Detection of 13,19-didesmethyl C spirolide by fluorescence polarization using Torpedo electrocyte membranes

作者：Fonfria Eva S; Vilarino Natalia; Molgo Jordi; Araoz Romulo; Otero Paz; Espina Begona; Carmen Louzao M; Alvarez Mercedes; Botana Luis M^*

来源：Analytical Biochemistry, 2010, 403(1-2): 102-107.

DOI：10.1016/j.ab.2010.04.006

• Cyclic imines
• Spirolide
• Phycotoxin
• Fluorescence polarization
• Detection method
• Nicotinic acetylcholine receptors
• Shellfish
• alpha-Bungarotoxin

Fluorescence polarization (FP) is a powerful tool for studying molecular interactions by monitoring changes in the apparent size of fluorescent molecules. In this paper, a previously described fluorescence polarization assay was used to detect 13,19-didesmethyl C spirolide. The assay is based on the competition of cyclic imine marine biotoxins with alpha-bungarotoxin for binding to nicotinic acetylcholine receptor-enriched membranes of Torpedo marmorata. The 13,19-didesmethyl C spirolide was detected in buffer and mussel matrix. The sensitivity of the assay for the 13,19-didesmethyl C spirolide and the 13-desmethyl C spirolide was similar. After an acetone/chloroform extraction of spiked mussel meat, the average recovery rate of 13,19-didesmethyl C spirolide was 77.7 +/- 1.9%. The quantification range for this toxin in mussel was 40-200 mu g/kg of shellfish meat. This assay can be used to detect the spirolides 13,19-didesmethyl C spirolide and 13-desmethyl C spirolide, in shellfish as a screening assay.

• 出版日期2010-8