CN1243443A - Fresh Rehmannia glutinosa extract and its extraction method for propagating bifidobacteria - Google Patents

Abstract

A rehmannia root extract for proliferating bifidobacteria is prepared from fresh rehmannia root or its dried slices through extracting in solvent, decoloring by adding activated carbon, centrifugal separation, filtering, vacuum concentrating, cold storage or depositing in alcohol, collecting deposit, and adding water.

Description

for the propagation of bifidobacteria

Fresh rehmannia glutinosa extract and its extraction method

The technical field of the invention

The present invention relates to a rehmannia extract used to proliferate bifidobacteria and an extraction method thereof, and more specifically relates to a fresh rehmannia extract extracted from fresh rehmannia that can multiply the bifidobacteria in the human body and its A method for extracting fresh rehmannia glutinosa with high yield.

Background technique

It is now known that among the bacteria living in the human intestinal tract, the most closely related to human health is Bifidobacterium bifidobacterium, which colonizes in the deep layer of the intestinal mucosa and forms a biological barrier to prevent the invasion of foreign bacteria , inhibit intestinal corruption, enhance human immunity, promote the absorption of nutrients, synthesize B vitamins and vitamin K, reduce the source of intestinal endotoxin, and thus play a beneficial role in protecting the liver, preventing tumors and anti-aging. However, due to certain reasons, such as long-term use of large amounts of antibiotics, use of immunosuppressants, chemotherapy or radiotherapy, surgery, mental and physical stimulation, or improper diet, etc., the number of bifidobacteria will decrease, and other harmful bacteria will invade , cause flora imbalance and metabolic abnormalities, various toxins, spoilage substances, and carcinogens increase, resulting in various diseases, such as diarrhea, constipation, digestive disorders, and some chronic diseases that are difficult to cure, such as arteriosclerosis, liver damage, intestinal Road cancer, etc., wherein diarrhea is the most obvious and the highest incidence of disease among the above-mentioned diseases.

At present, antibiotics or antimotility drugs are mostly used in the treatment of diarrhea, and the usage rate of antibiotics is as high as 84%, and the abuse situation is very serious. Abuse of antibiotics can lead to flora imbalance, which can lead to superinfection in serious cases, but in fact 70% of infectious diarrhea does not need antibiotic treatment. In addition, improper use of anti-motility drugs will inhibit the excretion of viruses and toxins, making the condition worse.

The results of the study showed that diarrhea caused intestinal flora imbalance, especially the greatly reduced number of bifidobacteria. Therefore, maintaining and restoring the dominant position of bifidobacteria in the intestine is an important condition for preventing and treating diarrhea. Based on the above understanding, in the last ten years, in-depth research has been carried out on the microecological preparations and treatment methods represented by bifid clubella and their proliferating agents. However, so far many countries have focused on the development of Bifidobacterium live bacteria preparations. The technical requirements for the preparation of live bifidobacteria are quite high. If the conditions are not strictly controlled, the quality is difficult to guarantee, and there are certain potential dangers. In addition, there are special and strict requirements for the use and preservation of live bacterial preparations. At the same time, only one kind of bifidobacterium strain is generally used in live bacterial preparations, and there are some strains that are single and cannot fully meet the needs of the human body. The issue of bifidobacteria species composition.

In view of the above situation, there is an urgent need to develop a new type of bifidobacterium proliferation preparation that can proliferate bifidobacteria and overcome the shortcomings of live bacteria preparations.

In the article "Development of Soybean Oligosaccharides and Its Product Characteristics" written by Japanese scientist Teruhisa Masai, it is recorded that stachyose and raffinose in soybeans have an excellent proliferation effect on Bifidobacteria, confirming that stachyose Sugar and raffinose are very good bifidobacteria proliferators. However, as recorded in this article, the content of stachyose and raffinose in soybeans is low, of which the content of stachyose is 24%, the content of raffinose is 8%, the total is 32%, and sucrose, fructose, glucose, etc. can be Absorbed sugar content is very high, reaching more than 55% (the contents are all calculated as dry products). Therefore, it is not suitable for diabetics, obese people and the elderly to take, and the scope of use is limited.

