Research Article
BibTex RIS Cite

Determination of Grayanotoxin-III from in Rhodendron Ponticum And Mad Honey Samples by Liquid Chromatography−Mass Spectrometry

Year 2019, Volume: 1 Issue: 1, 1 - 21, 25.07.2019

Abstract

Grayanotoxin-III and its glycoside derivatives were isolated from their natural sources. Grayanotoxin-III was extracted from Rhododendron Ponticum L. by liquid-liquid extraction and was isolated by prep-HPLC. Grayanotoxin-III glycoside was extracted from mad honey by SPE method. For isolation of grayanotoxin-III, a reverse phase C-18 column has been used in gradient conditions starting with 40/60 % (Methanol (A) /Water (B)) following 0 min. 34/66 (A/B), 11 min. 53/47 (A/B), 19 min. 70/30 (A/B), 22 min 100/0 (A/B) at 25 oC. Isolated fragments were examined with LC-MS analysis. The main ion of grayanotoxin-III was observed at m/z 370 with 100 % abundance with some of its fragments. Grayanotoxin-III was detected at 204 nm with Diode Array Detector at 7.5 min. with the above-mentioned conditions. Grayanotoxin-III glycoside from mad honey was identified by LC-MS and NMR analysis. 

GTX-III can be transferred to honey from Rhododendron Ponticum plant according to obtained results. As a conclusion, because of the toxicity of the GTX-III compound the honey from these regions should be carefully consumed.



 

Thanks

The research is one part of MS thesis of Talha Siddik Akkaya at Ege University, Graduate School of Natural and Applied Sciences.

