CN101015574A - Preparing method of traditional Chinese medicine pills for treating hepatitis - Google Patents

Abstract

The invention discloses a preparation method of a Chinese medicinal pill. The preparation method comprises decocting Penthorum chinense in water for 2-3 times for 1-3h each time, mixing decoctions, filtering, concentrating, cooling, adding ethanol to 55-70% for precipitating, stirring, standing, filtering to obtain filtrate and filter residue, cleaning the filter residue with ethanol, mixing the cleaning solutions and filtrate, recovering ethanol, concentrating to obtain an extract with relative density of 1.30-1.35, mixing with starch, drying, pulverizing, adding 65-85% ethanol, granulating, and packaging. The invention has the advantages of low dose, good dissolubility, and convenient administration. The inventive pill can be used to treat hepatitis, such as chronic active hepatitis, hepatitis B, and acute viral hepatitis.

Description

A kind of preparation method who is used for the treatment of the medicine pill of hepatitis

Technical field

The present invention relates to a kind of enzyme of falling that has, protect the liver, the preparation method of the medicine pill of removing jaundice, especially a kind of preparation method who is used for the treatment of the medicine pill of CAH, hepatitis B, acute viral hepatitis.

Background technology

This product prescription is from " Drug Standard of Ministry of Public Health of the Peoples Republic of China Traditional Chinese medicine historical preparation " the 13rd GANSU KELI, and standard No. be [WS3-B-2526-97], and its preparation method is for " get and pull root vegetables 1000g; chopping, filter boiling three times, each 2 hours; collecting decoction, and filtrate is condensed into the clear cream of relative density 1.15～1.18 (60～65 ℃); cooling; add ethanol and make and contain the alcohol amount and reach 60%, stir, leave standstill; filtration, precipitation is with 60% ethanol washing three times, merging washing lotion and filtrate, Recycled ethanol and to be condensed into relative density be 1.30～1.32 thick paste, add an amount of sucrose and starch, mixing, particle processed, drying, make 540g, and get final product ". Former side is used for for many years clinical with the granule form, determined curative effect. But former formulation (GANSU KELI) is oral 9g once, and dose is large; Contain sugar in the particle, the use crowd is limited, and the diabetic should not use. Therefore we to consider to reform formulation be ball (concentrated micropill) agent. Simultaneously also provide more more options for clinical application. Because concentrated micropill particle diameter is less, easily swallow, and auxiliary material does not contain sugar; Adopt Modern New Technology, the concentrated micropill of new technology preparation, can significantly reduce supplementary product consumption and reduce dose, be easier to accept into the patient, and applicable crowd is extensive. Concentrated micropill also has fast and leaches, and carries, transports, takes all advantages such as convenient.

Summary of the invention

The method that the purpose of this invention is to provide a kind of easy for patients to accept, dose is little, leach fast, taking convenience, applicable crowd are wide medicine pill that is used for the treatment of CAH, hepatitis B, acute viral hepatitis.

For realizing goal of the invention, the inventor has filtered out the method for the medicine pill that is used for the treatment of CAH, hepatitis B, acute viral hepatitis on the basis of test of many times, and described method comprises following steps:

1. will pull root vegetables boiling 2-3 time, each 1-3 hour, collecting decoction filtered, filtrate is condensed under 60-65 ℃ of condition and surveys relative density is the clear cream of 1.10-1.20, and cooling adds ethanol and makes and contain the alcohol amount and reach 55-70%, stirs, leave standstill, filter, get filter residue and filtrate A;

2. filter residue washs 2-3 time with the ethanol of 50-90%, merges washing lotion and filtrate A, and it is the thick paste of 1.30-1.35 that Recycled ethanol also is condensed into 40 ℃ of condition downside relative densities, adds appropriate amount of starch, mixes, and drying is pulverized, and gets medicinal powder; Add the 65-85% appropriate amount of ethanol, pill, packing, packing, and get final product.

1. described step is to pull the root vegetables boiling 3 times, and each 2 hours, collecting decoction filtered, and filtrate is condensed into and surveys relative density under 60-65 ℃ of condition is the clear cream of 1.15-1.18.

Described step amount of water 1. is respectively: for the first time 12 times of medicinal material amounts, for the second time 10 times of medicinal material amounts, for the third time 8 times of medicinal material amounts.

1. described step adds ethanol makes and contains alcohol amount and reach 60%.

Described step 2. filter residue merges washing lotion and filtrate A with 60% ethanol washing 3 times, Recycled ethanol and to be condensed into 40 ℃ of condition downside relative densities be the thick paste of 1.30-1.32.

Details are as follows to the preparation technology of described pill for the inventor:

One, process rationality research and explanation

1, technical study explanation

The primary standard method for making for " get and pull root vegetables 1000g, chopping, filter boiling three times, each 2 hours; collecting decoction, filtrate is condensed into the clear cream of relative density 1.15～1.18 (60～65 ℃), cooling; add ethanol and make and contain the alcohol amount and reach 60%, stir, leave standstill, filter; precipitation is with 60% ethanol washing three times, merging washing lotion and filtrate, Recycled ethanol and to be condensed into relative density be 1.30～1.32 thick paste; add an amount of sucrose and starch, mixing, particle processed; drying, make 540g, and get final product. "

Since the clear and definite extraction time of primary standard method for making and time, the amount of water when clearly not extracting, and the amount of water when therefore needing extraction is studied; The clear and definite alcohol precipitation process condition of primary standard, washing of precipitate solvent and concentration, therefore not clear and definite cleaning solvent consumption needs the cleaning solvent consumption is studied; Change pill into by granule, also tackle preparations shaping technique and study.

