A VALIDATED RP-HPLC-UV-VISIBLE METHOD FOR QUANTIFICATION OF SWERTIAMARIN USING SPE METHOD AND ITS APPLICATION TO PHARMACOKINETIC STUDY USING SPARSE SAMPLING METHODOLOGY

Kanchan Gandhi*, Dr. Sasikumar Menon and Dr. Sunita Shailajan

ABSTRACT

Swertiamarin (SWM), a secoiridoid glucoside from Swertia densifolia (Griseb.) Kashyap, has shown biological activities like antidiabetic, antihelmenthic, antinociceptive, hepatoprotective, antioxidant, etc. The present paper describes a selective and sensitive Reverse Phase – High Performance Liquid Chromatography method (RP-HPLC) using fixed wavelength UV detector for quantification of SWM in rat plasma. Briefly, the plasma samples were pre-treated using a simple solid-phase extraction method. The separation of SWM and the internal standard, Hydrochlorothiazide (HCTZ), was achieved on Cosmosil C18 column (150 mm X 4.6 mm i. d., 5 μ) using 1.0 mL/min isocratic mobile phase flow. Also, the developed bioanalytical method was validated as per the US FDA bioanalytical guidelines over the concentration range of 50.0 to 40,000.0 ng/mL (r2 > 0.9980) for SWM from rat plasma. The mean percentage recovery (n=6) for the low, middle and high quality control samples was 91.23 ± 1.88, 90.36 ± 2.69 and 87.52 ± 1.50 %, respectively. The applicability of the validated RP-HPLC method was demonstrated by successful measurement of SWM from plasma following oral administration of Swertia densiflora (Griseb.) Kashyap leaf extracts to female Albino Wistar rats.

Keywords: Swertiamarin, RP-HPLC, Method validation, Pharmacokinetics, Swertia densifolia (Griscb.) Kashyap