Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
권호 표지
DOI QR코드

DOI QR Code

Determination of Domoic Acid in Seafood Matrices using HPLC-UV with Solid Phase Extraction Cleanup

고체상 추출 전처리 및 HPLC-UV를 이용한 수산물 중 domoic acid의 분석

  • Si Eun Kim (School of Food Science and Technology. Chung-Ang University) ;
  • Sang Yoo Lee (School of Food Science and Technology. Chung-Ang University) ;
  • Ji Eun Park (School of Food Science and Technology. Chung-Ang University) ;
  • Hyunjin Jung (School of Food Science and Technology. Chung-Ang University) ;
  • Hyang Sook Chun (School of Food Science and Technology. Chung-Ang University)
  • 김시은 (중앙대학교 생명공학대학 식품공학과) ;
  • 이상유 (중앙대학교 생명공학대학 식품공학과) ;
  • 박지은 (중앙대학교 생명공학대학 식품공학과) ;
  • 정현진 (중앙대학교 생명공학대학 식품공학과) ;
  • 전향숙 (중앙대학교 생명공학대학 식품공학과)
  • Received : 2023.10.09
  • Accepted : 2023.10.18
  • Published : 2023.10.30
Download PDF
⟨ Previous Next ⟩

Abstract

Domoic acid (DA), a neurotoxin produced naturally by diatoms, is responsible for incidents of amnesic shellfish poisoning. In this study, a modified analytical method was established to determine domoic acid in seafood using solid phase extraction cleanup and optimizing the amount of sample and extraction solvent to reduce interference effects. The modified method using high-performance liquid chromatography with ultraviolet detection was validated using three seafood matrices (mussel, red snow crab, and anchovy) at three concentrations (1, 2, and 4 mg/kg) and compared to the Food Code method. Compared to the Food Code method, the modified method showed better performance in terms of linearity (R2>0.999), detection limit (0.02-0.03 mg/kg), quantification limit (0.05-0.09 mg/kg), intra-/inter-day accuracy (86.2-100.4%), and intra-/inter-day precision (0.2-4.0%). Furthermore, the method was successfully applied for the analysis of 87 seafood samples marketed in Korea, and DA was detected at a low concentration of 140 ㎍/kg in one anchovy sample. These results suggest that the modified method can be used for routine determination of DA in seafood.

본 연구는 패류 독소 중 기억상실성 독성을 유발하는 domoic acid의 분석법 개선, 검증 및 분석적용성을 살펴보았다. SAX 카트리지 정제, 검체 및 추출용매 양의 변경과 더불어 이동상을 용매구매 조건으로 변경시킨 분석법은 지중해담치, 홍게 그리고 멸치의 세 가지 매트릭스를 대상으로 세 농도에 대하여 식품공전법과 비교하여 유효성을 검증하였다. 그 결과 변경된 분석법은 LOD 0.02-0.03 mg/kg, LOQ 0.05-0.09 mg/kg, 일내 및 일간 정확도 86.2-100.4%와 일내 및 일간 정밀도 0.2-4.0%로 CODEX가이드라인을 만족하는 우수한 분석능을 나타내었다. 특히 변경된 시험법에서는 domoic acid와 유사한 머무름 시간을 갖는 방해물질이 검출되지 않아 위양성 결과를 방지할 수 있을 것으로 생각된다. 더 나아가 본 분석법이 국내 유통 중인 수산물을 대상으로 적용될 수 있는지 확인하고자 식품공전의 기억상실성 패독 분석법과 함께 수산물에 적용하여 분석하였다. 그 결과 5종의 수산물 87건 중 공전시험법으로 분석했을 때 domoic acid가 검출된 시료는 없었으나 변경된 분석법을 통하여 멸치 1건에 대하여 0.14 mg/kg의 domoic acid이 미량 검출되었다. 따라서 본 연구에서 확립된 분석법은 수산물 중 domoic acid 분석에 활용이 가능할 것으로 보인다.

Keywords

Acknowledgement

본 연구는 2023년도 식품의약품안전처의 연구개발비(20163MFDS641, 21153MFDS605)로 수행되었으며 이에 감사드립니다.

