PARENT SESSION

3. Algal Toxins
Hall 6
8:30 AM - 5:30 PM, Wednesday, 30 April 2003
Chair: Pflugmacher, S.¹, ¹
Co-chair: Wiegand, C.², ²

(WE4/7) Use of commercial ELISA plate kits for analysis of total hepatotoxin concentrations.

Kankanpää , Harri ¹, ¹ Finnish Institute of Marine Research, Helsinki, Finland

ABSTRACT- Enzyme-linked immunosorbent assay (ELISA) kits have been originally and primarily designed for analysis of cyanobacterial hepatotoxins in water samples. However, these kits are also well suited for the analysis of total hepatotoxin burden in tissues. The main advantages in use of ELISA include 1) little or no need for sample pre-treatment, 2) straightforward and rapid analysis and 3) low detection limits. The main disadvantages are cost (approx. 10 EUR/sample) and lack of specific information on toxins detected. The procedure in use allows detection limits of 1-20 g/kg dry tissue, depending on tissue type. We have employed the ELISA kits in the analysis of flounder, salmon, trout, herring, three-spine stickleback, blue mussel, Macoma balthica clams, crayfish and prawns (Penaeus monodon). Tissues from 5-10 specimens have been pooled and extracted simply using methanol/ultrasonication and extracts analysed in triplicate. Typically, the following concentration ranges have been found in Baltic Sea organisms: salmon (liver) <1 to 5 g/kg; herring (liver): <1 to 7 g/kg; flounder (liver) 140 to 410 g/kg; sticklebacks (whole) 35-170 g/kg; blue mussel <20 to 2150 g/kg and clams 100-130 g/kg. With field samples, the precision of tissue analysis (based on an extract analysed in triplicate) is 0.5-10%. In addition to field samples, the ELISA method has been used successfully in analysis of tissues of animals exposed to nodularin in laboratory conditions. The presence of hepatotoxins in Baltic Sea organisms has been confirmed by HPLC/MS, which indicated that the concentration of total extractable hepatotoxin in our Baltic Sea samples originates from nodularin only. The combined use of ELISA and chromatographic methods has indicated that bioaccumulation of nodularin in at least mussels and flounder is a constant phenomenon in the Baltic Sea.

Key words: ELISA, hepatotoxin , Baltic Sea