In view of the above situation, the present inventors conducted in-depth research for the purpose of developing a new bifidobacterium proliferative agent with high stachyose and raffinose content and low absorbable sugar content. The total content of sugar and raffinose is very high, and the extract extracted from it with an appropriate extraction method can meet the above requirements, thus completing the present invention.

Summary of the invention

The present invention provides a fresh rehmannia extract used to proliferate bifidobacteria in the intestinal tract, and a method for extracting the traditional Chinese medicine fresh rehmannia or dried fresh rehmannia slices as a raw material. The method can be used for rapid and high-yield extraction. During the process, there will be no adverse phenomena such as growth of bacteria, and the decline in yield will not be caused thereby.

Detailed Description of the Invention

The fresh rehmannia glutinosa extract of the present invention contains 55%-70% by weight of active ingredients stachyose and 2.5-5% by weight of raffinose, and the total amount of absorbable sucrose and fructose is 5-10% by weight (both calculated as dry products), The remainder is other sugars and unavoidable impurities.

The fresh rehmannia extract of the present invention uses the natural Chinese medicine fresh rehmannia or fresh rehmannia dried slices as a raw material, and its extraction method consists of the following steps:

(1) Extraction: Take fresh rehmannia or dried fresh rehmannia slices and add a water-miscible solvent to extract twice, and combine the two extracts;

(2) Decolorization: add activated carbon to the extract to decolorize, and obtain a light yellow or grayish yellow liquid after decolorization;

(3) Filtration: Centrifuge the decolorized liquid, then filter it with a sand core, and filter it with a hollow fiber after vacuum filtration;

(4) Concentration and cold storage: Concentrate the filtrate obtained above under reduced pressure, and store the concentrate in cold storage; (5) Separation: The refrigerated supernatant is separated by one of the following four methods. Method 1: The refrigerated solution is subjected to activated carbon column chromatography, and the first 2/3 of the eluate is retained;

Method 2: subject the refrigerated liquid to macroporous adsorption resin column chromatography, and retain the first 2/3 of the eluent; Method 3: subject the refrigerated liquid to C18 column chromatography, and retain the first 2/3 of the eluent;

Method 4: Precipitate the refrigerated solution with alcohol precipitation, and collect the precipitated part;

(6) Preparation: Concentrate the first 2/3 of the eluate obtained by any one of the above separation methods 1 to 3 under reduced pressure, or volatilize the alcohol precipitated part of method 4 until there is no alcohol smell, and then add water to prepare a solution.

The water-miscible solvent used in the above step (1) is preferably an ethanol solution with a concentration of 95% or less. The first extraction preferably adds 10-20 times the amount of solvent, and the second extraction preferably adds 5-10 times the amount of solvent, and the extraction time is preferably 2-4 hours (the sum of the two times).

The amount of activated carbon used in the above step (2) is preferably 0.15-5% by weight relative to the extract, and the decolorization temperature is preferably 85-95*C.

The centrifugation performed in the above step (3) is preferably performed at a speed of 5000-20000 rpm for 5-10 minutes, the pore size of the sand core used is preferably 10-15μ, and the pore size of the hollow fiber used is preferably 0.15-0.20μ .

In the above step (4), the filtrate is preferably concentrated under reduced pressure to a specific gravity of 1.05-1.27 (25°C), and the concentrate is preferably refrigerated for 24-48 hours.

The macroporous adsorption resin used in the second separation method of the above step (5) is preferably ZTC-Type I and Type II, and the particle size of the C18 column used in the third separation method is preferably 50-100μ.

