Fluorescent probe for the detection of acetylcholinesterase inhibitors using high performance thin layer chromatography effect-directed assay in complex matrices

Abstract

In recent years Bioautography has become an efficient bioassay for finding active compounds in complex matrices including extracts of; bacteria, plants or fungi. High Performance Thin Layer Chromatography (HPTLC) is a technique which allows effect-directive analysis (EDA) through the separation and identification of biologically active substances on a Thin Layer Chromatography (TLC) plate and can be run as a high throughput screening assay for enzymes. This paper presents a new bioautography method using a novel fluorescent probe derived from coumarin and its validation with Acetylcholinesterase (AChE) inhibition. This method combines the advantages of bioautography and the high sensitivity of fluorescence for detection. Combining these advantages, the limit of detection (LOD) and limit of quantification (LOQ) limits usually obtained with traditional chromophores has been reduced by three times. Therefore our new method has been applied on 14 mushroom extracts and we highlight a compound (variegatic acid) as a potentially new AChE inhibitor in X. Chrysenteron.