Background:Grahaṇimihira tailam is an unexplored ayurvedic oil preparation which consists of 34 ingredients. The efficacy of this traditional ayurvedic medicine is undisputable. Proper clinical standardization of this formulation will go a long way in securing greater recognition for it. The main objective of this study was to develop standardization parameters for the formulation in a multidisciplinary way.Materials and Methods:A simple and efficient method for the quantification of umbelliferone by high performance thin layer chromatography was developed and validated. Presence of the major fatty acids and their percentage were assessed by using gas chromatography-mass spectrometry (GC-MS). Various physio-chemical parameters, microbiological load, aflatoxins and mineral oil were also evaluated. Spread plate method was used for checking microbial contamination.Results:The results were validated as per standard protocols. Quantitative estimation revealed the percentage of umbelliferone to be in the range of 0.88-0.98 (w/w). GC-MS analysis of sample led to the identification of 14 fatty acids, in which linoleic acid was obtained as the major fatty acid. Microbes, aflatoxins and mineral oils were found to be absent in the tailam.Conclusion:The results which give the quantitative estimates of various physico-chemical parameters can be adopted to establish new standards for analysis of batch-to-batch variation and this data will facilitate shelf life studies in the future.