In the above step (6), the eluate is preferably concentrated under reduced pressure to a specific gravity of 1.05-1.45 (room temperature 25), and the specific gravity of the solution prepared by adding water after volatilizing the alcohol precipitation part until it has no alcohol smell is preferably 1.05-1.45 (room temperature 25 "). By using the extraction method of the present invention, active ingredients can be extracted from fresh rehmannia or fresh rehmannia dried slices quickly and with high yield, and the bacteria growth phenomenon that occurs in the common filtration methods in the past can be avoided, thereby avoiding as The decomposition of the active ingredients stachyose and raffinose greatly improves the yield.

The extract of the present invention can be made into oral liquid and taken. In addition, the above-mentioned extracts of the present invention can also be made into powders by common methods such as freeze-drying or spray-drying, or added with pharmacologically acceptable carriers or excipients to make any common oral solid preparations, these Preparations include, for example, powders, tablets, capsules, granules, powders, and the like. Of course, any formulation form that can be imagined by those skilled in the art is included within the scope of the present invention. The oral dosage form of the extract of the present invention is preferably an oral liquid, and the oral liquid is prepared by adding water to the above-mentioned extract to prepare a solution containing stachyose and raffinose at a total of 150-300 mg/ml, and bottled after sterilization Instantly.

Examples and pharmacological test examples are given below to describe the present invention in more detail.

Example

Example 1

Take 5000 grams of fresh rehmannia glutinosa, add 15 liters of 20% ethanol, heat to reflux for 2 hours, filter, and use the filtrate for later use. Add 10 liters of 20% ethanol to the filtered medicinal dregs, heat to reflux for 1 hour, filter, combine the filtrates twice, recover the ethanol until it has no alcohol smell, add 1% by weight (relative to the weight of the solution) of the recovered ethanol solution Activated carbon decolorization, the temperature is 85 to obtain a light yellow liquid, then centrifuge at 15000 rpm for 5 minutes, the supernatant is concentrated under reduced pressure to a specific gravity of 1.20 (25 °C), the concentrated solution is refrigerated for 24 hours, and the supernatant is poured out , carry out activated carbon column chromatography, collect 25,000 ml, concentrate the eluent under reduced pressure to a specific gravity of 1.19 (25 "C), store the concentrated solution in cold storage for 24 hours, decant the supernatant, and further concentrate to a specific gravity of 1.39 ( 25€ ), that is, 600 grams of yellow-brown fresh rehmannia extract.

According to high performance liquid chromatography analysis, it contains 57.5% stachyose, 3.6% raffinose, 4.2% sucrose, and 3.6% fructose (both calculated as dry products).

Example 2

Take 1,000 grams of fresh rehmannia dried slices (made by roasting fresh rehmannia slices), add 15 liters of water, heat to reflux for 1.5 hours, filter, and use the filtrate for later use. Add 10 liters of water to the filtered medicinal residue, heat to reflux for 1 hour, filter, and combine the two filtrates. Add 1.0% by weight (relative to the weight of the filtrate) of activated carbon to the filtrate, decolorize at 95 ° C to obtain a light yellow liquid, centrifuge at 10,000 rpm for 5 minutes, and then filter the supernatant with a sand core (15μ) , then carry out hollow fiber filtration (0.2μ), the filtrate is concentrated under reduced pressure to a specific gravity of 1.15 (room temperature 25 "), the concentrated solution is refrigerated for 48 hours, the supernatant is poured out, and the macroporous adsorption resin (ZTC-Type I and Type II The mixture (produced by Shanghai Reagent No. 1 Factory) was chromatographed, the first 20,000 ml eluate was collected, and the eluate was concentrated to a specific gravity of 1.25 (room temperature 25") to obtain 650 grams of yellow-brown fresh rehmannia extract.

According to high performance liquid chromatography analysis, it contains 52.3% stachyose, 2.5% raffinose, 3.7% sucrose, and 4.2% fructose (calculated as dry product).

