Planta Med 2008; 74 - PC129
DOI: 10.1055/s-0028-1084647

Analysis of Coptis sp., for the purpose of establishing a monograph draft proposal for the German Pharmacopoeia

T Brem 1, A Ankli 2, B Klier 3, J Heilmann 1, G Franz 1
  • 1University of Regensburg, Institute of Pharmazie, Pharmazeutische Biologie, Universitätsstraße 31, 93040 Regensburg, Germany
  • 2CAMAG, Sonnenmattstrasse 11, 4132 Muttenz, Switzerland
  • 3PhytoLab GmbH & Co KG, Dutendorfer Str. 5–7, 91487 Vestenbergskreuth, Germany

The development of appropriate analytical procedures is proposed for the TCM-herbal drug Coptis sp., which is planned for an implementation in the German Pharmacopoeia DAB. The quantification method described by the respective monograph of the Chinese Pharmacopoeia (English Edition, [1]), suggesting a TLC based densitometric procedure for the quantification of berberine, was improved to the simultaneous quantification of the alkaloids coptisine, epiberberine, berberine and palmatine. Using a twin trough chamber with toluene/ethyl acetate/isopropanol/MeOH/H2O (60:30:15:15:3) as solvent system and aqueous NH3 (32%) for conditioning, the alkaloids showed Rf values of 0.62 (coptisine), 0.51 (epiberberine), 0.42 (berberine) and 0.35 (palmitine) on HPTLC silica plates and were quantified without derivatisation by fluorescence measurement at 366nm with a TLC scanner.

As a first step for the implementation of an appropriate quantification with HPLC, the method suggested by Wagner [2] for the fingerprint analysis of Coptis was investigated. Its major disadvantage is the use of a complex buffer system consisting of acetic acid, sodium acetate, SDS, diethylamine and H2O, which is not stable and resulted in varying and not reproducible retention times for the alkaloids. We propose the use of a H2O/acetonitril/acetic acid gradient on a RP-18e column (Purospher Star, 5µm, 250×4) which allows the quantification of the alkaloids at a detection wavelength 348nm within 35 minutes. For the densitometric TLC and the HPLC quantification method a validation protocol is established. In addition, identification methods for this herbal drug (macroscopic, microscopic, TLC/HPTLC) and appropriate tests for purity are proposed [2]. All the newly developed and controlled methods will be the basis of a monograph draft suitable for the DAB.

Acknowledgements: The BfArM (Kurt-Georg-Kiesinger-Allee3, 53175 Bonn, Germany) is gratefully acknowledged for financial support.

References: 1. Anonymous, Pharmacopoeia of the People's Republic of China Engl. Edit. 2003. 2. Wagner, Chinese Drug Monographs and Analysis 2004. 3. Anonymous, European Pharmacopoeia 2005 5. Edit.