From Pharmacognosia to DNA-Based Medicinal Plant Authentication - Pharmacognosy through the Centuries.

For centuries, pharmacognosy was essential for the identification, quality, purity, and, until the end of the 18th century, even for the efficacy of medicinal plants. Since the 19th century, it concentrated on authenticity, purity, quality and the analysis of active substances, and was established as an academic branch discipline within pharmacy and continuously developed into a modern, highly sophisticated science. Even though the paradigm in pharmacy changed in the 19th century with the discovery of morphine and concentrated on single substances that could be synthesized fast by the upcoming industry, medicinal plants always remained an important element of the Materia medica, and during the last decades, medicinal plants continue to be a source of remedies, and natural products are an inspiration for new medicine. In this research, pharmacognostic skills remain an essential element, both with regards to identity, quality assurance of botanicals (both herbal medicines and supplements), and the discovery and development of new medicines. Over the years, the specific pharmacognostical tools have changed dramatically, and most recently, DNA-based techniques have become another element of our spectrum of scientific methods.

The relevance of pharmacognosy in pharmacological research on herbal medicinal products.

As all medicines, herbal medicinal products are expected to be safe, effective, and of appropriate quality. However, regulations on herbal medicinal products vary from country to country, and herbal preparations do occur not only in the form of medicinal products but also as less strictly regulated product groups like dietary supplements. Therefore, it is not always easy for the consumers to discriminate high-quality products from low-quality products. On the other hand, herbal medicines have many special features that distinguish them from conventional medicinal products. Plants are complex multicomponent mixtures; in addition, their phytochemical composition is not constant because of inherent variability and a plethora of external influences. Therefore, the production process of an herbal medicinal product needs to be strictly monitored. First of all, the starting materials need to be correctly authenticated and free of adulterants and contaminants. During plant growth, many factors like harvest season and time, developmental stage, temperature, and humidity have a strong impact on plant metabolite production. Also, postharvest processing steps like drying and storage can significantly alter the phytochemical composition of herbal material. As the production of many phytopharmaceuticals includes an extraction step, the extraction solvent and conditions need to be optimized in order to enrich the bioactive constituents in the extract. The quality of finished preparations needs to be determined either on the basis of marker constituents or on the basis of analytical fingerprints. Thus, all production stages should be accompanied by appropriate quality assessment measures. Depending on the particular task, different methods need to be applied, ranging from macroscopic, microscopic, and DNA-based authentication methods to spectroscopic methods like vibrational spectroscopy and chromatographic and hyphenated methods like HPLC, GC-MS and LC-MS. Also, when performing pharmacological and toxicological studies, many features inherent in herbal medicinal products need to be considered in order to guarantee valid results: concerning in vitro studies, difficulties are often related to lacking knowledge of ADME characteristics of the bioactive constituents, nuisance compounds producing false positive and false negative results, and solubility problems. In in vivo animal studies, the route of administration is a very important issue. Clinical trials on herbal medicinal products in humans very often suffer from a poor reporting quality. This often hampers or precludes the pooling of clinical data for systematic reviews. In order to overcome this problem, appropriate documentation standards for clinical trials on herbal medicinal products have been defined in an extension of the CONSORT checklist. This article is part of a Special Issue entitled "Botanicals for Epilepsy".

[Study on quality standard for mori cortex].

