Unknown

Mycobacterium tuberculosis were used for activity evaluation. Optical bottom black 96 well plates were used for the screening of the compounds. The Cetylamine was tested in triplicate and tested over three serial dilutions, starting at a concentration of 500 mg/mL, with repeated serial dilutions at a lower concentrations being performed on compounds showing initial inhibitory activity. Ethambutol was used as a positive control. To determine if the compounds had any inherent redox potential which might affect readouts, 200 lL solutions of compounds and PBS were added to individual control wells. Control wells of bacteria only, and PBS medium only were also prepared. 100 lL of BCG solution was added to all wells except the PBS only and compound only wells, and the plate was incubated at 37 C and 5% CO2 for 7 days. Alamar Blue dye (20 lL) was then added to all test wells on day 7, and the plate was incubated for another 24 h before fluorescence was measured (excitation at 544 nm, emission at 590 nm) using a FLUOstar OPTIMA plate reader. The results were graphed to determine the MICs. The lowest drug concentration effecting 100% growth inhibition was considered the MIC.