Huberantha senjiana (Annonaceae) - an endemic tree of pakkamalai reserve forest Gingee hills, Tamil Nadu, India was subjected to preliminary phytochemical tests and HPLC fingerprint chromatogram was established for use in future research. Subsequently, the HPLC-DAD-ESI (+) MS analysis to identify the common biomarkers revealed the presence of Quercetin, a polyphenolic flavonol in the ethyl acetate extract of the leaves. An isocratic RP-HPLC method for the quantification of quercetin was developed with phenomenex, RP C18 column using mobile phase [(H2O (0.1% Formic acid, A); (MeOH: ACN, (40:15v/v), B), 40:60 (v/v)] with a flow rate of 0.8 mL/min, detection max at 254 nm with a retention time of 7.58 min was validated according to the ICH Q2B guidelines. Linearity was achieved, with the concentration range of 5.0-17.5μg/ml with R2 value 0.996. Sensitivity was demonstrated with LOD 2.28µg/mL and LOQ 6.92 µg/mL. The robustness was estimated from purposeful changes in the composition of the mobile phase, wavelength and flow rate and the limits were NMT 4%. The foliar concentration of quercetin was found to be 0.0055 % w/w. The structural confirmation was done by isolation and characterization by divergent spectral analysis. This is the first published report on the identification, quantification and isolation of quercetin in the leaves of H. senjiana species.
Key words: Huberantha senjiana, high-performance liquid chromatography, HPLC fingerprints, isolation, quercetin.
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