Example 3

Take 1000 grams of fresh rehmannia dry slices, add 12 liters of water, heat and reflux for 1.5 hours, filter, add 10 liters of water to the dregs, heat and reflux for 1 hour, combine the two filtrates, add 1.5% by weight of activated carbon (relative to the weight of the filtrate) in the filtrate, Decolorize at 90°C to obtain light yellow liquid, separate at 10000 rpm Heart for 5 minutes, filter with sand core (10-15μ), filter with hollow fiber (0.15μ), concentrate the filtrate under reduced pressure to a specific gravity of 1.15 (room temperature 25), refrigerate for 48 hours, pour out the supernatant, use C18 (50-100μ) Perform chromatography, elute with water, collect the first 10,000 ml of eluate, concentrate the eluate to a specific gravity of 1.25 (25), and obtain 700 grams of yellow-brown fresh rehmannia extract.

According to HPLC analysis, the extract contains 53% stachyose, 2% raffinose, 2% sucrose, and 3% fructose (calculated as dry product).

Example 4

Take 5000 grams of fresh rehmannia glutinosa, add 15 liters of 60% ethanol, heat to reflux for 2 hours, filter, add 15 liters of 60% ethanol to the dregs, and heat to reflux for 1. After 5 hours, filter, combine the two extracts, recover ethanol under reduced pressure until there is no alcohol smell, adjust the volume of the extract to 2000 ml, add 3% by weight of activated carbon (relative to the weight of the filtrate), and decolorize at 90 ° C to obtain light The yellow liquid was centrifuged at 10,000 rpm for 5 minutes, filtered through a sand core (10-15μ), filtered through a hollow fiber (0.15μ), the filtrate was concentrated under reduced pressure to 1500 ml, heated at 80°C, and 95% ethanol was added to make the The alcohol content reaches 90%, let it stand overnight, collect the precipitate, and dry it to obtain 520 grams of dry extract.

According to the analysis by high performance liquid chromatography, the extract contains 65% stachyose, 3% raffinose, 2% sucrose, and 4% fructose (both calculated as dry products). Preparation Example 1

Take 1,000 grams of fresh rehmannia extract obtained in Example 1 above, add 2,200 ml of distilled water to prepare a solution containing stachyose and ladder seed sugar at a total of 200 mg/ml, pour it into small bottles, 10 ml per bottle, Sterilize and seal to obtain the oral liquid.

Preparation Example 1

Take 1000 grams of fresh rehmannia extract obtained in Example 2 above, and use a freeze dryer (produced by Jiangsu Freeze Drying Equipment Factory) to freeze-dry to obtain 830 grams of extract powder.

Take 100 grams of the extract powder obtained above, add 10 grams of carboxymethylcellulose, mix, granulate, and prepare granules containing 45.5% stachyose. Pharmacological test case

Test Example 1. In vitro Proliferation Effect on Bifidobacteria

(1) Materials and methods

Culture medium: Use the special ΧY medium that selectively promotes the growth of bifidobacteria, mainly containing two nutrients such as peptone, so that bifidobacteria can grow in this medium without flourishing, so as to highlight the water The role of threose. Pure stachyose was added to the medium to make the stachyose concentration reach 1.5% by weight (relative to the weight of the medium).

After opening the freeze-dried bacterial smear by the fragmentation method, inoculate it in the corresponding medium, and after 48 hours of anaerobic culture at 37 ° C, pick 3-4 colonies into the corresponding liquid medium, and then anaerobic culture, take out as After passing the pure bacteria inspection, dilute 1000 times, inoculate 10 ⁵ - 10 ⁶ CFU/ml into the corresponding medium containing stachyose, and count the viable bacteria on the plate to record the growth of each bacteria. The experiment was repeated 3 times and processed statistically.

(2) Test results

See Table 1 for stachyose to promote the growth of intestinal Bifidobacterium species in vitro. Table 1 Growth of stachyose-promoting bifidobacteria (CFU/ml in LoglO)

与空白培养基活菌数相比， *表示 PO.05; **表示 PO.01

Compared with the number of live bacteria in the blank medium, * indicates PO.05; ** indicates PO.01

The results showed that stachyose had growth-promoting effects on Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium angle, and Bifidobacterium infantis, but had almost no promoting effect on Bifidobacterium bifidum.

