CN104224926A - Traditional Chinese medicine extract for removing cigarette and wine toxins as well as preparation method and application thereof - Google Patents
Traditional Chinese medicine extract for removing cigarette and wine toxins as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine extract for removing cigarette and wine toxins. The traditional Chinese medicine extract is prepared from raw medicinal materials such as abrus cantoniensis hance and lotus leaf and is extracted from the abrus cantoniensis hance and lotus leaf according to a ratio of 27:9.5, and the main component of the traditional Chinese medicine extract is general flavone. The invention also relates to application of the traditional Chinese medicine extract in preparation of medicines or foods for treating and preventing cigarette and wine toxins. The pharmacological experiment proves that the traditional Chinese medicine extract disclosed by the invention can effectively prevent injury of cigarette and wine toxins to liver cells, is prepared into tablets, chewing gums and other oral dosage forms and can be convenient to carry and convenient to take.
Description
Technical field
The present invention relates to a kind of Chinese medicine extract, specifically, a kind ofly separate Chinese medicine extract of tobacco and wine poison and its preparation method and application.
Background technology
Comprise women, harm that child suffers passive smoking every day, plant containing harmful chemical more than 3 000 in medicated cigarette, carcinogen is kind more than 30, has nearly million people to die from and smoking relevant disease every year, cancer especially pulmonary carcinoma and toxicity of cigarette closely related.People is in smoking process, and no matter be passive smoking or initiatively smoking, the water-soluble portion of smoke from cigarette, more easily by the mucolysis of alveolar surface and absorption, causes damage to each tract of health.Therefore, except ban on opium-smoking and the opium trade, outside few smoking, medicine or the food of actively finding tobacco poisoning are significant.
China has long spirits culture, but along with the raising of people's living standard, fatty liver, patients with alcoholic liver disease are more and more, especially alcoholic liver number of patients rises year by year, and in rejuvenation trend, because the living habit that long-term heavy drinking etc. are bad, cause the number of patients of alcoholic liver to rise year by year and be rejuvenation trend.30 to 45 years old treat many business people, salesman, businessman etc. with courtesy is high-risk group.Ethanol has become the chief culprit causing liver cirrhosis patient to be fallen ill obviously, in low age trend.Excessive consumption of alcohol especially Chinese liquor damages liver.Alcoholic liver disease, comprises the liver injury that a series of degree such as alcoholic fatty liver, alcoholic hepatitis, alcoholic fibrosis and liver cirrhosis are different, most caused by long-term heavy drinking.How asymptomatic slight alcoholic liver is, in, heavy alcohol liver can present the performance of similar chronic hepatitis, as slight general malaise, asthenia, fatiguability, nausea and vomiting, inappetence, abdominal distention etc.Due in actual life, inevitably drink, before ethanol is taken in positive reduction, adopt the health of medicament protection alcohol user, there is important social meaning.
Inventor is by early-stage Study, and Herba Abri and Folium Nelumbinis, apart from detoxify or outside expelling summer-heat effect, both are mixed with extract in suitable ratio significantly can reduce the damage of tobacco poisoning to liver cell, alleviate smoke from cigarette and mistake manages the malaise symptoms brought of drinking.