References

  • Avci M. 2004. Rhododendron and their natural occurrences in Turkey. İstanbul Üniversitesi, Edebiyat Fak. Coğrafya Bölümü, Coğrafya Dergisi, 12: 13-29. (in Turkish)
  • Booth N.L., Kruger C.L., Hayes A.W., Clemens R. 2012. An innovative approach to the safety evaluation of natural products: Cranberry (Vaccinium macrocarpon Aiton) leaf aqueous extract as a case study. Food and Chemical Toxicology. 50: 3154-3165.
  • Cho H.E., Ahn S.Y., Kim D.W., Woo S.H., Park S.H., Hwang K., Moon D.C., Kim S. 2014. Development of a liquid chromatography–tandem mass spectrometry method for the determination of grayanotoxins in rat blood and its application to toxicokinetic study. Biomedical Chromatography, 28: 1624–1632.
  • Cucer N., Eroz R. 2010. Investigation of mutagenic effects of grayanotoxins II and III on cultured human lymphocytes. Al Ameen Journal of Medical Sciences. 4: 293-299.
  • Delgado D.D., Cuenca C.E.N., Sanchez L.P.R., Kushalappa A., Vasquez, M. (2015). Development and validation of a liquid chromatographic method to quantify sucrose, glucose, and fructose in tubers of Solanumtuberosum Group Phureja. The Journal of Chromatography. 975: 18-23.
  • Gunduz A., Eraydin I., Turkmen S., Kalkan O.F., Turedi T., Eryigit U., Ayar A. 2014. Analgesic effects of mad honey (grayanotoxin) in mice models of acute pain and painful diabetic neuropathy. Human and Experimental Toxicology. 33: 130–135.
  • Holstege D.M., Francis T., Puschner B., Booth M.C., Galey F.D. 2000. Multiresidue screen for cardiotoxins by two-dimensional thin-layer chromatography. Journal of Agricultural and Food Chemistry. 48: 60-64.
  • Holstege D.M., Pushchner B., Le T. 2001. Determination of grayanotoxins in biological samples by LC-MS/MS. Journal of Agricultural and Food Chemistry. 49: 1648-1651.
  • Hough R.L., Crews C., White D., Driffield M., Campbell C.D., Maltin C. 2010. Degradation of yew, ragworth and rhododendron toxins during composting. Science of the Total Environment. 408: 4128-4137.
  • Hwang T., Noh E., Jeong JH., Park S-K., Shin D., Kang H. 2018. Determination of Grayanotoxins from Rhododendron brachycarpum in Dietary Supplements and Homemade Wine by Liquid Chromatography-Quadrupole Time-of-Flight-Mass Spectrometry and Liquid Chromatography-Tandem Mass Spectrometry, J. Agric. Food Chem., 66: 1935-1940.
  • Kaplan M., Olgun E.O., Karaoglu O. 2014. Determination of grayanotoxins in honey by liquid chromatography tandem mass spectrometry using dilute-and-shoot sample preparation approach. J. Agric. Food Chem., 62: 5485-5491.
  • Karkacier M., Erbas M., Uslu M.K., Aksu M. 2003. Comparison of different extraction and detection methods for sugars using amino-bonded phase HPLC. The Journal of Chromatographic Science. 41: 331-333.
  • Kim S.E., Shin M.C., Akaike N., Kim C.J. 2010. Presynaptic effects of grayanotoxin III on excitatory and inhibitory nerve terminals in rat ventromedial hypothalamic neurons. NeuroToxicology. 31: 230–238.
  • Kurtoglu A.B., Yavuz R., Evrendilek G.A. 2014. Characterization and fate of grayanatoxins in mad honey produced from Rhododendron ponticum nectar. Food Chemistry. 161: 47-52.
  • Masutani T., Kawazu K., Uneyama K., Torii S., Iwasa J. 1979. Assignment of 13C-NMR Spectra of Grayanotoxin-I and –III. Agricultural and Biological Chemistry. 43: 631-635.
  • Meguri H., Katakawa J., Tetsumi T., Uchida S., Sumi M., Katai M., Terai T., Fukumato K. 1993. Simple and Rapid Determimation of Grayanotoxins by High−Performance Liquid Chrornatography. The Japanese Society of Pharmacognosy. 47: 429-433.
