PMID: 9177752Jun 1, 1997Paper

A fluorescent microplate assay for diarrheic shellfish toxins

Analytical Biochemistry
M R VieytesL M Botana

Abstract

A fluorescent enzyme inhibition assay for okadaic acid using 4-methylumbelliferyl phosphate and fluorescein diphosphate as substrates for the enzyme phosphatase 2A was developed. In the inhibition assay, performed in a microtiter plate, the PP2A was inhibited by adding okadaic acid and the resulting fluorescence enhancement derived from enzymatic hydrolysis of the substrate was quantified in a fluorescence plate reader. The measurable range of okadaic acid was 3.2 to 3200 pg/ml with an IC50 = 0.1 nM. The detection limit of okadaic acid was 2.56 pg/well in buffer solutions and 12.8 ng/g hepatopancreas in shellfish extracts. The coefficient of variation (CV, n = 22) for each point ranged from 18.80 to 37.90% (mean 28.35%). The proposed method is very convenient, rapid, and sensitive by using the enzyme inhibition assay system and fluorescent reaction as a detection system. This work demonstrates that the fluorescent assay can be used to quantify the amount of okadaic acid in shellfish samples and also is valid for very dilute samples, such as phytoplankton samples.

References

Nov 1, 1992·Toxicon : Official Journal of the International Society on Toxinology·Z AmzilP Masselin
Nov 1, 1992·Toxicon : Official Journal of the International Society on Toxinology·W S ShestowskyH M Sikorska
Jan 1, 1991·Toxicon : Official Journal of the International Society on Toxinology·O B StabellK Pedersen
Jan 1, 1989·Annual Review of Biochemistry·P Cohen
Jan 1, 1989·Toxicon : Official Journal of the International Society on Toxinology·T UsagawaT Yasumoto
Dec 1, 1988·Journal of Muscle Research and Cell Motility·A Takai
Jan 1, 1988·Toxicon : Official Journal of the International Society on Toxinology·L LevineH Van Vunakis
May 1, 1982·Journal of Clinical Microbiology·R H Yolken, F J Leister
Dec 1, 1995·Trends in Pharmacological Sciences·W P Bowen, J C Jerman
Jul 1, 1996·Toxicon : Official Journal of the International Society on Toxinology·A TubaroT Yasumoto

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Citations

Jan 22, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Maria-Teresa Fernández-SánchezAntonello Novelli
Oct 3, 2013·Marine Drugs·Bich-Thuy L EberhartVera L Trainer
Jan 23, 2014·Marine Drugs·Beatriz RegueraJuan Blanco
Nov 21, 2009·Toxicon : Official Journal of the International Society on Toxinology·Humberto J DominguezJosé J Fernández
Aug 13, 2009·Toxicon : Official Journal of the International Society on Toxinology·Clara AlbanoGian Paolo Rossini
Jan 25, 2005·Toxicology·Gian Paolo Rossini
Feb 21, 2006·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·A N SayerJ E Gibson
May 21, 2005·Toxicon : Official Journal of the International Society on Toxinology·Michael J TwinerGregory J Doucette
Oct 25, 2012·Bioengineered·Juan A RubioloLuis M Botana
Dec 3, 2014·Marine Environmental Research·Moira McCarthyFrank F N A M van Pelt
Jul 8, 2008·Chemistry : a European Journal·Patricia G CruzAntonio Hernández Daranas
May 12, 2010·Analytical and Bioanalytical Chemistry·Natalia VilariñoLuis M Botana
Jan 12, 2012·The Analyst·Mònica CampàsBeatriz Prieto-Simón
Jul 4, 2013·Analytical and Bioanalytical Chemistry·Natalia VilariñoLuis M Botana
May 18, 2006·Alternatives to Laboratory Animals : ATLA·Philipp HessUNKNOWN ECVAM/DG SANCO Workshop
Jul 17, 2007·Organic Letters·Patricia G CruzManuel Norte
May 1, 1999·Analytical Biochemistry·O I FontalL M Botana
Jun 7, 2002·The Science of the Total Environment·Mark E Hahn
Sep 9, 2000·Toxicon : Official Journal of the International Society on Toxinology·D O MountfortP Truman
May 22, 2002·Cellular Signalling·Yolanda RománLuis M Botana
May 26, 1999·Toxicon : Official Journal of the International Society on Toxinology·D O MountfortD J Hannah
Jan 10, 2002·Journal of Agricultural and Food Chemistry·José C GonzálezLuis M Botana
Mar 27, 2003·Bioorganic & Medicinal Chemistry Letters·María L SoutoJosé J Fernández
Jun 12, 2003·Journal of Immunological Methods·David A Rider, Stephen P Young
Aug 23, 2011·Environmental Science & Technology·T F H BoveeL A P Hoogenboom
Jun 5, 2012·Chemical Research in Toxicology·Luis M Botana

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