After a through phytochemical screening of the Carica papaya Leaf extract for the constituents present, the tannins namely tannic acid and gallic acid were analyzed and quantified in the crude methanolic extract and herbal formulations. Both the RP-HPLC and HPTLC methods were developed and validated according to the ICH Q2 (R1) guidelines. The tannic acid was quantified by the RP-HPLC method and gallic acid was quantified by the HPTLC method. The linearity of tannic acid and gallic acid was found to be in the range of 20-50 µg/ml (y=2121.99 x-29648.1) and 400-1200 ng/spot (y=6.4325 x-2016) respectively with a correlation coefficient of 0.999 for both calibration graphs. The % RSD values calculated for the intraday, inter day precision repeatability of measurement, repeatability of sample application and accuracy studies which was found to be below 2. The LOD and LOQ were found to be 1 and 5 µg/ml respectively for tannic acid. The LOD and LOQ were found to be 100 and 300 ng/band respectively for gallic acid. The percentage recovery of tannic acid was found to be between 95 to 97% and between 94-97% for gallic acid at 80%, 100% and 120% level of recovery. The amount of tannic acid and gallic acid quantified in the herbal formulation was found to be 11 µg/ml and 0.378 µg/ml respectively. Both the chromatographic methods are simple, precise, accurate and sensitive which can be applied for the routine analysis of the two constituents in the Carica papaya leaf extract.