2, Study on extraction

(1) water absorption rate is measured:

Take by weighing that to pull the root vegetables medicinal material an amount of, add 15 times of water gagings and soak, to the saturating heart of medicinal material, leach unabsorbed water liquid, trying to achieve water absorption rate is 289.7%.

(2) extract the amount of water screening

Take by weighing respectively medicinal material 100g, boiling three times, each 2 hours, decocting liquid filtered, and merges three times filtrate, is condensed into medicinal extract, weighs. Survey quercetin content, dried cream yield in the medicinal extract. Result of the test sees Table 1.

Because the water absorption rate of this product medicinal material is higher, and this product medicinal material is herb, and volume is large, is 12 times of medicinal material amounts so intend extracting the highest amount of water.

1 ^#: for the first time 12 times of amounts, for the second time 10 times of amounts, for the third time 8 times of amounts.

2 ^#: for the first time 10 times of amounts, for the second time 8 times of amounts, for the third time 6 times of amounts.

3 ^#: for the first time 8 times of amounts, for the second time 6 times of amounts, for the third time 4 times of amounts.

Table 1 extracts as a result table of amount of water screening test

Tested number	Medicinal extract amount (g)	Quercetin content in the medicinal extract (%)	Dried cream yield (%)
				1 #	43.2	0.0725	13.5
2 #	40.8	0.0720	12.3
					44.2	0.0611	11.1

From experimental result as can be known, 1^#Best, so the amount of water that determine to decoct extracts is: for the first time 12 times of medicinal material amounts, for the second time 10 times of medicinal material amounts, for the third time 8 times of medicinal material amounts.

(3) alcohol precipitation process research

The material of getting it filled is an amount of, boiling three times, each 2 hours, for the first time 12 times of medicinal material amounts, for the second time 10 times of medicinal material amounts, for the third time 8 times of medicinal material amounts, decocting liquid filters, and merges three times filtrate, is concentrated into the clear cream of relative density 1.15～1.18 (60～65 ℃), be cooled to room temperature, add ethanol and make and contain alcohol amount and reach 60%, accelerate slowly to stir, leave standstill, filter, get precipitation. Precipitation is divided equally three parts, and according to the form below adds the washing of 60% ethanol, and the residue after the washing is dried to constant weight, gets dry residue. Quercetin content is as index in the dry residue, and experimental result sees Table 2.

Table 2 washing of precipitate ethanol consumption screening table

Precipitation capacity (g)	60% total ethanol consumption (doubly)	Washing times (inferior)	Quercetin content in the dry residue (%)
				50	0	0	0.223
50	3	3	0.030
				50	6	3	0.011
50	9	3	0.010

From experimental result as can be known, the alcohol precipitation postprecipitation adds the washing of 6 times, 9 times amount 60% ethanol, and quercetin content is low in the residue after the washing. And Mongolian oak content does not have significant difference in the residue after 6 times, 9 times amounts are washed, and considers from saving the cost aspect, determines this product alcohol precipitation postprecipitation, with 6 times of amount 60% ethanol washing 3 times, each 2 times of amounts.

The extraction process checking

Get and pull root vegetables 2kg, boiling three times, each 2 hours, for the first time 12 times of medicinal material amounts, for the second time 10 times of medicinal material amounts, for the third time 8 times of medicinal material amounts, decocting liquid filters, merge three times filtrate, be concentrated into the clear cream of relative density 1.15～1.18 (60～65 ℃), be cooled to room temperature, adding ethanol makes and contains alcohol amount and reach 60%, slowly accelerate to stir, leave standstill, filter, clear cream (relative density 1.16,65 ℃), be cooled to room temperature, add ethanol and make and contain alcohol amount and reach 60%, slowly accelerate to stir, leave standstill, filter, filtrate is deposited in addition. Precipitation is washed 3 times with 6 times of amount 60% ethanol, and each 2 times of amounts merge washing lotion and filtrate, and Recycled ethanol also is condensed into medicinal extract, weighs. Survey quercetin content, dried cream yield in the medicinal extract. (repeated test 3 times) result of the test sees Table 12.3.

Table 12.3 extraction process demonstration test is table as a result

The experiment number	Medicinal extract amount (g)	Quercetin content (%) in the clear cream	Dried cream yield (%)	Average dried cream yield (%)
					1	725.5	0.0838	8.29	8.13
2	735.1	0.0832	7.76
				3	728.6	0.0837	8.34

Experimental result shows that this extraction process is reasonable, feasible, has reappearance, operability.

3, separation, purifying, concentrated, drying process research

(1) separation, purifying process research.

After extract merged, 200 mesh sieves filtered, and get filtrate, and are concentrated. Behind the alcohol precipitation, the filter paper suction filtration gets pure filtrate. Behind the Washing of Filter Cake, suction filtration gets pure filtrate, merges pure filtrate, and Recycled ethanol is concentrated.

