Authentication of the Panax genus plants used in Traditional Chinese Medicine (TCM) using Randomly Amplified Polymorphic DNA (RAPD) analysis

    Research output: ThesisMaster's Thesis

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    [Truncated abstract] Traditional medicines are used by millions of people throughout the world as their primary source of medical care. A range of materials are in used traditional medicines including plant and animal parts. Even though the traditional medicine trade is estimated to be worth sixty billion dollars annually the trade remains largely unregulated. Unscrupulous practices by vendors to increase their profit margins such as substituting and adulterating expensive material with cheaper varieties go unchecked. This can be dangerous to consumers because some substitutions involve poisonous material. Also, animal parts from endangered species can find their way into traditional medicines, therefore there needs to be a way to identify them in traditional medicines to prosecute poachers. The traditional techniques used for the identification of material used in Traditional Chinese Medicine (TCM) include, morphological, histological, chemical and immunological analysis. However, these techniques have their limitations. This makes applying multiple techniques essential to provide thorough authentication of the material. DNA profiling provides a technique well suited to analysing material used in TCM. DNA profiling is advantageous over other techniques used to authenticate material used in TCM because it requires only a small sample amount, can determine the cultivator, be used on all forms of TCM and potentially distinguish the components of mixtures. ... Therefore, profiles of different species/individual are different and species? can be distinguished. Commercially sold traditional medicines are processed which is likely to degrade the DNA of the sample making extraction and amplification difficult. Here an organic Phenol:Chloroform extraction technique extracted DNA from commercial dried root samples. The extracted DNA was amplifiable using RAPD primers. The RAPD primers used here produced enough polymorphic bands to distinguish different plant species. They were used to distinguish commercial samples that were sold as three different species within the Panax genus, Panax ginseng, Panax quinquefolium and Panax notoginseng and genetically unrelated plant material; Potato and Eleutherococcus senticosus. Liquid samples and mixtures were also profiled with the RAPD primers to determine whether the RAPD primers provide enough distinguishing ability to analyse these forms of TCM. DNA was extracted from the liquid samples, one a ginseng drink and the other an ginseng extractum. However, there was no reliability in the production of PCR products. The analysis of the mixture samples found that not enough polymorphic bands were produced by the RAPD primers used here to identify Panax species within mixtures of two Panax species. While when P. ginseng was mixed with a genetically unrelated sample there was enough polymorphism to differentiate the two samples in the mixture. The results of this research show that RAPD analysis provides a simple and inexpensive technique to begin analysis of materials used in TCM. Using RAPD analysis it is possible to distinguish Panax plant species from each other. However, the RAPD primers used here did not provide enough reproducibility or polymorphism to analyse liquid and mixtures of Panax species plants.
    Original languageEnglish
    Publication statusUnpublished - 2007


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