Selective Extraction and Purification of Azaspiracids from Blue Mussels ( Mytilus edulis) Using Boric Acid Gel.

Azaspiracids belong to a family of more than 50 polyether toxins originating from marine dinoflagellates such as Azadinium spinosum. All of the azaspiracids reported thus far contain a 21,22-dihydroxy group. Boric acid gel can bind selectively to compounds containing vic-diols or α-hydroxycarboxylic acids via formation of reversible boronate complexes. Here we report use of the gel to selectively capture and release azaspiracids from extracts of blue mussels. Analysis of the extracts and fractions by liquid chromatography-tandem mass spectrometry (LC-MS) showed that this procedure resulted in an excellent cleanup of the azaspiracids in the extract. Analysis by enzyme-linked immunoasorbent assay (ELISA) and LC-MS indicated that most azaspiracid analogues were recovered in good yield by this procedure. The capacity of boric acid gel for azaspiracids was at least 50 µg/g, making this procedure suitable for use in the early stages of preparative purification of azaspiracids. In addition to its potential for concentration of dilute samples, the extensive cleanup provided by boric acid gel fractionation of azaspiracids in mussel samples almost eliminated matrix effects during subsequent LC-MS and could be expected to reduce matrix effects during ELISA analysis. The method may therefore prove useful for quantitative analysis of azaspiracids as part of monitoring programs. Although LC-MS data showed that okadaic acid analogues also bound to the gel, this was much less efficient than for azaspiracids under the conditions used. The boric acid gel methodology is potentially applicable to other important groups of natural toxins containing diols including ciguatoxins, palytoxins, pectenotoxins, tetrodotoxin, trichothecenes, and toxin glycosides.

Liver concentrations of copper, cobalt, and selenium in wild Norwegian red deer (Cervus elaphus).

Liver samples from 245 wild red deer (Cervus elaphus) collected during the licensed hunting season in 2001 from five different locations in western Norway were analyzed for copper (Cu), cobalt (Co), and selenium (Se). The associations between these trace elements and geographical location, age group, and sex were studied. The median (and range of) liver concentrations (microg/g wet weight) for all the examined deer were: Cu 20 (1.7-103), Co 0.08 (<0.01-0.18), and Se 0.09 (0.04-1.0). The results indicate a generally low status of Cu and Se. In total, 15 (6%) red deer had deficient Cu levels (< 4 (microg/g). For all three elements, the liver concentrations showed a significant geographic variation. The geographic difference was most distinct for Cu. The lowest median Cu concentration was found in deer from the island Hitra, where 13% of the animals had deficient Cu levels. Significant differences between age groups were found for all elements, and generally, the adults (> or =2.5 yr) had the highest levels. No significant sex differences within the various age groups were found, with three exceptions: female calves and adults had significantly higher Co levels than male deer, and adult males had significantly higher Se levels than adult females. The Cu and Se status of wild red deer in parts of Norway is low; however, the significance of this needs to be explored further.