Isolation, characterization, and validation of RP-HPLC method for the quantification of quercetin in Huberantha senjiana leaf extract
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Date
2020-05
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Open Science Publishers LLP
Abstract
Huberantha senjiana (Annonaceae) – an endemic tree of Pakkamalai Reserve Forest Gingee Hills, Tamil Nadu, Indiawas subjected to preliminary phytochemical tests and high-performance liquid chromatography (HPLC) fingerprintchromatogram was established for use in future research. Subsequently, the HPLC-DAD-ESI (+) MS analysis toidentify the common biomarkers revealed the presence of Quercetin, a polyphenolic flavonol in the ethyl acetateextract of the leaves. An isocratic reversed-phase (RP)-HPLC method for the quantification of quercetin was developedwith 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 minutes and wasvalidated according to the International Conference on Harmonization Q2B guidelines. Linearity was achieved withthe concentration range of 5.0–17.5 μg/ml with a R2 value of 0.996. Sensitivity was demonstrated with the limit ofdetection of 2.28 µg/ml and the limit of quantification of 6.92 µg/ml. The robustness was estimated from purposefulchanges in the composition of the mobile phase, wavelength, and flow rate and the limits were Not More Than (NMT)4%. The foliar concentration of quercetin was found to be 0.0055% w/w. The structural confirmation was done by theisolation and characterization by the divergent spectral analysis. This is the first published report on the identification,quantification, and isolation of quercetin in the leaves of H. senjiana species
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Keywords
Huberantha senjiana, high-performance liquid chromatography, HPLC fingerprints, isolation, quercetin
Citation
Pandiyan Rajakannu, Ilango Kaliappan. Isolation, characterization, and validation of RP-HPLC method for the quantification of quercetin in Huberantha senjiana leaf extract. Journal of Applied Pharmaceutical Science. 2020 May; 2020 May: 110-118