Simultaneous Estimation of Luteolin and Apigenin in Methanol Leaf Extract of Bacopa monnieri Linn by HPLC

Aims: To develop, validate and quantify the content of flavonoids luteolin and apigenin in aerial parts of methanol leaf extract of Bacopa monnieri (B. monnieri) by reverse phase liquid chromatography.
Study Design: High Performance Liquid Chromatography
Place and Duration of Study: Department of Pharmaceutical Analysis, KMCH College of Pharmacy, Coimbatore and Indian Pharmacopoeia Commission, Ghaziabad, India from 1.7.2010 to 30.6.2011.
Methodology: Separation and quantification of flavonoids was performed on a C18 column (5µm, 200mm×4.6mm, id.) using potassium dihydrogen phosphate buffer (20mM, pH 3.5 adjusted with ortho phosphoric acid, v/v) and acetonitrile as mobile phase with a flow rate of 1ml/min. The column effluents were monitored at 348nm with column temperature kept at 30±1ºC.
Results: A validated method for simultaneous estimation of luteolin and apigenin was developed, where limits of detection and quantification was found to be 0.03 and 0.91µg/ml for luteolin, 0.041 and 0.13µg/ml for apigenin respectively. The percentage of these phytoconstituents recovered was in the range of 98.07-99.71 (%RSD<2%).
Conclusion: The developed validated HPLC method was found to be accurate, precise and robust and may be used for analysis of luteolin and apigenin in the extracts of B. monnieri.