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Mar. Drugs 2011, 9(3), 466-477; doi:10.3390/md9030466
Article

Preparation of Calibration Standards of N1-H Paralytic Shellfish Toxin Analogues by Large-Scale Culture of Cyanobacterium Anabaena circinalis (TA04)

1,* , 1
 and
2
1 National Research Institute of Fisheries Science, 2-12-4 Fukuura, Kanazawa-ku, Yokohama 236-8648, Japan 2 School of Marine Biosciences, Kitasato University, 160-4 Okirai-uto, Sanriku-cho, Ofunato 022-0101, Japan
* Author to whom correspondence should be addressed.
Received: 9 February 2011 / Revised: 3 March 2011 / Accepted: 18 March 2011 / Published: 22 March 2011
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Abstract

Mouse bioassay is the official testing method to quantify paralytic shellfish toxins (PSTs) in bivalves. A number of alternative analytical methods have been reported. Some methods have been evaluated by a single laboratory validation. Among the different types of methods, chemical analyses are capable of identifying and quantifying the toxins, however a shortage of the necessary calibration standards hampers implementation of the chemical analyses in routine monitoring of PSTs in bivalves. In our present study, we studied preparation of major PST analogues as calibrants by large-scale cultivation of toxic freshwater cyanobacteria Anabaena circinalis TA04. The cells were steadily grown in 10 L bottle for 28 days. The primary N1-H toxins, C1/C2, were produced at a concentration of 1.3 ± 0.1 µmol/L. The intracellular and extracellular toxins occupied 80% and 20%, respectively. Over 220 µmol of the toxins was obtained from approximately 200 L of the culture over six months, demonstrating that it is sufficient to prepare saxitoxin analogues. The toxins were chemically converted to six N1-H analogues. Preparation of the analogues was carried out at relatively high yields (50–90%). The results indicate that our preparation method is useful to produce N1-H toxins. In our present study, detailed conditions for preparation of one of the rare N1-H analogues, gonyautoxin-5, were investigated.
Keywords: cyanobacteria; Anabaena circinalis; paralytic shellfish toxin; saxitoxin; chemical derivatization; standard cyanobacteria; Anabaena circinalis; paralytic shellfish toxin; saxitoxin; chemical derivatization; standard
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Watanabe, R.; Suzuki, T.; Oshima, Y. Preparation of Calibration Standards of N1-H Paralytic Shellfish Toxin Analogues by Large-Scale Culture of Cyanobacterium Anabaena circinalis (TA04). Mar. Drugs 2011, 9, 466-477.

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