Herba Abri is Lingnan features Chinese medicine, derive from the dry herb of pulse family Abrus plant abrus cantoniensis Hance (Abrus cantoniensis Hance), have another name called Caulis et Folium Desmodii Gangetici, crossdrift bank, yellow food grass, spire Herba Titanotrichi oldhamii, Radix Et Rhizoma Rhei, for the genuine medicinal materials of the southern provinces and regions such as Guangdong, Guangxi, it is also one of Chinese herbal medicine of China's export." south of the Five Ridges gather medicinal herbs record ", " south of the Five Ridges herbal " etc. are all on the books, and Herba Abri heat clearing away, soothing liver-QI, and spleen, use treating acute and chronic hepatitis.2010 editions " Chinese Pharmacopoeias " record, and Herba Abri has dampness removing jaundice eliminating, heat-clearing and toxic substances removing, effect of dispersing the depressed liver-QI for alleviating pain, and for jaundice due to damp-heat, side of body rib does not relax, and epigastric distending pain, acute mastitis swells and ache.Among the people being widely used in treats acute hepatitis, chronic hepatitis and cirrhotic ascites, satisfactory for result, there is the pharmacological action (Chen Xiaobai such as antioxidation, antiinflammatory, immunity simultaneously, Mo Zhixian. Chinese medicine chicken GUCAO chemical composition and Pharmacological Advancement. time precious traditional Chinese medical science traditional Chinese medicines .2008,19 (7): 1781-1782.）。Herba Abri is commonly used it and carrys out Baoshang nourishing in In Guangdong Province.At present except magistral medicines is used, most of export trade outlet.Herba Abri is mainly containing composition (Fu X. such as alkaloid, flavonoid, saponins, Anthraquinones, Wu S.W., Meng H.C., Li C.Isolation and antioxidant activity of polysaccharides purified from Abrus cantoniensis Hance .Modern Food Science and Technology.2013,29 (7): 1559-1564; History hamming, Wen Jing, Tu Pengfei. the chemical constitution study of Herba Abri. Chinese herbal medicine .2006,37 (11): 1610-1613), there is no Herba Abri up to now for separating the report of tobacco and wine poison.
Folium Nelumbinis is the dried leaves of nymphaeaceae plant lotus Nelumbo nucifera Gaertn..Hardship, flat, return liver, spleen, stomach warp, there is clearing away summer-heat, sending up the lucid YANG, effect of cooling blood for hemostasis.For summer-heat excessive thirst, diarrhea due to summer heat and dampness, diarrhea due to hypofunction of the spleen, heat in blood tells nosebleed, metrorrhagia of having blood in stool.Folium Nelumbinis is put into the list that Ministry of Public Health " is food and medicine ", its edible and medicinal two in all have and apply more widely, it is mainly containing alkaloid, flavone, volatilization wet goods composition.Pharmacological research shows, Folium Nelumbinis have adjust fat, fat-reducing, antioxidation, defying age, bacteriostasis (Wang Fugang, Cao Juan, Liu Bin, etc. the chemical composition of Folium Nelumbinis and Advance on Pharmacological Activities thereof. time precious traditional Chinese medical science traditional Chinese medicines, 2010,21(9): 2339-2340).
Therefore develop a kind of can available protecting tobacco poisoning to the damage of liver cell, and to be convenient for carrying the medicine taken be very necessary.
Summary of the invention
The object of the invention is for deficiency of the prior art, a kind of Chinese medicine extract separating tobacco and wine poison.
Of the present invention again one object be that the preparation method of above-mentioned Chinese medicine extract is provided.
Another object of the present invention provides the application of above-mentioned Chinese medicine extract.
For achieving the above object, the technical scheme that the present invention takes is:
Separate a Chinese medicine extract for tobacco and wine poison, the active constituents of medicine of described Chinese medicine extract is extracted by the crude drug of following weight portion: Herba Abri 27 parts, 9.5 parts, Folium Nelumbinis.
For realizing above-mentioned second object, the technical scheme that the present invention takes is:
The preparation method of described Chinese medicine extract, comprises the following steps:
(1) take each crude drug by weight portion according to claim 1, described crude drug carried out decoct, concentrated, add ethanol or methanol carries out precipitate with ethanol, after precipitate with ethanol, collect supernatant concentration;
(2) cross macroporous resin column, the water elution removal of impurity is the ethanol elution of 50 ~ 85% by concentration, collects the eluent at 30-70% position, and concentrating under reduced pressure is dry obtains extract.
