Preparation, Phytochemical Analysis, Antimicrobial, Antioxidative, and Cellular Damage Protective Activity of Gomutra Haritaki

Introduction:Gomutra Haritaki (GH) is a well-known classical Ayurvedic formulation widely used in various disorders such as anemia, edema, disorders of the oral cavity, liver disorders, and piles. Gomutra (cow urine) and Terminalia chebula fruits are the main ingredients used to formulate this potential formulation. The present work was aimed to develop standardization protocols for this formulation based on the marker phytoconstituents present, total phenolic and flavonoid contents, and essential elements/heavy metals as well as establish its antioxidant and antimicrobial potential. Methods:GH was prepared as per the procedure described in the Ayurvedic text. The total phenolic and total flavonoid contents (TFCs) were determined by Folin-Ciocalteau colorimetric method and colorimetric method, respectively. 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, nitric oxide scavenging activity, superoxide dismutase activity, and total antioxidant capacity by phosphomolybdenum method were adopted for antioxidant and free radical scavenging potential analysis. Antimicrobial ability was tested by the agar-well diffusion method against bacterial and fungal pathogens. High-performance thin-layer chromatography (HPTLC) and inductively coupled plasma mass spectrometry were used for the quantification of phenolic compounds and elemental analysis, respectively. Result:The total phenolic and flavonoid contents were detected in significant amounts in GH. The highest polyphenolic (44.85 ± 1.06 mg gallic acid equivalent/g) as well as flavonoid (13.82 ± 1.50 mg quercetin equivalent/g) contents were detected in methanol extract (Gomutra Haritaki methanolic extract [GHME]). It also showed promising antioxidant and antimicrobial activities through different assay methods. HPTLC quantification of phytoconstituents suggested that appreciable amounts of phenolic constituents are present in GH. GHME was found to contain the highest amount of all the three phenolic compounds viz. gallic acid (25.00 μg/mg), ellagic acid (8.5 μg/mg), and ferulic acid (2.44 μg/mg). It was found to have high content of essential elements/minerals viz. Fe, Mg, Ca, and K. Conclusion:GH is a good source of bioactive phytoconstituents as well as essential nutrient elements. It contains significant amounts of total phenolic content, TFC, and phenolic compounds and exhibits potent antioxidant and cellular damage protective activity. GH also has the potential to be used as an antimicrobial agent.