Application of MABA (Micro plate alamar Blue Assay) in determining antitubercular activity of Phellandrium in comparison with streptomycin - An in vitro study

Abstract

Tuberculosis (TB) remains a major global public health problem, particularly in developing countries such as India, where the burden of multidrug-resistant tuberculosis (MDR-TB) continues to rise. The limitations of existing anti-tubercular drugs, including long treatment duration, toxicity, and increasing resistance, highlight the need for alternative or adjunct therapeutic approaches. Homoeopathy, derived from medicinal plants, has been explored for its potential therapeutic effects with minimal side effects. The present in vitro laboratory study was conducted to evaluate the antitubercular activity of Phellandrium aquaticum in different homoeopathic potencies (6CH, 30CH, and 200CH) using the Microplate Alamar Blue Assay (MABA) against Mycobacterium tuberculosis H37Rv strain (ATCC 27294). Serial dilutions ranging from 100 µg/ml to 0.8 µg/ml were prepared in 96-well microplates containing Middlebrook 7H9 broth medium. Streptomycin served as the standard control drug. Antimycobacterial activity was determined based on colour change of Alamar Blue reagent, where blue indicated inhibition of bacterial growth and pink indicated growth. The minimum inhibitory concentration (MIC) values observed were 12.5 µg/ml for Phellandrium aquaticum 200CH, 6.25 µg/ml for 30CH, and 12.5 µg/ml for 6CH. Among the tested potencies, Phellandrium aquaticum 30CH showed comparatively higher antimycobacterial activity. The findings suggest that Phellandrium aquaticum possesses potential antitubercular properties in vitro and may be considered as a supportive or adjunct therapeutic option in tuberculosis management. However, further in vivo studies and large-scale clinical trials are required to validate its efficacy and therapeutic application.