(2) concentration technology research

1. extract is concentrated

For improving as far as possible thickening efficiency, reducing production costs, shorten concentration time, avoid long-time heating to the destruction of active ingredient, adopt triple effect reduced pressure concentration method to concentrate. Actual conditions is: temperature: 80 ℃ of effects; 70 ℃ of two effects; 60 ℃ of triple effects. Vacuum: an effect 0.04Mpa; Two effect 0.06Mpa; Triple effect 0.08Mpa.

2. pure filtrate is concentrated

Alcohol precipitation rear filtrate and washing of precipitate rear filtrate merge, and first decompression recycling ethanol is concentrated again, gets clearly cream (relative density is about 1.15～1.17,70 ℃ of surveys). Actual conditions is: temperature: 70 ℃～80 ℃; Vacuum: 0.06Mpa～0.08Mpa;

3. clear cream is concentrated

Because decompression concentrator is the totally enclosed type inspissator, it is too large that medicinal extract is concentrated into relative density, gets cream not to the utmost, and the medicinal extract loss is large. So liquid is concentrated into clearly cream (about 1.15～1.17, the 70 ℃ of surveys of relative density) with decompression concentrator behind the alcohol precipitation, select again steam concentrated. For making things convenient for next drying process, determine that clear cream is concentrated into the thick paste of relative density 1.30～1.32 (60 ℃ of surveys) with steam.

(3) extract dry technical study

This product medicinal extract is water extract-alcohol precipitation medicinal extract, has stronger viscosity. Once adopted spray drying process that material is carried out drying, the sticking wall of material, bonding are seriously lost greatly as a result, so adopt bake drying commonly used. Adopt bake drying, temperature is larger to the quality influence of material, so the temperature when needing drying is screened.

1. extract dry temperature screening

Get same thick medicinal extract (1.30,60 ℃ of relative densities), bake drying under 70 ℃, 80 ℃, 90 ℃ conditions respectively, in the different time sections sampling, the moisture in the test sample product, thick skin cellulose content and the broken situation of dried cream powder are as index in the constant weight sample. Result of the test sees Table 4.

Table 4 extract dry temperature screening table

Baking temperature (℃)	Dry 6 hours moisture (%)	Dry 8 hours moisture (%)	Quercetin content in the constant weight sample (%)	The broken situation of dried cream powder
					70	10	7	0.345	Moisture absorption is fast, pulverizes difficulty
80	7	5	0.335	Moisture absorption is fast, pulverizes difficulty
					90	5	/	0.299	Moisture absorption is fast, pulverizes difficulty

According to table 4 result of the test, determine that this product extract dry temperature is 80 ℃.

2. auxiliary material and amount screening during extract dry

From extract dry temperature screening test as can be known, the dried cream hygroscopicity of this product is strong, shatters difficulty, so it is dry to consider to add in the medicinal extract auxiliary material. Intend adding starch commonly used.

Get respectively 0.1 times of thick medicinal extract of recipe quantity (1.30,60 ℃ of relative densities), according to the form below adds starch and mixes, and is strong and weak with the hygroscopicity of dried cream after the drying 80 ℃ of dryings, pulverizes difficulty or ease for investigating index, and result of the test sees Table 5.

Auxiliary material and amount screening table during table 5 extract dry

Sample	Amount of starch (g)	Dried cream hygroscopicity	Pulverize difficulty or ease
				1	17.0	----	+
2	32.5	--	+++
				3	37.0	--	+++

Annotate: dried cream hygroscopicity: "-" expression hygroscopicity is strong.

Pulverize difficulty or ease: "+" expression is pulverized easily.

Table 5 experimental result shows: 2, a little less than No. 3 dried cream hygroscopicity of sample, easily pulverize. Consider from reducing dose and cost aspect, select No. 2. Namely one times of thick medicinal extract of recipe quantity adds about 325g starch.

3. determine extract dry technique

Get the thick paste of relative density 1.30～1.32 (60 ℃ of surveys), add appropriate amount of starch, mix 80 ℃ of bake dryings.

5, preparations shaping Journal of Sex Research

(1) recipe quantity design

1. dose is determined

Each serving consumption, be equivalent to primary dose 16.7g according to primary standard (GANSU KELI standard), determine that this product (liver Soviet Union ball) dose is consistent with primary standard (GANSU KELI standard), namely each serving consumption, be equivalent to primary dose 16.7g.

By experiment as can be known, this product paste-forming rate is about 8.1%, and namely each serving being about 1.35g with the dry extract amount, auxiliary material is about 0.65g, amounts to 2.00g, namely each serving using about 2.0g.

2. preparation prescription design

A, Extraction parts medicinal material are 8350g, and dried cream amount is about 676g.

B, the about 325g of auxiliary material amount.

Amount to 1001g, make 500 bottles, every bottle of heavily about 2.0g.

(2) preparations shaping research:

1. medicinal powder granularity

Because this product pill is less, so higher to the medicinal powder fineness requirement. According to experience in the past, determine that this product medicinal powder is fine powder.

2. wetting agent and consumption screening

Because this product is the medicinal powder pill, so need to add wetting agent. According to this product dried cream powder character, intending ethanol is the wetting agent of this product.

A ethanol consumption

Get one times of recipe quantity medicinal powder 1000g, add 30% ethanol, to the medicinal powder complete wetting, softwood processed. Calculate ethanol percentage consumption about 28%.

