Molecular Identification Of The Cockroach Eupolyphaga Sinensis In A Chinese Prescription, Huoxuezhitong Capsule

Huoxuezhitong capasules is one of the state's basic drugs. It is the national protected variety of traditional Chinese medicines. Its prescriptions for:Angelica Sinensis Radix, Notoginseng Radix et Rhizoma, Olibanum (vinegar), Eupolyhaga, Pyritum and Borneolum syntheticum. This prescription were reported in the first "ZhaoBingNa clinical experience set". It is the classic recipe treating bruises, bleeding sore.it is not only relive pain and disperse biood stasis, but sife effects.Eupolyphaga sinensis, another name TuYuan etc, insect, containing many kinds of amino acid, naphtha, fat aldehydes and aromatic aldhydes, alkaloids, beta sitosterol, shark liver alcohol, uracil and allantoin, still contains a variety of trace elements. In our country has a long medicinal history.Because contain animal medicine in preparations, and animal medicine quality standards are now mostly for microscopic identification organizational structure, such as the bristles of this method in biology appraisal marks are the genetic phenotype for species, with a great variability and plasticity, and greatly influenced by appraiser subjective judgment in compound, complex, medicinal materials composing microstructure form under the condition of the damaged more, microscopic identification much restricted. And TLC, high performance liquid chromatography is only identification methods such as identification of one or several medicinal chemical composition, if to extract medicine or directly to join a appraisal composition, the substitution materials by second-stage medicine to identify such methods. The development of traditional Chinese medicine, raise the standard basic drugs action the brunt. Therefore, it is urgent to explore a kind of objectivity, intuitive and reliable, strong specificity and has to invigorate the inspection methods of analgesic capsule to identify a terrapin worm.This topic is to identify Eupolyphaga sinensis by the way of moleclar biology. We succeed in identifying it from huoxuezhitong capsule.Established a new way to DNA long-term preservation and a genomic DNA extraction method.In the study basing on 16S rRNA sequencing technologies, for the inadequate of traditional methods that genomic DNA of Eupolyphaga sinensis has serious degradation, we explored the three methods containing hot waterand ethanol,95% ethanol to death, after drying preserve 0 months, months and 6 months. The results showed that after saved 6 months Eupolyphaga sinensis through 95% alcohol, genomic DNA is good, without degradation. As the features of complex composition for the Chinese herbal compound, genomic DNA extracted is a difficult problem, this experiment separately inspected the proteinase K method, proteinase K modified with adsorption column method and improved method of proteinase K modified adsorption column to extract genomic DNA of huoxuezhitong capsules and Eupolyphaga sinensis. The results showed that three methods all can extract good genome DNA of Eupolyphaga sinensis, but only improved method proteinase K modified adsorption column can succeed extracting with huoxuezhitong capsules.