ISOLATION AND DEVELOPMENT OF AN HPLC METHOD FOR THE QUANTIFICATION OF A BIOMARKER IN THE ROOTS OF PAULLINIA PINNATAAbstract
An RP-HPLC method to be used to quantitate the amount of a biomarker in the roots of Paullinia pinnata was developed. The root bark was milled and extracted and the dry powder obtained taken through column chromatography (CC). The biomarker was then isolated. The melting point of the biomarker was determined and found to be 117-119 oC. A RP-HPLC method was successfully developed and used to quantify the biomarker in the chloroform extract. The mobile phase used was Methanol:Water (9:1) with an injection volume of 100µl and a range of 1.000. The method employs a Phenomenex Kromasil C-8, 5µ, 250mm × 4.6mm, 100Ǻ reverse phase column at a flow rate of 1.3ml/min, and a wavelength of 210nm. The method gave a retention time of 2.9687 ± 0.2657 (n=15). The percentage content of the biomarker in the root of Paullinia pinnata was determined to be 0.052009±0.004321% w/w (n=12). The HPLC method was validated for linearity, repeatability, intermediate precision, and robustness. The limits of detection and quantitation were also determined.
E. K. Cudjoe , John N. A. Addotey *, Nathaniel N. A. Okine , Reimmel K. Adosraku, Kofi Annan
Department of Pharmaceutical Chemistry *, KNUST, Kumasi, Ghana. KNUST, Kumasi, Ghana.
04 March, 2016
08 April, 2016
31 May, 2016
01 August 2016