Test example 2. Proliferation of bifidobacteria in mice

(1) Experimental equipment and materials

(1) Animals: Kunming white mice, weighing 18-20 grams, half male and half male.

(2) Medicine: fresh rehmannia extract.

(3) Instrument: American VITEK-AMS automatic bacteria identification instrument, AN1 identification card.

(2) Experimental methods and results

The animals were randomly divided into 4 groups. Except for the blank group, each mouse was orally administered once a day. The dosage is shown in Table 2. The administration was continued for one month. After the last administration, 12 hours after the last administration, the feces of the mice were taken and placed immediately in a prepared anaerobic bottle. The quantitative fecal specimens were homogenized with a tissue homogenizer, and serially diluted 10 times with 0.1% agar-peptone aqueous solution to make a suspension of ^10_1 ~10_1 ^(>) , and various diluted suspensions were inoculated with a graduated dropper. Dry the surface of the Bib medium, 3 drops for each dilution, and quickly place the inoculated plate in an anaerobic environment, and cultivate it at 35°C for 48 hours, and then check the colonies on the Bib medium according to the shape, size, appearance, and transparency , color, edge, and whether hemolysis and other characteristics are separated, identified, and counted. The identification uses the American VITEK-AMS automatic bacterial identification instrument, the identification card is AN1, and the similar screening value and standardized percentage probability are calculated through 30 specific biochemical reactions. and Biochemical Digital, the bacteria reported as A.bifidobacterium were quantitatively calculated to obtain the number of bifidobacteria (cfu/g), and the results are shown in Table 2. Table 2 The effect of fresh rehmannia extract on the proliferation of Mufidobacterium in mice

***Ρ<0·001 *Ρ<0.05

The experimental results showed that oral administration of fresh rehmannia extract to mice for one month could effectively increase the number of bifidobacteria in the intestines, and compared with the blank control group, PO.001, there was a very significant difference.

Test example 3, antagonism to acute diarrhea caused by senna

(1) Experimental materials

(1) Animals: Kunming white mice, weighing 20-22g, can be either male or female.

(2) Medicine: fresh rehmannia extract.

(2) Experimental methods and results

Add 5 times the amount of water to 50 g of senna leaves, boil on low heat for 10 minutes, filter, add 3 times the amount of water to the medicinal residues, boil for 10 minutes, filter, and concentrate the filtrate under reduced pressure to 1g/ml for later use.

The mice were randomly divided into 4 groups according to body weight, administered once a day by intragastric administration, and the model group was administered with an equal volume of normal saline for three consecutive days, and 30 minutes before the last administration, the mice in each group were administered with 20 g/kg Stomach senna decoction, mouse is placed on alone in the cage that pad has 12.5 * 23.0cm filter paper after administration, every The filter paper was replaced every 1 hour, observed for 4 hours, and the number of wet fecal particles on the filter paper was recorded at each time, and the accumulated diarrhea times at each time between the groups were tested and compared. The results are shown in Table 1. Table 3 The antidiarrheal effect of fresh rehmannia glutinosa extract in mice

与空白组比较 A P<0.01, A A A P<0.001

Compared with blank group A P<0.01, AAA P<0.001

Compared with the model group *P<0.05, **P<0.01, ***P<0.001

The experimental results show that the fresh rehmannia extract has a significant antagonistic effect on the acute diarrhea caused by senna leaves, and there is a very significant difference by mathematical statistics PO.001.

Test example 4. Protective effect against antibiotic enteritis and reduction of bifidobacteria caused by lincomycin hydrochloride

(1) Experimental materials

(1) Animals: 50 Kunming mice, weighing 18-20g, both male and female.

(2) Drugs: fresh rehmannia glutinosa extract; lincomycin hydrochloride, batch number: 9601152, produced by Jiangsu Xinyi Pharmaceutical Factory.

