Cimicifuga racemosa as an alternative to hormone therapy is the most extensively studied medicinal herb used for the alleviation of menopausal symptoms. Cimicifuga/Hypericum film-coated tablets (CHF) is a fixed combination containing extracts from the herbal substances Hypericum perforatum and C. racemosa. The absorption of constituents from H. perforatum has already been demonstrated after oral intake of monopreparations. Only few data are available for the absorption of Cimicifuga-components. No data at all exist for the fixed combination [1,2,3]. The purpose of this study was to prove the absorption of CHF after oral administration, using valid analytical methods. Hypericin was selected as biomarker for H. perforatum and caffeic acid derivatives (ferulic acid, cimicifugic acid B) as marker substances for C. racemosa. Wistar rats received single, 10-fold und 100-fold human equivalent dosages (HED) of CHF-granulate via gavage. At defined time points blood plasma samples were taken and analysed by HPLC with fluorescence detection. The absorption of the Hypericum-specific marker hypericin was demonstrated after application of the single and 10-fold HED of CHF. Ferulic acid, a marker compound of C. racemosa, could be quantified in the plasma samples in the case of the 10- and 100-fold HED. An enzymatic pretreatment of the plasma samples enabled the determination of cimicifugic acid B, another Cimicifuga-specific compound. The resulting plasma concentrations of all three marker compounds were dependent from the administered dosage. These results prove the absorption of CHF by the detection of specific marker compounds in the plasma of rats after oral application of CHF.

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.

Zubereitungen aus Bärentraubenblättern (Uvae ursi folium) (Abb. 1) werden zur Behandlung von entzündlichen Erkrankungen der ableitenden Harnwege eingesetzt. Das in Deutschland meistverkaufte Fertigarzneimittel aus Bärentraubenblättern (Cystinol akut(r) Dragees) basiert auf einem ethanolischen (60% v/v) Trockenextrakt und ist auf den Gehalt an Hydrochinonde-rivaten, berechnet als Arbutin, standardisiert. Dieses Phenolglykosid wird enzyma-tisch in Hydrochinon (HQ) und Glukose ([1]) gespalten. Durch Konjugation wird HQ in seine wasserlösliche Form überführt und damit nierengängig.

For the herbal immunomodulator Esberitox® containing an aqueous-ethanolic extract of a mixture of Thujae occidentalis herba, Baptisiae tinctoriae radix, Echinaceae purpureae radix and E. pallidae radix, immunological activity has been proven in vitro and in vivo [1]. In comparative in vitro investigations on the activity profiles of the single drugs, it was shown that Baptisia has its most striking activities in the activation of neutrophilic granulocytes and the stimulation of antibody production, especially IgM antibodies [1].

For the herbal immunomodulator Esberitox® containing an aqueous-ethanolic extract of a mixture of Thujae occiden-talis herba, Baptisiae tinctoriae radix, Echinaceae purpureae radix and Echinaceae pallidae radix, immunological activity has been proven in vitro and in vivo [1]. In comparative in vitro investigations on the activity profiles of the single drugs used for the preparation of the Esberitox® extract, it was shown that Thuja has its most striking activities in the activation of macrophages and CD4+ T-helper cells, resulting in an enhanced production of IL-2 [1,2].

Cimicifuga racemosa (CR) as an alternative to hormone therapy is the most extensively studied medicinal herb used for the alleviation of menopausal symptoms. In the last two decades a considerable amount of investigations in research and clinic documented for the special isopropanolic CR rootstock extract (iCR, Remifemin®) significant efficacy and safety [1]. Up to now the active principle of CR is not fully understood, however, the two major groups of constituents, triterpene glycosides and polyphenolic compounds are under discussion, represented by actein respectively caffeic acid as well-established analytical marker substances. In this investigation iCR was compared with 5 US-marketed CR containing products regarding (i) their amounts of both marker substances per recommended daily dose and (ii) their effects on MCF-7 cell proliferation according to [2]. Using standard analytical methods for medicinal preparations, actein amounts ranging from 0 to 230µg (170µg for iCR) were determined. Polyphenolic compounds calculated as caffeic acid varied from 148 to 2198µg (148µg for iCR). In the MCF-7 assay none of the CR products stimulated the proliferation of human estrogen-receptor positive breast cancer cells.