The preparation method of described Chinese medicine extract, comprises the following steps:
(1) take each crude drug by weight portion according to claim 1, cleaned by described crude drug, pulverize, crossing 60 mesh sieves, is 30-95% ethanol percolation by concentration, to total flavonoid assay reactions is negative;
(2) extracting solution is collected, concentrated, regulate proportion to 1.02 ~ 1.05g/ml, PH5 ~ 6 with distilled water, filtrate passes through macroporous resin column, and discard effluent, wash macroporous resin column with water, discard eluent, continue by 30-70% ethanol elution macroporous resin column, collect 30-70% position eluent, the drying of eluent concentrating under reduced pressure obtains extract.
The preparation method of described Chinese medicine extract, comprises the following steps:
Take each crude drug by weight portion according to claim 1, described crude drug is clean, pulverizing, crosses 60 mesh sieves, and with Extraction solvent 30-95% alcohol reflux, filtrate concentrates, and crosses macroporous resin column, collection 30-70% position, and concentrating under reduced pressure is dry obtains extract.
Described macroporous resin refers to AB-8 or D-101.
In described extract, the content of total flavones is 35 ~ 85%.
For realizing above-mentioned 3rd object, the technical scheme that the present invention takes is:
The application of described Chinese medicine extract in preparation treatment and prevention tobacco and wine cytotoxic drug or food.
The dosage form of described medicine is tablet, capsule, granule, oral liquid, drop pill, pill, chewable tablet, buccal tablet, chewing gum, unguentum, mixture, patch or gel.
The invention has the advantages that:
The extract that the present invention is prepared for primary raw material with Herba Abri, Folium Nelumbinis, experiment proves that Chinese medicine extract energy available protecting tobacco poisoning of the present invention is to the damage of liver cell, and is prepared into the oral formulations such as tablet and chewing gum and conveniently can carries with and take.
Accompanying drawing explanation
Accompanying drawing 1 is that the extract prepared of different Herba Abri and Folium Nelumbinis ratio is on the impact of V79 cell survival rate.(B: blank group, M: model group J1-J7: Herba Abri, Folium Nelumbinis different proportion extract).
Accompanying drawing 2 is that the extract prepared of same Herba Abri and Folium Nelumbinis ratio is on the impact of alcoholic liver injury mice ALT, AST.(B: blank group; M: model group; J1-J7: Herba Abri, Folium Nelumbinis different proportion extract).
Accompanying drawing 3 is the impacts on V79 cell survival rate of extract consumption.(P: smog water extract processed group; M: model group; A0-L: Herba Abri low concentration group; A0-M: concentration group in Herba Abri; A0-H: the low high concentration group of Herba Abri; A1-L: Herba Abri Folium Nelumbinis (27:9.5) low concentration group; A1-M: concentration group in Herba Abri Folium Nelumbinis (27:9.5); A1-H: the low high concentration group of Herba Abri Folium Nelumbinis (27:9.5)).
Accompanying drawing 4 is the impacts on alcoholic liver injury mice ALT, AST of extract consumption.(B: blank group; M: model group; J6-L: low concentration group; J6-M: middle concentration group; J6-H: high concentration group; X ± S, n=10,
vsmodel group, * * P<0.01, * P<0.05).
Detailed description of the invention
Below in conjunction with embodiment, detailed description of the invention provided by the invention is elaborated.
embodiment 1
1. the preparation of Chinese medicine extract
Get Herba Abri, Folium Nelumbinis 27:9.5 cleans, soak after 6 hours and insert the interior decoction of extraction vessel 3 times, each time decocted is 0.5 hour.The medicinal liquid extracted for several times is merged, is condensed into extractum, adds Extraction solvent methanol and carry out precipitate with ethanol, the supernatant concentration after precipitate with ethanol, cross AB-8 macroporous resin column.Resin column blade diameter length ratio 1:8, adsorption rate 3.2BV/h, resolve flow velocity 1.6BV/h, regulates sample solution proportion 1.02 ~ 1.05, collects 30-70% position, and concentrating under reduced pressure is dry obtains extract.
