Planta Med 2008; 74 - P-130
DOI: 10.1055/s-2008-1075326

A Fast and Sensitive UPLC/MS Method for the Quantitation of Bioactive Bacosides of Bacopa monnieri for Bioanalytical Applications

R Earla 1, B Avula 1, Y-H Wang 1, RS Pawar 1, LMV Madgula 1, T Smillie 1, SI Khan 1, IA Khan 1, 2
  • 1National Center for Natural Products Research and
  • 2Department of Pharmacognosy, School of Pharmacy,The University of Mississippi, University, MS, 38677, USA

Bacopa monnieri (L.) Pennell. (Scrophulariaceae) has been used for its memory enhancing properties and as a nerve tonic for treatment of epilepsy and insanity in Ayurvedic medicine. In the Western world several commercial products are available claiming adaptogenic and cognitive enhancing effects [1]. The presence of dammarane saponins (bacosides) is considered to be responsible for the pharmacological properties of Bacopa. monnieri [1]. A fast and sensitive UPLC-MS method was developed and validated for the quantification of bacosides in buffer medium (HBSS buffer pH 7.4). The best results were obtained with Acquity UPLC™ BEH C8 column (50 mm × 2.1 mm, i.d., 1.7 µm) system using a gradient elution with a mobile phase consisting of 0.05% formic acid in water and 0.05% formic acid in an acetonitrile-methanol mixture (5:95) at a constant flow rate of 0.210 mL/min. The compounds were detected with single quadrupole mass detector. This method involved the use of the [M+Na]+ ions of bacosides in the positive ion mode with selective ion recording (SIR). The mass transition of bacoside A3, bacopaside–II, bacopasaponin C and bacoside X were detected at m/z 951.2, 951.3, 921.2 and 921.3, respectively. The LOD and LOQ for all four bacosides were in the range of 4.8–5.6 ng/mL and 10.3–12.7 ng/mL respectively. The standard calibration curve was linear over the concentration range of 10–800 ng/mL. The percentage accuracy of intra-day and inter-day batches varied between 95.3 to 109.8% and the coefficient of variation (precision) of intra-day and inter-day batches were in the range of 0.5–7.7%. The analytical method described has been successfully applied to study the bioavailability of these bacosides in a Caco-2 model. Acknowledgements: This research is funded in part by “Science Based Authentication of Dietary Supplements” funded by the Food and Drug Administration grant number 2 U01 FD 002071-07 and authors like to thank Dr. Vishalia C Joshi for the species authentication. References: [1] Ganzera M, et al. (2004) Analytica Chimica Acta 516: 149–154.