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乙醇提取龙眼核总黄酮及鉴别
作者:  黄锁义,唐玉莲,黎海妮

《时珍国医国药》 2006年 第10期

  多个检索词,请用空格间隔。
       【关键词】  乙醇提取;,,龙眼核;,,总黄酮;,,鉴别
       摘要:目的探讨龙眼核中总黄酮的提取及鉴别方法,以充分利用龙眼核植物资源,避免资源的浪费。方法 采用乙醇提取法从龙眼核中提取黄酮类物质,对所提取的黄酮类物质进行验证,并用分光光度法测定其含量。结果测得样品中总黄酮的含量C=1.634mg/ml,回收率为99.65%,其纯度和产率均较高。结论该方法采用全物理过程,无任何污染,是提取龙眼核黄酮类物质的有效途径。
       关键词:乙醇提取;  龙眼核;  总黄酮;  鉴别
         (Department of Chemistry, Youjiang Medical College for Nationalities,Baise, Guangxi ,533000,China  )
       Abstract:ObjectiveIn order to make use of the resources of  longan seed  , avoid  wasting and approach the extraction of total flavanone of longan seed.MethodsThe flavonids were extracted from longan seed  with ethanol .Using spectrophotometry to extract and check the flavanone of longan seed. Results By this method getting the contents of the total flavanone of longan seed  is C=1.634mg/ml and the rate of  recovery is 99.65%. ConclusionThis text provides the extraction and purifying methods to get the outcome and the purity of the flavanone is all very high .This method is a purely physical process and has not any pollution. It is an ideal way to extract the flavanone of longan seed.
       Key words:extracted by ethanol;  longan seed;  total flavanone;  identification
       Is often the green fruit tree for the sapindaceae longan,Longan"s fruit common name dried longan, also names Long Mu, compared to item, the longan, the Asian litchi chinensis, the longan, the Sichuan billard ball, the pure black horse bead, Yan Luan, the honey spleen,the jiaolei, the wooden bomb, embroiders the wooden group and so on,For our country famous subtropics fruit[1].The longan originally produces south our country with Vietnam"s north South Asia tropics region, had more than 2000 years cultivation history in our country, is the important South Asia tropics often the green longevity fruit tree, is one of four Lingnan good fruits.Li Shi Zhen said in "Native Chinese Plants" that, "Food take litchi chinensis as expensive, but profits take the longan as is good."Longan"s nutritional value is very high, is south one of famous good fruits. The longan is the important raw material for medicine.Su Song composes "Chart after Book on Chinese medicine", said longan "Gansu even non-toxic, mainly treats the five internal organs perverse trend, satisfied the mind and pevented the food, long having can strong soul intelligent, enlivens is not old, feel enerthing good".Li Shi Zhen "Native Chinese Plants" also said the longan has "whets the appetite is good for the spleen, makes up the empty longan" the effect.The longan meat may treat the debility of long illness which makes the people feel weakly,Loses sleep , easy to forget,to palpitate with fear, as well as spleen weakness diarrhea, post-natal edema and so on sickness.In addition, the longan shell, the longan nucleus, dragon vertigo and the dragon eyelid, the root may take blends Chinese native medicine.Longan nuclear nature of a medicine astringent, even, is mad micro, taste pale but micro painstakingly, has stops bleeding, stops, regulates the flow of vital energy is wet and so on the effect, uses in treating the wound hemorrhage, the hernia, the mange, the skin disease attributed to dampness and so on[2,3].The longan nucleus contains the alkaloid and the fatty oil and so on,Except black seed shell,At the end of grinding or fries black at the end of grinding to externally apply,May treat the wound hemorrhage;Or spreads the treatment for burn with the tea oil accent[4]."Compendium Appropriates lost articles" said that, "Every family has the young children, this cannot be unprepared, wounded by cutting, spreads it by this, stops  bleeding and lives the muscle, after can produce again , other  medicine cannot be copared , not as good as it does."The longan nucleus weight accounts for the longan fruit fresh weight 17%, because has not carried on the comprehensive utilization, abandons every year the longan nuclear weight amounts to several hundred thousand tons, causes the massive longans nucleus to waste in vain[5].But,At present the total flavanone withdraws the research to the longan nucleus in not to see the report.Therefore, we withdraw the total flavanone for the first time from the longan nucleus, and determines its content, discovered in the longan nucleus includes the massive flavanone ingredient. Now reports as follows.