Arbor vitae (Thuja occidentalis L.) is a native European tree widely used in homeopathy and evidence-based phytotherapy. Many reviews and monographs have been published on the herbal substance's description, mode of action and clinical use. However, no comprehensive evidence-based review is available. Therefore, our aim was to search MEDLINE databases and survey manufacturers for further details or unpublished data. This review presents the botany, ethnobotany and phytochemistry, especially the different contents of essential oil (Thujone) in relation to different extraction procedures of this medicinal plant. Thuja's antiviral action and immunopharmacological potential, such as stimulatory and co-stimulatory effects on cytokine and antibody production and activation of macrophages and other immunocompetent cells, have been evaluated in numerous in vitro and in vivo investigations. Although no controlled trials have been conducted on Thuja occ alone, many clinical studies have been performed with a herbal medicinal product containing a special extract of Thuja occ and other immunostimulants, demonstrating its therapeutic efficacy and safety in respiratory tract infections.

The influences of different arabinogalactan-proteins (AGPs) on proliferation and IgM-production of mouse lymphocytes as well as nitrite- and IL6-production of mouse macrophages were investigated in vitro. AGPs have been isolated and purified from roots of Baptisia tinctoria and Echinacea pallida and suspension culture of Echinacea purpurea. Comparing the AGPs, there are differences with regard to fine structure as well as to activities. AGPs from roots of B. tinctoria and E. pallida show high activity in all test systems. AGP from cell culture of E. purpurea shows no influence on proliferation of mouse lymphocytes, only weak influence on the IgM-production of mouse lymphocytes and weak stimulation of nitrite- and IL6-production in alveolar mouse macrophage culture.

The influence of the peroral administration of the macromolecular components of a herbal immunomodulator isolated from an aqueous-ethanolic extract of the mixed herbal drugs Thujae summitates, Baptisiae tinctoriae radix, Echinaceae purpureae radix and Echinaceae pallidae radix on the function of Peyer’s patches cells was investigated in mice. Peyer’s patches cells isolated from mice which had received oral administration of the macromolecular fraction of the plant extract developed a significantly enhanced plaque-forming cell (PFC) response to sheep red blood cells after incubation with these cells in the presence of lipopolysaccharide or the extract fraction for 7 days in vitro. These results show that after oral administration of the herbal immunomodulator, the immunologically active macromolecules can contact the cells of the gut-associated lymphoid tissue and modulate the mucosal immune response.

UNLABELLED Cystinol akut containing arbutin was developed as an antiseptic since it liberates hydroquinone in the urine. The in vivo release of hydroquinone from arbutin with or without addition of glusulase or an E. coli suspension was investigated in 4 volunteers. They ingested 6 dragees Cystinol akut (420 mg arbutin equivalent to 168 mg hydroquinone), urine was sampled and assayed by a validated HPLC method. RESULTS In comparison to incubation with glusulase the E. coli-suspension resulted in a 2.3 fold higher increase in free hydroquinone. When separating bacteria from the urine, the hydroquinone concentration in bacteria was 20 fold higher than in the supernatant. CONSEQUENCES Glucuronic acid or sulfuric acid conjugates of hydroquinones obviously are taken up, enriched and metabolized to hydroquinone by bacteria. Deconjugation of hydroquinone likely is catalyzed by intracellular enzymes presumably present in bacterial cytoplasm; comparable activities in eucaryotic cells usually are localized in lysosomes. Alkalization of the urine seems not to be a prerequisite to improve the antiseptic properties of hydroquinone released from arbutin.

The influence of the oral administration of an aqueous-ethanolic extract of a mixture of Thujae occidentalis herba, Baptisiae tinctoriae radix, Echinaceae purpureae radix and Echinaceae pallidae radix, on the course of Influenza A virus infection in Balb/c mice was investigated. The extract was administered to mice via the drinking water for 14 days starting 6 days before intranasal infection with Influenza A virus. The progress of infection was recorded during a time range of 21 days. Parameters for the evaluation of antiviral activity were survival rate and mean day to death. In a further set of experiments infected mice were sacrificed on defined days. Determination of consolidation score and virus titer were performed for each lung. The data show that the oral treatment with the extract induced a statistically significant increase in the survival rate, prolonged the mean survival time and reduced lung consolidation and virus titer. The experiments demonstrate that the plant immunomodulator given 6 days before exposure is a potent inhibitor of Influenza A virus pathology in vivo.

