Structure Elucidation of a New Toxin from the Mushroom Cortinarius rubellus Using Gas Chromatography-Mass Spectrometry (GC-MS)

Ilia Brondz

Department of Biosciences, University of Oslo, Oslo, Norway 2R & D Department of Jupiter Ltd., Ski, Norway.

DOI: 10.4236/ijamsc.2013.12014                

Abstract

Cortinarius orellanus (Fries) and C. rubellus (Cooke),which were formerly also known as C. speciosissimus, are poisonous mushrooms containing the toxin orellanine and several degradation products of orellanine,includingorelline and orellinine. Mass intoxication by poisonous mushrooms was observed in Poland in 1952-1957 [1]. In 1957, the cause of these outbreaks was described by Grzymala as poisoning by a member of the Cortinarius family. The toxin orellanine was first isolated from C. orellanusby Grzymala in 1962; the chemical structure of orellanine was later determined to be 3,3',4,4'-tetrahydroxy-2,2'-bipyridine-N,N'-dioxide. Poisoning with C. orellanus and C. rubellus has a very specific character. The first symptoms of intoxication usually do not appear until 2-3 days after ingestion, but in some cases intoxication appears after three weeks. The target organ for the toxin is the kidney. Histologically, it is easy to record the specific damage. The presence of degradation products of orellanine in kidney can be confirmed chromatographically, suggesting that the cause of poisoning is orellanine. However, the presence of orellanine in the blood of intoxicated persons has not been directly detected. A specific model was developed by Brondz et al. for the detection of orellanine, orelline, and orellininein animal stomach fluids [2-4]. The hypothesis that the fungal toxin orellanine as a diglucoside can be transported from the digestive system by the blood to the kidney could not be supported. The toxin orellanine as a diglucoside is very unstable in an aqueous acidic environment.[i1] However, in the present study, it was possible to record an additional substance in animal stomach fluids using GC-MSafter ingestion ofC. rubellus. This substance, which has been namedrubelline, is part of a toxic mixture inC. orellanusandC. rubellusand is closely related to orellanine. The structure of rubelline is more suitable than orellanine for absorptionfromthe digestive tract and for transport in the blood. The presented hypothesis is that rubellineis absorbed in the digestive tract and transportedin the blood to the kidney, where it is biotransformed to orellanine and accumulatedto toxic levels. The process of biotransformationis in itself also damaging for the kidney and liver.[i2] GC-MS instrumentation enables the separation of substances in biological samples and in the extract fromC. rubellus. The GC-MS with SMB technique was used to record the mass ion and to record a detailed fragmentation picture.

