The quality and safety of the Unani formulations have to be scientifically validated in order to produce quality medicines and protect the health of people suffering from various kinds of diseases. For the validation or standardization of Unani formulations a systematic scientific protocol is very much needed. Though the Unani medicines are an important healthcare option, but due to lack of scientific standards, these traditional medicines are hindering us from capitalizing on the global opportunities provided by increasing interests all over the world. The drug ‘Majoon-e-Hamal Ambari Alwi Khani’ is one of the ancient most commonly used Unani formulations in the ailments of atony to the uterus and habitual abortion. For the validation of this drug the parameters such as Standard Operating Procedure (SOP’s) for the preparation of drug, powder microscopy to find out the characteristic botanical characters of raw drugs added in the formulation were worked out. Evaluation of physico-chemical, TLC/HPTLC, finger prints, analysis of WHO parameters like heavy metals, microbial loads, aflotoxins and pesticide residues were also carried out to ascertain the quality of drug. The evaluated data will help to lay down the scientific standards for the drug studied to contribute mateial for in Unani pharmacopoeia of India.