Quantitative Determination of Bergenin in Callus, Twig and Root of Ficus racemosa L. Extract by High Performance Liquid Chromatography

Abstract

Root of Ficus racemosa is the ingredient of Ya-Ha-Rak remedy or Benjalokawichien use for antipyretics in Thai Traditional Medicine. Previous studies show that bergenin can be found in the root bark of F. racemosa. Bergenin is a polyphenol compound which have various biological activities such as anti-inflammatory, anti-bacteria, and anti-pyretic.

 The objectives of this study were validate the reverse phase high performance liquid chromatography (RP-HPLC) for quantitation the amount of bergenin in calluses, twigs and roots of F. racemosa. by using the acetonitrile: water (10:90 v/v) as mobile phase at a flow rate of 0.8 mL/min. All of samples were analyzed by using UV detector with the wavelength at 272 nm. The concentration of bergenin range at 10-320 µg/mL was analyzed. The r-squared data of linearity was 0.9999. The accuracy values measured as % recovery with three concentration of bergenin, 30, 60 and 120 µg/mL were 98.009 ± 0.168, 102 ± 0.621 and 98.807 ± 0.188 % respectively. The precision of intraday and interday show as % relative standard deviation of 0.703- 1.822% and 0.251- 1.982%, respectively. The LOD and LOQ can be detected and quantified at 5.23 and 15.84 µg/mL respectively. The aqueous extract of F. racemosa twig showed high percentage of bergenin content at 2.06±0.00% dry weight followed by root extract at 0.646±0.00% dry weight, and an aqueous extract of callus had the least of bergenin content at 0.018±0.00% dry weight. The RP-HPLC method for determine of bergenin was met the AOAC Guidelines for Single Laboratory Validation of Chemical Methods for Dietary Supplements and Botanicals (2002). It was reliable for determination of bergenin content in calluses, twigs and roots of F. racemosa.