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Molecular Identification of the Cockroach Eupolyphaga Sinensis in a Chinese Prescription, Huoxuezhitong Capsule

Author: WangYunLing
Tutor: ZuoCunHai
School: Yanbian University
Course: Pharmacy
Keywords: Huoxuezhitong capsules Eupolyphaga DNA extraction 16S rRNA Sequencing
CLC: R286.0
Type: Master's thesis
Year: 2011
Downloads: 72
Quote: 1
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Abstract


Huoxuezhitong capsule is included in the National Essential Drugs List as one of the drugs, the State Protected Chinese Medicine. Its pharmaceutical formulations as: Angelica, Panax, frankincense (vinegar), Eupolyphaga, natural copper (calcined), borneol. The prescription was first reported in \Eupolyphaga prescription drugs for the minister, the country has a long medicinal history. Eupolyphaga, also known as land turtle worms, earth element, etc., containing a variety of amino acids, essential oils, aliphatic aldehydes and aromatic aldehydes, alkaloids, β-sitosterol, shark liver alcohol, uracil and allantoin, still contain a variety of trace element. As preparations containing animal drugs, animal drugs and quality standards are mostly microscopic identification of its bristles and other organizational structure, the method identifies the identification of species in biology are genetic phenotype, with great variability and plasticity and influenced by subjective judgments Determined in complex compound Smell, herbs Morphology many cases destroyed, microscopic identification and more restricted. The thin-layer chromatography, high performance liquid chromatography and other methods of identification can only be identified in one or more medicinal chemical composition, terms of extracts of medicinal ingredients or added directly to an identification of the original powder medicine alternative medicine, the use of such methods are difficult to identify. To carry out actions to improve the standard of Chinese medicine, essential drugs bear the brunt. Therefore, the need to explore a strong objectivity, intuitive and reliable, and has a specific test method Huoxuezhitong capsules Eupolyphaga were identified. The topic for Huoxuezhitong capsules Eupolyphaga molecular identification, successful capsules from the compound were identified Huoxuezhitong Eupolyphaga and explore a suitable long-term preservation of DNA Eupolyphaga processing method establishes a genomic DNA was extracted The extraction method, was amplified by PCR to obtain Eupolyphaga 16S rRNA sequences of related species and build phylogenetic trees. Based on 16S rRNA sequencing technology Eupolyphaga molecular identification studies Eupolyphaga traditional treatment methods for DNA degradation serious shortcomings discussed hot burnt to death, ethanol, 95% ethanol were sacrificed after drying save 0 months , three months and six months after the DNA preservation, the results show 95% ethanol processing Eupolyphaga place six months after the DNA preservation integrity, and no degradation. For compound Chinese medicine Huoxuezhitong capsules Smell complex genomic DNA extraction difficult puzzles in this experiment were investigated proteinase K, proteinase K in the adsorption column and modified proteinase K in the adsorption column extraction Eupolyphaga and Huoxuezhitong capsules genomic DNA, results display, three methods are of good quality can be extracted Eupolyphaga genomic DNA, but only modified proteinase K adsorption column method can be successfully combined extract capsules Huoxuezhitong Eupolyphaga genomic DNA.

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CLC: > Medicine, health > Chinese Medicine > Of Pharmacy > In drugs > Drug identification
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