【Oviductus,Ranae,method,of,extraction,and,isolation,of,active,ingredients,progress】 ingredients是什么意思

【www.zhangdahai.com--转正述职报告】

　　【Abstract】 Oviductus Ranae is one of valuable traditional chinese medicine, with a variety of medicinal effects. For further research and application, this paper reviewed the technologies of the extraction and isolation of efficient components in Oviductus Ranae and analyzed the advantages and disadvantages.
　　【Key words】Oviductus Ranae; active ingredients; extraction andisolation
　　
　　Oviductus Ranae is the dry oviduct of female Rana temporariachensinensis David,which is also called Rana chensinensis oil. Themedicine was recorded in chinese pharmacopoeia from edition 1985 to edition 2005, and it has effects of invigorating the spleen and the stomach, nourishing yin and the kidney, moistening the lung and promoting the production of the body fluid[1]. Oviductus Ranae is a valuable chinese medicine, which is also known as soft gold. Oviductus Ranae is applied to cure weakness, insomnia, night-sweat, neurasthenia, climacteric
　　syndrome and so on. Oviductus Ranae contains proteins, amino acids, fatty acids and esters, steroids, nucleic acids, sterols, amides, hydantions, vitamins and many other active ingredients. This paper reviewed the development of the extraction and isolation technologies of efficient components in Oviductus Ranae, in order to facilitate further research and development use.
　　1Fatty acids and esters
　　These conponents have similar effect as prednisone stimulating the syntheses of protein, thus improve the powers of endurance and have anti-fatigue effect[1、2].
　　Lu Li ming[3、4] got hexadecanoic acid, 2,3- dihydroxypropyl ester, hexadecanoic acid, 2-hydroxy-1,3- propanediyl ester, 9-octadecenoic acid,2,3-dihydroxypropyl ester, octadecanoic acid, hexadecanoic acid ethyl ester and octadecanoic acid ethyl ester from Oviductus Ranae by supercritical carbon dioxide fluid extraction technology combining column chromatographies on silica gel（mobile phase: petrol ether-chloroform, petrol ether- ethyl acetate ）, gel and AgNO3(mobile phase: chloroform- acetone). Wang Yong sheng[5] got hexadecylic acid cholesterol ester from petroleum ether extraction of Oviductus Ranae with silica gel column chromatography（mobile phase: petroleum ether-ethyl acetate）and recrystallization(petroleum ether-ethanol).
　　Comparing with other traditional methods, supercritical carbon dioxide fluid extraction technology has many special advantages, such as high extraction rate, strong extraction power, low-temperature operation, protecting the active ingredients from damage. This method applies to lipid components extraction, especially fats oils. The silica gel column chromatography is distinct in isolation, but presents high adsorption rate to sample, so to Oviductus Ranae, rare medicinal herbs, this method should be used as little as possible.
　　2 Steroids and sterols
　　These conponents also have similar effect as prednisone stimulating the syntheses of protein, thus improve the powers of endurance and have anti-fatigue effect[1、2].
　　Some reserchers obtained estradiol, vitamin VA and cholesterol from Oviductus Ranae[6-8], which were extracted with acetone, and thenisolated with Al2O3 column chromatography and preparative silica thin-layer chromatography. Zhao Wen ying[9] got cholest-3,6-dione, cholesterol and cholest-4-en-3-one from Oviductus Ranae, the method was that Oviductus Ranae was seccessively extracted with petroleum ether, chloroform, acetone and methanol with soxhlet extractor, then followed through a column of silica(mobile phase: petroleum ether-ethyl acetate). Wang Yong sheng[5] obtained cholest-4-en-3-one, cholesterol from Oviductus Ranae by extractiong Oviductus Ranae with petroleum ether, isolating and purifing with silica column chromatography(mobile phase: petrol ether-ethyl acetate）, recrystallization(petroleum ether-ethanol, acetone ).
　　Wang Li li[10] obtained cholest-5-en-3β,7β-diol and 3β-hydroxy-cholest-5- en-7-one from Oviductus Ranae for the first time. The process was that Oviductus Ranae were pre-extracted with supercritical carbon dioxide (for 4.5h, at 45℃, under 35MPa, 55mL/100g ) to remove the lipin components, then the degreasing Oviductus Ranae were extracted with 85% ethanol. Subsequently, the dried ethanol extracted residue was dissolved in deionised water and extractedwith ethyl acetate, and then the ethyl acetate extraction was concentrated and applied onto columns of sephadex LH-20(mobile phase:chloroform- methano 1:1) and ODS(mobile phase: 90% (v/v) methano) repeately to gave crude products, which were purified by preparative HPLC.
