A reversed-phase compatible thin-layer chromatography autography for the detection of acetylcholinesterase inhibitors.
J Sep Sci
; 38(21): 3788-94, 2015 Nov.
Article
en En
| MEDLINE
| ID: mdl-26489065
ABSTRACT
A dual readout autographic assay to detect acetylcholinesterase inhibitors present in complex matrices adsorbed on reversed-phase or normal-phase thin-layer chromatography plates is described. Enzyme gel entrapment with an amphiphilic copolymer was used for assay development. The effects of substrate and enzyme concentrations, pH, incubation time, and incubation temperature on the sensitivity and the detection limit of the assay were evaluated. Experimental design and response surface methodology were used to optimize conditions with a minimum number of experiments. The assay allowed the detection of 0.01% w/w of physostigmine in both a spiked Sonchus oleraceus L. extract chromatographed on normal phase and a spiked Pimenta racemosa (Mill.) J.W. Moore leaf essential oil chromatographed on reversed phase. Finally, the reversed-phase thin-layer chromatography assay was applied to reveal the presence of an inhibitor in the Cymbopogon citratus (DC.) Stapf essential oil. The developed assay is able to detect acetylcholinesterase inhibitors present in complex matrixes that were chromatographed in normal phase or reversed-phase thin-layer chromatography. The detection limit for physostigmine on both normal and reversed phase was of 1×10(-4) µg. The results can be read by a change in color and/or a change in fluorescence.
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Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Fisostigmina
/
Inhibidores de la Colinesterasa
/
Cromatografía en Capa Delgada
/
Cromatografía de Fase Inversa
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
J Sep Sci
Año:
2015
Tipo del documento:
Article
País de afiliación:
Argentina