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Home / Equine Research Bank / Toxins (Basel) / Co-Occurrence of Hypoglycin A and Hypoglycin B in Sycamore a...
Toxins2022; 14(9); doi: 10.3390/toxins14090608

Co-Occurrence of Hypoglycin A and Hypoglycin B in Sycamore and Box Elder Maple Proved by LC-MS/MS and LC-HR-MS.

Abstract: Hypoglycin A (HGA) and methylenecyclpropylglycine (MCPrG) are formed by some maple trees (Acer species) and have been associated with incidences of atypical myopathy among horses in pastures. In this work, a simple and sensitive ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method without derivatization was developed for the quantification of HGA and MCPrG in maple samples and validated according to EU guidelines. The LOQ presented here for HGA (16.4 µg/kg) is considerably lower than the lowest published LOQ (500 µg/kg). This method confirms that sycamore and box elder maple contain considerable amounts of HGA and MCPrG. In addition, the presence of the dipeptides hypoglycin B and γ-glutamyl-MCPrG in these two maple species is shown using high-resolution MS. This is the first report on the presence of these dipeptides in maple since 1973. The presence of HGB and γ-glutamyl-MCPrG could change the way we understand animal intoxication following the ingestion of maple.
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Publication Date: 2022-09-01 PubMed ID: 36136546PubMed Central: PMC9504185DOI: 10.3390/toxins14090608Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research work focuses on the presence of harmful components hypoglycin A (HGA) and methylenecyclpropylglycine (MCPrG) in some maple tree species which has been linked to atypical myopathy in horses. Using a new method, the study verified the presence of these components in sycamore and box elder maples that could be influential in understanding why animals get intoxicated after consuming maple.

Development of a New UPLC-MS/MS Method

  • The researchers developed an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method, which is a technique used to measure the mass and concentration of molecules. This allows for the quantification of HGA and MCPrG in maple samples.
  • This new method is simple and sensitive and was validated according to EU guidelines. Importantly, it did not require the process of derivatization which often adds complexity and uncertainty to these sorts of analyses.

Improved Detection of Harmful Components

  • With this new method, the researchers were able to detect HGA at a much lower concentration (16.4 µg/kg) than what was previously possible with the lowest published limit of quantification (LOQ) (500 µg/kg). This allows for better and more precise measurement of these harmful compounds.
  • They confirmed that the sycamore and box elder maple trees contain substantial amounts of HGA and MCPrG, which are likely responsible for incidences of atypical myopathy among horses that consume these trees.

Discovery of Dipeptides in Maple

  • In addition to identifying HGA and MCPrG, the investigation also found the presence of the dipeptides hypoglycin B and γ-glutamyl-MCPrG in these maple species using high-resolution mass spectrometry.
  • This is the first documented instance of the identification of these dipeptides in maple trees since 1973, signaling a significant development in the understanding of the components of maple trees.
  • The existence of these dipeptides could potentially change our understanding of how animals get intoxicated by ingesting maple.

Cite This Article

APA
El-Khatib AH, Engel AM, Weigel S. (2022). Co-Occurrence of Hypoglycin A and Hypoglycin B in Sycamore and Box Elder Maple Proved by LC-MS/MS and LC-HR-MS. Toxins (Basel), 14(9). https://doi.org/10.3390/toxins14090608

Publication

Toxins
ISSN: 2072-6651
NlmUniqueID: 101530765
Country: Switzerland
Language: English
Volume: 14
Issue: 9

Researcher Affiliations

El-Khatib, Ahmed H
  • German Federal Institute for Risk Assessment (BfR), Department for Safety in the Food Chain, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
Engel, Anna Maria
  • German Federal Institute for Risk Assessment (BfR), Department for Safety in the Food Chain, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
Weigel, Stefan
  • German Federal Institute for Risk Assessment (BfR), Department for Safety in the Food Chain, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.

