CN104666740A - Preparation method of pepsin and trypsin treated Liuwei Dihuang composite micro-ecological oral liquid - Google Patents

Abstract

The invention relates to a preparation method of pepsin and trypsin treated Liuwei Dihuang composite micro-ecological oral liquid. The preparation method comprises the following steps: (1) crushing Liuwei Dihuang pill raw materials, adding water, and mixing uniformly to prepare raw material serous fluid; (2) adding acid into the raw material serous fluid to adjust the pH value, and adding pepsin for treatment, thereby preparing pepsin treated serous fluid; (3) adding trypsin into the pepsin treated serous fluid for treatment to prepare the pepsin and trypsin treated serous fluid; and (4) inoculating bifidobacteria and lactobacillus plantarum to obtain the pepsin treated Liuwei Dihuang composite micro-ecological oral liquid. The pepsin and trypsin treated Liuwei Dihuang composite micro-ecological oral liquid prepared by the preparation method provided by the invention is relatively small in side effect, and is palatable in sweet and sour tastes; and by virtue of practical application, after being taken for a long time, the pepsin and trypsin treated Liuwei Dihuang composite micro-ecological oral liquid not only can achieve a treatment effect of Liuwei Dihuang pills but also greatly reduces the side effects causing stomach cold and digestion obstruction.

Description

The preparation method of the six drugs containing rehmanniae compound microecological oral liquid of a kind of stomach, trypsin treatment

Technical field

The present invention relates to the preparation method of six drugs containing rehmanniae compound microecological oral liquid of a kind of stomach, trypsin treatment, particularly a kind of method utilizing the classical LIUWEI DIHUANG WAN prescription of biotechnology process, belongs to biomedicine technical field.

Background technology

LIUWEI DIHUANG WAN stems from the JINGUI SHENQI WAN in the large physician's Zhang Zhongjing " Medical Treasures of the Golden Chamber " of the Eastern Han Dynasty, also be " shenqi pill ", " Rehmanniae Bolus of Eight Ingredients ", be originally eight taste Chinese crude drugs, afterwards to the Northern Song Dynasty, by national imperial physician assistant officer Qian Yi, former side is deducted Cortex Cinnamomi, Radix Aconiti Lateralis Preparata two taste Chinese medicine, defines by Radix Rehmanniae Preparata, Fructus Corni, Rhizoma Dioscoreae (three taste tonics), Poria, Rhizoma Alismatis, the LIUWEI DIHUANG WAN of Cortex Moutan (three taste cathartics) ingenious compatibility; Three mend and three to rush down, and mend and not dry.LIUWEI DIHUANG WAN, from Song dynasty Cheng Fang, continues more than 1000 year till now always, lasting through successive dynasties doctor and undergoing of patient, can be described as " volt cattle prescription changes, and within thousand, medicine does not wane ".

But because LIUWEI DIHUANG WAN belongs to sweet warm nature, the relatively medicine of nourishing YIN, cure mainly deficiency of the kidney yin, in its formula, cloudy soft medicine is more, so sometimes can antidigestive side effect after eating, therefore not only spleen and stomach function is weak, dyspepsia person is careful eats, and long-term taking can cause the malaise symptoms such as abdominal distention, loose stool, inappetence.