2. the mensuration of general flavone content
According to the content of " Chinese Pharmacopoeia " 2010 editions one annex V A spectrophotometry total flavones.Taking Quercetin is reference substance (content 98%, laboratory is made by oneself) 35mg, with anhydrous alcohol solution, is settled to 100ml, is configured to the reference substance solution that concentration is 0.35mg/ml.Take the extract 25mg prepared in 1 and add anhydrous alcohol solution in 25ml measuring bottle, dehydrated alcohol is settled to scale, and shake up and get in 1ml to 10ml measuring bottle, ethanol is settled to scale, shakes up to obtain test sample liquid.Pipette ethanol (blank), contrast liquid and each 2ml of test sample liquid respectively, in conical flask, respectively add 5% sodium nitrite solution 0.5ml and place 5min, add 10 aluminum nitrate solution 0.5ml, place 5min, then add 1 sodium hydroxide solution 5ml and shake up, place 15min, measure at 431nm wavelength and absorb ripple.Recording general flavone content in extract is 51.2%.
embodiment 2
1. the preparation of Chinese medicine extract
Get Herba Abri, Folium Nelumbinis 27:9.5 cleans, soak after 24 hours and insert the interior decoction of extraction vessel 5 times, each time decocted is 3 hours.The medicinal liquid extracted by several merges, and is condensed into extractum, adds Extraction solvent ethanol and carry out precipitate with ethanol, the supernatant concentration after precipitate with ethanol, crosses D-101 macroporous resin column, collects 30-70% position, and concentrating under reduced pressure is dry obtains extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 35.0%.
embodiment 3
1. the preparation of Chinese medicine extract
Get Herba Abri, Folium Nelumbinis 27:9.5 cleans, soaks to insert in extraction vessel after 6-24 hour and decoct with water 4 times, the time of each decoction is 2 hours.The medicinal liquid extracted for several times is merged, is condensed into extractum.After being left standstill by extractum, add ethanol and carry out precipitate with ethanol, get the supernatant after precipitate with ethanol, concentrated, cross AB-8 macroporous resin column, collect 30-70% position, concentrating under reduced pressure is dry obtains extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 47.1%.
embodiment 4
1. the preparation of Chinese medicine extract
Get crude drug Herba Abri in 27:9.5 ratio, Folium Nelumbinis is pulverized, with 95% alcohol reflux 3 times of 8 times amount, each 2 hours, filtrate concentrated, and crossed AB-8 macroporous resin column, resin column blade diameter length ratio 1:8, adsorption rate 3.2BV/h, resolves flow velocity 1.6BV/h, adjusts sample solution proportion 1.02 ~ 1.05, collect 30-70% position, concentrating under reduced pressure is dry obtains extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 56.3%.
embodiment 5
1. the preparation of Chinese medicine extract
Get crude drug Herba Abri in 27:9.5 ratio, Folium Nelumbinis is pulverized, with 95% alcohol reflux 3 times of 8 times amount, each 2 hours, filtrate concentrated, and crossed D-101 macroporous resin column, collected 30-70% position, concentrating under reduced pressure dry extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 37.2%.
embodiment 6
1. the preparation of Chinese medicine extract
Get Herba Abri in proportion, Folium Nelumbinis 27:9.5 pulverizes, add Extraction solvent 95% ethanol, percolation, to total flavonoid assay reactions is negative, is collected percolate, is obtained extracting solution; Concentrated by said extracted liquid, distilled water regulates proportion to be 5 to 1.05g/ml, PH, and filtrate by D-101 macroporous resin column, and discards effluent; Wash macroporous resin column with water, discard eluent; Continue by 70% ethanol elution macroporous resin column, collect 30-70% position eluent, the drying of eluent concentrating under reduced pressure obtains extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 43.2%.