MeSH Terms

  • Acer / chemistry
  • Animals
  • Chromatography, Liquid
  • Dipeptides
  • Horse Diseases
  • Horses
  • Hypoglycins / analysis
  • Hypoglycins / toxicity
  • Tandem Mass Spectrometry / methods

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 49 references
  1. Manchester K. Biochemistry of hypoglycin. FEBS Lett 1974;40:S128–S132.
    doi: 10.1016/0014-5793(74)80696-4pubmed: 0google scholar: lookup
  2. Gaillard Y, Carlier J, Berscht M, Mazoyer C, Bevalot F, Guitton J, Fanton L. Fatal intoxication due to ackee (Blighia sapida) in Suriname and French Guyana. GC–MS detection and quantification of hypoglycin-A. Forensic Sci. Int. 2011;206:e103–e107.
    pubmed: 21324617
  3. Bowen-Forbes C.S., Minott D.A.. Tracking hypoglycins A and B over different maturity stages: Implications for detoxification of ackee (Blighia sapida K.D. Koenig) fruits. J. Agric. Food Chem. 2011;59:3869–3875.
    doi: 10.1021/jf104623cpubmed: 21410289google scholar: lookup
  4. Isenberg S.L., Carter M.D., Hayes S.R., Graham L.A., Johnson D., Mathews T.P., Harden L.A., Takeoka G.R., Thomas J.D., Pirkle J.L.. Quantification of toxins in soapberry (Sapindaceae) arils: Hypoglycin A and methylenecyclopropylglycine. J. Agric. Food Chem. 2016;64:5607–5613.
    doi: 10.1021/acs.jafc.6b02478pmc: PMC5098216pubmed: 27367968google scholar: lookup
  5. Sanford A., Isenberg S.L., Carter M.D., Mojica M.A., Mathews T.P., Harden L.A., Takeoka G.R., Thomas J.D., Pirkle J.L., Johnson R.C.. Quantitative HPLC–MS/MS analysis of toxins in soapberry seeds: Methylenecyclopropylglycine and hypoglycin A. Food Chem. 2018;264:449–454.
    doi: 10.1016/j.foodchem.2018.04.093pmc: PMC5985522pubmed: 29853400google scholar: lookup
  6. Yang J., Zhu X., Zhang P., Wang Y., Xiao Y., Yang B., Qu H., Jiang Y.. Detection of toxic methylenecyclopropylglycine and hypoglycin A in litchi aril of three Chinese cultivars. Food Chem. 2020;327:127013.
    doi: 10.1016/j.foodchem.2020.127013pubmed: 32454275google scholar: lookup
  7. Unger L., Nicholson A., Jewitt E., Gerber V., Hegeman A., Sweetman L., Valberg S.. Hypoglycin A Concentrations in Seeds of Acer pseudoplatanus Trees Growing on Atypical Myopathy-Affected and Control Pastures. J. Vet. Intern. Med. 2014;28:1289–1293.
    doi: 10.1111/jvim.12367pmc: PMC4857957pubmed: 24863395google scholar: lookup
  8. Baise E., Habyarimana J.A., Amory H., Boemer F., Douny C., Gustin P., Marcillaud-Pitel C., Patarin F., Weber M., Votion D.M.. Samaras and seedlings of Acer pseudoplatanus are potential sources of hypoglycin A intoxication in atypical myopathy without necessarily inducing clinical signs. Equine Vet. J. 2016;48:414–417.
    doi: 10.1111/evj.12499pubmed: 26278545google scholar: lookup
  9. Westermann C., Van Leeuwen R., van Raamsdonk L., Mol H.. Hypoglycin A Concentrations in Maple Tree Species in the Netherlands and the Occurrence of Atypical Myopathy in Horses. J. Vet. Intern. Med. 2016;30:880–884.
    doi: 10.1111/jvim.13927pmc: PMC4913566pubmed: 26995161google scholar: lookup
  10. Medina S.G., Hyde C., Lovera I., Piercy R.J.. Detection of equine atypical myopathy-associated hypoglycin a in plant material: Optimisation and validation of a novel LC-MS based method without derivatisation. PLoS ONE 2018;13:e0199521.
