| Oviductus Ranae is the dry oviductus of the thely-Rana temporaria chensinensis David, which is also called Rana chensinensis oil. The medicine was recorded in Chinese pharmacopoeias from edition 1985 to edition 2005, it has the effectiveness of invigorating the spleen and stomach, nourishing the yin and invigorating the kidney, moistening lung and promoting the production of body fluid. Oviductus Ranae is the precious and famous Chinese crude drug which is animality and support-healthy energy, and it has widespread application foundation in our civilian. Rana temporaria chensinensis David is mainly produced in the northeast Changbai Mountainous Areas, and Jilin province is the main place of production, which has its history origin, and the product is well-known as the excellent quality. At present, Oviductus Ranae has been an endemic and genuine medical material for medicine in the northeast Changbai Mountainous Areas. Because of its steep economic value and the geographic setting of Jilin province, Jilin province, which is our country's depot of the northern medicine, has list Oviductus Ranae as one of ten varieties which were first approved in GAP. Oviductus Ranae is the valuable raw material in the trade of medicine, light industry, foodstuff and so on, it has excellent economic value, nutritive value and health care effect. Now there are various kinds of dosage forms of health care products on the market, at present probably eleven dosage forms and sixty kinds of products have gone on sale. There are not only some frequent dosage forms, for example oral solution, powder preparation for infusion, soft capsule drug granule and so on, but also many deep processing products whose raw material is Oviductus Ranae, for example Xuehadabu amoxicillin, Xueha radix spirit, Dabu wine, Zhuangyangchun wine, Rana temporaria chensinensis David wine, frog fondants and so on. These products sell briskly internal, furthermore many of these are fist products which can export and create profits. As one of three most distinguished treasures of the traditional Chinese medicine in the northeast Changbai Mountainous Areas, Oviductus Ranae and its correlated products have become one of important products in the local medicine industry.Although Oviductus Ranae obtains extensive application, at present because the production of Oviductus Ranae is not standard, the quality is not unification, the results are the low technology contents and the instable quality, which have seriously affected the elevation of economic returns and restricted the development of Oviductus Ranae. In April 2003,"Jilin changbai forest frog's oviduct", as the patria product of Jilin province, has been approved to practice the protection of patria product by State General Administration of the People's Republic of China for Quality Supervision and Inspection and Quarantine, the government standard of its products has also been published carried out by AQSIQ. However, the index of physics and chemistry proposed by this standard has not powerful specificity and sufficient science nature, so it can't represent and reflect the internal quality of Oviductus Ranae, which are disadvantages to develop the industry of wood frog overall. This phenomenon has restricted the elevation of the quality and credit of Oviductus Ranae, and also affected the products'export. At present although studies and reports about Oviductus Ranae are a good many, a few ones can reflect its drug action substances truly. The main reason is that the fundamental study of Oviductus Ranae is weak to a great extent.The foundation of study of Oviductus Ranae's quality control method is the study of drug efficacy ingredient. As the animality Chinese crude drug, its study has considerable difficulty, and its fundamental study is very weak compared to other valuable Chinese crude drug. At present the profound study of its index nature, specificity or characteristic chemical constituent is not clear-cut enough. And because the underfined characteristic chemical constituent, the pharmacologic action which can reflect the traditional effectiveness of Oviductus Ranae is short of foundation, and this in turn leads to the result that the systematic and penetrating quality study of Oviductus Ranae is short of sufficient technical foundation. This is a problem that needs to be solved urgently. Therefore we need to develop the fundamental study of Oviductus Ranae's pharmacodynamic action substances profoundly and search drug efficacy ingredients or specificity ingredients to reveal the internal quality rule of Oviductus Ranae overall, which have great significance. Now there are many studies and reports about identification of Oviductus Ranae, the numbr of adulterants and spurious breeds reported is at least ten, which leads to the result that there exists differences among the internal qualities of Oviductus Ranae. However at present a universal phenomenon is that with the popular of Oviductus Ranae, some common adulterants and spurious breeds, for example Heilung-kiang wood frog, pond frog oil, toad oil and so on, their purchase quantity and market price are higher increasingly. Where are all these adlterants sold? Obviously, if there isn't a reliable quality control index of Oviductus Ranae and its related products, its internal quality isn't guaranteed at all, which will affect the credit and economic returns of Oviductus Ranae ultimately. So a practicable, scientific and reasonable quality criteria is an important scientific foundation for Oviductus Ranae to develop, utilize and produce for sale, and the quality control system which consists with the desire of international market is also the important guarantee to raise its added value and export to create profits.In this article, we, connecting the basic theory of traditional Chinese physician and medicine on the basis of inheriting sufficiently our nation's precious heritage for several thousand years, and combining study achievements about modern animal medicine chemical composition, proposed a new idea to study animal medicine chemical composition firstly, the new idea is that the study of animal medicine chemical composition is attached to the traditional Chinese medicine's study system. Therefore, its study should connect the basic theory of traditional Chinese physician and medicine and personify the peculiarity of traditional Chinese physician and medicine. The view about main drug efficacy ingredients in the traditional Chinese medicine is illuminating those micromolecule drug efficacy compounds in the traditional Chinese medicine, which establishes the guiding ideology for our study.In the study of drug efficacy ingredient of Oviductus Ranae, we, on the basis of Oviductus Ranae's traditional drug action which is strengthening with tonics and stopping coughing, filtered the drug efficacy ingredients of Oviductus Ranae to ensure that the extract by prtroleum benzene is its effective spot of strengthening with tonics, and the extract by alcohol is its effective position