        1   Instrument and reagent
       1.1   Instruments KQ5200DB numerical control ultrasonic wave purifier (Kunshan supersonic instrument limited company); UV-755B ultraviolet visible spectrophotometer (Shanghai analysis instrument main plant); ZF-I three uses the ultraviolet analyzer (Shanghai to attend to village photoelectricity instrument plant); SHB-III water-circulation type multifunction vacuum pump (Zhengzhou great wall scientific industry and trade co.ltd ). 800B centrifuge (on Hai"an pavilion scientific instrument factory).
       1.2  Reagents  95% Ethyl alcohol; Sodium nitrite AR; Aluminium nitrate AR; Sodium hydroxide AR; Aluminum trichloride AR; Hydrochloric acid AR; Ammonia water AR; Glacial acetic acid AR; Ethyl acetate AR; Magnesium acetate AR; Methyl alcohol AR; Magnesium powder AR; Normal butyl alcohol AR; Standard rutin (Chinese drugs biological preparations examination); Hydrogen peroxide solution AR; Ferrous sulfate AR; Salicylic acid AR. longans nucleus originates from the Guangxi Baise.
         2  Methods and result
       2.1  Total flavanone ingredients withdraw  Take the fresh longan seed, drying, the smashing. The name takes longan seed powder approximately 6g, adds the 80ml 95% ethyl alcohol, the ultrasonic wave withdraws 2.5 h, pulls out filters. The filter residue adds the 40ml 95 % ethyl alcohol again, the ultrasonic wave withdraws 2.5h, pulls out filters, merges two times of filtrate, the reduced pressure recycling ethyl alcohol to filtrates only remains 5~7ml up to, lays aside in the 250ml measuring flask, dilutes with 60% ethyl alcohol to the scale division, results in the sample fluid.
       2.2  Determinations methods restOn take the grass cutter as the comparison determination longan seed in the total flavanone content, joins the aluminum ion reagent, at the same time controls is suitable the pH value, causes the flavanone compound and the aluminum salt forms the complex compound, can obtain the stable characteristic absorption peak in the visible light area.
       2.3  Quantitative experiments total flavanone content determination
  
  2.3.1  Wave lengths choiceSample fluids right amount, in under the 0.30ml 5% sodium nitrite solution existence alkalinity condition, after the aluminium nitrate coloration, attracts the luminosity take the reagent as the spatial dried ginseng compared to the fluid in the 420~700nm wavelength range determination chelate, the chelate has the biggest absorption in the 510nm wave length place, when determines selects this wave length.
       2.3.2  Standard curves plan distinctionsPrecisely absorb the grass cutter to compare the fluid (0.10mg/ml) 0.00, 0.50, 1.00, 2.00, 3.00, 4.00, 5.00ml in the 10.00ml measuring flask, joins 5% sodium nitrite solution 0.30ml separately, swings uniform, settles 6min; Again adds 10% aluminium nitrate solution 0.30ml, swings uniform, settles 6min; Again adds 4% caustic soda solution 4.00ml, dilutes with 60% ethyl alcohol to the scale division, swings uniform, settles 12min, makes the blank by the reagent, absorbance was determinated at 510nm place. determination result of absorbance to see Table 1.
       Table 1 determination of absorbance by ultraviolet visible spectrophotometer (略)
       Results in the regression equation according to on table: A=-2.208×10-2+13.7578C,r=0.9999.
       2.3.3  Withdrawing content determinationPrecise absorption sample fluid 1.00ml, sets at the 10.00ml measuring flask, absorbance A=1.102, according to the standard curve preparation method determination, according to regression equation: A=-2.208×10-2+13.7578C, in computation sample total flavanone content C=1.634mg/ml.
       2.3.4  Recyclings experimentsPrecise quantity sample fluid 1.00ml, joins standard grass cutter comparison 2.00ml (0.10mg/ml), attracts the luminosity with the front sample determination method operation determination, extracts the returns-ratio is 99.65%.
