Journal of Pharmaceutical and Biomedical Analysis
Analytical methods for determination of magnoflorine and saponins from roots of Caulophyllum thalictroides (L.) Michx. Using UPLC, HPLC and HPTLC
Introduction
Caulophyllum thalictroides (L.) Michx. (Berberidaceae), known as blue cohosh, is an indigenous perennial herb commonly found in eastern North America where the roots are used in diuretic, uterine antispasmodic and laxative preparations [1]. Traditionally the roots and rhizomes of C. thalictroides are used for the treatment of menstrual difficulties and inducing uterine contractions [2]. Earlier studies have revealed blue cohosh rhizomes and roots contain two types of saponins, caulosaponins and caulophyllosaponins [3], [4], [5], [6]. The caulosaponins (caulosides A, C, D, G) contain caulosapogenin (later corrected to hederagenin [3]) as aglycon [4], [5] whereas caulophyllosaponins (caulosides B, H, leonticin D) show caulophyllosapogenin as aglycon [4], [5], [6]. The saponins in blue cohosh are considered to be responsible for the uterine stimulant effects together with teratogenic alkaloids [2]. Between 1882 and 1905, blue cohosh was listed in the United States Pharmacopoeia as a labor inducer [6] and sold as an herbal supplement that can aid in childbirth. There is considerable concern about the safety of blue cohosh with reports of new born babies having heart attacks or strokes after the mother consumed blue cohosh to induce labor [6], [7], [8], [9]. Blue cohosh is avoided during the first trimester (three months) of pregnancy [6]. In addition to the quinolizidines, the aporphine alkaloid magnoflorine is found in substantial quantities [10], [11] which have been implicated as teratogens. Based on available scientific evidence, there is no proven safe or effective dose for blue cohosh [6].
Few published analytical methods have been reported for the analysis of alkaloids alone or for alkaloids and saponins using GC, HPLC and densitometry for blue cohosh [1], [2], [12], [13]. Levels of the major quinolizidine alkaloids in herbal preparations have been determined by gas chromatography, HPLC and densitometry [1], [12]. Betz et al. [12], developed a GC methodology for the quantitative determination of alkaloids from the roots of C. thalictroides L. Woldemariam et al. [1], developed a TLC densitometric method and an HPLC method for the quantification of alkaloids from extracts of C. thalictroides L. roots. The studies by Woldemariam et al. [1], and Betz et al. [12], focused on isolation and determination of alkaloids including N-methylcystisine, baptifoline, anagyrine and magnoflorine. The saponins present in C. thalictroides L. have received less attention than the alkaloids. Ganzera et al. [2], developed an HPLC method for the quantitative analysis of primary alkaloids and saponins from C. thalictroides L. roots. Subramaniam et al. [13], also developed a HPLC method for the separation and quantification of three alkaloids and three saponins from extracts of blue cohosh roots and dietary supplements. The newly developed UPLC method for quantitative determination of one major alkaloid [magnoflorine (1)] and eight triterpene saponins [cauloside H (2), leonticin D (3), cauloside G (4), cauloside D (5), cauloside B (6), cauloside C (7), cauloside A (8) and saponin PE (9)] (Fig. 1) from the roots of C. thalictroides is found to be capable of giving shorter retention times while maintaining good resolution and sensitivity. Detection of the saponins was achieved with the use of an ELS detector. The compounds were numbered by the order of elution using LC–UV–ELSD method. A comparison of chromatographic performance of HPLC and UPLC was performed. A simple and fast HPTLC method was also developed for the chemical fingerprint analysis of alkaloid (magnoflorine) and saponins. The highly sensitive UPLC–MS method was used to identify and confirm the compounds in blue cohosh root samples and dietary supplements that claim to contain C. thalictroides. These methods are useful in establishing the quality and safety of herbal products claiming to contain C. thalictroides.
Section snippets
UPLC–UV–ELSD
All analyses were performed on a Waters Acquity UPLC™ system (Waters Corp., Milford, MA, USA) including binary solvent manager, sampler manager, column compartment, PDA (Waters Acquity model code UPD), ELS detector (Waters Acquity model code UPE), and MS detector (Waters Acquity model code SQD), all connected to Waters Empower 2 data station. An Acquity UPLC™ BEH Shield RP18 column (50 mm × 2.1 mm I.D., 1.7 μm) also from Waters was used. The column and sample temperature were maintained at 35 °C and
Chromatographic conditions optimization
Optimal chromatographic conditions were obtained after running different mobile phases with a reversed phase C18 column. The different columns tried for UPLC were Acquity UPLC BEH C18 (100 mm × 2.1 mm I.D., 1.7 μm), Acquity UPLC BEH C18 (50 mm × 2.1 mm I.D., 1.7 μm) and Acquity UPLC BEH Shield RP18. The best results were observed with BEH shield RP18 column (50 mm × 2.1 mm I.D., 1.7 μm) using ammonium acetate (pH 7.2) and acetonitrile as the mobile phase. Acetonitrile was preferred over methanol as the mobile
Conclusion
The newly developed UPLC method for the chemical analysis of nine compounds from blue cohosh was found to be capable of providing short retention times while maintaining good resolution as compared to conventional HPLC. The new UPLC technique allowed for the reduction in the mobile phase flow rate and an increase in acquisition rate with the benefit of decreased injection volumes to achieve good peak shapes. The method is suitable for rapid analysis of magnoflorine and saponins and for chemical
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 5U01FD002071-10 and 1U01FD003871-02, and the United States Department of Agriculture, Agricultural Research Service, Specific Cooperative Agreement No. 58-6408-2-0009 and the authors would like to thank Annette Ford for samples preparation.
References (15)
- et al.
Analysis of aporphine and quinolizidine alkaloids from Caulophyllum thalictroides by densitometry and HPLC
J. Pharm. Biomed. Anal.
(1997) - et al.
A pharmacological study of a crystalline glycoside of Caulophyllum thalictroides
J. Am. Pharm. Assoc.
(1954) - et al.
Profound neonatal congestive heart failure caused by maternal consumption of blue cohosh herbal medication
J. Pediatr.
(1998) - et al.
Isolation and characterization of alkaloids from Caulophyllum thalictroides
J. Pharm. Sci.
(1967) - et al.
Alkaloids and saponins from blue cohosh
Phytochemistry
(2008) Determination of saponins and alkaloids in Caulophyllum thalictroides (blue cohosh) by high-performance liquid chromatography and evaporative light scattering detection
Phytochem. Anal.
(2003)- et al.
Caulosapogenin and its identity with hederagenin
J. Chem. Soc.
(1956)
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