B, concentration of alcohol are determined

Get one times of recipe quantity medicinal powder, the adding different concentration ethanol is fully moistening, softwood processed, pill. Take the softwood quality with become the ball situation as investigating index, the results are shown in Table 12.6.

Table 12.6 wetting agent screening table

Tested number	The medicinal powder amount	Wetting agent	The softwood quality	Become the ball situation
					1	1000g	28%30% ethanol	Viscosity is strong	Difficult one-tenth ball
2	1000g	28%50% ethanol	Viscosity weakens to some extent	Difficult one-tenth ball
					3	1000g	28%70% ethanol	A little less than the viscosity	Can become ball
4	1000g	28%80% ethanol	A little less than the viscosity	Easily become ball

From result of the test as can be known: 4^#The softwood quality is better, easily pill. So determine that this product wetting agent concentration of alcohol is 80%.

3. pill piece

The powder of getting it filled adds 28% 80% ethanol pill piece, gets the uniform ball piece of color and luster. A large amount of production adopted kneading machine, and limited production and medicine can carry out in basin.

4. pill bar, gradation and stranding are round:

Large production adopted the machine pill, and limited production is with rubbing ball wrench worker preparation with the hands.

The pill bar is most important to moulding the pill quality. The fineness and the ball method of double differences that directly have influence on pill are different. Cause that shaping is not bright and clean, the inconsistent main cause of thickness is the temperature of softwood. Therefore need the softwood temperature is screened.

Get the ball piece that makes, the temperature of according to the form below is carried out the pill bar, and gradation take pill bar situation and gradation situation as investigating index, the results are shown in Table 7.

Table 7 pill bar, gradation temperature investigation table

Tested number	Temperature	Pill bar situation and gradation situation
			1	10℃±2	The ball bar is even, is difficult for gradation
2	5℃±2	The ball bar is even, easily gradation
			3	0℃±2	The ball bar is inhomogeneous, easily gradation

As known from Table 7, this product pill bar, gradation temperature are 5 ℃ ± 2.

5. wet ball is dry:

The factor of the wet ball drying of impact mainly is temperature. Therefore need wet ball baking temperature is screened.

Get respectively the upper wet ball that makes, the temperature of according to the form below is carried out drying, and it is moisture below 9% to be dried to a ball. Take the quality of a ball as investigating index, the results are shown in Table 8.

The wet ball baking temperature screening of table 8 table

Tested number	Baking temperature	Ball quality after dry
			1	60 ℃ of dryings of elder generation 1.5 hours are warming up to 80 ℃ of dryings 8 hours more gradually	Moisture content 5.0% has the aberration phenomenon
2	50 ℃ of dryings of elder generation 1.5 hours are warming up to 70 ℃ of dryings 8 hours more gradually	Moisture content 6.0% ball is matt
			3	40 ℃ of dryings of elder generation 1.5 hours are warming up to 60 ℃ of dryings 8 hours more gradually	Moisture content 6.5%, a ball is glossy

As known from Table 8,3^#Baking temperature is better, so this product selects 3^# Namely 40 ℃ of dryings of wet ball elder generation are 1.5 hours, are warming up to gradually 60 ℃ of dryings again.

6. select ball

The pill of screening 2.5mm～1.0mm.

7. determine pill making craft

Get dried cream, be ground into fine powder, add about 28% 80% ethanol, softwood processed, on be equipped with the pellet processing machine pill of ball bar, gradation temperature, wet ball is 40 ℃ of dryings 1.5 hours first, be warming up to gradually again 60 ℃ of dryings (will stir in the dry run), be dried to moisture below 9.0%, get dried ball, the pill of screening 2.5mm～1.0mm.

6, packing

(1) critical relative moisture is measured

Get respectively 5 parts of concentrated micropills, every part of about 1g puts into respectively relative humidity 33%, 42.8%, 59.7%, 75.3%, 81% environment, places 72 hours in 25 ℃ of incubators, takes out weighing, calculates moisture absorption weightening finish percentage, the results are shown in Table 9. Take moisture absorption weightening finish percentage as ordinate, relative humidity is that abscissa is made curve, the results are shown in Figure 1.

Table 9 critical relative moisture investigation table

The saturated aqueous solution of salt	Relative humidity (%)	Moisture absorption weightening finish percentage (%)
			MgCl.6H 2O	33	0.30
K 2CO 3.2H 2O	42.8	0.61
			NaBr.2H 2O	59.7	1.24
NaCl	75.3	2.16
			(NH 4) 2SO 4	81	3.05

Result of the test shows: this kind critical relative moisture is 67%. So this product is 25 ℃ of temperature, relative humidity is carried out packing below 65%, can not affect the quality of product.

(2) packing condition

In 300,000 grades of clean areas (18 ℃～25 ℃ of room temperatures, relative humidity is below 65%), carry out packing.

7, packaging material determines

For being convenient for carrying, transporting, taking, determine this product oral stable medicinal polythene bottle with high density packing.

