Evaluation of extraction methods for analysis of domoic acid in naturally contaminated shellfish from Portugal |
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| Institution: | 1. University of Vigo, Department of Analytical and Food Chemistry, Campus Universitario de Vigo, 36310 Vigo, Spain;2. European Union Reference Laboratory for Marine Biotoxins, CITEXVI, Campus Universitario de Vigo, 36310 Vigo, Spain;3. Japan Food Research Laboratories, 6-11-10 Nagayama, Tama, Tokyo 206-0025, Japan;4. University of Texas at Austin Marine Science Institute, Port Aransas, TX 78373, United States;1. Centre for Ocean Life, National Institute of Aquatic Resources, Technical University of Denmark, DK-2920 Charlottenlund, Denmark;2. Marine Biological Section, University of Copenhagen, Strandpromenaden 5, DK-3000 Helsingør, Denmark;3. Alfred-Wegener-Institute for Polar and Marine Research, Am Handelshafen 12, D-27570 Bremerhaven, Germany;1. Marine Biotoxins Program, Center for Coastal Environmental Health and Biomolecular Research, NOAA-National Ocean Service, Charleston, SC 29412, USA;2. UMR 241-EIO, Laboratoire des microalgues toxiques, Institut Louis Malardé, BP30, 98713 Papeete Tahiti, French Polynesia;1. Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2. Laboratory of Marine Ecology and Environmental Science, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China;3. University of Chinese Academy of Sciences, Beijing 100049, China;4. Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao 266071, China;1. Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;2. Carbon-sink Fisheries Laboratory, Qingdao 266071, China;3. National Center for Quality Supervision and Test of Aquatic Products, Qingdao 266071, China;4. AB SCIEX, Analytical Instrument Trading Co., Shanghai 200233, China |
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| Abstract: | Domoic acid (DA) is a marine neurotoxin that is somewhat unstable, particularly in acidic media. Several protocols were used to extract DA from naturally contaminated tissues of shellfish harvested in Portugal. A modified version of AOAC method no. 991.26, with a simplified 10 g extraction, was used and compared with an aqueous 50% methanol extraction. Mean recoveries were between 81 and 85% when extracts were analysed by LC on the same day of extraction. When acid extracts were frozen for 1 or 2 days recoveries lowered to 72%, and if injection was repeated on the following 3rd or 4th days only 57–65% was recovered. Relative standard deviation of recovery for these miscellaneous samples, which was between 10 and 13% on the day of extraction, increased approximately 10% each day the extract was reanalysed. On our regulatory monitoring work, we employ an aqueous 80% methanol extraction that is common with the lipid-soluble DSP toxins. We report here a mean recovery of 90±6% for this methodology. Our long-term stability studies of domoic acid in shellfish extracts showed that slow decomposition of this compound occur in filtered aqueous methanol extracts. Additionally, we also found that in frozen tissues slow decomposition is clearly observable over a time span of 1 month. |
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