    pmc: PMC6029767pubmed: 29969503
  11. Valberg S.J., Sponseller B.T., Hegeman A.D., Earing J., Bender J.B., Martinson K.L., Patterson S.E., Sweetman L.. Seasonal pasture myopathy/atypical myopathy in North America associated with ingestion of hypoglycin A within seeds of the box elder tree. Equine Vet. J. 2012;45:419–426.
    doi: 10.1111/j.2042-3306.2012.00684.xpubmed: 23167695google scholar: lookup
  12. Vashishtha V.M.. Outbreaks of hypoglycemic encephalopathy in Muzzaffarpur, India: Are these caused by toxins in litchi fruit?. Indian Pediatr. 2016;53:399–402.
    pubmed: 27254049
  13. Shrivastava A., Kumar A., Thomas J.D., Laserson K.F., Bhushan G., Carter M.D., Chhabra M., Mittal V., Khare S., Sejvar J.J.. Association of acute toxic encephalopathy with litchi consumption in an outbreak in Muzaffarpur, India, 2014: A case-control study. Lancet Glob. Health 2017;5:e458–e466.
    doi: 10.1016/S2214-109X(17)30035-9pubmed: 28153514google scholar: lookup
  14. Scott H.H.. On the ‘Vomiting Sickness’ of Jamaica. Ann. Trop. Med. Parasitol. 1916;10:1–78.
    doi: 10.1080/00034983.1916.11684104google scholar: lookup
  15. Trauner D.A., Nyhan W.L., Sweetman L., Tanaka K.A.Y.. Jamaican Vomiting Sickness and Reye’s Syndrome. N. Engl. J. Med. 1976;295:1481–1482.
    pubmed: 995147
  16. Golden K.D., Kean E.A., Terry S.I.. Jamaican vomiting sickness: A study of two adult cases. Clin. Chim. Acta. 1984;142:293–298.
    doi: 10.1016/0009-8981(84)90265-1pubmed: 6488562google scholar: lookup
  17. Joskow R., Belson M., Vesper H., Backer L., Rubin C.. Ackee fruit poisoning: An outbreak investigation in Haiti 2000–2001, and review of the literature. Clin. Toxicol. 2006;44:267–273.
    doi: 10.1080/15563650600584410pubmed: 16749544google scholar: lookup
  18. Gordon A. Biochemistry of Hypoglycin and Toxic Hypoglycemic Syndrome. 2015;pp. 47–61.
  19. Hörügel U., Simon H., Dressel A., Birke H., Matz K., Klunker M.. Atypical myopathy in horses: Review and investigation results from Saxony and Thuringia. Prakt. Tierarzt. 2014;95:444–457.
  20. Aboling S., Schliephake A., Cavalleri J.-M.V., Kamphues J.. Proof of sycamore maple in the content of the digestive tract in a horse on suspect of atypical myopathy. Pferdeheilkunde Equine Med. 2015;31:135–139.
    doi: 10.21836/PEM20150204google scholar: lookup
  21. Bochnia M., Ziegler J., Sander J., Uhlig A., Schaefer S., Vollstedt S., Glatter M., Abel S., Recknagel S., Schusser G.F.. Hypoglycin A Content in Blood and Urine Discriminates Horses with Atypical Myopathy from Clinically Normal Horses Grazing on the Same Pasture. PLoS ONE 2015;10:e0136785.
    doi: 10.1371/journal.pone.0136785pmc: PMC4574941pubmed: 26378918google scholar: lookup
  22. González-Medina S., Ireland J., Piercy R.J., Newton J.R., Votion D.M.. Equine atypical myopathy in the UK: Epidemiological characteristics of cases reported from 2011 to 2015 and factors associated with survival. Equine Vet. J. 2017;49:746–752.
    doi: 10.1111/evj.12694pubmed: 28445006google scholar: lookup
