Optimization of fermentation variables for Ayurvedic formulation, Drakshasava by Response Surface Methodology and its marker based validation studies

Abstract

The concept of fermentation has been part of various Ayurvedic formulations since ages. Drakshasava is one of the self fermented Ayurvedic medicine containing Vitis vinifera L. as major ingredient. However, lack of modern technology and long processing time of these classical formulations may interfere with growing needs of industries. To cope up with the current health care standards, optimization of existing Ayurvedic formulations with multi-marker approach is required. A statistical model has been developed to optimise the fermentation process of Drakshasava (without prakshepa dravya). The selected fermentation process parameters i.e., pH, incubation temperature and fermentation time were optimized by BBD of Response Surface Methodology. The combination of parameters was selected to achieve a required alcohol percentage. The method was validated by HPTLC with respect to non established biomarker piperine, a bioavailability enhancer required to prevent early metabolism of resveratrol (therapeutic active constituent of Drakshasava). The quadratic model was found significant with F and P value, 2.20 and 2.302, respectively. The optimum conditions obtained in the batch fermentation process were incubation temperature (30.59°C); pH (4.84) and fermentation period of 9 days with predicted concentration of 9.82% alcohol. The method was validated in accordance to ICH guidelines and found statistically reproducible and selective for the quantitative estimation of piperine in Drakshasava. The in-house fermented samples were found to have significant increase in piperine concentration (2.42 μg/mL), compared to decoction (0.11 μg/mL). The optimized method reduced the manufacturing time without interefering the Pharmacopoeial limits. It leads to standardization, better quality and consistency of Drakshasava.