Keywords

Gas Chromatography; Mass Spectrometry

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	S. Grzymala, “Massenvergiftungen durch den Orangefuch- sigen Hautkopf,” Zeitschriftfür Pilzkunde, Vol. 23, 1957, pp. 139-142.
[2]	I. Brondz, E. Nevo, S. P. Wasser, K. H?iland, F. P. St?rmer and A. C. Brondz, “Method Development for in Vivo Analyses of Toxin Orellanine from the Toad Mush- room Cortinarius orellanus,” Separation Science Europe 2011, 10-11 October 2011, The Royal Institution of Great Britain, London, Abstract 9.
[3]	I. Brondz, E. Nevo, S. Wasser and A. Brondz, “A Direct Gas Chromatography-Mass Spectrometry (GC-MS) Method for the Detection of Orellanine Present in Stomach Con- tent. Part I,” Journal of Biophysical Chemistry, Vol. 3, No. 1, 2012, pp. 29-34. http://dx.doi.org/10.4236/jbpc.2012.31003
[4]	I. Brondz and A. Brondz, “A High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) Quali- tative Detection Method Developed for in Vivo Analyses of Toxin Orellanine from the Cortinarius orellanus Fr. —Part II,” ISRN Chromatography, Vol. 2012, 2012, Ar- ticle ID 293830.
[5]	I. Brondz, “Fungi, Classification of the Basidiomycetes,” In: C. Batt and M. L. Tortorello, Eds., Encyclopedia of Food Microbiology, 2nd Edition, Elsevier, Oxford, 2014, in Press.
[6]	T. Schumacher and K. H?iland, “Review Mushroom Poisoning Caused by Species of the Genus Cortinarius Fries,” Archives of Toxicology, Vol. 53, 1983, pp. 87-106.
[7]	I. R. Tebbett and B. Caddy, “Mushroom Toxins of Genus Cortinarius,” Experientia, Vol. 40, No. 5, 1984, pp. 441- 446. http://dx.doi.org/10.1007/BF01952379
[8]	L. Matthies and H. Laatsch, “Cortinarins in Cortinarius speciosissimus? A Critical Revision,” Experientia, Vol. 47, No. 6, 1991, pp. 634-640. http://dx.doi.org/10.1007/BF01949895
[9]	M. Rohrmoser, M. Kirchmair, E. Feifel, A. Valli, R. Corradini, E. Pohanka, A. Rosenkranz and R. P?der, “Orellanine Poisoning: Rapid Detection of the Fun-Gal Toxin in Renal Biopsy Material,” Journal of Toxicology—Clinical Toxicology, Vol. 35, No. 1, 1997, pp. 63-66. http://dx.doi.org/10.3109/15563659709001167
[10]	S. Rapior, N. Delpech, C. Andary and G. Huchhard, “Intoxication by Cortinarius orellanus: Detection and Assay of Orellanine in Biological Fluids and Renal Biopsies,” Mycopathologia, Vol. 108, No. 3, 1989, pp. 155-161. http://dx.doi.org/10.1007/BF00436220
[11]	W. Z. Antkowiak, R. Antkowiak, E. Wyrzykiewicz and G. Czerwiński, “Mass Spectral Fragmentation of Orellanine and Its Tetramethyl Ether with Regard to Their Facile Thermal and Photochemical Deoxygenation,” Heterocycles, Vol. 39, No. 2, 1994, pp. 477-484. http://dx.doi.org/10.3987/COM-94-S(B)72
[12]	J. Holmdahl, J. Ahl?en, S. Bergek, S. Lundberg, S.-A. Persson, “Isolation and Nephrotoxic Studies of Orellanine from the Mushroom Cortinarius speciosissmus,” Toxicon, Vol. 25, No. 2, 1987, pp. 195-199. http://dx.doi.org/10.1016/0041-0101(87)90241-8
[13]	P. Spiteller, M. Spiteller and W. Steglich, “Occurrence of the Fungal Toxin Orellanine as a Diglucoside and Investigation of Its Biosynthesis,” Angewandte Chemie Inter- national Edition, Vol. 42, No. 25, 2003, pp. 2864-2867. http://dx.doi.org/10.1002/anie.200351066
[14]	A. Herrmann, H. Hedman, J. Rosén, D. Jansson, B. Ha- raldsson and K.-E. Hellen?s, “Analysis of the Mashroom Nephrotoxin Orellanine and Its Glucosides,” Journal of Natural Products, Vol. 75, No. 10, 2012, pp. 1690-1696. http://dx.doi.org/10.1021/np300135k
[15]	I. Brondz, A. B. Fialkov and A. Amirav, “Analysis of Quinocide in Unprocessed Primaquine Diphosphate and Primaquine Diphosphate Tablets Using Gas Chromatography-Mass Spectrometry with Supersonic Molecular Beams,” Journal of Chromatography A, Vol. 1216, No. 5, 2009, pp. 824-829. http://dx.doi.org/10.1016/j.chroma.2008.11.043
[16]	A. Amirav, “What Can Be Improved in GC-MS—When Multi Benefits Are Transformed into a GC-MS Revolution,” International Journal of Analytical Mass Spectrometry and Chromatography (IJAMSC), Vol. 1, No 1, 2013, pp. 31-47. http://dx.doi.org/10.4236/ijamsc.2013.11005
[17]	H. Prast, E. R. Wemer, W. Pfaller and M. Moser, “Toxic Properties of the Mushroom Cortinarius orellanus,” Archives of Toxicology, Vol. 62, No. 1, 1988, pp. 81-88. http://dx.doi.org/10.1007/BF00316263
[18]	I. Brondz, D. Mantzeilas, U. Klein, M. N. Lebedeva, F. S. Mikhailitsyn, G. D. Souleimanov and D. Ekeberg, “The Main Contaminant of the Anti-Malarial Drug Primaquine is Its Positional Isomer,” 3rd International Symposium on Separation in BioSciences SBS 2003 A 100 Years of Chromatography, 13-18 May 2003, Moscow, Abstract p. 57, 165.
[19]	I. Brondz, D. Mantzilas, U. Klein, D. Ekeberg, E. Hvattum, M. N. Lebedeva, F. S. Mikhailitsyn, G. D. Souleimanov and J. R?e, “Nature of the Main Contaminant in the Antimalaria Drug Primaquine Di-Phosphate: A Qualitative Isomer Analysis,” Chromatography B: Biomedical Sciences and Applications, Vol. 800, No. 1-2, 2004, pp. 211- 223.
[20]	G. G. Coté and R. C. Crain, “Why Do Plants Have Phosphoinositides?” BioEssays, Vol. 16, No. 1, 1994, pp. 39- 46. http://dx.doi.org/10.1002/bies.950160106
[21]	I. Brondz and A. Brondz, “Supercritical Fluid Chromatography-Mass Spectrometry (SFC-MS) and MADI-TOF- MS of Heterocyclic Compounds with Trivalent and Pentavalent Nitrogen in Cough Relief Medical Forms Tuxi and Cosylan,” American Journal of Analytical Chemistry, Vol. 3, No. 12A, 2012, pp. 870-876. http://dx.doi.org/10.4236/ajac.2012.312A115
[22]	I. Brondz, “Mass Spectrometric Structure Elucidation of the Trivalent and Pentavalent Nitrogen Contaminants of Pholcodine in the Cough Relief Medical Form Tuxidrin,” International Journal of Analytical Mass Spectrometry and Chromatography, Vol. 1, No. 1, 2013, pp. 5-10. http://dx.doi.org/10.4236/ijamsc.2013.11002
[23]	G. M. Jones and J. A. Vale, “Mechanisms of Toxicity, Clinical Features, and Management of Diquat Poisoning: A Review,” Journal of Toxicology-Clinical Toxicology, Vol. 38, No. 2, 2000, pp. 123-128. http://dx.doi.org/10.1081/CLT-100100926