An Improved Purification Method for Removing Colour Interference from 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl Tetrazolium Bromide (MTT) Antibacterial Assays

Abstract

The MTT antibacterial assay is an important method in natural antibacterial component discovery. Researchers can use the MTT antibacterial assay to quickly and efficiently evaluate the antibacterial activity of natural compounds. The aim of this research was to find a method to reduce background colour and bacterial cell interference when using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to study the antibacterial effect of phytochemicals. This study used NZ ‘Hass’ ripe avocado seed extract as an example to develop a silica gel column chromatography method that could combine with the MTT assay to remove the background colour and bacterial cells before the formazan measuring stage. This method is particularly suitable for MTT-based antibacterial inhibition studies when the tested phytochemical extracts have an interfering colour.

Keywords

• background colour
• bacterial cell
• MTT
• silica gel column chromatography method
• formazan