Determination and Monitoring of Grayanotoxins in Honey Using LC-MS/MS

LC-MS/MS를 이용한 벌꿀 중 grayanotoxin 분석법 연구 및 실태조사

  • Lee, Sook-Yeon (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Choi, Youn-Ju (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Lee, Kang-Bong (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Cho, Tae-Yong (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Kim, Jin-Sook (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Son, Young-Wook (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Park, Jae-Seok (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Im, Sung-Im (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Choi, Hee-Jung (Food & Risk Standardization Team, Korean Food and Drug Administration) ;
  • Lee, Dong-Ha (Food & Risk Standardization Team, Korean Food and Drug Administration)
  • 이숙연 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 최윤주 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 이강봉 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 조태용 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 김진숙 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 손영욱 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 박재석 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 임성임 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 최희정 (식품의약품안전청 유해물질관리단 위해기준팀) ;
  • 이동하 (식품의약품안전청 유해물질관리단 위해기준팀)
  • Published : 2008.02.28

Abstract

This study was performed to establish analysis methods, and evaluated for grayanotoxin in domestic/foreign honey and wild honey. The molecular weight of grayanotoxins I, II and III, excluding grayanotoxin III that has been commercialized, were analyzed by LC-MS/MS. Then, the molecular structure of grayanotoxins I and II were analyzed by NMR. A total 111 samples (25 Korean honey, 21 Korean wild honey, 13 Korean honeycomb honey, 44 foreign honey, 8 foreign wild honey) were examined to determined whether or not each sample contained grayanotoxins I, II, and III. The honey samples were mixed with methanol and loaded into a tC18 cartridge, the filtrate was diluted with water, and the mixture was then analyzed by ESI triple-quadrupole LC-MS/MS. Grayanotoxins were only found in the foreign wild honey and were not detected in Korean honey, Korean honeycomb honey, or Korean wild honey. Three of the samples contained grayanotoxin I, II, and III, and one sample contained only grayanotoxins I and III. The lowest level for grayanotoxin I was 3.13 ${\pm}$ 0.00 mg/kg, and the highest level was 12.93 ${\pm}$ 0.01 mg/kg. The levels of grayanotoxin II were 0.84 ${\pm}$ 0.01 mg/kg, 0.92 ${\pm}$ 0.00 mg/kg and 1.08 ${\pm}$ 0.01 mg/kg, respectively. The lowest level of grayanotoxin III was 0.25 ${\pm}$ 0.01 mg/kg and the highest level was 3.29 ${\pm}$ 0.74 mg/kg. Through this study, safety management for foreign wild honey has been enabled.

Keywords

grayanotoxin;wild honey;honey;LC-MS/MS;NMR

References

  1. Sutlupinar N, Mat A, Akkus I, Erkul I. Poisoning by toxic honey in Turkey. Arch. Toxicol. 67: 148-150 (1993) https://doi.org/10.1007/BF01973687
  2. Bruneton J. Toxic plants: Dangerous to Humans and Animals. Intercept Ltd., Hampshire, UK. 250-257: 250-257 (1999)
  3. Gossinger H, Hruby K, Haubenstock A. Cardiac arrhythmias in a patient with grayanotoxin-honey poisoning. Vet. Hum. Toxicol. 25: 328-329 (1983)
  4. Kinghorn AD, Jawad FH, Doorenbons NJ. Thin-layer chromatographic and spectroscopic determination of some diterpenes of the grayanotoxin type. J. Chromatogr. 147: 229-308 (1978)
  5. Terai T, Katai M. Meguri H. Quantitative analysis of grayanotoxins. I. Spectrometric determination of grayanotoxin I, II, and III. Yakuga. Zasshi 103: 686-689 (1983) https://doi.org/10.1248/yakushi1947.103.6_686
  6. Terai T, Uda T, Katakawa J. Tetsumi T. Assay of Grayanotoxins by gas chromatography. Nippon Nogeik. Kaishi 68: 979-981 (1994) https://doi.org/10.1271/nogeikagaku1924.68.979
  7. Choi JS, Young HS, Park JC, Choi JH. Woo WS. Further study on the constituents of Rhododendron brachycarpum. Arch. Pharm. Res. 10: 169-172 (1987) https://doi.org/10.1007/BF02861908
  8. von Malottki K. Wiechmann HW. Acute life-threatening bradycardia : Food poisoning by Turkish wild honey. Deut. Med. Wochenschr. 121: 936-938 (1996) https://doi.org/10.1055/s-2008-1043090
  9. Kim AJ, Kim JS, Shin DW, Baek KJ, Han SB, Lee YJ. Grayanotoxin Intoxication. J. Korean Soc. Emerg. Med. 11: 373-377 (1987)
  10. Plugge PC, de Zaayer HG. Andromedotoxin. Am. J. Pharm. 61: 6-8 (1889)
  11. Frohne D, Pfander HJ. A Colour Atlas of Poisonous Plants. Wolfe Publishing Ltd., London, England. 172-182 (1983)
  12. Kim HK, Jeong TO, Jin YH, Lee JB. Korean rosebay intoxication. J. Korean Soc. Clin. Toxicol. 3: 52-55 (2005)
  13. Dilber E, Kalyoucu M, Yaris N. Okten A. A case of mad honey poisoning presenting with convulsion: Intoxication instead of alternative therapy. Turkish. J. Med. Sci. 32: 361-362 (2002)
  14. Kerkvliet JD. Analysis of a toxic rhododendron honey. J. Apicult. Res. 20: 249-253 (1981) https://doi.org/10.1080/00218839.1981.11100505
  15. Sohn CH, Kim W, Ahn S, Oh BJ, Kim WY, Lim KS. Three cases of Mad-honey poisoning presenting with cardiovascular emergencies. J. Korean. Soc. Emerg. Med. 16: 322-325 (2005)
  16. Holstege DM, Puschner B. Le, T. Determination of grayanotoxins in biological samples by LC-MS/MS. J. Agric. Food Chem. 49: 1648-1651 (2001) https://doi.org/10.1021/jf000750s
  17. Terai T. Determination of grayanotoxins I, II, and III by thinlayer chromatography. Nippon Nogeik. Kaishi 58: 1117-1122 (1984) https://doi.org/10.1271/nogeikagaku1924.58.1117
  18. FDA/CFSAN. Bad Bug Book. Foodborne Pathogenic Microorganisms Natural Toxins Handbook. International Medical Publishing Inc., McLean, VA, USA. pp. 119-120 (2004)
  19. Kaiya T, Sakakibara J. Diterpenoids from enicaceous plants. Annual Report of the Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan, 30: 1-34 (1982)
  20. Sutlupinar N, Mat A, Satganoglu Y. Poisoning by toxic honey in Turkey. Arch. Toxicol. 67: 148-150 (1993) https://doi.org/10.1007/BF01973687