The validity of two HPLC methods and a colorimetric PP2A assay related to the mouse bioassay in quantification of diarrhetic toxins in blue mussels (Mytilus edulis).
Validity of two HPLC methods and a PP2A assay in relation to the mouse bioassay for diarrhetic shellfish poisoning (DSP) toxins was evaluated. The mouse bioassay for DSP toxins was performed on a total of 177 mussel samples from the Sognefjord, Norway, using diethyl ether in the final step of extraction. For fluorimetric HPLC analyses, either 4-bromomethyl-7-methoxycoumarin (BrMMC) or 9-anthryl diazomethane (ADAM) were used for analysis of 48 and 118 of the samples, respectively. The colorimetric PP2A inhibition assay was performed on all 177 samples that were analysed with the mouse bioassay. When comparing the HPLC-BrMMC, the HPLC-ADAM and the PP2A assays with the mouse bioassay, cut off values of < or =4, 5 and 6 microg okadaic acid (OA) equivalents (eq.)/5 g digestive gland (DG) was used. With reference to the results from the mouse bioassay, the total number of failure and correct classification by HPLC-ADAM and the PP2A method was compared for the three cut off values. No significant differences between the methods were detected. However, all differences were found in favour of HPLC-ADAM. All three methods could replace the mouse bioassay in detecting levels of diarrhetic toxins approved internationally for safe consumption of mussels. However, HPLC-ADAM seems to be the method of choice.
Ramstad
H
H
Department of Pharmacology, Microbiology and Food Hygiene, Norwegian School of Veterinary Science, PO Box 8146, Oslo, Norway. hmramstad@hotmail.com
Shen
J L
JL
Larsen
S
S
Aune
T
T
D000818:Animals; D001681:Biological Assay; D049872:Bivalvia / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry; D003967:Diarrhea / Q000139:chemically induced*; D005260:Female; D051379:Mice; D010749:Phosphoprotein Phosphatases / Q000032:analysis* / Q000037:antagonists & inhibitors; D015203:Reproducibility of Results; D014118:Toxins, Biological / Q000032:analysis*
D014118:Toxins, Biological; D010749:Phosphoprotein Phosphatases