Abstract
Objective: The objective of the present work was to establish a simple, precise, accurate and robust method for simultaneous estimation of gallic acid, curcumin and piperine from the marketed ayurvedic formulation by liquid chromatography.
 Methods: The separation was carried out on Hemochrom C18 Column (250 mm × 4.6 mm ID, 5 µm pore size) with a mobile phase methanol: acetonitrile: water (pH 3.2adjusted by using orthophosphate acid) in the ratio 70:20:10v/v by isocratic elution modeat 25 °C and the flow rate was setat0.8 ml/min. The analysis was carried out atisoabsorptive wavelength of 295 nm.
 Results: The retention time of gallic acid, curcumin and piperine was found to be 3.3(±0.2), 4.7 (±0.2) and 5.6 (±0.2) min, respectively. The linearity range for gallic acid, curcumin and piperine was found to be 10-70 μg/ml, 20-80 µg/ml and 2-14 µg/ml, respectively with the coefficient of linear regression greater than 0.99 for all markers. Mean percent recoveries for gallic acid, curcumin, and piperine were found within the limit of acceptance (99-100%). The percent relative standard deviation (%RSD) for precision and robustness was found less than 2%, which indicates the method is precise and robust. The developed method applied for quantification of these markers from the marketed ayurvedic formulation of Dekofcyn tablet.
 Conclusion: The developed method was found to be simple, rapid, precise and reproducible for standardization of Dekofcyn tablet and can be useful for other formulations containing these three markers.
Highlights
The advance chromatographic separation techniques are used for qualitative and quantitative analysis of multiple constituents present in medicinal plants and herbal formulations.In the present study, the Reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed for simultaneous estimation of curcumin, gallic acid and piperine from the Dekofcyn tablet
A UV spectrum of gallic acid, curcumin and piperine of methanolic solutions was scanned in the range of 200-400 nm
The developed method was optimized under the chromatographic conditions mobile phase methanol: acetonitrile: water in the ratio of 70:20:10v/v
Summary
The RP-HPLC method was developed for simultaneous estimation of curcumin, gallic acid and piperine from the Dekofcyn tablet. It is used against chronic cough, cold, sore throat and chronic oral allergies. It consists of haldi (Curcuma longa), Talispatra (Taxus buccata), Shatavari (Asparagus racemosus), Piper(Piper longum), Ashwagandha (Withania somnifera), Amla (Emblica officinalis), Kachura (Curcuma zedoria), Vasaka (Adhatoda vasica), Galo ghan (Tinospora cordifolia),Dagdi Prashanbhed (Bergenia ciliate), Suvarna mashik bhasma Gallic acid which is one the constituent of Amlapossess antioxidant, anti-cancer, anti-inflammatory and anti-diabetic properties. Structure-activity relationship (SAR) analysis showed that the odihydroxy group of gallic acid is important for the inhibitory effect in inflammation [2,3,4]
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