High-throughput enzyme inhibition screening of 44 Iranian medicinal plants via piezoelectric spraying of planar cholinesterase assays.

A rapid and straightforward approach was developed for screening the acetyl- and butyrylcholinesterase (ChE) inhibitory activity of 44 Iranian medicinal plant extracts at laboratory scale. After a fast ChE inhibitory pre-testing of samples applied as band pattern, 40 out of the 44 Iranian medicinal plant extracts were selected. These were adjusted in the application volume depending on their inhibition activity, applied on both plate sides and simultaneously developed in a horizontal developing chamber. Different mobile phases were studied to achieve maximum separation of ChE inhibitors and minimum co-elution with matrix. Contrary to immersion, the piezoelectric spraying reduced the consumption of assay solutions, prevented zone tailing, zone shift and cross-contamination, and homogeneously covered the entire plate surface with the assay solutions. The ChE inhibitors of the six most bioactive plant extracts were tentatively assigned by high-resolution mass spectrometry in combination with the spectral and chromatographic information obtained.