OBJECTIVE: To establish a new evaluation model and compare the differences of Mori Cortex from different habitats at different harvest time. METHODS: TLC method was used to identify sanggenon D in the sample with silica gel G plate and a mixture of chloroform-methanol-formic acid (5: 1:0. 3) as a developing solvent. UV was used for determination the centent of total flavonoids of Mori Cortex with sanggenon C as the reference substance. HPLC was applied for determination the contents of sanggenons C, D and DNJ of Mori Cortex. RESULTS: In the TLC chromatogram, sanggenon D showed a distinct fluorescence spot under UV 365 nm with good separation. In UV, sanggenon C calibration curve showed a good linear relationship at the range of 146.8 - 734.0 microg/mL, the average recovery was 97.4% (RSD = 2.1%); For the HPLC quantitation method, sanggenons C, D and DNJ showed good linear within the scope of 700 - 4 000 microg, 500 - 3 000 microg and 4.8 - 96 microg, respectively. The average recovery of sanggenons C, D and DNJ were 98.8% (RSD = 2.6%), 99.1% (RSD = 2.2%) and 98.9% (RSD = 1.7%), respectively. The contents of index components in samples from different habitats at different harvest time were different. CONCLUSION: The established method is reliable and accurate. It can be used for quality control of Mori Cortex.

[The standard operating procedure of the standardization breeding for Poecilobdella manillensis].

OBJECTIVE: To prepare Standard Operating Procedure (SOP) for the artificial breeding technology of Poecilobdella manillensi. METHODS: The environment of producing area and breeding technology, the control of diseases, reserving seeds for breeding, harvesting and processing,the standard of the product quality and supervision for the standardization breeding of Poecilobdella Manillensis was studied. RESULTS: The average yield of vital specimen of Poecilobdella Manillensis was 420.88 kg/mu, the natural hirudin in 1kg of the living specimen of Poecilobdella Manillensis contained 430,000 AT-U, and the quality index and hygienic standard of all products complied with the regulations and standards of the State. CONCLUSION: SOP is applicable for the breeding of Poecilobdella manillensi for Guangxi district.

[Determination of the total content of astragaloside IV in Radix Astragali].

OBJECTIVE: To determine the total content of astragaloside IV in Radix Astragali. METHODS: The measurement conditions were used as follows: Irregular C18 (4.6 mm x 250 mm, 5 microm) column; mobile phase: acetonitrile-water (32:68); flow rate: 1.0 ml/min; detector: ELSD 2000; the temperature of drift tube: 100 degrees C; gas flow: 2.7 L/min. RESULTS: Eight batches of Radix Astragali from different sources were determined. The stability, precision and reproducibility of the method were studied, RSD <3%. CONCLUSION: There is great difference between the content of astragaloside IV in Radix Astragali by different processing methods.

[Study on quality specification of Rhizoma et Radix Valeriana Jatamansi].

OBJECTIVE: To provide scientific basis for the utilization and development of Valeriana jatamansi by setting up the quality control specification of V. jatamansi. METHOD: The pharmacognostical methods were applied. The extract of V. jatamansi was examined. Moisture and ash were determined. And the bioactive constituents were analyzed by TLC and HPLC. RESULT: The morphological and histological characters of V. jatamansi were observed. Content of total ash, acid-insoluble ash, and moisture of 15 samples from different habitats and times were determined. The qualitative and quantitative analysis of valtrate and acevaltrate by TLC and HPLC were preformed respectively. CONCLUSION: The established method can be used for the quality control of V. jatamans.

Pharmacognostic standardization of Turnera aphrodisiaca Ward.

Turnera aphrodisiaca Ward (Turneraceae) has been used traditionally as an aphrodisiac, stimulant, nerve tonic, and laxative and in kidney, menstrual, and pregnancy disorders. Despite a long tradition of use in the treatment of various ailments, no systematic phytochemical and pharmacological work has ever been carried out on T. aphrodisiaca. The authors suggest that the major stumbling block in systematic exploration of the plant is non-availability of authentic plant material. In the present investigation, various pharmacognostic standards for the plant have been generated so that authentic T. aphrodisiaca could be explored for its traditional claims. Microscopically, T. aphrodisiaca leaf showed the presence of abundant unicellular, warty, non-glandular trichomes, anomocytic stomata, and a large number of calcium oxalate crystals along the veins. Powdered stem of the plant showed lignified spiral and pitted tracheidal vessels, and pericyclic fibers were observed in powder microscopy of stem. Total ash of the aerial parts of T. aphrodisiaca was approximately eight and four times more than acid-insoluble and water-soluble ash, respectively. The water-soluble extractive value of the plant was slightly higher than its ethanol-soluble extractive value. Volatile oil content of T. aphrodisiaca was found to be 0.44% (wt/vol), the thin-layer chromatography of which exhibited seven spots using toluene:ethyl acetate (93:7 vol/vol) as mobile phase. Thin-layer chromatography of the petroleum ether extract showed nine spots using hexane:dichloromethane (1:1 vol/vol), while the chloroform extract showed 11 spots using toluene:ethyl acetate:glacial acetic acid (35:4:1 by volume). Phytochemically, the plant was found to contain alkaloids, cyanogenic glycosides, steroids, saponins, flavonoids, tannins, carbohydrates, and proteins.

