CHARACTERIZATION AND RP-HPLC METHOD DEVELOPMENT OF A BIOMARKER (BERGENIN) FROM THE ROOTS OF Securinega virosa

The quality of the roots of Securinega virosa (Euphorbiaceae) commonly marketed in Ghana for medicinal purposes needs to be assured through standard quality control procedures. This work has isolated and characterized a biomarker to be used in assessing the quality of products and herbal preparations containing the roots of Securinega virosa. Using column chromatography of 20 g ethanolic roots extract; 0.02% w/w biomarker was obtained and characterized by melting point determination, UV-Vis, IR, 1D and 2D NMR spectroscopic analysis and comparison to reported literature. The biomarker was identified as Bergenin. A RP-HPLC method has been developed and validated for the quality control of S. virosa root preparations with Bergenin as standard. The most appropriate resolution was attained by a gradient elution of methanol: 0.05% v/v trifluoroacetic acid in water using Waters Bondapak C18 (3.9×300 mm, 5 μ) column at a flow rate of 1.0 ml/min and a wavelength of 220 nm. The retention time of the biomarker was found to be 14.20 ± 0.01 mins. The method was subjected to ICH guidelines for validation and showed a good linearity with a correlation coefficient (r) of 0.9993. The LOD and LOQ were found to be 15.23 mg/L and 46.16 mg/L respectively. Tolerable levels of accuracy, precision, specificity and robustness were observed. The developed RP-HPLC method estimated 0.465%w/w Bergenin in the roots of S. virosa and may therefore be employed in the quality control of product containing roots of S. virosa.