12.5 pilot scale research

By the process conditions of determining, manufacture experimently ten batches in test agent (lot number: 031201,031202,031203; 040801,040802,040803,040804; 050101,050102,050103) three batches of pilot experiments of 12.5.1 (031201,031202,031203) the results are shown in Table 10, table 11

Test agent creation data table in the table 10

Lot number	031201	031202	031203
				Pull root vegetables (kg)	83.5	83.5	83.5
Medicinal extract amount (kg)	16.490	15.866	13.982
				Medicinal extract relative density (60 ℃ of surveys)	1.30	1.30	1.32
Dried cream yield (%)	8.0	7.9	7.9
				Amount of starch (kg)	3.328	3.404	3.404
Medicinal powder weight (kg)	9.892	9.857	9.805
				Theoretical yield (kg)	10.000	10.000	10.000
Actual production (kg)	9.563	9.533	9.498
				Product yield (%)	95.6	95.3	95.0

Table 11 pilot scale sample quality inspection knot

	Lot number	031201	031202	031203
					Identification check is little	Proterties	The sepia condensed pill; Bitter is little puckery.	The sepia condensed pill; Bitter is little puckery.	The sepia condensed pill; Bitter is little puckery.
Differentiate (1)	Be positive reaction	Be positive reaction	Be positive reaction
				Differentiate (2)		Up to specification	Up to specification	Up to specification
Moisture (%)≤9.0%	5.32	5.56	5.03
				Content uniformity		Up to specification	Up to specification	Up to specification
Difference limit ± 10%
				Dissolve scattered time limit (minute)		8	8	9
Bacterial population (individual/gram)≤1000	＜10	10	10

Biological content	Fungi count (individual/gram)≤100	＜10	＜10	＜10
					Escherichia coli must not detect	Do not detect	Do not detect	Do not detect
	Every bottle contains Quercetin and must not be less than 4.0mg	5.30	5.28	5.28
					Pull root vegetables content (%)		0.069
	The rate of transform (%)	46.0	45.8	45.8

12.5.2 (040801,040802,040803,040804) four batches of pilot experiments the results are shown in Table 12, table 13

Test agent creation data table in the table 12

Lot number	040801	040802	040803	040804
					Pull root vegetables (kg)	83.5	83.5	83.5	83.5
Medicinal extract amount (kg)	17.565	17.779	15.990	16.922
					Medicinal extract relative density (60 ℃ of surveys)	1.30	1.30	1.31	1.30
Dried cream yield (%)	8.1	8.0	7.9	8.0
					Amount of starch (kg)	3.236	3.320	3.404	3.320
Medicinal powder weight (kg)	9.827	9.809	9.881	9.866
					Theoretical yield (kg)	10.000	10.000	10.000	10.000
Actual production (kg)	9.563	9.488	9.603	9.550
					Product yield (%)	95.6	94.9	96.0	95.5

Table 13 pilot scale sample quality inspection knot

	Lot number	040801	040802	040803	040804
						Differentiate	Proterties	The sepia condensed pill; Bitter is little puckery.	The sepia condensed pill; Bitter is little puckery.	The sepia condensed pill; Bitter is little puckery.	The sepia condensed pill; Bitter is little puckery.
Differentiate (1)	Be positive reaction	Be positive reaction	Be positive reaction	Be positive reaction
					Differentiate (2)		Up to specification	Up to specification	Up to specification	Up to specification
Moisture (%)≤9.0%	5.61	5.83	5.39	5.80
					Check		Content uniformity difference limit ± 10%	Up to specification	Up to specification	Up to specification	Up to specification
Dissolve scattered time limit (minute)	5	6	6	6
						Microorganism	Bacterial population (individual/gram)≤1000	＜10	＜10	10	＜10
Fungi count (individual/gram)≤100	＜10	＜10	＜10	＜10
					Escherichia coli must not detect		Do not detect	Do not detect	Do not detect	Do not detect
Content	Every bottle contains Quercetin and must not be less than 4.0mg	5.45	5.68	5.31							5.52
					Pull root vegetables content (%)	0.069
	The rate of transform (%)	47.3	49.3	46.1						47.9

12.5.3 (050101,050102,050103) three batches of pilot experiments the results are shown in Table 14, table 15

Test agent creation data table in the table 14

Lot number	050101	050102	050103
				Pull the root Lay	83.5	83.5	83.5
Medicinal extract amount (kg)	17.655	18.206	16.683

Medicinal extract relative density (60 survey)	1.30	1.30	1.32
				Dried cream yield (%)	8.2	8.3	8.3
Amount of starch (kg)	3.153	3.070	3.070
				Medicinal powder weight (kg)	9.806	9.622	9.639
Theoretical yield (kg)	10.000	10.000	10.000
				Actual production (kg)	9.560	9.511	9.468
Product yield (%)	95.6	95.1	94.7

Table 15 pilot scale sample quality inspection knot

	Lot number	050101	050102	050103
					The identification check content of microorganisms	Proterties differentiate (1) differentiate (2) moisture (%)≤9.0% content uniformity difference limit ± 10% dissolve scattered time limit (minute) bacterial population (individual/gram)≤1000 fungi counts (individual/gram)≤100 Escherichia coli must not detect every bottle and contain Quercetin and must not be less than 4.0mg	The sepia condensed pill; Bitter is little puckery. Be positive reaction up to specification 5.82 up to specification 5 10＜10 and do not detect 8.02	The sepia condensed pill; Bitter is little puckery. Be positive reaction up to specification 6.28 up to specification 6＜10＜10 and do not detect 8.24	The sepia condensed pill; Bitter is little puckery. Be positive reaction up to specification 6.12 up to specification 5＜10＜10 and do not detect 7.92

Pull root vegetables content (%) rate of transform (%)

44.26

0.069     45.99

42.52

Experimental result shows, every regulation that pilot scale samples met pill is relevant, and this preparation process is stablized feasible.

