Specific and dynamic detection of palytoxins by in vitro microplate assay with human neuroblastoma cells.
Palytoxin is one of the most complex and biggest molecules known to show extreme acute toxicity. The dinoflagellate Ostreopsis spp., the producer organism of palytoxin, has been shown to be distributed worldwide, thus making palytoxin an emerging toxin. Rat-derived hepatocytes (Clone 9) and BE (2)-M17 human neuroblastoma cells were used to test palytoxin or palytoxin-like compounds by measuring the cell metabolic rate with Alamar Blue. The dose-dependent decrease in viability was specifically inhibited by ouabain in the case of BE (2)-M17 neuroblastoma cells. This is a functional, dynamic and simple test for palytoxins with high sensitivity (as low as 0.2 ng/ml). This method was useful for toxin detection in Ostreopsis extracts and naturally contaminated mussel samples. A comparative study testing toxic mussel extracts by LC (liquid chromatography)-MS/MS (tandem MS), MBA (mouse bioassay), haemolysis neutralization assay and a cytotoxicity test indicated that our method is suitable for the routine determination and monitoring of palytoxins and palytoxin-like compounds.
Espiña
Begoña
B
Departamento de Farmacologia, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27002 Lugo, Spain.
Cagide
Eva
E
Louzao
M Carmen
MC
Fernandez
Maria M
MM
Vieytes
Mercedes R
MR
Katikou
Panagiota
P
Villar
Adriano
A
Jaen
David
D
Maman
Luz
L
Botana
Luis M
LM
D000178:Acrylamides / Q000032:analysis* / Q000378:metabolism*; D000818:Animals; D049872:Bivalvia / Q000737:chemistry; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D003064:Cnidarian Venoms; D006801:Humans; D051381:Rats; D012680:Sensitivity and Specificity; D000095702:Polyether Toxins
D000178:Acrylamides; D003064:Cnidarian Venoms; C010272:palytoxin; D000095702:Polyether Toxins