoglycins / toxicity
- Muscular Diseases / chemically induced
- Muscular Diseases / diagnosis
- Muscular Diseases / veterinary
- Plant Poisoning / veterinary
- Reagent Kits, Diagnostic
- Tandem Mass Spectrometry / veterinary
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