[Discussion on criterions of endanger and protection levels of traditional Chinese medicine herbs].

To efficiently protect the endangered traditional Chinese medicine herbs is essential for the sustainable development of traditional Chinese medicine. On the bases of present species endanger and protection levels, problems in the traditional Chinese medicine herbs are analyzed. The endangered levels of traditional Chinese medicine herbs should refer to the standard of IUCN, and the protection levels should adopt qualitative and quantitative analysis based on the characteristic of traditional Chinese medicine herbs. Some qualitative and quantitative factors are discussed, some useful information was also provided for the establishment of protection levels of traditional Chinese medicine herbs.

[A study on quality standard for Herba Siegesbeckidae].

OBJECTIVE: To establish the qualitative and quantitative methods of Herba Siegesbeckiae. METHOD: A TLC method was used for qualitative identification and a HPLC analysis was applied for quantitative determination of Herba Siegesbeckiae with kirenol as the reference substances. RESULT: Chloroform-methanol-formic acid (25:5:1) as a mobile phase of TLC, the spot of kirenol can be easily detected; Methanol extracts of Herba Siegebeckiae were separated on a Polaris C18 column with acetonitrile-water (25:75) as mobile phase and kirenol was separated well. The average content of kirenol in Herba Siegebeckiae was 0.14%. A good linear relationship between the peak areas and injected amounts of kirenol in the range of 0.19-14.9 microg and the average recovery was 100.0% (RSD = 2.4%). CONCLUSION: The method can be used for qualitative identification and quantitation determination of Herba Siegesbeckiae.

[Chemical pattern recognition of traditional Chinese medicine xixin (I)].

In this paper, three species of traditional Chinese medicine Xixin (Asarum heterotropoides Fr. var. mandshuricum (Maxim.) kitag, Asarum sieboldii Miq. var. seoulense Nakai and Asarum sieboldii Miq.) were identified by means of chemical pattern recognition, with 26 samples as training set and 19 samples as test set. The numerical characteristic features for identification were obtained from GC-MS analysis of volatile oils, all of obtained data were treated with PRIMA (Pattern Recognition by Independent Multivariate Analysis), thus computerized classification of Xixin samples was accomplished. The results agree with those from pharmacognosy. As a new method, chemical pattern recognition is especially suitable for identification of a large number of traditional Chinese medicine samples.

Academic research: policies and practice.

The Bayh-Dole Act of 1980 allowed universities in the US to own and manage inventions obtained using federal funds. This Act laid the foundation for university technology transfer activities in most major research universities of the country. Consequently, the interaction between universities and industry has increased, and so has the sophistication in university intellectual property management. UIC's model for managing its intellectual property is efficient and successful, and is the one increasingly used by university technology transfer offices. The model deals with all aspects of UIC's intellectual property: disclosure, protection, marketing, negotiating and licensing, as well as intellectual property provisions in UIC's research agreements, material transfer agreements, option agreements, licensing agreements and others. In a pioneer effort, UIC has developed a policy on contracts for collecting natural product samples for drug discovery which includes royalty sharing and other important provisions. Accompanying this policy we have also drafted a standard contractual agreement for collectors.