Chinese patent literature CN102920882A(application number 201110338130.2) disclose a kind of preparation method of concentrated pill of six ingredients with rehmannia, it is characterized in that comprising the steps: that (1) takes raw material by following weight portion: Radix Rehmanniae Preparata 120 parts, wine Fructus Corni 60 parts, Cortex Moutan 45 parts, Rhizoma Dioscoreae 60 parts, 45 parts, Poria, Rhizoma Alismatis 45 parts; (2) Cortex Moutan is dropped in extraction pot, extract paeonol with steam distillation; First open condensed water, open straight-through steam valve again, steam regulation pressure is at 0.1Mpa ~ 0.3Mpa, in tank, temperature remains on 100 ~ 110 DEG C, and pressure is ﹤ 0.05Mpa, and regulating condensate flow to make material pipe above crystallizer flow out paeonol distillate temperature is 40 DEG C ~ 60 DEG C, collect paeonol distillate, cold preservation, crystallize, collects paeonol crystal; (3) get Radix Rehmanniae Preparata 120 parts, wine Fructus Corni 20 parts, 45 parts, Poria, Rhizoma Alismatis 45 parts, adopt dynamic extraction mode, decoct with water 1 ~ 3 time, each 1 ~ 1.5 hour, decocting liquid filters, and merges, and is concentrated into the thick paste (55 ~ 60 DEG C of mother-in-law prunus mume (sieb.) sieb.et zucc. proportion measurements) that proportion is 1.35 ~ 1.40; (4) after Rhizoma Dioscoreae 60 parts and wine Fructus Corni 40 parts pulverized 80 mesh sieves, mixing, obtains fine powder, obtains sterilized powder after moist heat sterilization; (5) by the paeonol crystal of step (2) (3) (4) gained, thick paste, fine powder mixing, add pharmaceutically acceptable adjuvant, obtained wet ball, dry, polishing, to obtain final product.

Although the active ingredient in LIUWEI DIHUANG WAN can be extracted concentrated obtained concentrated pill of six ingredients with rehmannia by above-mentioned patent, the antidigestive side effect of LIUWEI DIHUANG WAN does not weaken.

Bacillus bifidus Bifidobacterium is the Gram-positive bacillus by French scholar Tissier isolated a kind of anaerobism from the feces of breast milk nutrition youngster in 1899, and end is bifurcated usually, therefore named bacillus bifidus.Bacillus bifidus has following therapeutic effect: 1, safeguard intestinal normal bacteria colony balance, suppresses the growth of pathogen, prevents constipation, dysentery and gastrointestinal disorders etc.; 2, antitumor; 3, in intestinal synthetic vitamin, aminoacid and raising body to the absorption of calcium ion; 4, reduce Blood Cholesterol level, prevent and treat hypertension; 5, improve the tolerate lactose of milk product, improve digestibility; 6, strengthen human immunologic function, prevent antibiotic side effect, defying age, life lengthening.

Lactobacillus plantarum (Lactobacillus plantarum) is the one in lactobacillus, in the vegetable being often present in fermentation and fruit juice.Lactobacillus plantarum, as the beneficial flora of human gastrointestinal tract, has and maintains intestinal flora balance, raising immunity of organisms and promote the several functions such as absorption of nutrient ingredients.Lactobacillus plantarum is applied very extensive in the food industry.Owing to having good toleration to Sal, nitrite, to protein, fat without features such as obvious directly decompositions, in fermentation meat product, application is a lot.Meanwhile, be also widely used in fermenting plant food, utilize Lactobacillus plantarum to produce conjugated linoleic acid in addition.

Summary of the invention

The present invention is directed to the deficiencies in the prior art, the preparation method of the six drugs containing rehmanniae compound microecological oral liquid of a kind of stomach, trypsin treatment is provided.

Term explanation

Pepsic unit of activity (U): adopt the unit of activity that the detection method of Chinese Pharmacopoeia version formulation in 1977 detects.

Tryptic unit of activity (U): the unit of activity that the detection method adopting Chinese Pharmacopoeia nineteen ninety version to formulate detects.

Technical solution of the present invention is as follows:

A preparation method for the six drugs containing rehmanniae compound microecological oral liquid of stomach, trypsin treatment, step is as follows:

(1) by the Six-element crude drug in LIUWEI DIHUANG WAN raw material after crushed, obtained material powder, then adds 200 ~ 600 grams of water and refined honey by every hectogram material powder, mix homogeneously, obtained raw slurry;

(2) in the obtained raw slurry of step (1), adding sour adjust pH is 1.5 ~ 2.2, add the pepsin of 20 ~ 60 unit of activity by every hectogram LIUWEI DIHUANG WAN raw material, temperature 36 ~ 38 DEG C, process 2 ~ 3 hours, adjusting PH with base value is 7.8 ~ 8.2, the serosity of obtained pepsin;

(3) in the serosity of the obtained pepsin of step (2), add the trypsin of 20 ~ 50 unit of activity by every hundred milliliters, temperature 36 ~ 38 DEG C, process 2 ~ 3 hours, sour adjust pH to 5.5 ~ 6, obtain the serosity of stomach, trypsin treatment;

(4) in the serosity of the obtained stomach of step (3), trypsin treatment, by every hundred milliliters of inoculation activated bacillus bifiduss of 0.5 ~ 2g and the activated Lactobacillus plantarum of 1.5 ~ 3g, the six drugs containing rehmanniae compound microecological oral liquid of obtained stomach, trypsin treatment.

