Suitability of the MTT-based cytotoxicity assay to detect okadaic acid contamination of mussels.
The suitability of a cytotoxicity assay based on the MTT colorimetric method has been evaluated for the detection of okadaic acid in mussels. On KB cells, okadaic acid exhibited a dose-dependent cytotoxic effect, the IC50 being inversely related to the exposure time (IC50 = 6.3 ng/ml, 4.0 ng/ml and 1.1 ng/ml after 24, 48 and 72 hr of contact, respectively). Using a contact time of 24 hr, the MTT cytotoxicity assay is suitable for revealing okadaic acid concentrations in mussel samples as low as 50 ng/g of digestive glands, with a sensitivity higher than that of the commercially available kits for enzyme-linked immunosorbent assay (ELISA). In the okadaic acid concentration range from 50 to 1500 ng/g of digestive glands the MTT cytotoxicity assay showed satisfactory accuracy and reproducibility. A high degree of correlation was found between the okadaic acid content of 16 naturally contaminated samples measured by the MTT cytotoxicity assay and by an ELISA.
Dipartimento di Scienze Biomediche, Università di Trieste, Italy.
A
A
Tubaro
C
C
Florio
E
E
Luxich
R
R
Vertua
R
R
Della Loggia
T
T
Yasumoto
D000818:Animals; D049872:Bivalvia / Q000378:metabolism*; D003124:Colorimetry; D004396:Coloring Agents / Q000737:chemistry / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007274:Injections, Intraperitoneal; D007624:KB Cells / Q000166:cytology / Q000187:drug effects; D007928:Lethal Dose 50; D051379:Mice; D019319:Okadaic Acid / Q000032:analysis / Q000378:metabolism* / Q000633:toxicity; D010084:Oxidation-Reduction; D015203:Reproducibility of Results; D057096:Shellfish Poisoning; D013778:Tetrazolium Salts / Q000737:chemistry / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D019319:Okadaic Acid; C022616:thiazolyl blue