Open AccessRP-HPLC and HPTLC Identification and Quantification of Flavonoids in the Leaves Extract of Bauhinia acuminata Linn. (Caesalpiniaceae)
Author(s) -
Sudipta Chakraborty,
Nripendra Nath Bala,
Sudipta Das
Publication year - 2021
Publication title -
journal of pharmaceutical research international
Language(s) - English
Resource type - Journals
ISSN - 2456-9119
DOI - 10.9734/jpri/2021/v33i44a32622
Subject(s) - chromatography , chemistry , high performance liquid chromatography , repeatability , quercetin , silica gel , phosphoric acid , formic acid , organic chemistry , antioxidant
Aims: The present investigation is aimed at the identification and quantification of flavonoids in the methanol extract of leaves of Bauhinia acuminata Linn. (Caesalpiniaceae) by a validated HPTLC method and confirmation of the same by RP-HPLC.
Methodology: CAMAG HPTLC system with VisionCATS software and HPLC Agilent Infinity 1260 were employed for the study. Pre-coated silica gel 60 F254 plates were used as stationary phase and Toluene : Ethyl acetate : Formic acid (5:4:0.2, v/v/v) was used as mobile phase in HPTLC while in HPLC Thermo hypersil BDS C18 column and Methanol and 0.4% Phosphoric acid (65 : 35) were used as stationary and mobile phase respectively.
Results: Among different standards used in HPTLC, only quercetin was found to be present and further quantitatively analyzed. The method shows good correlation coefficient (R2 ≥ 0.99) and linearity in the range of 0.5 to 5.0 µg/band. The concentration of quercetin in the test extract was found to be 0.514 µg/mg of the extract. The LOD and LOQ values were 0.84 µg/band and 2.59 µg/band respectively. The RP-HPLC study also represented good % RSD of Retention time (0.025) and Area (0.09) which signifies the high precision and repeatability. The assay result (4.99 %) also indicated good presence of the quercetin in the extract.
Conclusion: The methods were rapid, cost effective and may be employed for quality-control and standardization of quercetin in different plant extracts.