References

  1. Lefebvre, K.A., Robertson, A., Domoic acid and human exposure risks: a review. Toxicon., 56, 218-230 (2010).  https://doi.org/10.1016/j.toxicon.2009.05.034
  2. Zabaglo, K., Chrapusta, E., Bober, B., Kaminski, A., Adamski, M., Bialczyk, J., Environmental roles and biological activity of domoic acid: A review. Algal Res., 13, 94-101 (2016).  https://doi.org/10.1016/j.algal.2015.11.020
  3. Ben-Gigirey, B., Solino, L., Bravo, I., Rodriguez, F., Casero, M. V., Paralytic and amnesic shellfish toxins impacts on seabirds, analyses and management. Toxins., 13, 454 (2021). 
  4. Kershaw, J.L., Jensen, S.K., McConnell, B., Fraser, S., Cummings, C., Lacaze, J.P., Hermann, G., Bresnan, E., Dean, K. J., Turner, A.D., Davidson, K., Hall, A.J., Toxins from harmful algae in fish from Scottish coastal waters. Harmful Algae., 105, 102068 (2021). 
  5. Gibble, C.M., Kudela, R. M., Knowles, S., Bodenstein, B., Lefebvre, K. A., Domoic acid and saxitoxin in seabirds in the United States between 2007 and 2018. Harmful Algae., 103, 101981 (2021). 
  6. Jeffery, B., Barlow, T., Moizer, K., Paul, S., Boyle, C., Amnesic shellfish poison. Food Chem. Toxicol., 42, 545-557 (2004).  https://doi.org/10.1016/j.fct.2003.11.010
  7. Wright, J.L.C., Boyd, R.K., Freitas, A.D., Falk, M., Foxall, R.A., Jamieson, W.D., Laycock, M.V., McCulloch, A.W., Mclnnes, A.G., Odense, P., Pathak, V.P., Quilliam, M.A., Ragan, M.A., Sim, P.G., Thibault, P., Walter, J.A., Gilgan, M., Richard, D.J.A., Dewar, D., Identification of domoic acid, a neuroexcitatory amino acid, in toxic mussels from eastern Prince Edward Island. Can. J. Chem., 67, 481-490 (1989).  https://doi.org/10.1139/v89-075
  8. Choi, K.D., Lee, J.S., Lee, J.O., Oh, K.S., Shin, I.S., Investigation of domoic acid in shellfish collected from Korean fish retail outlets. Food Sci Biotechnol., 18, 842-848 (2009). 
  9. Petroff, R., Hendrix, A., Shum, S., Grant, K.S., Lefebvre, K.A., Burbacher, T.M., Public health risks associated with chronic, low-level domoic acid exposure: A review of the evidence. Pharmacol. Ther., 227, 107865 (2021). 
  10. AOAC., AOAC Official Method 959.08. Paralytic shellfish poison, biological method. Official methods of analysis of AOAC International, AOAC International, Gaithersburg, Maryland, USA (2000). 
  11. AOAC., AOAC Official Method 991.26. Domoic acid in mussels, liquid chromatographic method. Official methods of analysis of AOAC International. AOAC International, Gaithersburg, Maryland, USA (2000). 
  12. AOAC., AOAC Official Method 2006.02. Domoic acid toxins in shellfish. Biosense ASP. Official methods of analysis of AOAC International. AOAC International, Gaithersburg, Maryland, USA (2006). 
  13. Furey, A., Lehane, M., Gillman, M., Fernandez-Puente, P., James, K.J., Determination of domoic acid in shellfish by liquid chromatography with electrospray ionization and multiple tandem mass spectrometry. J. Chromatogr. A., 938, 167-174 (2001).  https://doi.org/10.1016/S0021-9673(01)01385-1
  14. KFDA., Korean Food Standard. Chapter 8. Korea Food and Drug Administration, Cheongju, Korea (2023). 
  15. Hess, P., Gallacher, S., Bates, L.A., Brown, N., Quilliam, M. A., Determination and confirmation of the amnesic shellfish poisoning toxin, domoic acid, in shellfish from Scotland by liquid chromatography and mass spectrometry. J. AOAC Int., 84, 1657-1667 (2001).  https://doi.org/10.1093/jaoac/84.5.1657