  23. Bochnia M., Sander J., Ziegler J., Terhardt M., Sander S., Janzen N., Cavalleri J., Zuraw A., Wensch-Dorendorf M., Zeyner A.. Detection of MCPG metabolites in horses with atypical myopathy. PLoS ONE 2019;14:e0211698.
    doi: 10.1371/journal.pone.0211698pmc: PMC6363182pubmed: 30721263google scholar: lookup
  24. Bunert C., Langer S., Votion D.M., Boemer F., Müller A., Ternes K., Liesegang A.. Atypical myopathy in Père David’s deer (Elaphurus davidianus) associated with ingestion of hypoglycin A. J. Anim. Sci. 2018;96:3537–3547.
    doi: 10.1093/jas/sky200pmc: PMC6095259pubmed: 29762728google scholar: lookup
  25. Bochnia M., Ziemssen E., Sander J., Stief B., Zeyner A.. Methylenecyclopropylglycine and hypoglycin A intoxication in three Pére David’s Deers (Elaphurus davidianus) with atypical myopathy. Vet. Med. Sci. 2021;7:998–1005.
    doi: 10.1002/vms3.406pmc: PMC8136943pubmed: 33314647google scholar: lookup
  26. Hirz M., Gregersen H.A., Sander J., Votion D.M., Schänzer A., Köhler K., Herden C.. Atypical myopathy in 2 Bactrian camels. J. Veter. Diagn. Investig. 2021;33:961–965.
    doi: 10.1177/10406387211020721pmc: PMC8366255pubmed: 34092152google scholar: lookup
  27. Renaud B., François A.-C., Boemer F., Kruse C., Stern D., Piot A., Petitjean T., Gustin P., Votion D.-M.. Grazing Mares on Pasture with Sycamore Maples: A Potential Threat to Suckling Foals and Food Safety through Milk Contamination. Animals 2021;11:87.
    doi: 10.3390/ani11010087pmc: PMC7824825pubmed: 33466424google scholar: lookup
  28. Sander J., Terhardt M., Janzen N.. Detection of maple toxins in mare’s milk. J. Vet. Intern. Med. 2021;35:606–609.
    doi: 10.1111/jvim.16004pmc: PMC7848382pubmed: 33336854google scholar: lookup
  29. Bochnia M., Ziegler J., Glatter M., Zeyner A.. Hypoglycin A in Cow’s Milk—A Pilot Study. Toxins 2021;13:381.
    doi: 10.3390/toxins13060381pmc: PMC8230099pubmed: 34073628google scholar: lookup
  30. Westermann C.M., Dorland L., Votion D.M., De Sain-van der Velden M.G.M., Wijnberg I.D., Wanders R.J.A., Spliet W.G.M., Testerink N., Berger R., Ruiter J.P.N.. Acquired multiple Acyl-CoA dehydrogenase deficiency in 10 horses with atypical myopathy. Neuromuscul. Disord. 2008;18:355–364.
    doi: 10.1016/j.nmd.2008.02.007pubmed: 18406615google scholar: lookup
  31. Sander J., Terhardt M., Sander S., Janzen N.. Quantification of Methylenecyclopropyl Compounds and Acyl Conjugates by UPLC-MS/MS in the Study of the Biochemical Effects of the Ingestion of Canned Ackee (Blighia sapida) and Lychee (Litchi chinensis). J. Agric. Food Chem. 2017;65:2603–2608.
    doi: 10.1021/acs.jafc.7b00224pubmed: 28290200google scholar: lookup
  32. Barceloux D.G.. Akee Fruit and Jamaican Vomiting Sickness (Blighia sapida Köenig). Dis. Mon. 2009;55:318–326.
    doi: 10.1016/j.disamonth.2009.03.002pubmed: 19446675google scholar: lookup
  33. Seeff L., Stickel F., Navarro V.J.. Chapter 35—Hepatotoxicity of Herbals and Dietary Supplements. 2013;pp. 631–657.
  34. Hassall C.H., Reyle K., Feng P.. Hypoglycin A, B: Biologically active polypeptides from Blighia sapida. Nature 1954;173:356–357.
    doi: 10.1038/173356b0pubmed: 13144762google scholar: lookup