of stopping coughing. We abstracted and indentified compounds of hexadecanoic acid cholesterol ester, hexadecanoic acid, hexadecane amide, cholest-4-ene-3-ketone, cholest-3,5,6-tri-itol, choleterin and so on by the method of gel silica column chromatography which is also integrated with the preparation TLC. And we identified thirty-four kinds of fatty acids by GC-MS; we abstracted a prism crystallization from the extract by methanol which is Oviductus Ranae's effective position of stopping coughing. We definited this compound is 1-methylhydantion through the method of elementary analysis, IR, NMR and X-ray diffraction, IR, NMR and X-ray diffraction. This compound is abstracted from Oviductus Ranae for the first time. The initial pharmacodynamics experiments confirm that hexadecanoic acid cholesterol ester has a better function of strengthening with tonics and 1-methylhydantion has a better function of stopping coughing, so they can be listed as drug efficacy ingredients of Oviductus Ranae, which initially solved the status quo that the drug action substances of Oviductus Ranae are epinephelus constantly. And we successfully synthesized 1-methylhydantoin by the method of chemical synthesis, and we also verified its drug action through the further pharmacodynamics experiments. We, on the basis of these works, declared the application patent of 1-methylhydantoin in the preparation of relieving cough and reducing sputum medicine, which can supply the safeguard to study and develop this compound profoundly. Generally speaking, all these findings can establish the foundation to set up a scientific and reasonable quality control method of Oviductus Ranae.In the study of quality control method of Oviductus Ranae, we made a full-scale study for the qualitation identification of Oviductus Ranae by the methods of color spectrum differential method (TLC, HPLC ) and light spectrum differential method to establish a stable qualitation differential method. Through the study of TLC, we comprehensively inspected its TLC condition for the first time, and established a quick, direct and convenient TLC condition to identify the Oviductus Ranae raw material for medicine own, which can supply the basis to control prerequisite to further study its TLC finger printing and the multiple techniques with other color spectrum and light spectrum. Especially through the study of UV and HPLC finger printings, we not only formulated the standard atlas of the quality goods Oviductus Ranae, but also obtained the feasible means to identify Oviductus Ranae through the contrast with many kinds of Oviductus Ranae. And we, on the basis of these works, undertook the full-scale study to the quality control method of Oviductus Ranae. In the aspect of qualitation identification, we estabished the qualitation identification methods of its TLC, UV, HPLC and so on. Through the contrast study of the quality goods Oviductus Ranae and spurious breed Oviductus Ranae, we detected the quallity goods Oviductus Ranae's supersonic wave extracts in different polarity organic solvents by ultraviolet spectra repeatedly, their peak shapes were similar extremely, which indicated the main components of the quality goods Oviductus Ranae are analogical, and the different contents in some compounds resulted in the differences of their absorption coefficients. We can also found that the locations of characteristic absorption peak wave lengths of the ultraviolet spectras and derivative spectrums of the quality goods Oviductus Ranae were similar through deternimed repeatly, and he reproducibility was also excellent. We compared the characteristic absorption peaks of the ultraviolet spectras and derivative ultraviolet spectras in different polarity organic solvent extracts, through this work we can distinguish the quality goods Oviductus Ranae and spurious breed Oviductus Ranae successfully. Therefore we can obtain the excellent identify effect through comparing its ultraviolet absorption peak shape, ultraviolet characteristic absorption peak and derivate characteristic absorption peak by the ultraviolet spectraline method. And we , on the basis of traditional identification methods, enriched the identification method of Oviductus Ranae in some degree. In the HPLC finger printing study of Oviductus Ranae, we applied a new poly-steps gradient elution method to optimize chromatographic seperation conditions. We, through the contrast of different tipes of chromatographic columns, discovered that the reproductibility of the same tipe chromatographic column was excellent in the finger printing detection of Oviductus Ranae, even the chromatographic columns were different batches. This discover is profit to choose chromatographic column in further generalization and application of HPLC finger printing , and also raises its applicability and feasibility strikingly. The mark and specificity of eleven batches of samples in this experiment are powerful, and the stability of twelve common peaks is excellent, all these consist with the related definition of《The Specification of Finger Printing Study of the Traditional Chinese Medicine Parenteral Injection》[104]. One of the features of the traditional Chinese medicine finger printing is the identification of samples. The samples in this experiment mainly come from the main place of production of Oviductus Ranae-Northeast of China, and also considered other several provinces, we undertook the shape identification, semblance direct-viewing analysis and cluster analysis of these samples, these three identification results were concident. Therefore the HPLC finger printing of Oviductus Ranae established in this article possesses excellent application value, which can supply the scientific basis to identify Oviductus Ranae and also provide a new means to control its quality effectively.In the aspect of establishing the method of content assaying, we utilized cholesteryl palmitate and 1-methylhydantoin extracted as control article. And through the study of HPLC technology, we also established the content assay method of Oviductus Ranae for the first time. This fills a blank that there is not the item of content assaying in the quality control method of Oviductus Ranae continuously, which lays the foundation for establishing a scientific quality control system of Oviductus Ranae.Generally speaking, this article, under the instruction of the basic theory of traditional Chinese physician and traditional Chinese medicine, undertook the systemic study of the drug efficacy ingredients of Oviductus Ranae and its quality control method, and we also found the drug efficacy components of cholesteryl palmitate and 1-methylhydantoin which can represent the traditional drug action of Oviductus Ranae. Finally we combined the means of TLC, UV, HPLC and so on to establish the scientific and reasonable quality control method on this basis, which lays the foundation for establishing a scientific quality control system of Oviductus Ranae.
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