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Mar. Drugs 2012, 10(6), 1360-1382; doi:10.3390/md10061360
Article

Production and Isolation of Azaspiracid-1 and -2 from Azadinium spinosum Culture in Pilot Scale Photobioreactors

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Received: 15 May 2012; in revised form: 1 June 2012 / Accepted: 4 June 2012 / Published: 13 June 2012
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Abstract: Azaspiracid (AZA) poisoning has been reported following consumption of contaminated shellfish, and is of human health concern. Hence, it is important to have sustainable amounts of the causative toxins available for toxicological studies and for instrument calibration in monitoring programs, without having to rely on natural toxin events. Continuous pilot scale culturing was carried out to evaluate the feasibility of AZA production using Azadinium spinosum cultures. Algae were harvested using tangential flow filtration or continuous centrifugation. AZAs were extracted using solid phase extraction (SPE) procedures, and subsequently purified. When coupling two stirred photobioreactors in series, cell concentrations reached 190,000 and 210,000 cell·mL−1 at steady state in bioreactors 1 and 2, respectively. The AZA cell quota decreased as the dilution rate increased from 0.15 to 0.3 day−1, with optimum toxin production at 0.25 day−1. After optimization, SPE procedures allowed for the recovery of 79 ± 9% of AZAs. The preparative isolation procedure previously developed for shellfish was optimized for algal extracts, such that only four steps were necessary to obtain purified AZA1 and -2. A purification efficiency of more than 70% was achieved, and isolation from 1200 L of culture yielded 9.3 mg of AZA1 and 2.2 mg of AZA2 of >95% purity. This work demonstrated the feasibility of sustainably producing AZA1 and -2 from A. spinosum cultures.
Keywords: solid phase extraction; photobioreactor; chemostat; dinoflagellate; micro-algae; LC-MS/MS; tangential flow filtration; azaspiracid; HP-20 solid phase extraction; photobioreactor; chemostat; dinoflagellate; micro-algae; LC-MS/MS; tangential flow filtration; azaspiracid; HP-20
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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MDPI and ACS Style

Jauffrais, T.; Kilcoyne, J.; Séchet, V.; Herrenknecht, C.; Truquet, P.; Hervé, F.; Bérard, J.B.; Nulty, C.; Taylor, S.; Tillmann, U.; Miles, C.O.; Hess, P. Production and Isolation of Azaspiracid-1 and -2 from Azadinium spinosum Culture in Pilot Scale Photobioreactors. Mar. Drugs 2012, 10, 1360-1382.

AMA Style

Jauffrais T, Kilcoyne J, Séchet V, Herrenknecht C, Truquet P, Hervé F, Bérard JB, Nulty C, Taylor S, Tillmann U, Miles CO, Hess P. Production and Isolation of Azaspiracid-1 and -2 from Azadinium spinosum Culture in Pilot Scale Photobioreactors. Marine Drugs. 2012; 10(6):1360-1382.

Chicago/Turabian Style

Jauffrais, Thierry; Kilcoyne, Jane; Séchet, Véronique; Herrenknecht, Christine; Truquet, Philippe; Hervé, Fabienne; Bérard, Jean Baptiste; Nulty, Cíara; Taylor, Sarah; Tillmann, Urban; Miles, Christopher O.; Hess, Philipp. 2012. "Production and Isolation of Azaspiracid-1 and -2 from Azadinium spinosum Culture in Pilot Scale Photobioreactors." Mar. Drugs 10, no. 6: 1360-1382.


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