  35. Golden K.D., Williams O.J., Bailey-Shaw Y.. High-Performance Liquid Chromatographic Analysis of Amino Acids in Ackee Fruit with Emphasis on the Toxic Amino Acid Hypoglycin A. J. Chromatogr. Sci. 2002;40:441–446.
    doi: 10.1093/chromsci/40.8.441pubmed: 12387335google scholar: lookup
  36. Fowden L., Pratt H.M.. Cyclopropylamino acids of the genus Acer: Distribution and biosynthesis. Phytochemistry 1973;12:1677–1681.
    doi: 10.1016/0031-9422(73)80387-5google scholar: lookup
  37. Kean E.A.. Hypoglycin. 1989;pp. 229–262.
  38. Brown M., Bates R., McGowan C., Cornell J.. Influence of Fruit Maturity on the Hypoglycin a Level in Ackee (Blighia sapida). J. Food Saf. 1991;12:167–177.
    doi: 10.1111/j.1745-4565.1991.tb00075.xgoogle scholar: lookup
  39. McGowan C., Wiley V.A., Bates R.P.. Application of methodology for RP-HPLC amino acid analysis to the measurement of hypoglycin A. BioChromatography 1989;4:161–164.
  40. Whitaker T.B., Saltsman J.J., Ware G.M., Slate A.B.. Evaluating the Performance of Sampling Plans to Detect Hypoglycin A in Ackee Fruit Shipments Imported into the United States. J. AOAC Int. 2007;90:1060–1072.
    doi: 10.1093/jaoac/90.4.1060pubmed: 17760344google scholar: lookup
  41. Sander J., Terhardt M., Sander S., Janzen N.. Quantification of hypoglycin A as butyl ester. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 2016;1029–1030:169–173.
    doi: 10.1016/j.jchromb.2016.07.005pubmed: 27433981google scholar: lookup
  42. Żuraw A., Dietert K., Kühnel S., Sander J., Klopfleisch R.. Equine atypical myopathy caused by hypoglycin A intoxication associated with ingestion of sycamore maple tree seeds. Equine Vet. J. 2016;48:418–421.
    doi: 10.1111/evj.12460pubmed: 25970235google scholar: lookup
  43. González-Medina S., Bevin W., Alzola-Domingo R., Chang Y., Piercy R.J.. Hypoglycin A absorption in sheep without concurrent clinical or biochemical evidence of disease. J. Vet. Intern. Med. 2021;35:1170–1176.
    doi: 10.1111/jvim.16077pmc: PMC7995363pubmed: 33675130google scholar: lookup
  44. Kean E.A., Hare E.R.. Gamma-Glutamyl-Transferase Transpeptidase of the Ackee Plant. Phytochemistry 1980;19:199–203.
    doi: 10.1016/S0031-9422(00)81960-3google scholar: lookup
  45. Young M.. Studies on the Growth in Culture of Plant Cells: XVI. Nitrogen Assimilation during Nitrogen-Limited Growth of Acer pseudoplatanus L. Cells in Chemostat Culture. J. Exp. Bot. 1973;24:1172–1185.
    doi: 10.1093/jxb/24.6.1172google scholar: lookup
  46. Chen K.K., Anderson R.C., McCowen M.C., Harris P.N.. Pharmacologic action of hypoglycin A and B. J. Pharmacol. Exp. Ther. 1957;121:272–285.
    pubmed: 13481850
  47. Persaud T.V.. Effect of intra-amniotic administration of hypoglycin B on foetal development in the rat. Exp. Pathol. 1972;6:55–58.
    pubmed: 5020320
  48. European-Commission, Directorate-General for Health and Food Safety SANTE. Guidance Document on Analytical Quality Control and Method Validation Procedures for Pesticide Residues Analysis in Food and Feed. 2019.
  49. European-Commission, Wenzl T., Haedrich J., Schaechtele A., Robouch P., Stroka J.. Guidance Document on the Estimation of LOD and LOQ for Measurements in the Field of Contaminants in Feed and Food. 2016.
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