High performance thin layer chromatography based chemo profiling of Ashvagandharishta and its antidepressant activity.

Withania somnifera (L.) has long been used as a traditional rasayana herb against a variety of human ailments. This research presents a high performance thin layer chromatography based chemo profiling of Ashvagandharishta and its antidepressant activity. The in-house formulation was made using a fermentation process according to the Indian Pharmacopoeia. Physiochemical standardization of the formulation was performed using different quality control parameters such as total ash, acid insoluble ash, alcohol soluble extract value and water soluble extract value. A column chromatography and high performance thin layer chromatography method was used to isolate and estimation of withanolide-A, withaferin-A & ß-sitosterol from the root of W. somnifera. In addition. The antidepressant effect of different formulations were carried out by force swimming test in albino mice. The thiobarbituric acid reactive substances (TBARS) and Glutathione (GSH) assay was used to find out the oxidative stress. W. somnifera root has been standardized macroscopically, microscopically, physico-chemically according to the guidelines of the Ayurvedic Pharmacopoeia. The qualitative and quantitative analysis was performed using high performance thin layer chromatography and it was performed on each formulation and found the content of withanolide-A and -sitosterol in the in-house formulation is higher while withaferin-A is rather contained in the decoction. The antidepressant effect showed that the immobility time was lowest in the case of the standard formulation followed by in house formulation, while the increase in glutathione and the reduction in thiobarbituric acid reactive substances levels revealed the antioxidant nature of the formulation. In conclusion, based on the above results, we can conclude that Ashvagandharishta could be a breakthrough for the treatment of depression in the future.

'Ashvagandharishta' prepared using yeast consortium from Woodfordia fruticosa flowers exhibit hepatoprotective effect on CCl4 induced liver damage in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: [corrected] Consortium of yeasts sourced from traditionally used Woodfordia fruticosa flowers proved to be beneficial for fermenting Ashvagandharishta. It resulted in faster fermentation, acceptable organoleptic properties and demonstrable hepatoprotective potential in CCl4 induced hepatotoxicity. To formulate Ashvagandharishta using consortium of yeasts and to investigate its physiochemical parameters. Standardize the formulation with the help of standard withaferin-A and withanolide-A and to evaluate its hepatoprotective potential in CCl4 induced hepatotoxicity in the rat model. MATERIAL AND METHODS: Ashvagandharishta was prepared using a 5% consortium of yeasts and ascertained its quality through physiochemical and phytochemical investigation. Withaferin-A and withanolide-A was simultaneously estimated by HPLC for standardization. Hepatoprotective potential was evaluated by administering 2.31 and 1.15 ml/kg doses while considering biochemical parameters like serum AST, ALT, ALP and lipid profile. Gene expression study was carried out for the expression of antioxidant and inflammatory genes such as CAT, GPx and proinflammatory gene IL-6. Histopathology of liver was also studied with the help of H&E staining. RESULTS: Ashvagandharishta was found organolepticaly acceptable with optimized physiochemical parameters. Withaferin-A and withanolide-A in Ashvagandharishta estimated as 0.3711, 0.7426 (%w/v), respectively. In the CCl4 induced hepato-toxicity model, Ashvagandharishta-2.31ml/kg dose showed significant decrease in elevated hepatic level of AST(p<0.001), ALT(p<0.01) and ALP(p<0.001). Both doses of Ashvagandharishta showed significant reduction of TG, Cholesterol, VLDL and LDL in serum, with corresponding reduction of (p<0.001) serum-HDL. Ashvagandharishta also showed increased serum protein (p<0.05) and albumin (p<0.01) with decrease in bilirubin (p<0.01). Additionally, Ashvagandharishta administration revealed up-regulation in antioxidant genes such as CAT and GPx in liver with concomitant down-regulation in proinflammatory IL-6gene (p<0.01). Histopathological parameters revealed restoration of normal tissue architecture by both doses of Ashvagandharishta. CONCLUSIONS: Consortium of yeasts from Woodfordia fruticosa flowers showed better fermentation pattern for Ashvagandharishta produced with acceptable organoleptic properties. Hepatoprotection shown by Ashvagandharishta was mainly through prevention of oxidative damage. Up-regulation of CAT and GPx genes and corresponding down regulation of proinflammatory IL6 gene was revealed as possible mechanism of its action.