  • Michie D., Litterick A., Crews C. 2011. The influence of outdoor windrow composting on the concentration of grayanotoxins in Rhododendron leaves. Compost Science & Utilization. 19: 44-51.
  • Milne R., Abbott R.J. 2000. Origin and evolution of invasive naturalized material of Rhododendron ponticum L. in the British Isles. Molecular Ecology. 9: 543-556.Nishida R., Fukami H., Iriye R., Kumazawa Z. 1990. Accumulation of highly toxic Ericaceous diterpenoids by the Geometrid Moth, Arichanna gaschkevitchii. Agricultural and Biological Chemistry, 54: 2347-2352.
  • Niu C.S., Li Y., Liu Y.B., Ma S.G., Li L., Qu J., Yu S.S. 2016. Analgesic diterpenoids from the twigs of Pieris Formosa. Tetrahedron, 72: 44-49.Popescu R., Krupitza G., Kopp B. 2009. Novel insights into the mechanism of action of grayanotoxin III. Planta Medica, 75: 970-970.
  • Popescu R.K. 2013. The genus Rhododendron: An ethnopharmacological and toxicological review. The Journal of Ethnopharmacology. 147: 42-62.
  • Sahin H., Turumtay E.A., Yildiz O., Kolaylı S. 2015. Grayanotoxin-III dedection and antioxidant activity of mad honey. International Journal of Food Properties, 18: 2665-2674.
  • Sakata K., Hattori M., Sakurai A., Hosotsuji, T. (1977). Isolation and identification of biologically active constituents of Leucothoe catesbaei A. Gray. Journal of Pesticide Science, 2: 453-456.
  • Silici S., Atayoglu A.T. 2015. Mad honey intoxication: A systematic review on the 1199 cases. Food and Chemical Toxicology. 86: 282–290.
  • Silici S., Yonar M.E., Huseyin S., Atayoğlu A.T., Ozkok D. 2014. Analysis of grayanotoxin in rhododendron honey and effect on antioxidant parameters. The Journal of Ethnopharmacology. 156: 155-161.
  • Terai T., Araho D., Osakabe K., Katai M., Narama I., Matsuura T., Katakawa J., Tetsumi T. Sato M. 2000. Isolation of iso-grayanotoxin II from leaves of Leucothoe grayana MAX. Its X-Ray crystallographic analysis and acute toxicity in mice. Chemical & Pharmaceutical Bulletin, 48: 142-144.
  • These A., Bodi D., Uecker S., Reimers K, Ronczka S., Weigert A.P., Wiederholt M.L. 2015. A case of human poisoning by grayanotoxins following honey ingestion: elucidation of the toxin profile by mass spectrometry. Food Additives & Contaminants Part A, 32: 1679-1683.
  • Tiedeken E.J., Stout J.C., Stevenson P.C., Wright G.A. 2014. Bumblebees are not deterred by ecologically relevant concentrations of nectar toxins. Journal of Experimental Biology. 217: 1620-1625.
  • Victorita B., Margihitas L.A., Stanciu O., Laslo L., Dezmiream D., Bobis O. 2008. High-performance liquid chromatographic analysis of sugars in Transylvanian honeydew honey. The Bulletin of University of Agricultural Sciences and Veterinary Medicine. 65: 229-232.
  • Wong J., Youde E., Dickinson B., Hale M. 2002. Report of the rhododendron feasibility study. Prepared for the Beddgelert Rhododendron Management Group by the School of Agricultural and Forest Sciences, University of Wales, Bangor, 64-64.
  • Zhang M., Xie Y., Zhan Lei L., Shu P., Chen Y., Xue Y., Luo Z., Wana Q., Yao G., Zhang Y. 2015. Grayanane and leucothane diterpenoids from the leaves of Rhododendron micranthum. Phytochemistry. 117: 107–115.
  • Zhou C., Li X., Li H., Li R. 2012. Chemical constituents from the leaves of Craibiodendron yunnanense. Biochemical Systematics and Ecology, 45: 179–182.
  • Zielinski A.A.F., Braga C.M., Demiate I.M., Beltrame F.L., Nogueira A., Wosiacki G. 2014. Development and optimization of an HPLC-RI method for the determination of major sugars in apple juice and evaluation of the effect of the ripening stage. Food Science and Technology, 408: 4128-4137.
Year 2019, Volume: 1 Issue: 1, 1 - 21, 25.07.2019

