A validated method for the simultaneous quantitation of bioactive alkaloid markers in the leaf ethanolic extract of Cissampelos sympodialis Eichl.: a phenological variation study.

摘要

Introduction - The leaf hydroalcoholic extract of Cissampelos sympodialis Eichl. (Menispermaceae) has shown promising activity in different animal models of asthma. Several alkaloids have been identified in the extract, including warifteine and methylwarifteine (bisbenzylisoquinoline), as well as milonine (morphinandienone). Objective - To develop and validate an analytical method for the simultaneous quantitation of the bioactive markers of C. sympodialis hydroalcoholic leaf extract and to apply the method to a seasonal (phenological) study of the concentration of the alkaloid markers. Methodology - The method used reversed phase high performance liquid chromatography with UV detection and calibration by standard addition. Separation was achieved using a C₁₈-column (250x4.6 mm, 5 micro m) and a mobile phase consisting of a mixture of 0.05% aqueous (Et)₃NH₂(A):MeOH(B) in gradient mode at a flow rate of 1.0 mL/min. Results - The method proved to be linear in the concentration range tested (2-100 micro g/mL, r2