Preferred according to the present invention, the LIUWEI DIHUANG WAN raw material of described step (1) is as follows, is weight portion:

Radix Rehmanniae Preparata 160 parts, Fructus Corni (processed) (wine Fructus Corni) 80 parts, Cortex Moutan 60 parts, Rhizoma Dioscoreae 80 parts, 60 parts, Poria, Rhizoma Alismatis 60 parts, refined honey 7 ~ 10 parts.

Preferred according to the present invention, the LIUWEI DIHUANG WAN raw material of described step (1) is as follows, is weight portion:

Radix Rehmanniae Preparata 160 parts, Fructus Corni (processed) (wine Fructus Corni) 80 parts, Cortex Moutan 60 parts, Rhizoma Dioscoreae 80 parts, 60 parts, Poria, Rhizoma Alismatis 60 parts, refined honey 16 ~ 22 parts.

Preferred according to the present invention, the acid in described step (2), (3) is hydrochloric acid.

Preferred according to the present invention, the alkali of described step (1) is sodium carbonate.

Preferred according to the present invention, in described step (4), activated bacillus bifidus refers to and is inoculated in TPY fluid medium by bacillus bifidus, is cultured to the bacillus bifidus of exponential phase at 36 ~ 38 DEG C.

Preferred further according to the present invention, it is as follows that described TPY fluid medium often rises component:

Peptone 15.0g, yeast powder 2.0g, glucose 20.0g, soluble starch 0.5g, sodium chloride 5.0g, 5wt% cysteine solution 10.0mL, Fructus Lycopersici esculenti leachate 400.0mL, Tween 80 1.0g, liver extracting solution 80.0mL, distilled water 520.0mL, pH7.0.

Described Fructus Lycopersici esculenti leachate be prepared as this area conventional method, step is as follows:

New fresh tomato is cleaned chopping after weighing, add equivalent distilled water and heat in 100 DEG C of water-baths, stir 90min, then use filtered through gauze, adjust pH7.0.

Described liver extracting solution be prepared as this area conventional method, step is as follows:

Take fresh liver 1000g, be cut into small pieces or rub, adding distil water is to 2000mL, and mixing, puts refrigerator overnight, within second day, boil 15min, and filtered through gauze also extrudes collection filtrate, adds water and is settled to 2000mL.

Described bacillus bifidus is beneficial bacteria of intestinal tract, those skilled in the art can select as required, as the bacillus bifidus that the culture presevation in Chinese patent literature CN102946891A number is NCIMB41676, the culture presevation number in Chinese patent literature CN101679938A is the bifidobacterium longum etc. of NCIMB41387.

Preferred according to the present invention, in described step (4), activated Lactobacillus plantarum refers to and is inoculated in MRS fluid medium by Lactobacillus plantarum, is cultured to the Lactobacillus plantarum of exponential phase at 30 ~ 35 DEG C.

Preferred further according to the present invention, it is as follows that described MRS fluid medium often rises component:

Peptone 10.0g, Carnis Bovis seu Bubali cream 10.0g, yeast extract 5.0g, diammonium hydrogen citrate [(NH ₄) ₂hC ₆h ₅o ₇] 2.0g, glucose (C ₆h ₁₂o ₆h ₂o) 20.0g, Tween 80 1.0mL, sodium acetate (CH ₃cOONa3H ₂o) 5.0g, dipotassium hydrogen phosphate (K ₂hPO ₄3H ₂o) 2.0g, magnesium sulfate (MgSO ₄7H ₂o) 0.58g, manganese sulfate (MnSO ₄h ₂o) 0.25g, agar 18.0g, distilled water are settled to 1000mL, pH6.2 ~ 6.6.

