A Validated Method for the Simultaneous Quantitation of Bioactive Alkaloid Markers in the Leaf Ethanolic Extract of Cissampelos sympodialis Eichl.: a Phenological Variation Study
Article first published online: 18 NOV 2011
Copyright © 2011 John Wiley & Sons, Ltd.
Volume 23, Issue 5, pages 426–432, September/October 2012
How to Cite
Marinho, A. F., Barbosa-Filho, J. M. and Oliveira, E. J. (2012), A Validated Method for the Simultaneous Quantitation of Bioactive Alkaloid Markers in the Leaf Ethanolic Extract of Cissampelos sympodialis Eichl.: a Phenological Variation Study. Phytochem. Anal., 23: 426–432. doi: 10.1002/pca.1376
- Issue published online: 10 AUG 2012
- Article first published online: 18 NOV 2011
- Manuscript Accepted: 19 OCT 2011
- Manuscript Revised: 5 SEP 2011
- Manuscript Received: 1 JUL 2011
- bisbenzylisoquinoline alkaloids;
- Cissampelos sympodialis;
- seasonal variation
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).
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.
The method used reversed phase high performance liquid chromatography with UV detection and calibration by standard addition. Separation was achieved using a C18-column (250 × 4.6 mm, 5 µm) 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.
The method proved to be linear in the concentration range tested (2–100 µg/mL, r2 > 0,99), precise (RSD ≤ 15%), accurate (85–115%), selective and robust. Detection limits for warifteine, methyl-warifteine and milonine were 0.39, 1.10 and 1.77 µg/mL respectively. The highest concentration of total alkaloids (determined as the sum of the three alkaloids) in the hydroalcoholic extract of the leaves was 2.9 ± 0.2 mg/g extract (n = 3), prior to fruit development. Both warifteine and methylwarifteine were detected in the total alkaloid fraction of the ripened fruits.
The results demonstrated that significant variations in the concentration of the biomarkers occurred throughout the vegetative cycle. The lowest concentration of the alkaloids in the leaves coincided with their appearance in the ripened fruits. Copyright © 2011 John Wiley & Sons, Ltd.