　　 Column chromatographies of gel and ODS have been widely used in studies on the chemical constituents of traditional chinese medicine.Gel column chromatography rests on varying molecule and fits to the chemical components isolation of vare medicines because of easy operation, fine separation, low adsorption rate to sample. The basis of separating and purifying chemical components of ODS column chromatography is varing molecular polarity. The main features of this method are easy operation and low adsorption rate to sample, so this method is also applied to research of chemical ingredients of vare medicines.
　　3Amides
　　Wang Yong sheng[5] got hexadecanamide from petroleum ether extraction of Oviductus Ranae by silica column chromatography(mobile phase: petroleum ether-ethyl acetate）, recrystallization(petroleum ether-ethanol and acetone ). Lu Li ming[3] also obtained hexadecanamide with supercritical carbon dioxide combining column chromatographies of silica gel（mobile phase: petroleum ether-chloroform and petroleum ether- ethyl acetate ）, gel, AgNO3 gel(mobile phase: chloroform- acetone).
　　4Polysaccharide
　　Polysaccharide can strengthen the immunity, it can not only enhance meshy endodermis system to gobble up a function, but slao increase the amount of immuglobulin. Polysaccharide is effective on tumor and cardiovascular diseases[11].
　　Polysaccharide were separated form Oviductus Ranae showing positive reactions to Fehling and Molish. The method was thatOviductus Ranae was extracted with water decocting and separated with ion-exchange process. The result of quantitative determination was that the content of reducing suger and polysaccharide were 6.8% and 6.18% in Oviductus Ranae.
　　5Hydantions
　　1-methylhydantion had the extremelyeffect ofrelieving cough[13].
　　Wang Yong sheng[13] got 1-methylhydantion from Oviductus Ranae for the first time. The method was that Oviductus Ranae was successively extracted by petrol ether, ethyl acetate and ethanol, subsequently, the ethanol extraction was separated with silica column chromatography（mobile phase: petrol ether- ethyl acetate） and purified with recrystallization(petroleum ether- chloroform).
　　6Conclusion
　　At present, the stuty on Oviductus Ranae has many difficulties, such as the complex chemical ingredients, the low content of active components and the rarely reports about effective ingredients in Oviductus Ranae. So the basic research of Oviductus Ranae is more weaker than that of other valuable chinese medicines. But the wide application of gel column chromatography, ODS column chromatography, microwave extraction, semi-bionic extraction (SBE), CO2-SFE, ultrafiltration and high-speed countercurrent chromatography provides advantage for obtaining high purity active ingredients form Oviductus Ranae, so the extraction and isolation of the active ingredients in Oviductus Ranae is yet to be further studied.
　　
　　References
　　[1]Pharmacopoeia of the People,s Republic of China[S]. One volume 2010.
　　[2]Zhang S. Effect of frog oil in eliminating sports fatigue and its mechanism[J]. Chinese Journal of Clinical Rehabilitation, 2004, 8(12): 2340.
　　[3]Lu L M, Song S J, Wu Y, et al. Study on the chemical constituents of the Ovum Oil of Rana temporaria chensinensis David Ⅱ[J]. Journal of Shenyang Pharmaceutical University, 2002, 19（1）: 25-26.
　　[4]Lu L M, Song S J, Xu S X, et al. Study on the chemical constituents of the Ovum Oil of Rana temporaria chensinensis David[J]. Chinese Journal of Medicinal Chemistry, 2001, 11(4): 224.
　　[5]Wang Y S, Jiang D C, Wang E S. Studies on fat-soluble chemical constituents of Oviductus Ranae[J]. Chinese Traditional Patent Medicine, 2006, 28(1): 93-94.
　　[6]Wang L L, Wang Z G, Jiang Y G. Analysis of constituents of Oviductus Ranae[J]. Chinese Traditional and Herbal Drugs, 1982, 13(9): 5.
　　[7]Lin J X, Zhou Z S, Yu Z Z, et al. Analysis of ovduct testosterone of Oviductus Ranae and Bufo bufo gargartzans[J]. Chinese Traditional and Herbal Drugs, 1989, 20(7): 11 -14.
　　[8]Bai Y, Yang X Y. Extraction and assay of testosterone in Ranae temporaria chensinensis David[J]. Heilongjiang Medical Journal, 2003, 16（2）: 97.
　　[9]Zhao W Y, Hou Z R, Li J X, et al. Studies on chemical constituents of Oviductus Ranae[J]. Journal of Shenyang Pharmaceutical University,1997, 14(1): 24.
　　[10]Wang L L. Studies on the chemicalconstituents of the active fraction with relieving cough of Oviductus Ranae[D]. Changchun: Changchun University of Chinese Medicine, 2009
　　[11]Gao Y L. The effects of Oviductus Ranae from Gansu province on immunity and irritability of organism in mice[J]. Journal of Chinese Medicinal Materials, 1996, 19(2): 90.
　　[12]Fan Y L, Cui X S, Yao Y X, et al. Research advance on Oviductus Ranae composition[J]. Journal of Jilin Agricultural University, 1996, 18(3): 105.
　　[13]Wang Y S. The study on drug efficacy ingredients of Oviductus Ranae and its quality control method[D]. Changchun: Jilin university, 2007.
　　
　　作者简介： 王丽丽（1982-），女，吉林松原人，石家庄医学高等专科学校，助教，硕士学位， 研究方向：天然药物有效成分的研究。
　　*通讯作者：王永生（1962-），男，吉林长春人，吉林大学，博士，硕士生导师，研究方向：天然药物有效成分的研究。

推荐访问:method extraction Oviductus Ranae