Described Lactobacillus plantarum is beneficial bacteria of intestinal tract, those skilled in the art can select as required, as the Lactobacillus plantarum CJLP133 that the culture presevation in Chinese patent literature CN102597214A number is KCTC11403BP, the culture presevation number in Chinese patent literature CN102597215A is the Lactobacillus plantarum CJLP55 etc. of KCTC11401BP.

Beneficial effect

The present invention passes through traditional LIUWEI DIHUANG WAN raw material through stomach, trypsin resolution process, obtain the six drugs containing rehmanniae compound microecological oral liquid of the less stomach of side effect, trypsin treatment, this product sour and sweet palatability, and through practical application, the six drugs containing rehmanniae compound microecological oral liquid of this stomach of long-term taking, trypsin treatment can not only reach takes LIUWEI DIHUANG WAN therapeutic effect, and cause cold syndrome of the stomach, hinder digestion side effect also reduced greatly.

Detailed description of the invention

Below in conjunction with embodiment, technical scheme of the present invention is described further, but institute of the present invention protection domain is not limited thereto.

It is as follows that TPY fluid medium in embodiment often rises component:

Peptone 15.0g, yeast powder 2.0g, glucose 20.0g, soluble starch 0.5g, sodium chloride 5.0g, 5wt% cysteine solution 10.0mL, Fructus Lycopersici esculenti leachate 400.0mL, Tween 80 1.0g, liver extracting solution 80.0mL, distilled water 520.0mL, pH7.0;

The preparation process of described Fructus Lycopersici esculenti leachate is as follows:

New fresh tomato is cleaned chopping after weighing, add equivalent distilled water and heat in 100 DEG C of water-baths, stir 90min, then use filtered through gauze, adjust pH7.0;

The preparation process of described liver extracting solution is as follows:

Take fresh liver 1000g, be cut into small pieces or rub, adding distil water is to 2000mL, and mixing, puts refrigerator overnight, within second day, boil 15min, and filtered through gauze also extrudes collection filtrate, adds water and is settled to 2000mL.

It is as follows that MRS fluid medium in embodiment often rises component:

Peptone 10.0g, Carnis Bovis seu Bubali cream 10.0g, yeast extract 5.0g, diammonium hydrogen citrate [(NH ₄) ₂hC ₆h ₅o ₇] 2.0g, glucose (C ₆h ₁₂o ₆h ₂o) 20.0g, Tween 80 1.0mL, sodium acetate (CH ₃cOONa3H ₂o) 5.0g, dipotassium hydrogen phosphate (K ₂hPO ₄3H ₂o) 2.0g, magnesium sulfate (MgSO ₄7H ₂o) 0.58g, manganese sulfate (MnSO ₄h ₂o) 0.25g, agar 18.0g, distilled water are settled to 1000mL, pH6.2 ~ 6.6.

Embodiment 1

A preparation method for the six drugs containing rehmanniae compound microecological oral liquid of stomach, trypsin treatment, step is as follows:

(1) by the Six-element crude drug in LIUWEI DIHUANG WAN raw material after crushed, obtained material powder, then adds 200 grams of water and refined honey by every hectogram material powder, mix homogeneously, obtained raw slurry;

(2) in the obtained raw slurry of step (1), adding hydrochloric acid adjust pH is 1.5, add the pepsin of 20 unit of activity by every hectogram LIUWEI DIHUANG WAN raw material, temperature 36 DEG C, process 3 hours, sodium carbonate adjust pH is 8, the serosity of obtained pepsin;

(3) in the serosity of the obtained pepsin of step (2), add the trypsin of 50 unit of activity by every hundred milliliters, temperature 37 DEG C, process 2 hours, hydrochloric acid adjust pH to 5.5, obtain the serosity of stomach, trypsin treatment;

(4) in the serosity of the obtained stomach of step (3), trypsin treatment, by every hundred milliliters of inoculation activated bacillus bifiduss of 0.5g and the activated Lactobacillus plantarum of 1.5, the six drugs containing rehmanniae compound microecological oral liquid of obtained stomach, trypsin treatment.