embodiment 7
1. the preparation of Chinese medicine extract
Get Herba Abri in 27:9.5 ratio, Folium Nelumbinis is pulverized, add Extraction solvent 30% ethanol, percolation, to total flavonoid assay reactions is negative, is collected percolate, is obtained extracting solution; Concentrated by said extracted liquid, distilled water regulates proportion to be 6 to 1.05g/ml, PH, and filtrate by AB-8 macroporous resin column, and discards effluent; Wash macroporous resin column with water, discard eluent; Continue by 30% ethanol elution macroporous resin column, collect 30-70% position eluent, the drying of eluent concentrating under reduced pressure obtains extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 48.4%.
embodiment 8
1. the preparation of Chinese medicine extract
Get Herba Abri in 27:9.5 ratio, Folium Nelumbinis is pulverized, add Extraction solvent 55% ethanol, percolation, to total flavonoid assay reactions is negative, is collected percolate, is obtained extracting solution; Concentrated by said extracted liquid, distilled water regulates proportion to be 5 to 1.05g/ml, PH, and filtrate by AB-8 macroporous resin column, and discards effluent; Wash macroporous resin column with water, discard eluent; Continue by 30% ethanol elution macroporous resin column, collect 30-70% position eluent, the drying of eluent concentrating under reduced pressure obtains extract.
2. the mensuration of general flavone content
Adopt the content of spectrophotometry total flavones.Take Quercetin as reference substance, specific experiment method is with embodiment 1, and recording general flavone content is 85.0%.
the preparation of embodiment 9 pharmaceutical preparation
Extract of the present invention can also be prepared into the oral formulations such as tablet, capsule, granule, oral liquid according to the known method of pharmaceuticals industry and auxiliary element, wherein preferably be applicable to carrying the tablet and chewing gum taken, described auxiliary element can be that lactose, starch, dextrin, stearate etc. are as carrier or excipient.
one, the preparation of granule
The arbitrary described extract of Example 1-8 adds the ethanol of 2 times amount, stirs precipitates overnight.Get supernatant, be concentrated into thick extractum; Add suitable pharmaceutical aids, granulate, dry, cross 300 mesh sieves, granulate, obtain 20g granule, subpackage 10g/ bag.
two, the preparation of tablets/capsules
The arbitrary described extract 50g of Example 1-8, starch 40g, lactose 30g, microcrystalline Cellulose 30g, 7% starch slurry is appropriate, magnesium stearate 1%, by said ratio, extract powder, starch, lactose, microcrystalline Cellulose are crossed 80 mesh sieves respectively, mixing, after 40 mesh sieve 3 times, by above-mentioned mixed powder 7% starch slurry soft material, cross 24 mesh sieve grains, 50 DEG C of dryings 2 hours, dried particles crosses 30 mesh sieve granulate, adds magnesium stearate, mixing, tabletting, obtains 1000, tablet.
three, the preparation of teabag
The arbitrary described extract of Example 1-8
100g, adds 1% white sugar, is distributed into 1000 wraps with teabag packaging material.
four, the preparation of oral liquid/mixture
The arbitrary described extract of Example 1-8, adds 2 times amount ethanol, stirs precipitates overnight.Get supernatant, be concentrated into thick extractum; Add suitable pharmaceutical aids, make oral liquid, mixture.
five, the preparation of chewing gum agent
The arbitrary described extract of Example 1-8 is dissolved in water, adds the acetic starch of 18%, 80% corn syrup, the glycerol of 0.3%, the Oleum Menthae Rotundifoliae of 0.5% at 55 DEG C of mix homogeneously, is divided into the spherolite that 3g is heavy while hot, pack.
embodiment 10. cell toxicant method measures the anti-tobacco poisoning effect of extract
1. anti-tobacco poisoning effect
1.1 materials and reagent commercial cigarettes smog hydrotrope CSE, tetrazolium bromide (MTT, Sigma), hamster pneumonocyte (V79 cell).
