Determination of Alkaloids and Flavonoids from Aerial Parts of P. incarnata, Various Species and Dietary Supplements containing P. incarnata using UPLC-UV-MS and HPTLC Methods

Passion flower plants belong to the genus Passiflora, and are inhabitants of the tropical and semi-tropical zones of North, Central and South America[1]. Leaves, aerial parts and flowers of Passiflora have been traditionally used for treatment of stress, anxiety, depression, insomnia, cardiac arrthymia and asthma [1,2]. C-glycosyl flavonoids are used as markers in the quality control of Passiflora and until now, the attributed harmane β-carbolinic alkaloids have not been found in the Passiflora incarnata at well-known quantities [3].

A rapid UPLC method was developed for the simultaneous analysis of five indole alkaloids (harmalol (1), harmol (2), harmane (3), harmaline (4) and harmine (5)) and four flavonoids (iso-orientin (6), orientin (7), iso-vitexin (8), and vitexin (9)) from the leaves of Passiflora incarnata L. (Passifloracea), different species of Passiflora (P. violacea, P. capsularis, P. edulis) and dietary supplements that claim to contain Passiflora. The separation was achieved within 8.0 minutes by using C-18 column material, a water/acetonitrile mobile phase, both containing 0.05% formic acid using a gradient system and a temperature of 35°C. The method was validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). The wavelength used for quantification with the diode array detector was 340nm for flavonoids and alkaloids. The developed method is simple, economic, fast and especially suitable for quality control analysis of flavonoids and alkaloids in plant samples and dietary supplements. The compounds (including isoschaftoside (10) and schaftoside (11)) in plant samples and commercial products of Passiflora were identified and confirmed by UPLC-MS. An HPTLC method was also developed for the chemical fingerprint analysis of Passiflora samples.

Fig.1: UPLC Chromatograms of a mixture of standard (A) and leaves of Passiflora incarnata L. (B)

Fig.2: Comparison of P. incarnata samples by HPTLC method. Tracks: 1, 2–6; leaves of Passiflora incarnata L., 2, standard mix-4 [isoorientin (1), isovitexin (2), orientin (3), vitexin (4)] at 366nm

Acknowledgements: This research is supported in part by „Science Based Authentication of Dietary Supplements“ and „Botanical Dietary Supplement Research“ funded by the Food and Drug Administration grant numbers 5U01FD002071–09 and 1U01FD003871–01, and the United States Department of Agriculture, Agricultural Research Service, Specific Cooperative Agreement No. 58–6408–2-0009. References: [1] Bisset NG, (2001) Herbal Drugs and Phytopharmaceuticals, CRC Press, New York [2] Soulimani R, et al. (1997)J Ethnopharmacol, 57: 11–20. [3] Muller SD, et al. (2005)J Pharm Biomed Anal, 37: 399–403.