Tuberculosis is a life threatening disease and second to HIV in terms of deaths due to infectious diseases. Drug resistance development of the first-line drugs is a major concern in the treatment of this disease. There is no comprehensive and critical review in the literature of the bioanalytical methods for the determination of anti-tubercular agents from last two decades. This work offers a detailed account on the liquid chromatographic methods reported in the literature for the estimation of various anti-tubercular drugs. Major emphasis is given to sample preparation process, sensitivity of method, chromatographic separation conditions and detection systems used in their bioanalysis.