(2) Experimental methods and results

The mice were randomly divided into 4 groups. Except for the blank group, the mice in each group were orally administered lincomycin hydrochloride aqueous solution (concentration 100mg/ml) 0.5ml/time, 2 times/day, for 3 days. The fresh rehmannia extract group was administered according to the dosage in the table, once a day, for a total of 10 days. On the 4th and 10th days, the defecation frequency of mice in each group within 2 hours was observed 1 hour after administration, and the results are shown in Table 4. On the 10th day, the feces of mice in each group were taken for culture of Bifidobacteria and counted. The results are shown in Table 5. Table 4 Comparison of defecation times of mice in each group (X±SD)

Compared with the blank group: A p<0.05 Compared with the model group: *P<0.05, **P<0.01 Table 5 Comparison of the number of bifidus club bacteria in the feces of mice in each group (X±SD)

Compared with the model group: ***P<0.001, compared with the blank group: A A A p<0.001

The experimental results show that the fresh rehmannia extract has obvious protective effect on lincomycin-induced antibiotic enteritis in mice, and fecal points were recorded on the 4th day of administration. Compared with the model group, P<0.01, there is a very significant difference. The feces spot results after 10 days of administration showed that the symptoms of enteritis in mice in each administration group had completely recovered. At the same time, fresh rehmannia glutinosa extract had a significant proliferation effect on the reduction of bifidobacteria in vivo caused by lincomycin hydrochloride, reaching 23 orders of magnitude, P<0.01.

Claims (7)

1. for the fresh rehmannia root extract for breeding bifid pestle bacterium, wherein the weight % of stachyose containing active ingredient 55-70 and weight % of raffinose 2.5-5, absorbable sucrose and fructose are added up to 5-10 weight % and (calculated with dry product）, remaining is other sugared and inevitable impurity, and the fresh rehmannia root extract is raw material using fresh rehmannia root or fresh rehmannia root dry plate, is made using the method being made up of following steps：

(1) extract：Take fresh rehmannia root or power fresh rehmannia root dry plate to add solvent extraction miscible with water 2 times, merge the extract solution of 2 times；

Standing grain

(2) decolourize：Added in extract solution and faint yellow or pale yellow liquid is obtained after decolorizing with activated carbon, decolouring；

(3) filter：Liquid after decolouring is centrifuged, is then filtered, is asked in use using core

Hollow fiber is filtered；

(4) concentration refrigeration：Filtrate decompression obtained above is concentrated, concentrate is stored refrigerated；

(5) separate：Supernatant after refrigeration is separated using one of following method,

Method 1:Refrigeration liquid is subjected to carbon column chromatography, preceding 2/3 eluent is left and taken；

Method 2:Cold liquid is subjected to macroporous adsorbent resin column chromatography, preceding 2/3 eluent is left and taken；Method 3:Refrigeration liquid is subjected to C18 column chromatographies, preceding 2/3 eluent is left and taken；

Method 4:The refrigeration liquid alcohol precipitation method are precipitated, sediment fraction is collected；

(6) prepare：2/3 eluent before any one method is obtained in above-mentioned separation method 1 to 3 is concentrated under reduced pressure, or the alcohol sediment fraction that separation method 4 is obtained is evaporated into add water after no alcohol taste is modulated into solution.

2. fresh rehmannia root extract as claimed in claim 1, wherein

The solvent miscible with water used in step (1) is below the % of concentration 95 ethanol solution, and the 1st extraction adds the solvents of 10-20 times of amounts, and the 2nd extraction adds 5-10 times of solvents measured, and extraction time is 2-4 hours（2 sums）；

Step（2) amount of the activated carbon used in is 0.15-5 weight % relative to extract solution, and bleaching temperature is 85-95 *;

Bu Sudden（3) centrifugation carried out in is carried out 5-10 minutes with 5000-20000 revs/min of speed, and the aperture of core used is 10 1 15 μ, and the aperture of doughnut used is 0.15 μ;

