Extraction and Analysis of Alkaloids from Roots of Goldenseal and Dietary Supplements by Using UPLC-UV-MS Methods

The roots of Hydrastis candense (goldenseal) are popular phytomedicines for the treatment of gastrointestinal disorders and upper respiratory tract infections [1–2]. Simple and fast UPLC-UV-MS methods were developed for the quantification of the major constituents, berberine and hydrastine from roots of Hydrastis canadensis L. and dietary supplements containing goldenseal and Echinacea purpurea/goldenseal combination formulations. The extraction (with acidified water and methanol) and analysis were applied to several other alkaloids including canadine, hydrastinine, palmatine, coptisine, and jatrorrhizine by a UPLC method with PDA and MS, C18 column. The mobile phase consisted of solvent A (50 mM ammonium formate, pH 3.3) and solvent B (acetonitrile with 0.05% formic acid). The developed method was validated for all the parameters tested and successfully applied to the identification of seven alkaloids in plant sample and ten dietary supplements. The plant material and ten dietary supplements were found to contain major alkaloids, hydrastine and berberine. One commercial product also contained palmatine, coptisine and jatrorrhizine, thus indicating that the material was not pure goldenseal. LC-mass spectrometry coupled with electrospray ionization (ESI) method is described for the identification of seven alkaloids in plant sample and dietary supplements. This method involved the use of the [M]⁺ ions for coptisine, jatrorrhizine, palmatine and berberine, [M+H]⁺ ions for hydrastine and canadine, [M+H-18]⁺ ions for hydrastinine in the positive ion mode with selective ion recording (SIR).

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. The authors would like to thank Annette Ford, University of Mississippi for extraction of samples. References: [1] Brown PN, Roman MR (2008), J. AOAC International, 91(4): 694–701. [2] Abourashed EA, Khan IA, (2001), J. Pharm. Sceinces, 90(7): 817–822.

Fig. 1 UPLC Chromatograms of a mixture of standards (A), roots of goldenseal (B) and dietary supplements (C–D) at 290 nm. 1 hydrastinine, 2 hydrastine, 3 coptisine, 4 jatrorrhizine, 5 canadine, 6 palmatine and 7 berberine.