Description of drawings

The critical relative moisture figure of Fig. 1 pill of the present invention

Pharmacodynamic study

Medicine of the present invention pulls the root vegetables single by genuine traditional Chinese medicine and forms, and has the enzyme of falling, and protects the liver removing jaundice, the effect of invigorating the spleen. Be used for CAH, hepatitis B, also can be used for acute viral hepatitis. This pharmaceutically dosage form also has granule on market at present, records in " Drug Standard of Ministry of Public Health of the Peoples Republic of China Traditional Chinese medicine historical preparation) " the 13rd. Former granule clinical practice for many years, determined curative effect, but oral 9g once, dose is large, reduces patient's compliance. The present invention adopts concentrated pellet preparations, and particle diameter is less, easily swallow, and auxiliary material does not contain sugar, therefore can reduce dose, and concentrated micropill also has fast and leach, and carries, transports, stores the advantages such as convenient.

This research is by causing the impact of acute liver model on D-Gal and carbon tetrachloride, the impact that the mouse hemolytic antibody is generated is on the observation of the impact of mouse macrophage system phagocytic function, with the drug effect of checking medicine of the present invention.

1 experiment material

Animal: small white mouse, the Kunming kind, one-level is qualified, male and female half and half, body weight 18-22g is provided by The Fourth Military Medical University's Experimental Animal Center.

Medicine and reagent: the used preparation of test group is that namely the lab scale medicine is provided by Jiangxi Yaodu Renhe Pharmaceutical Co., Ltd, lot number 060519 according to the concentrated pellet preparations of the embodiment of the invention 1 preparation. Control drug: GANSU KELI, Sichuan doctor pharmaceutcal corporation, Ltd produces, lot number 051223. D-Gal is produced by SIGMA CHEMICAL CO., and all tested medicines all are made into desired concn for experiment with distilled water.

Reagent: serum glutamic pyruvic transminase (ALT) kit, lot number 060710; Glutamic-oxalacetic transaminease (AST) kit, lot number 060710; Co., Ltd provides by Bai Ding bioengineering (Beijing). Malondialdehyde Kit (MDA), superoxide dismutase (SOD) kit, building up bio-engineering research by Nanjing is provided.

Instrument: AUT0TECH-128 type automatic clinical chemistry analyzer.

2 experimental techniques and result

2.1 D-Gal is caused the impact of acute liver model

Get 60 of body weight 18～22g mouse, male and female half and half are divided into blank group, model control group, lab scale medicine high dose, middle dosage, low dosage and GANSU KELI group at random. Each group is pressed the dosage gastric infusion of table 1, successive administration 3 days, 50min after the last administration, except the blank group, all the other respectively organize lumbar injection 650mg/kgD-amine-galactose, get blood from the mouse orbit veniplex behind the 24h, survey Serum ALT, AST, SOD and MDA content; Get the part mouse liver through 10% formaldehyde fixedly more than the 48h, FFPE, section, HE dyeing, Microscopic observation each treated animal liver organization structure and lesion characteristic. The result of variations that each organizes mice serum ALT, AST sees Table 16; Mice serum SOD vigor and MDA content influence see Table 17.

Table 16 causes acute hepatic injury mice Serum ALT and AST impact to D-Gal

Group	Dosage (g/kg)	Number of animals (only)	ALT(IU/L)	AST(IU/L)
					Dosage group lab scale medicine low dose group in the blank control group model control group lab scale medicine high dose group lab scale medicine	--   --   1.248   0.624   0.312	10   10   10   10   10	39.59±6.54 **   69.54±8.49 △△   45.80±10.65 **   51.27±7.33 **   57.71±9.10 **	125.32±14.65 **   174.27±16.73 △△   132.12±12.53 **   146.95±13.46 **   155.28±15.37 **
The GANSU KELI group	3.900	10	50.10±10.51 **	147.98±1371 **

Annotate: compare with model control group,^*P＜0.05， ^**P＜0.01; With blank control group ratio,^△△P＜0.01。

Experimental result shows that the lab scale medicine has obvious prevention D-Gal to cause that chmice acute liver injury model Serum ALT, AST activity increase effect, has compared conspicuousness difference (P＜0.01) with model control group.

Table 17 pair D-Gal causes acute hepatic injury mice serum activity of SOD and MDA content influence

Group	Dosage (g/kg)	Number of animals (only)	SOD(NU/mL)	MDA(μmol/L)
					Dosage group lab scale medicine low dose group in the blank control group model control group lab scale medicine high dose group lab scale medicine	-- -- 1.248 0.624 0.312	10 10 10 10 10	450.61±24.04   264.98±85.79 △△   437.89±40.81   416.45±36.66   324.59±64.69	125.32±14.65   174.27±16.73 △△   132.12±12.53   146.95±13.46   155.28±15.37
The GANSU KELI group	3.900	10	377.66±52.62	147.98±1371

Annotate: compare with model control group,^*P＜0.05， ^**P＜0.01; With blank control group ratio,^△△P＜0.01。

Experimental result shows, compares with blank control group, and the MDA content rising in the model group mice serum is (P＜0.01) significantly; MDA content in the high, medium and low dosage group of lab scale medicine and the GANSU KELI group mice serum is starkly lower than model group, P＜0.05 and P＜0.01.