Abstract

References

  • Avci M. 2004. Rhododendron and their natural occurrences in Turkey. İstanbul Üniversitesi, Edebiyat Fak. Coğrafya Bölümü, Coğrafya Dergisi, 12: 13-29. (in Turkish)
  • Booth N.L., Kruger C.L., Hayes A.W., Clemens R. 2012. An innovative approach to the safety evaluation of natural products: Cranberry (Vaccinium macrocarpon Aiton) leaf aqueous extract as a case study. Food and Chemical Toxicology. 50: 3154-3165.
  • Cho H.E., Ahn S.Y., Kim D.W., Woo S.H., Park S.H., Hwang K., Moon D.C., Kim S. 2014. Development of a liquid chromatography–tandem mass spectrometry method for the determination of grayanotoxins in rat blood and its application to toxicokinetic study. Biomedical Chromatography, 28: 1624–1632.
  • Cucer N., Eroz R. 2010. Investigation of mutagenic effects of grayanotoxins II and III on cultured human lymphocytes. Al Ameen Journal of Medical Sciences. 4: 293-299.
  • Delgado D.D., Cuenca C.E.N., Sanchez L.P.R., Kushalappa A., Vasquez, M. (2015). Development and validation of a liquid chromatographic method to quantify sucrose, glucose, and fructose in tubers of Solanumtuberosum Group Phureja. The Journal of Chromatography. 975: 18-23.
  • Gunduz A., Eraydin I., Turkmen S., Kalkan O.F., Turedi T., Eryigit U., Ayar A. 2014. Analgesic effects of mad honey (grayanotoxin) in mice models of acute pain and painful diabetic neuropathy. Human and Experimental Toxicology. 33: 130–135.
  • Holstege D.M., Francis T., Puschner B., Booth M.C., Galey F.D. 2000. Multiresidue screen for cardiotoxins by two-dimensional thin-layer chromatography. Journal of Agricultural and Food Chemistry. 48: 60-64.
  • Holstege D.M., Pushchner B., Le T. 2001. Determination of grayanotoxins in biological samples by LC-MS/MS. Journal of Agricultural and Food Chemistry. 49: 1648-1651.
  • Hough R.L., Crews C., White D., Driffield M., Campbell C.D., Maltin C. 2010. Degradation of yew, ragworth and rhododendron toxins during composting. Science of the Total Environment. 408: 4128-4137.
  • Hwang T., Noh E., Jeong JH., Park S-K., Shin D., Kang H. 2018. Determination of Grayanotoxins from Rhododendron brachycarpum in Dietary Supplements and Homemade Wine by Liquid Chromatography-Quadrupole Time-of-Flight-Mass Spectrometry and Liquid Chromatography-Tandem Mass Spectrometry, J. Agric. Food Chem., 66: 1935-1940.
  • Kaplan M., Olgun E.O., Karaoglu O. 2014. Determination of grayanotoxins in honey by liquid chromatography tandem mass spectrometry using dilute-and-shoot sample preparation approach. J. Agric. Food Chem., 62: 5485-5491.
  • Karkacier M., Erbas M., Uslu M.K., Aksu M. 2003. Comparison of different extraction and detection methods for sugars using amino-bonded phase HPLC. The Journal of Chromatographic Science. 41: 331-333.
  • Kim S.E., Shin M.C., Akaike N., Kim C.J. 2010. Presynaptic effects of grayanotoxin III on excitatory and inhibitory nerve terminals in rat ventromedial hypothalamic neurons. NeuroToxicology. 31: 230–238.
  • Kurtoglu A.B., Yavuz R., Evrendilek G.A. 2014. Characterization and fate of grayanatoxins in mad honey produced from Rhododendron ponticum nectar. Food Chemistry. 161: 47-52.
  • Masutani T., Kawazu K., Uneyama K., Torii S., Iwasa J. 1979. Assignment of 13C-NMR Spectra of Grayanotoxin-I and –III. Agricultural and Biological Chemistry. 43: 631-635.
  • Meguri H., Katakawa J., Tetsumi T., Uchida S., Sumi M., Katai M., Terai T., Fukumato K. 1993. Simple and Rapid Determimation of Grayanotoxins by High−Performance Liquid Chrornatography. The Japanese Society of Pharmacognosy. 47: 429-433.
  • Michie D., Litterick A., Crews C. 2011. The influence of outdoor windrow composting on the concentration of grayanotoxins in Rhododendron leaves. Compost Science & Utilization. 19: 44-51.
