Quantification of Baicalein, Chrysin, Biochanin-A and Ellagic Acid in Root Bark of Oroxylum indicum by RPHPLC with UV Detection

Abstract

Oroxylum indicum (Syonakh), belonging to the family Bignoniaceae, has been used for
the present study. Preliminary phytochemical screening revealed the presence of
phytocostituents like flavonoids, alkaloids, tannins and anthraquinone in the root bark of
Oroxylum indicum. The preliminary screening using TLC technique reflected the presence of
four phytoconstituents such as baicalein, chrysin, biochanin-A and ellagic acid in both petroleum
ether and hydrolysed n-butanol fractions. Modern phytochemical analysis in terms of
quantification of different bioactive phytoconstituents in the root bark of Oroxylum indicum was
performed using reverse phase high performance liquid chromatography (RP-HPLC) fingerprint.
To evaluate the quality of root bark of Oroxylum indicum, a simple rapid and accurate RP-HPLC
method was developed for the assessment of four bioactive phytoconstituents viz. chrysin,
baicalein, biochanin-A and ellagic acid. The components were quantified in different extracts
viz. alcohol, petroleum ether, ethyl acetate and n-buatnol successively. Quantification of
phytoconstituents was done by using RP-HPLC in petroleum ether and hydrolysed n-butanol
fraction. Standard baicalein, chrysin biochanin-A and ellagic acid (Sigma-Aldrich) were
employed for the development of the method. The RP-HPLC system used a base deactivated
C18 column with water, methanol, acetonitrile and orthophosphoric acid as the mobile phase
and detection was performed at 262 nm. The method was precise with relative standard
deviation for these constituents that ranged between 0.5-1.0% (interday). The content of four
phytoconstituents in the root bark of Oroxylum indicum was determined to establish the
effectiveness of the method.