With blank control group ratio, active obviously reduce (P＜0.05) of SOD in the model group mice serum; SOD activity in the high, medium and low dosage group of lab scale medicine and the GANSU KELI group mice serum obviously is higher than model group, P＜0.05.

2.2 D-Gal is caused the pathological observation of chmice acute hepatic injury.

Dissect and check: blank control group is without obvious pathology; Model control group Mouse Liver weight increases, and is yellowish-brown to aubergine, and tangent plane turns up, and sepage is more; Each treatment group all has in various degree similar form to model group, and lesion degree obviously alleviates. Micro-pathologic finding: blank control group mice: the lobuli hepatis structural integrity changes without pathology. The model control group mouse: widely water sample sex change of liver cell, looseization of endochylema and the visible point-like of muddy swelling liver cell and focal necrosis, Visible Core is cracked, has a liking for sour little body and fat change, and the scorching sexual cell in header district infiltrates. GANSU KELI group mouse: liver cell is turbid swollen, and endochylema looseization is obvious, visible spotty necrosis, and visible focal necrosis still, the header district is a small amount of scorching sexual cell infiltration as seen. Lab scale medicine high dose group mouse: indivedual liver cells occur slight turbid swollen, accidental slight spotty necrosis. Dosage group mouse in the lab scale medicine: slight hepatic cell is turbid swollen, and the less spotty necrosis of minority scope is arranged, and the visible a small amount of scorching sexual cell in header district infiltrates. Lab scale medicine low dose group mouse: the sex change of liver cell water sample, looseization of endochylema and muddy swelling, visible spotty necrosis, accidental slight focal necrosis, the visible scorching sexual cell in header district infiltrates.

2.3 carbon tetrachloride is caused the impact of chmice acute liver injury model

Get 60 of body weight 18～22g mouse, male and female half and half are divided into blank control group, model control group, lab scale medicine high dose, middle dosage, low dosage and GANSU KELI group at random. Each group is pressed the dosage gastric infusion of table 3, and administration is 3 days continuously, and 1h is except blank control group after the last administration, and all the other are respectively organized equal gavage and give 0.12%

CCl ₄Soybean oil solution 10ml/kg behind the 24h, gets its serum and surveys ALT and AST activity, and experimental result sees Table 18.

Table 18 on carbon tetrachloride cause the chmice acute liver injury model impact (

)

Group	Dosage (g/kg)	Number of animals (only)	ALT(IU/L)	AST(IU/L)
					Dosage group lab scale medicine low dose group in the blank control group model control group lab scale medicine high dose group lab scale medicine	--   --   1.248   0.624   0.312	10   10   10   10   10	51.05±9.82 **   81.92±9.70 △△   69.26±10.54 **   70.25±10.60 *   72.13±11.73	139.50±12.77 **   186.35±14.65 △△   145.06±13.11 **   165.21±15.46 **   170.79±13.88 *
The GANSU KELI group	3.900	10	69.51±8.30 **	162.01±11.39 **

Annotate: with the model control group ratio,^*P＜0.05， ^**P＜0.01; With blank control group ratio,^△△P＜0.01。

Experimental result shows that lab scale medicine group has obvious prevention carbon tetrachloride to cause that acute hepatic injury mice model Serum ALT, AST activity increase effect, has compared conspicuousness difference (P＜0.05) with model control group.

2.4 the impact on the generation of mouse hemolytic antibody

Get 50 of body weight 18～22g mouse, male and female half and half are got the chicken red blood cell and are made into 5% normal saline suspension, and every mouse lumbar injection 0.2ml immunity with mouse random packet after the immunity, sees Table 4. Each group is pressed table 4 dosage gastric infusion, and every day 1 time, administration is 7 days continuously, and blank control group gives with volume distilled water. 1h after the last administration, mouse eyeground vein clump is got blood, centrifuging and taking serum, with 100 times of normal saline dilutions, other gets the dilution in 1: 10 of 3 cavy serum physiological saline, and is for subsequent use as complement. Getting above-mentioned dilution liquid 1ml mixes with 5% chicken red blood cell 0.5ml, in 0 ℃ of ice bath, add complement 0.5ml, be incubated 30min in 37 ℃ of warm casees, take out, 0 ℃ stops reaction, centrifugal, get supernatant liquid in " 721 " spectrophotometer colorimetric, transfer " 0 " with the blank pipe of increase serum not, 540nm measures optical density, generate index with OD value as serum hemolysin, the results are shown in Table 19.

The impact that table 19 generates the mouse hemolytic antibody

Annotate: with blank control group ratio,^*P＜0.05， ^**P＜0.01。

Table 19 is the result show, the lab scale medicine can obviously promote the mice serum hemolytic antibody to generate, each administration group mice serum hemolysin OD value has conspicuousness difference (P＜0.05 or P＜0.01) with blank control group than all, illustrates that lab scale medicine and control drug GANSU KELI all have the function that improves humoral immunity of organism.

