Combined use of analytical high-performance liquid chromatography and cell morphology transformation assay to detect new protein phosphatase inhibitors of okadaic acid type.
The search for new protein phosphatase inhibitors in shellfish contaminated by toxin-producing dinoflagellates generally relies on preliminary separation techniques followed by biological tests. To detect such substances without purifying them initially, we developed an approach based on a correlation of the results of two different analytical techniques applied to toxic extracts: high-performance liquid chromatography after derivation of the toxins and the cell morphology transformation assay on KB cells. Application of this protocol to stored frozen mussels showed a decrease in okadaic acid concentration during storage, with formation of degradation derivatives, some of which possessed notable protein phosphatase inhibition activity.
Pouchus
Y F
YF
Université de Nantes, SMAB-URM11, ISOMER, Faculté de Pharmacie, France.
Dauvergne
S
S
Morel
D
D
Mondeguer
F
F
Marcaillou-Lebaut
C
C
Amzil
Z
Z
Verbist
J F
JF
D000818:Animals; D049872:Bivalvia; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004791:Enzyme Inhibitors / Q000302:isolation & purification*; D008387:Marine Toxins / Q000302:isolation & purification*; D019319:Okadaic Acid / Q000302:isolation & purification*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D008387:Marine Toxins; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases