Suitability of the MTT-based cytotoxicity assay to detect okadaic acid contamination of mussels

Toxicon. 1996 Sep;34(9):965-74. doi: 10.1016/0041-0101(96)00073-6.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bivalvia / metabolism*
  • Colorimetry
  • Coloring Agents / chemistry
  • Coloring Agents / metabolism*
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Injections, Intraperitoneal
  • KB Cells / cytology
  • KB Cells / drug effects
  • Lethal Dose 50
  • Mice
  • Okadaic Acid / analysis
  • Okadaic Acid / metabolism*
  • Okadaic Acid / toxicity
  • Oxidation-Reduction
  • Reproducibility of Results
  • Shellfish Poisoning*
  • Tetrazolium Salts / chemistry
  • Tetrazolium Salts / metabolism*
  • Thiazoles / chemistry
  • Thiazoles / metabolism*
  • Tumor Cells, Cultured

Substances

  • Coloring Agents
  • Tetrazolium Salts
  • Thiazoles
  • Okadaic Acid
  • thiazolyl blue