       2.4  Qualitative experimental total flavanone distinction
       2.4.1  Colors respondedUnder the ultraviolet ray assumes the color response: Takes this sample solution on the filter paper, assumes the yellow under the visible light, assumes the dust color under the ultraviolet ray and has the fluorescence spot.Strong ammonia water response that, Takes this sample solution on the filter paper, smokes the filter paper above the ammonia water 0.5min, observes immediately under the ultraviolet ray, assumes the extremely obvious tan fluorescence spot.Aluminum trichloride response that, The sample solution on the filter paper, adds by drops 1% aluminum trichloride ethyl alcohol solution, air-dries. Assumes the yellow under the visible light, assumes the decadent fluorescence spot under the ultraviolet ray.Magnesium acetate response that, The sample solution on the filter paper, adds by drops 1% magnesium acetate methyl alcohol solution, air-dries, under the ultraviolet ray assumes the decadent spot.The hydrochloric acid - magnesium powder responded that, Takes the ethyl alcohol to extract 1ml to add the magnesium powder in the test tube, again joins the thick hydrochloric acid number drop (1 time to join), assumes the purple red in the froth place.
       2.4.2  Paper chromatographicAnalyses sample solution 10 ml on filter paper. With normal butyl alcohol: Acetic acid: Water =4: 1: 5 for launches the medicinal preparation, previous line launches 5h, takes out air dries. Spurts 1% aluminum chloride ethyl alcohol solution. After air-dries under the ultraviolet ray, obviously fluorescence spot.
       3  Discussions
       3.1  Experimental results indicated that, this experimental technique craft simple, the returns-ratio is 99.65%, the extraction efficiency is higher than the traditional method, the ultrasonic wave which use this article provides withdraws, yellow ketones matter its purity which the purification method but obtains is high. The ultrasonic wave may strengthen the ethyl alcohol to soak the formulation, achieved time-saving, is highly effective, the energy conservation goal, this method uses the entire physical process, does not have any pollution, is ideal withdraws the yellow ketones matter the way, has the broad application prospect.
       3.2  Flavonoid are big kind of natural products, widely exists in the vegetable kingdom, is many Chinese medicine effective components, has the very high for medicinal purposes value. In the nature most common is the flavanone and the flavonol, other yellow (are mellow), the isoflavone including the double hydrogen, the double flavanone, the flavanol, looks up your alkone, the orange alkone, the variety glucoside and the new yellow ketones and so on[6]. The natural origin biological flavanone molecular weight is small, can by the human body rapid absorption, be able through the blood brain barrier, to be able to enter the fatty tissue, then manifests the following function: Eliminates weary, protects the blood vessel, guards against the arteriosclerosis, the expansion blood capillary, dredges the microcycle, the anti-fat oxidation, anti-is senile, activates the cerebrum and other internal organs cells functions.The flavonoid has the anti-cancer, the anti-tumor, anti-heart blood vessel of brain disease, the anti- inflammation analgesia, the immunity adjustment, falls the blood sugar, treatment osteoporosis, the bacteriostasis anti-virus, the anti-oxidation, anti-is senile, function and so on anti-radiation[7]. In recent years, in the world raised the plant medicine development upsurge, the plant medicine has doubled by its natural low poisonous characteristic receives favors, but the flavonoid was conspicuous by its Guang Pu pharmacological action, the longan seed distributed widely, this article tested the result to indicate, the longan seed center included a richer flavanone. it withdraws the thing total flavanone to have the very good scavenging action to the hydroxyl radical.Therefore, the longan seed has the widespread development and the use value, is worth the comprehensive utilization, strengthens the resources development.
       References:
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       [2]  Xu Jiankai. Longan high production technology question and answer[M]. Guangzhou: Guangdong Science and Technology Publishing Press,2001:56.
       [3]  Tan Shuhui, Zhan Reting. Chinese medicine open country recognition handbook (2)[M]. Guangzhou: Guangdong Science and Technology Publishing Press,2004:208.
       [4]  Nanjing college of pharmacy << Chinese medicine study >> compilation group Chinese medicine study (center book)[M]. Nanjing:Jiangsu People"s Publishing Press,1976:628.
       [5]  Xiao Gengsheng, Huang Ruqiang,Ceng Qingxiao,etc. Nutrition composition of Longan seeds[J]. Food Science and Technology,2004,(1):93.
       [6]  Xia Xingzhou, Zhang Hui, Wei Chuanwan. In the banyan tree leaf the flavonoid withdraws the condition research[J].Food Research and Developent.2002,23 (5):35.
       [7]  Huang Suoyi, Yang Wenhui, Li Weibin,etc. The total flavanone of murraya paniculata extraction and the identification by ultrasonic wave[J].shizhen Medicine and Materia Medica Research,  2006,17(3):395.
       (右江民族医学院化学教研室,广西 百色  533000)

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