Hormone replacement therapy, which is a common menopausal treatment, is contraindicated in women with breast cancers due to concerns regarding the potential for breast cell proliferation. As such, there is a need for alternative methods for treating menopausal symptoms. To determine the influence of one such alternative, black cohosh (Cimicifuga racemosa [CR]), on estrogen-dependent mammary cancers, we conducted an in vitro investigation of the effect of an isopropanolic CR-extract on the proliferation of estrogen receptor-positive breast cancer cells. The experiments were performed using the human breast adenocarcinoma (MCF-7) cell test system, an established in vitro model for estrogen-dependent tumors. The influence of CR-extract on the proliferation of the MCF-7 cells was determined by measuring the incorporation of radioactively labeled thymidine. Under estrogen-deprived conditions, the CR-extract (10−3–10−5 dilutions) significantly inhibited MCF-7 cell proliferation. Additionally, application of the CR-extract inhibited estrogen-induced proliferation of MCF-7 cells. Moreover, the proliferation-inhibiting effect of tamoxifen was enhanced by the CR-extract. Such data that suggest a non-estrogenic, or estrogen-antagonistic effect of CR on human breast cancer cells lead to the conclusion that CR treatment may be a safe, natural remedy for menopausal symptoms in breast cancer.

The influence of the oral administration of a herbal immunomodulator, consisting of an aqueous-ethanolic extract of the mixed herbal drugs Thujae summitates, Baptisiae tinctoriae radix, Echinaceae purpureae radix and Echinaceae pallidae radix, on cytokine induction and antibody response against sheep red blood cells was investigated in mice. The treatment of the animals with the extract caused no enhancement of the cytokine titers in the serum. Spleen cells isolated from the treated mice, however, produced higher amounts of IL-2, IFNgamma and GM-CSF ex vivo in comparison to spleen cells isolated from control animals, especially after additional stimulation by lipopolysaccharides or concanavalin A. The application of the extract also triggered the production of IL-1 and TNFalpha by peritoneal macrophages ex vivo. The influence of the herbal extract on the antibody response was examined by the plaque forming cell assay. The administration of the extract caused a significant enhancement of the antibody response against sheep red blood cells, inducing an increase in the numbers of splenic plaque forming cells and the titers of specific antibodies in the sera of the treated animals. In mice, immunosuppressed by old age or additional treatment with hydrocortisone, the therapy with the extract resulted in a normalization of the antibody response against sheep red blood cells.

Despite the wide use of extracts from the rhizome of black cohosh (Cimicifuga racemosa) for the treatment of menopausal complaints, surprisingly little is known on their potential estrogenic properties, e.g. on estrogen dependent gene transcription. In addition, available informations on the effects on cell proliferation are contradictory. We therefore, tested for estrogenic and antiestrogenic effects of Cimicifuga racemosa extracts on proliferation of MCF-7 cells and on gene expression using ethanolic and iso-propanolic extracts of this medical plant. Estrogenic properties of plant extracts could neither be detected in proliferation assays, nor on gene expression using an estradiol-inducible yeast assay or the estrogen-inducible MVLN cells. In contrast, in all three experimental systems Cimicifuga racemosa antagonized estradiol induced activities. Estradiol induced stimulation of proliferation was inhibited by a dosage >1 microg/ml of extract concentration, gene expression was suppressed by doses of 100-1000 microg/ml of Cimicifuga racemosa extracts. From these results we conclude, that extracts from the rhizome of Cimicifuga racemosa contain compounds with antiestrogenic properties.

Pharmacodynamic Effects and Clinical Efficacy by a Combination of Herbal Active Substances of Cone Flower, Wild Indigo and White Cedar Summary: The effect of the single active ingredients Echinaceae (purpureae et pallidae) radix, Baptisiae tinctoriae radix and Thujae occidentalis herba as well as of the combination Esberitox® N has been verified in vitro, in vivo, in animal experiments and in human pharmacology (5). The pharmaceutical has immunomodulating and antiviral characteristics, whereby the immune system is probably influenced unspecifically, especially by increasing the macrophage activity. The efficacy for viral respiratory tract infections has been proven yet again in recent GCP-compliant, clinical studies (15, 17). Under the influence of the phytotherapeutic the duration of the illness decreased with a responder rate that was significantly higher than under placebo. The therapeutic benefit was even more pronounced, if the pharmaceutical was applied as early as possible. It has a high drug safety.

The effect of the single active ingredients Echinaceae (purpureae et pallidae) radix, Baptisiae tinctoriae radix and Thujae occidentalis herba as well as of the combination Esberitox N has been verified in vitro, in vivo, in animal experiments and in human pharmacology (5). The pharmaceutical has immunomodulating and antiviral characteristics, whereby the immune system is probably influenced unspecifically, especially by increasing the macrophage activity. The efficacy for viral respiratory tract infections has been proven yet again in recent GCP-compliant, clinical studies (15, 17). Under the influence of the phytotherapeutic the duration of the illness decreased with a responder rate that was significantly higher than under placebo. The therapeutic benefit was even more pronounced, if the pharmaceutical was applied as early as possible. It has a high drug safety.