Ethambutol is 1,2-diamino alcohol, being a medication it is primarily used to treat tuberculosis. It plays an important role in antitubercular agent inhibiting the synthesis of spermidine in myobacteria and also inhibit transfer of mycolic acids into cell wall of tubercle bacillus. The route of administration is oral and is easily available in market. This is readily absorbed by gastrointestinal tract and 50% excreted unchanged by urine. Being a powerful chemotherapeutic agent it is specifically effective against actively growing microorganism of yobacterium and tuberculosis. Its analysis is usually done from biological samples such as blood and urine which are the main sample of choice in management and medicolegal cases. There are several techniques such as Gas chromatograph (GC), High Performance Liquid Chromatography (HPLC), Gas Chromatography-Mass spectrometry (GC-MS), Liquid Chromatography-Mass Spectrometry (LC-MS) for its analysis. These techniques are not only costly but also time consuming and require more sophisticated instruments handling. An attempt has been made to develop a new method for analysis of ethambutol in blood using High Performance Thin Layer Chromatography plate. Ethambutol was extracted from blood using liquid-liquid extraction method and analyzed by using High Performance Thin Layer Chromatographic plate. For chromatographic separation various solvent systems were used as mobile phase. After the development of plates they were examined under UV light followed by spray of chromogenic reagents such as ninhydrin and iodine fumes which successfully increase the sensitivity without dispensing with the simplicity of the method. Present method is simple, non destructive, reproducible and repeatable which can easily be performed in any laboratory.

Keywords: Ethambutol; Ninhydrin; Iodine Vapours; HPTLC Plate; Liquid-Liquid Extraction etc.