| Assay Method Information | |
| | Anticholinesterase Activity Assays |
| Description: | The substrates of the reaction were acetylthiocholine iodide and butyrylthiocholine iodide. 5,5'-dithio-bis(2-nitrobenzoic) acid (DTNB) was used for the measurement of the anticholinesterase activity. The solutions of the samples and galantamine in n-propanol were prepared at concentration of 4000 µg/mL. Aliquots of 150 µL of 100 mM phosphate buffer (pH 8.0), 10 µL of sample solution and 20 µL AChE (2.476 × 10^-4 U/µL) (or 3.1813 × 10^-4 U/µL BuChE) solution were mixed and incubated for 15 min at 25 °C. 10 µL of DTNB solution was prepared by adding 2.0 mL of pH 7.0 and 4.0 mL of pH 8.0 phosphate buffers to a mixture of 1.0 mL of 16 mg/mL DTNB and 7.5 mg/mL NaHCO3 in pH 7.0 phosphate buffers. The reaction was initiated by the addition of 10 µL (7.1 mM) acetylthiocholine iodide (or 0.79 mM butyrylthiocholine iodide). Propyl alcohol was used as a solvent to controls. The hydrolysis of these substrates was monitored at 412 nm. |
| Affinity data for this assay | |
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