The LIUWEI DIHUANG WAN raw material of described step (1) is as follows, is weight portion:

Radix Rehmanniae Preparata 160 parts, Fructus Corni (processed) (wine Fructus Corni) 80 parts, Cortex Moutan 60 parts, Rhizoma Dioscoreae 80 parts, 60 parts, Poria, Rhizoma Alismatis 60 parts, refined honey 7 parts.

Embodiment 2

A preparation method for the six drugs containing rehmanniae compound microecological oral liquid of stomach, trypsin treatment, step is as follows:

(1) by the Six-element crude drug in LIUWEI DIHUANG WAN raw material after crushed, obtained material powder, then adds 600 grams of water and refined honey by every hectogram material powder, mix homogeneously, obtained raw slurry;

(2) in the obtained raw slurry of step (1), adding sour adjust pH is 2.2, add the pepsin of 60 unit of activity by every hectogram LIUWEI DIHUANG WAN raw material, temperature 38 DEG C, process 2 hours, sodium carbonate adjust pH is 7.8, the serosity of obtained pepsin;

(3) in the serosity of the obtained pepsin of step (2), add the trypsin of 20 unit of activity by every hundred milliliters, temperature 37 DEG C, process 3 hours, hydrochloric acid adjust pH to 6, obtain the serosity of stomach, trypsin treatment;

(4) in the serosity of the obtained stomach of step (3), trypsin treatment, by every hundred milliliters of inoculation activated bacillus bifiduss of 2g and the activated Lactobacillus plantarum of 3g, the six drugs containing rehmanniae compound microecological oral liquid of obtained stomach, trypsin treatment.

The LIUWEI DIHUANG WAN raw material of described step (1) is as follows, is weight portion:

Radix Rehmanniae Preparata 160 parts, Fructus Corni (processed) (wine Fructus Corni) 80 parts, Cortex Moutan 60 parts, Rhizoma Dioscoreae 80 parts, 60 parts, Poria, Rhizoma Alismatis 60 parts, refined honey 22 parts.

Test example 1

Bai, man, people from Changqing District, Jinan, soreness of the waist and knees, two skelasthenia, vertigo and tinnitus, insomnia and dreamful sleep, spleen and stomach function is normal.Take the six drugs containing rehmanniae compound microecological oral liquid of the stomach described in embodiment 1, trypsin treatment, every day secondary, each 15 grams, take one month, symptom is obviously improved, and spleen and stomach function is normal.

Marquis, man, Weihai people from Wendeng City, long-term taking LIUWEI DIHUANG WAN causes the symptom of cold syndrome of the stomach, abdominal distention, loose stool, inappetence.Take the six drugs containing rehmanniae compound microecological oral liquid of the stomach described in embodiment 1, trypsin treatment, every day secondary, each 15 grams, take one month, the symptom of cold syndrome of the stomach, abdominal distention, loose stool, inappetence alleviates gradually.

Test example 2

Have through 50 the six drugs containing rehmanniae compound microecological oral liquid that remarkable deficiency of the kidney yin patient takes stomach, trypsin treatment described in embodiment 2, every day takes twice, each 35 can, take one month, be designated as group 1.Simultaneously 50 have remarkable deficiency of the kidney yin patient and take common commercially available LIUWEI DIHUANG WAN, and by specification is taken, and is designated as group 2.Result is as shown in table 1.

Table 1

	Deficiency of the kidney yin symptom is improved (people)	Deficiency and coldness of spleen and stomach symptom (people)
			Group 1	47	0
Group 2	49	12

Interpretation of result

As can be seen from Table 1, group 1 and group 2 all obviously can improve the symptom of deficiency of the kidney yin patient, but existing commercially available LIUWEI DIHUANG WAN can cause part to take the symptom that patient produces cold syndrome of the stomach, abdominal distention, loose stool, inappetence, and taking the six drugs containing rehmanniae compound microecological oral liquid of stomach described in embodiment 2, trypsin treatment, the patient that takes producing above-mentioned symptom obviously reduces.

