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A high performance liquid chromatography coupled with photodiode array detection method was developed for the identification and quantification of p-hydroxy benzoic acid and agnuside in the extracts of Vitex negundo and Vitex trifolia. The separation was achieved using acetonitrile and O-phosphoric acid–water (0.5%, v/v) as the mobile phase in an isocratic elution mode. Mean retention times of standard p-hydroxy benzoic acid and agnuside were 6.14 and 11.90 min respectively. The developed method was validated as per the ICH guidelines for limit of detection, limit of quantification, linearity, accuracy and precision. Good linearity (r2≥0.999) was observed for both the compounds in wide concentration range. Relative standard deviation values for intra-day and inter-day precision studies were less than 2%. The analytical recoveries of p-hydroxy benzoic acid and agnuside by the developed HPLC method were 93.07% and 106.11% respectively. Two compounds were identified and quantified in leaves and bar extracts of V. negundo and V. trifolia using the developed HPLC method.

A validated rapid HPLC-PDA method was developed for identification and quantification of five tannin-related constituents gallic acid (GA), corilagin (CL), chebulagic acid (CB), ellagic acid (EA) and chebulinic acid (CN) in the extracts prepared from the bark and fruits of four Terminalia species available in India. The separation of the five analytes was achieved on an RP-18 column (4.6 × 250 mm, 5 µm) at 25°C using a solvent mixture comprising of acetonitrile and (0.05%) trifluoroacetic acid-water in a gradient elution mode. Limit of detection was 1.0, 0.5, 1.0, 0.5 and 1.0 μg/mL for GA, CL, CB, EA and CN, respectively. Similarly, limit of quantification was 2.5, 1.0, 2.5, 1.0 and 2.5 μg/mL for GA, CL, CB, EA and CN, respectively. Good linearity (r(2) > 0.992) was observed for all the five compounds in wide concentration range. Using the developed HPLC method, the five analytes were identified and quantified in bark and fruit extracts of Terminalia chebula, Terminalia bellirica, Terminalia arjuna and Terminalia catappa. This is the first report of identification and quantification of the five tannin-related marker constituents in the bark and fruit extracts of T. chebula, T. bellirica, T. arjuna and T. catappa.

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