In step（4) proportion is concentrated filtrate in for 1.05-1.27 (25), stored refrigerated 24-48 hours of concentrate； In above-mentioned steps（5) macroporous absorbent resin used in separation method 2 uses ZTC-I types and II types, and the granularity of the C18 posts used in separation method 3 is 50-100 μ;

In above-mentioned steps（6) eluent is concentrated under reduced pressure into proportion for 1.05-1.45 (room temperatures 25 in.), C alcohol sediment fraction is evaporated into add water after no alcohol taste proportion is made for 1.05 1 1.45 (room temperatures 25

°C) solution.

3. the method for containing stachyose and raffinose as the extract of main component is extracted from fresh rehmannia root or fresh rehmannia root dry plate, it is characterised in that be made up of following steps：

(1) extract：Take fresh rehmannia root or fresh rehmannia root dry plate to add solvent extraction miscible with water 2 times, merge the extract solution of 2 times；

(2) decolourize：Added in extract solution and faint yellow or pale yellow liquid is obtained after decolorizing with activated carbon, decolouring；

(3) filter：Liquid after decolouring is centrifuged, is then filtered using core, uses hollow fibre filtering；

(4) concentration refrigeration：Filtrate decompression obtained above is concentrated, concentrate is stored refrigerated；

(5) separate：Supernatant after refrigeration is separated using one of following method,

Method 1:Refrigeration liquid is subjected to carbon column chromatography, preceding 2/3 eluent is left and taken；

Method 2:Cold liquid is subjected to macroporous adsorbent resin column chromatography, preceding 2/3 eluent is left and taken；Method 3:Refrigeration liquid is subjected to C18 column chromatographies, preceding 2/3 eluent is left and taken；

Method 4:The refrigeration liquid alcohol precipitation method are precipitated, sediment fraction is collected；

(6) prepare：2/3 eluent before any one method is obtained in above-mentioned separation method 1 to 3 is concentrated under reduced pressure, or the alcohol sediment fraction that separation method 4 is obtained is evaporated into add water after no alcohol taste is modulated into solution.

4. the method for containing stachyose and raffinose as the extract of main component is extracted from fresh rehmannia root or fresh rehmannia root thousand as claimed in claim 3, wherein

Step（1) solvent miscible with water used in is below the % of concentration 95 ethanol solution, and the 1st extraction adds the solvents of 10-20 times of amounts, and the 2nd extraction adds 5-10 times of solvents measured, and extraction time is 2-4 hours（1 sum）；

Step（2) amount of the activated carbon used in is 0.15-5 weight % relative to extract solution, and bleaching temperature is 85-95

Step（3) centrifugation carried out in is carried out 5-10 minutes with 5000-20000 revs/min of speed, and the aperture of core used is 10-15 μ, and the aperture of doughnut used is 0.15 μ;

In step（4) proportion is concentrated filtrate in for 1.05-1.27 (25 " C), it is dense Stored refrigerated 24-48 hours of contracting liquid；

In above-mentioned steps（5) macroporous absorbent resin used in separation method 2 uses ZTC-I types and II types, and the granularity of the C18 posts used in separation method 3 is 50-Ι Ο Ο μ;

In above-mentioned steps（6) eluent is concentrated under reduced pressure into proportion for 1.05-1.45 (25 °C of room temperatures) in, alcohol sediment fraction is evaporated into add water after no alcohol taste proportion is made for 1.05-1.45 (room temperatures 25.C solution).

5.-kind it is used for the oral formulations that promote bifid pestle bacterium in enteron aisle to breed, it is made up of the fresh rehmannia root extract described in claim 1.

6. oral formulations as claimed in claim 5, described oral formulations are oral liquid, granule, glue Nang agent, tablet, powder or pulvis.

7. oral formulations as claimed in claim 5, for treating the enterogastric diseases such as diarrhoea, constipation, intestinal flora imbalance, the disorders of digestion.