Planta Med 2010; 76 - P558
DOI: 10.1055/s-0030-1264856

Validation of TLC procedures for the identification of Cimicifuga racemosa and Hypericum per-foratum during quality control of a Cimicifuga/Hypericum fixed combination

A Hoppenheit 1, R Volk 1, H Gerke 1, C Bodinet 1
  • 1Schaper & Brümmer GmbH & Co. KG, Bahnhofstraße 35, 38259 Salzgitter, Germany

For quality control of pharmaceuticals, validated analytical procedures are required. Identity verification of herbal substances and phytopharmaceuticals is usually performed by thin layer chromatography (TLC). Cimicifuga/Hypericum film-coated tablets contain dry extract preparations of Cimicifuga racemosa and Hypericum perforatum. Two TLC procedures on silica gel 60 F254 HPTLC plates were validated for this fixed combination: (a) for detection of characteristic constituents of C. racemosa, such as triterpene glycosides, (b) for detection of marker compounds typical for H. perforatum, such as hypericin. Mobile phases were (a) ethyl acetate: methanol (85:15), (b) toluol: ethyl acetate: formic acid: water (5:15:2:1). The typical validation characteristic for identification tests is their specificity. A characteristic necessary for any analytical procedure is its robustness [1]. For verifying the specificity of both procedures, test solutions of the herbal medicinal product, of herbal substances and of individual placebos were analysed. The procedures were suitable to detect specifically characteristic constituents of both herbal substances as mentioned above. Identification of both herbal substances was possible without any restriction. Specificity was also tested by comparison of two different stationary phases. HPTLC plates were essential (Cimicifuga), respectively prefered (Hypericum) instead of aluminium foil plates. For verifying the robustness of both procedures, three parameters were modified: sample preparation: variation of numbers of extraction cycles during soxhlet extraction; chromatography: variation of the mobile phase; variation of detection (duration of heating, respectively drying conditions). All tested variations led to chromatograms comparable to those obtained under standard conditions. Therefore, robustness was proven, too.

References: 1. ICH guideline Q2(R1) Validation of analytical procedures: text and methodology; current step 4 version, Nov. 2005 (http://www.ich.org).