  16. de Souza, M.M.M., da Boa Morte, E.S., Cardoso, L.G., Nunes, D.V., de Souza, C.O., Druzian, J.I., Cardoso, R.D.C.V., Nutritional contribution of shellfish from the biodiversity of Todos os Santos Bay, Brazil. J. Food Compos. Anal., 102, 103999 (2021). 
  17. Wright, A.C., Fan, Y., Baker, G.L., Nutritional value and food safety of bivalve molluscan shellfish. J. Shellfish Res., 37, 695-708 (2018).  https://doi.org/10.2983/035.037.0403
  18. Kari, N.M., Ahmad, F., Ayub, M.N.A., Proximate composition, amino acid composition and food product application of anchovy: a review. Food Res., 6, 16-29 (2022).  https://doi.org/10.26656/fr.2017.6(4).419
  19. Lopez-Rivera, A., Suarez-Isla, B.A., Eilers, P.P., Beaudry, C.G., Hall, S., Fernandez Amandi, M., Furey, A., James, K. J., Improved high-performance liquid chromatographic method for the determination of domoic acid and analogues in shellfish: effect of pH. Anal. Bioanal. Chem., 381, 1540-1545 (2005).  https://doi.org/10.1007/s00216-005-3109-4
  20. Maroulis, M., Monemvasios, I., Vardaka, E., Rigas, P., Determination of domoic acid in mussels by HPLC with post-column derivatization using 4-chloro-7-nitrobenzo-2-oxa-1, 3-diazole (NBD-Cl) and fluorescence detection. J. Chromaatogr. B., 876, 245-251 (2008).  https://doi.org/10.1016/j.jchromb.2008.10.053
  21. Quilliam, M.A., Sim, P.G., McCulloch, A.W., McInnes, A.G., High-performance liquid chromatography of domoic acid, a marine neurotoxin, with application to shellfish and plankton. Int. J. Environ. Anal. Chem., 36, 139-154 (1989).  https://doi.org/10.1080/03067318908026867
  22. Kvrgic, K., Lesic, T., Dzafic, N., Pleadin, J., Occurrence and Seasonal Monitoring of Domoic Acid in Three Shellfish Species from the Northern Adriatic Sea. Toxins., 14, 33 (2022). 
  23. Codex Alimentarius Commission, Standard for Live and Raw Bivalve Molluscs CXS 292-2008. Rome, Italy, 1-9 (2015). 
  24. EURLMB., EU-Harmonised Standard Operating Procedure for determination of domoic acid in shellfish and finfish by RP-HPLC using UV detection. Vigo, Spain, 1-12 (2008). 
  25. Ujevic, I., Roje-Busatto, R., Ezgeta-Balic, D., Comparison of amnesic, paralytic and lipophilic toxins profiles in cockle (Acanthocardia tuberculata) and smooth clam (Callista chione) from the central Adriatic Sea (Croatia). Toxicon., 159, 32-37 (2019).  https://doi.org/10.1016/j.toxicon.2018.12.008
  26. Quilliam, M.A., Xie, M., Hardstaff, W.R., Rapid extraction and cleanup for liquid chromatographic determination of domoic acid in unsalted seafood. J. AOAC Int., 78, 543-554 (1995).  https://doi.org/10.1093/jaoac/78.2.543
  27. EFSA., Marine biotoxins in shellfish - Domoic acid. EFSA J., 1181, 1-61 (2009). 
  28. Zheng, G., Wu, H., Che, H., Li, X., Zhang, Z., Peng, J., Guo, M., Tan, Z., Residue analysis and assessment of the risk of dietary exposure to domoic acid in shellfish from the coastal areas of China. Toxins., 14, 862 (2022). 
  29. Lim, W.A., Park, K.W., Suh, Y.S., Seasonal changes in phytoplankton composition in Jinhae Bay, 2011. J. Korean Soc. Mar. Environ. Saf., 18, 520-529 (2012). https://doi.org/10.7837/kosomes.2012.18.6.520