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A Strategy for Sensitive, Large Scale Quantitative Metabolomics
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Quantification of hypoglycin A as butyl ester.

Johannes Sander1, Michael Terhardt1, Stefanie Sander1

  • 1Screening-Labor Hannover, Postbox 91 10 09, 30430 Hannover, Germany.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|July 20, 2016
PubMed
Summary

L-α-amino-methylenecyclopropyl propionic acid (Hypoglycin A, HGA) is the toxic compound in Sapindaceae fruits causing intoxication and symptoms of acquired multiple acyl-CoA dehydrogenase deficiency (MADD). A new method quantifies HGA and its metabolites for diagnosing poisoning.

Keywords:
Acquired multiple acyl-CoA dehydrogenase deficiencyAtypical myopathy of horsesHypoglycine AJamaican vomiting sicknessMethylenecyclopropyl acetate

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Area of Science:

  • Toxicology
  • Biochemistry
  • Analytical Chemistry

Background:

  • Hypoglycin A (HGA), a toxic compound in Sapindaceae fruits, causes acute intoxication and symptoms mimicking acquired multiple acyl-CoA dehydrogenase deficiency (MADD).
  • Accurate quantification of HGA and its metabolites is crucial for diagnosing poisoning and understanding MADD.

Purpose of the Study:

  • To develop and validate a sensitive analytical method for quantifying L-α-amino-methylenecyclopropyl propionic acid (Hypoglycin A, HGA).
  • To integrate HGA quantification into a broader method for analyzing MADD-related metabolites, including acyl glycines and acyl carnitines.

Main Methods:

  • Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was employed for HGA measurement after butylation.
  • A 7-point linear calibration curve using d3-leucine as an internal standard was utilized for quantification in serum and urine samples.
  • HGA measurement was integrated into a previously established method for acyl conjugates and HGA metabolites.

Main Results:

  • The method demonstrated high precision with coefficients of variation <15% at low concentrations (0.01 μmol/L) and ≤4.1% at higher concentrations (10 μmol/L).
  • Excellent linearity was achieved, with R(2) values ≥0.995 for the calibration curves.
  • The integrated method allows for the simultaneous quantification of non-metabolized HGA, its metabolites, and other acyl conjugates typical of acquired MADD.

Conclusions:

  • The developed UPLC-MS/MS method provides reliable quantification of HGA, essential for diagnosing poisoning.
  • This integrated approach facilitates the biochemical diagnosis of Ackee fruit poisoning, atypical equine myopathy, and forensic investigations of suspected HGA toxicity.