1.2 methods: get the V79 cell of exponential phase of growth with after 0.25% trypsinization, are mixed with 1O with the RPM1640 culture fluid containing 10% hyclone
6the cell suspension of individual/ml, adds 96 orifice plates, 5%CO by cell suspension
2, after 37 DEG C of cultivation 24 h, except blank group B, CSE (20mg/ml) is added in all culture plates, J1(extract: Herba Abri: Folium Nelumbinis=15:7.5) group, J2(extract: Herba Abri: Folium Nelumbinis=18:13.5) group, J3(extract: Herba Abri: Folium Nelumbinis=21:5.5) group, J4(extract: Herba Abri: Folium Nelumbinis=23:11.5) group, J5(extract: Herba Abri: Folium Nelumbinis=25:3.5) group, J6(extract: Herba Abri: Folium Nelumbinis=27:9.5) group, J7(extract: Herba Abri: Folium Nelumbinis=30:15.5) organize dosage and be 100mg/ml, model group M, matched group B is to the 0.5%CMC-Na aqueous solution of equivalent.Add 5 g/ml MTT solution 2O μ l after 24h to continue to cultivate.Discard culture fluid after 4 h, every hole adds 200 μ l DMSO, and vibration 5min, makes purple crystal thing in cell fully dissolve, measure each hole absorbance (A) in the microplate reader of wavelength 564nm.Each dosage does 4 hole Duplicate Samples, adopts absorbance values (A) to carry out the calculating of cell survival rate: cell survival rate=A
dosage group/ A
group of solvents× 100%.
1.3 result
As shown in Figure 1, as can be seen from the figure, when extract addition is identical, J6 group, i.e. Herba Abri: the extract prepared by Folium Nelumbinis=27:9.5 is obviously better than other administration groups to the protected effect of V79 cell.
the zoopery of the anti-alcohol damaged effect of embodiment 11. different proportion Herba Abri and the extract prepared by Folium Nelumbinis
1 laboratory animal and reagent
ICR mice, body weight 18-22g, male and female half and half, Shanghai Slac Experimental Animal Co., Ltd. provides, credit number SCXK(Shanghai) 2012-0002.56% Red Star strong, colourless liquor distilled from sorghum Chinese liquor is Hongxing Co., Ltd. Beijing's product; ALT, AST detection kit (biology is built up in Nanjing).
2 experimental techniques
After mice adaptability feeds 1 week, 9 groups are divided at random by sex body weight, often organize 10, male and female half and half, are respectively blank group B, model group M, J1(extract: Herba Abri: Folium Nelumbinis=15:7.5) group, J2(extract: Herba Abri: Folium Nelumbinis=18:13.5) group, J3(extract: Herba Abri: Folium Nelumbinis=21:5.5) group, J4(extract: Herba Abri: Folium Nelumbinis=23:11.5) group, J5(extract: Herba Abri: Folium Nelumbinis=25:3.5) group, J6(extract: Herba Abri: Folium Nelumbinis=27:9.5) group, J7(extract: Herba Abri: Folium Nelumbinis=30:15.5) group.The ig dosage of J1 ~ J7 administration group is 10g/kg, and blank group gives isopyknic 0.5%CMC-Na aqueous solution.Often organize mice gavage every day 1 time, continuous 10 days.3 h after last administration, except blank group gavage gives water, all the other respectively organize the Red Star strong, colourless liquor distilled from sorghum that gavage gives 0.15 mL/10g body weight, and fasting 12 h, eye socket gets blood, separation of serum.Detection kit measures Serum ALT, AST is active.