  • Milne R., Abbott R.J. 2000. Origin and evolution of invasive naturalized material of Rhododendron ponticum L. in the British Isles. Molecular Ecology. 9: 543-556.Nishida R., Fukami H., Iriye R., Kumazawa Z. 1990. Accumulation of highly toxic Ericaceous diterpenoids by the Geometrid Moth, Arichanna gaschkevitchii. Agricultural and Biological Chemistry, 54: 2347-2352.
  • Niu C.S., Li Y., Liu Y.B., Ma S.G., Li L., Qu J., Yu S.S. 2016. Analgesic diterpenoids from the twigs of Pieris Formosa. Tetrahedron, 72: 44-49.Popescu R., Krupitza G., Kopp B. 2009. Novel insights into the mechanism of action of grayanotoxin III. Planta Medica, 75: 970-970.
  • Popescu R.K. 2013. The genus Rhododendron: An ethnopharmacological and toxicological review. The Journal of Ethnopharmacology. 147: 42-62.
  • Sahin H., Turumtay E.A., Yildiz O., Kolaylı S. 2015. Grayanotoxin-III dedection and antioxidant activity of mad honey. International Journal of Food Properties, 18: 2665-2674.
  • Sakata K., Hattori M., Sakurai A., Hosotsuji, T. (1977). Isolation and identification of biologically active constituents of Leucothoe catesbaei A. Gray. Journal of Pesticide Science, 2: 453-456.
  • Silici S., Atayoglu A.T. 2015. Mad honey intoxication: A systematic review on the 1199 cases. Food and Chemical Toxicology. 86: 282–290.
  • Silici S., Yonar M.E., Huseyin S., Atayoğlu A.T., Ozkok D. 2014. Analysis of grayanotoxin in rhododendron honey and effect on antioxidant parameters. The Journal of Ethnopharmacology. 156: 155-161.
  • Terai T., Araho D., Osakabe K., Katai M., Narama I., Matsuura T., Katakawa J., Tetsumi T. Sato M. 2000. Isolation of iso-grayanotoxin II from leaves of Leucothoe grayana MAX. Its X-Ray crystallographic analysis and acute toxicity in mice. Chemical & Pharmaceutical Bulletin, 48: 142-144.
  • These A., Bodi D., Uecker S., Reimers K, Ronczka S., Weigert A.P., Wiederholt M.L. 2015. A case of human poisoning by grayanotoxins following honey ingestion: elucidation of the toxin profile by mass spectrometry. Food Additives & Contaminants Part A, 32: 1679-1683.
  • Tiedeken E.J., Stout J.C., Stevenson P.C., Wright G.A. 2014. Bumblebees are not deterred by ecologically relevant concentrations of nectar toxins. Journal of Experimental Biology. 217: 1620-1625.
  • Victorita B., Margihitas L.A., Stanciu O., Laslo L., Dezmiream D., Bobis O. 2008. High-performance liquid chromatographic analysis of sugars in Transylvanian honeydew honey. The Bulletin of University of Agricultural Sciences and Veterinary Medicine. 65: 229-232.
  • Wong J., Youde E., Dickinson B., Hale M. 2002. Report of the rhododendron feasibility study. Prepared for the Beddgelert Rhododendron Management Group by the School of Agricultural and Forest Sciences, University of Wales, Bangor, 64-64.
  • Zhang M., Xie Y., Zhan Lei L., Shu P., Chen Y., Xue Y., Luo Z., Wana Q., Yao G., Zhang Y. 2015. Grayanane and leucothane diterpenoids from the leaves of Rhododendron micranthum. Phytochemistry. 117: 107–115.
  • Zhou C., Li X., Li H., Li R. 2012. Chemical constituents from the leaves of Craibiodendron yunnanense. Biochemical Systematics and Ecology, 45: 179–182.
  • Zielinski A.A.F., Braga C.M., Demiate I.M., Beltrame F.L., Nogueira A., Wosiacki G. 2014. Development and optimization of an HPLC-RI method for the determination of major sugars in apple juice and evaluation of the effect of the ripening stage. Food Science and Technology, 408: 4128-4137.
There are 32 citations in total.

Details

Primary Language English
Journal Section Articles
Authors

Perihan Ünak 0000-0002-0172-257X

Talha Sıddık Akkkaya This is me 0000-0001-6201-1459

Publication Date July 25, 2019
Submission Date June 21, 2019
Acceptance Date July 25, 2019
Published in Issue Year 2019 Volume: 1 Issue: 1

Cite

APA Ünak, P., & Akkkaya, T. S. (2019). Determination of Grayanotoxin-III from in Rhodendron Ponticum And Mad Honey Samples by Liquid Chromatography−Mass Spectrometry. Journal of Spectroscopy and Molecular Sciences, 1(1), 1-21.