Evaluation of the Possibility That Free Fatty Acids Cause False-Positive Result in Diarrhetic Shellfish Poisoning Mouse Bioassay in Actual Use

Midgut glands of bivalves are used for the mouse bioassay of diarrhetic shellfish poison (DSP). A large quantity of free fatty acids (FFAs) causes a false positive outcome in the assay. To examine whether this is likely to occus under conditions of actual use, we analyzed the contents of the FFAs in the enlarged midgut glands during gametogenesis of Japanese scallops Patinopecten yessoensis that had been caught at two points in Hunka Bay on March 27, 2006, because the content of FFAs may increase with activation of lipogenesis for gametogenesis. Fatty acids (FAs) were measured with fluorometric high-performance liquid chromatography after derivatization with 9-anthryldiazomethane. The total FFAs (14 : 0, 16 : 0, 18 : 0, 16 : 1, 18 : 1, 18 : 4, 20 : 5 and 22 : 6) represented 3.3-4.2 wt% of the lipid. The toxic FFAs accounted for 40-43 wt% of the total FFAs. Content of each FFA (18 : 1, 2.7-5.0 mg/g lipid; 18 : 4, tr.-2.0 mg/g; 20 : 4, n.d.; 20 : 5, 8.0-9.1 mg/g and 22 : 6, 2.0-2.1 mg/g) was lower than the lethal dose tentatively calculated from the relative toxicity. It appears that the likelihood of FFAs causing false-positives in the mouse bioassay is low if the sample is fresh and is extracted immediately after homogenizing.