Background: Diakure is an indigenously prepared hypoglycemic polyherbal formulation, containing mixture of Vetiveria zizanoides (root), Hemidesmus indicus (rhizome), Strychnos potatorum (seed), Salacia reticulata (bark), Holarhena antidysenterica (seed), Cassia auriculata (bark), Trigonella graecum (seed) and Acacia catechu (bark) and each individual herb has scientific background in treating diabetes. This study aimed to perform pharmacognostical evaluation, preliminary phytochemical evaluation, standardization, heavy metal analysis, pesticidal residual analysis and antimicrobial assay of DiaKure. Materials and Methods: 100 g of the powder mixture of all eight herbs mentioned above was boiled with 900 ml water for 30 min to produce the polyherbal mixture and this mixture was used for performing various tests. Results: Organoleptic characters and macroscopic characters such as colour, odour, taste, size and shape are evaluated under pharmacognostical evaluations and ash values, extractive values and loss on drying are considered under standardisation parameters and are reported. Phytochemical screening of the extract revealed the presence of alkaloids, tannins, saponins, flavonoids, steroids, glycoside and sugars. The formulation has normal limits of heavy metal and pesticidal residue. Also, the aqueous mixture, during its antimicrobial assay, does not exhibit any antibacterial or anti-fungal activities. Conclusion: The result of this study validates the use of aqueous extract of DiaKure in ethnomedicine, favouring the isolation of antidiabetic agents from the extract of DiaKure. Since the study was conducted in a controlled and authenticated manner, all the evaluation measures used in the study can be used for the standardization of the above-said formulation.