Claims (9)

1. a preparation method for the six drugs containing rehmanniae compound microecological oral liquid of stomach, trypsin treatment, it is characterized in that, step is as follows:

(1) by the Six-element crude drug in LIUWEI DIHUANG WAN raw material after crushed, obtained material powder, then adds 200 ~ 600 grams of water and refined honey by every hectogram material powder, mix homogeneously, obtained raw slurry;

(2) in the obtained raw slurry of step (1), adding sour adjust pH is 1.5 ~ 2.2, add the pepsin of 20 ~ 60 unit of activity by every hectogram LIUWEI DIHUANG WAN raw material, temperature 36 ~ 38 DEG C, process 2 ~ 3 hours, adjusting PH with base value is 7.8 ~ 8.2, the serosity of obtained pepsin;

(3) in the serosity of the obtained pepsin of step (2), add the trypsin of 20 ~ 50 unit of activity by every hundred milliliters, temperature 36 ~ 38 DEG C, process 2 ~ 3 hours, sour adjust pH to 5.5 ~ 6, obtain the serosity of stomach, trypsin treatment;

(4) in the serosity of the obtained stomach of step (3), trypsin treatment, by every hundred milliliters of inoculation activated bacillus bifiduss of 0.5 ~ 2g and the activated Lactobacillus plantarum of 1.5 ~ 3g, the six drugs containing rehmanniae compound microecological oral liquid of obtained stomach, trypsin treatment.

2. preparation method as claimed in claim 1, it is characterized in that, the LIUWEI DIHUANG WAN raw material of described step (1) is as follows, is weight portion:

Radix Rehmanniae Preparata 160 parts, Fructus Corni (processed) (wine Fructus Corni) 80 parts, Cortex Moutan 60 parts, Rhizoma Dioscoreae 80 parts, 60 parts, Poria, Rhizoma Alismatis 60 parts, refined honey 7 ~ 10 parts.

3. preparation method as claimed in claim 1, it is characterized in that, the LIUWEI DIHUANG WAN raw material of described step (1) is as follows, is weight portion:

Radix Rehmanniae Preparata 160 parts, Fructus Corni (processed) (wine Fructus Corni) 80 parts, Cortex Moutan 60 parts, Rhizoma Dioscoreae 80 parts, 60 parts, Poria, Rhizoma Alismatis 60 parts, refined honey 16 ~ 22 parts.

4. preparation method as claimed in claim 1, it is characterized in that, the acid in described step (2), (3) is hydrochloric acid.

5. preparation method as claimed in claim 1, it is characterized in that, the alkali of described step (1) is sodium carbonate.

6. preparation method as claimed in claim 1, it is characterized in that, in described step (4), activated bacillus bifidus refers to and is inoculated in TPY fluid medium by bacillus bifidus, is cultured to the bacillus bifidus of exponential phase at 36 ~ 38 DEG C.

7. preparation method as claimed in claim 6, it is characterized in that, it is as follows that described TPY fluid medium often rises component:

Peptone 15.0g, yeast powder 2.0g, glucose 20.0g, soluble starch 0.5g, sodium chloride 5.0g, 5wt% cysteine solution 10.0mL, Fructus Lycopersici esculenti leachate 400.0mL, Tween 80 1.0g, liver extracting solution 80.0mL, distilled water 520.0mL, pH7.0.

8. preparation method as claimed in claim 1, it is characterized in that, in described step (4), activated Lactobacillus plantarum refers to and is inoculated in MRS fluid medium by Lactobacillus plantarum, is cultured to the Lactobacillus plantarum of exponential phase at 30 ~ 35 DEG C.

9. preparation method as claimed in claim 8, it is characterized in that, it is as follows that described MRS fluid medium often rises component:

Peptone 10.0g, Carnis Bovis seu Bubali cream 10.0g, yeast extract 5.0g, diammonium hydrogen citrate [(NH ₄) ₂hC ₆h ₅o ₇] 2.0g, glucose (C ₆h ₁₂o ₆h ₂o) 20.0g, Tween 80 1.0mL, sodium acetate (CH ₃cOONa3H ₂o) 5.0g, dipotassium hydrogen phosphate (K ₂hPO ₄3H ₂o) 2.0g, magnesium sulfate (MgSO ₄7H ₂o) 0.58g, manganese sulfate (MnSO ₄h ₂o) 0.25g, agar 18.0g, distilled water are settled to 1000mL, pH6.2 ~ 6.6.