Estimation of twelve bacopa saponins in Bacopa monnieri extracts and formulations by high-performance liquid chromatography.

A simple and sensitive reversed phase high performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of twelve bacopa saponins present in the extracts of the Indian Medicinal Plant, Bacopa monnieri. The separation was achieved on a reversed phase C(18) column (Luna C(18)), 5 microm by isocratic elution with 0.05 M sodium sulphate buffer (pH 2.3) and acetonitrile (68.5 : 31.5, v/v) as the mobile phase at a flow rate of 1.0 ml/min with an operating temperature of 30 degrees C. The method was validated for linearity, precision, intra- and inter-day precision and accuracy. Several Bacopa samples (plant materials, extracts and commercial formulations) were successfully analyzed. Major bacopasaponins were bacosides A(3) (3), bacopaside II (4), bacopaside I (5), bacopaside X (6), bacopasaponin C (7), bacopaside N2 (9) and the minor components were bacopasaponin F (1), bacopasaponin E (2), bacopaside N1 (8) bacopaside III (10), bacopaside IV (11) and bacopaside V (12). The total saponin content in the samples, plant materials and extracts varied from 5.1 to 22.17% and 1.47 to 66.03 mg/capsule or tablet in the commercial formulations.