3. experimental result
As shown in Figure 2, the work of as can be seen from the figure Herba Abri extract resisting alcoholic hepatic injury is better in order to J6 group effect, i.e. Herba Abri: the extract prepared by Folium Nelumbinis=27:9.5 is best to alcoholic liver injury effect for result.
anti-tobacco poisoning effect--the cell toxicant method of embodiment 12 Herba Abri extract
1. material and reagent commercial cigarettes smog hydrotrope CSE, tetrazolium bromide (MTT, Sigma), hamster pneumonocyte (V79 cell).
2. method: get the V79 cell of exponential phase of growth with after 0.25% trypsinization, is mixed with 1O with the RPM1640 culture fluid containing 10% hyclone
6the cell suspension of individual/ml, adds 96 orifice plates, 5%CO by cell suspension
2, after 37 DEG C of cultivation 24 h, except blank group B, CSE (20mg/ml) is added in all culture plates, A0-L (Herba Abri extract 50mg/ml) group, A0-M(Herba Abri extract 100 mg/ml) group, A0-H(Herba Abri extract 200 mg/ml) group, A1-L (Herba Abri: Folium Nelumbinis=27:9.5, 50mg/ml) group, A1-M(Herba Abri: Folium Nelumbinis=27:9.5, 100 mg/ml) group, A1-H(Herba Abri: Folium Nelumbinis=27:9.5, 200 mg/ml) group, smog water extract processed group P, matched group B is to the 0.5%CMC-Na aqueous solution of 200mg/ml.Add 5 g/ml MTT solution 2O μ l after 24h to continue to cultivate.Discard culture fluid after 4 h, every hole adds 200 μ l DMSO, and vibration 5min, makes purple crystal thing in cell fully dissolve, measure each hole absorbance (A) in the microplate reader of wavelength 564nm.Each dosage does 4 hole Duplicate Samples, adopts absorbance values (A) to carry out the calculating of cell survival rate: cell survival rate=A
dosage group/ A
group of solvents× 100%.
3. experimental result
As shown in Figure 3, as can be seen from the figure, A0-L, A0-M, A0-H group cell survival rate is less than 50%, A1-L, A1-L, A1-L group cell survival rate is greater than 60%, and extract anti-tobacco poisoning effect prepared by Herba Abri and Folium Nelumbinis is better than extract prepared by alone Herba Abri.
the anti-alcohol damage effect of embodiment 13 Herba Abri extract
1. laboratory animal and reagent
ICR mice, body weight 18-22g, male and female half and half, Shanghai Slac Experimental Animal Co., Ltd. provides, credit number SCXK(Shanghai) 2012-0002.56% Red Star strong, colourless liquor distilled from sorghum Chinese liquor is Hongxing Co., Ltd. Beijing's product; ALT, AST detection kit (biology is built up in Nanjing).
2. experimental technique
After mice adaptability feeds 1 week, random point 5 groups, often organize 10, experimental group is with Herba Abri and Folium Nelumbinis extract J6(Herba Abri: Folium Nelumbinis=27:9.5) point basic, normal, high 3 dosage group (50mg, 100mg, 200mg/10g body weight/day, by crude drug gauge), blank group gives the 0.5%CMC-Na aqueous solution of 200mg/10g.Often organize mice gavage every day 1 time, continuous 10 days.3 h after last administration, except blank group gavage gives water, all the other respectively organize the Red Star strong, colourless liquor distilled from sorghum that gavage gives 0.15 mL/10g body weight, and fasting 12 h, eye socket gets blood, separation of serum.Detection kit measures Serum ALT, AST is active.
3. experimental result
As shown in Figure 4, as can be seen from the figure, Herba Abri: Folium Nelumbinis carries=27:9.5 time prepared extract prodigiosin reduce Alt and AST level (P<0.01) very significantly, to Alcoholic Hepatic Injury, there is good protective effect.
In sum, experimental result shows, when Herba Abri: during Folium Nelumbinis=27:9.5, and prepared extract has significant anti-alcohol damage effect and anti-tobacco poisoning effect.
embodiment 12 acute toxicity test
1. experimental technique
Extract prepared by Example 8, adds the suspension that 0.5% CMC-Na preparation becomes 0.1g/ml.ICR mice, body weight 18-22g, male and female half and half, Shanghai Slac Experimental Animal Co., Ltd. provides, credit number SCXK(Shanghai) 2012-0002.Fasting 12h, 0.4ml/10g body weight gastric infusion, after 4h by above dosage repeat administration once, to observation post administration animal continuous 7 days.
2. experimental result
Result shows, experimental mice each side is showed no exception, and without dead, body weight increases good, increases to 25.1 ± 2.1g by giving front 20.2 ± 0.38g.This Chinese medicine extract gives 8g/kg/ day, is equivalent to crude drug amount 320g/kg/ day, and animal does not occur that serious toxicity reacts.This dosage is clinical people's consumption more than 2000 times.Prove that extract is safe.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the inventive method; can also make some improvement and supplement, these improve and supplement and also should be considered as protection scope of the present invention.
Claims (8)
1. separate a Chinese medicine extract for tobacco and wine poison, it is characterized in that, the active constituents of medicine of described Chinese medicine extract is extracted by the crude drug of following weight portion: Herba Abri 27 parts, 9.5 parts, Folium Nelumbinis.
2. the preparation method of Chinese medicine extract according to claim 1, is characterized in that, described method comprises the following steps:
(1) take each crude drug by weight portion according to claim 1, described crude drug carried out decoct, concentrated, add ethanol or methanol carries out precipitate with ethanol, after precipitate with ethanol, collect supernatant concentration;
(2) cross macroporous resin column, the water elution removal of impurity is the ethanol elution of 50 ~ 85% by concentration, collects the eluent at 30-70% position, and concentrating under reduced pressure is dry obtains extract.
3. the preparation method of Chinese medicine extract according to claim 1, is characterized in that, described method comprises the following steps:
(1) take each crude drug by weight portion according to claim 1, cleaned by described crude drug, pulverize, crossing 60 mesh sieves, is 30-95% ethanol percolation by concentration, to total flavonoid assay reactions is negative;
(2) extracting solution is collected, concentrated, regulate proportion to 1.02 ~ 1.05g/ml, PH5 ~ 6 with distilled water, filtrate passes through macroporous resin column, and discard effluent, wash macroporous resin column with water, discard eluent, continue by 30-70% ethanol elution macroporous resin column, collect 30-70% position eluent, the drying of eluent concentrating under reduced pressure obtains extract.
4. the preparation method of Chinese medicine extract according to claim 1, is characterized in that, described method comprises the following steps:
Take each crude drug by weight portion according to claim 1, described crude drug is clean, pulverizing, crosses 60 mesh sieves, and with Extraction solvent 30-95% alcohol reflux, filtrate concentrates, and crosses macroporous resin column, collection 30-70% position, and concentrating under reduced pressure is dry obtains extract.
5. preparation method prepared by the Chinese medicine extract according to Claims 2 or 3, is characterized in that, described macroporous resin refers to AB-8 or D-101.
6. Chinese medicine extract according to claim 2, is characterized in that, in described Chinese medicine extract, the content of total flavones is 35 ~ 85%.
7. the application of Chinese medicine extract according to claim 1 and 2 in preparation treatment and prevention tobacco and wine cytotoxic drug or food.
8., according to the arbitrary described application of claim 3, it is characterized in that, the dosage form of described medicine is tablet, capsule, granule, oral liquid, drop pill, pill, chewable tablet, buccal tablet, chewing gum, unguentum, mixture, patch or gel.
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| JPH02200640A (en) * | 1989-01-31 | 1990-08-08 | Nippon Mektron Ltd | Production of agent for preventing and improving hindrance of lever function |
| CN101653552A (en) * | 2009-09-07 | 2010-02-24 | 施彦雄 | Smoke-quitting alexeteric traditional Chinese medicine preparation and production method thereof |
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