2.5 mouse macrophage system (RES) is engulfed the impact of function

Get 50 of body weight 18～22g healthy mices, male and female half and half by the body weight random packet, see Table 5. Each group is pressed table 5 dosage gastric infusion, and every day 1 time, administration is 7 days continuously. 30min after the last administration, by mouse carbon particle clearance method, the India prepared Chinese ink 0.1ml/10g body weight of 8: 1 normal saline dilutions of tail vein injection, respectively after injection 2min, 6min with heparin solution moistening suction pipe in advance, the eyeground vein clump is got blood 20 μ l, is dissolved in 2ml 0.1% Na₂CO ₃Mixed even in the solution, " 721 " spectrophotometer 640nm wavelength place colorimetric is dissolved in 1% Na with the normal small white mouse blood of 20 μ l₂CO ₃Mixed even rear " 0 " point of transferring among the solution 2ml, photometry density (OD) value is calculated as follows and engulfs index K. With the t check method significance of difference between each administration group and blank control group relatively, the results are shown in Table 20.

$K=\frac{\mathrm{LogO}{D}_1-\mathrm{LogO}{D}_2}{T_2-T_1}$

Table 20 is engulfed the impact of function on mouse macrophage system (RES)

Annotate: with blank control group ratio,^*P＜0.05， ^**P＜0.01。

By table 20 result as seen, lab scale medicine group can obviously improve the index of engulfing of the netted endothelium of small white mouse system (RES), with blank control group ratio conspicuousness difference (P＜0.05) is arranged. Control drug GANSU KELI group also can improve the index of engulfing of the netted endothelium of small white mouse system (RES), but without the statistics meaning.

3 discuss

Glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST) content in liver cell are higher, the important index enzyme that raises in the serum when being hepar damnification. Detecting serum alt, AST activity level and can judge the serious degree of hepatic injury, is the sensitive indicator of hepatocellular damage. Pharmacodynamic experiment is the result show, medicine of the present invention has obvious prevention D-Gal hydrochloric acid salt, carbon tetrachloride to cause the effect that mice serum ALT, AST activity increase, and illustrates that this medicine has the effect of certain protecting liver, lowering enzymes.

The experimental results shows, free radical participates in many basic life processes, such as biological oxidation, cell propagation, body aging etc., the free radical reaction is to cause the molecular pathology of numerous disease and body injury basis, and it causes damage mechanism mainly is the biomembrane lipid peroxidating that free radical causes. The main mechanism that D-Gal causes hepatocellular injury is because the lipid peroxy chain reaction of free radical and startup thereof. In the peroxidatic reaction of lipid process, the unrighted acid oxidation Decomposition discharges reactive aldehyde such as MDA, 4-hydroxyl nonene (4-HNE) etc., the amount of measuring MDA often can reflect body inner lipid peroxidating degree, indirectly reflects body tissue, cellular damage degree. SOD is the removing agent of super oxygen cation free radical, can suppress the peroxidatic reaction of lipid that free radical starts, and when liver cell was subject to the free radical attack, SOD can reduce because of its exhaustion. SOD plays a part most important to the oxidative and anti-oxidative balance of body, this enzyme can be removed the ultra-oxygen anion free radical Cell protection and avoid damage. The height of SOD vigor has reflected the ability of body removing free radical indirectly, and MDA content and SOD vigor have reacted the degree of medicine to the body peroxide injury in the liver. Experimental result shows that also the SOD that medicine of the present invention can raise in the hepatic injury mouse blood is active, and reduces the MDA content in the blood. Illustrate that we can Anti-lipid peroxidation, improve the ability of body Anti-lipid peroxidation. Pathological examination shows that liver must be answered capsule can make mouse liver cell degeneration necrosis degree alleviate, and particularly only the liver cells appearance is turbid swollen individually for heavy dose of group, basic approaching normally. Pathology prompting medicine of the present invention can improve liver tissue injury.

Experiment by drug effect studies show that medicine of the present invention has ALT, AST effect in the liver injury model of the reduction mouse blood; Improve SOD vigor and reduction MDA content in the liver injury model mouse blood, have the effect of the mouse of raising immunity of organisms. Illustrate that medicine of the present invention has the effect of good protecting liver, lowering enzymes pharmacology, thereby be treatment chronic active hepatitis, hepatitis B, acute viral hepatitis provides the drug effect foundation.

Concrete enforcement mode

Embodiment 1

1. will pull root vegetables 8350g, boiling 3 times, each 2 hours, merge decocting liquid, filter, filtrate is condensed into and surveys relative density under 60-65 ℃ of condition is 1.15 clear cream, cooling adds ethanol and makes and contain the alcohol amount and reach 60%, stirs, and leaves standstill, and filters, and gets filter residue and filtrate A;

2. filter residue merges washing lotion and filtrate A with 60% ethanol washing 3 times, and Recycled ethanol and being condensed into is surveyed relative density under 40 ℃ of conditions be 1.32 thick paste, adds appropriate amount of starch, mix, and drying, pulverizing gets medicinal powder; It is an amount of to add 80% ethanol, ball processed, and packing, packing, and get final product.

Embodiment 2

1. will pull root vegetables 8400g, boiling 2 times 3 hours for the first time, 1 hour for the second time, merges decocting liquid, filter, filtrate is condensed into and surveys relative density under 60-65 ℃ of condition is 1.20 clear cream, and cooling adds ethanol and makes and contain the alcohol amount and reach 70%, stir, leave standstill, filter, get filter residue and filtrate A;

2. filter residue merges washing lotion and filtrate A with 50% ethanol washing 2 times, and Recycled ethanol and being condensed into is surveyed relative density under 40 ℃ of conditions be 1.35 thick paste, adds appropriate amount of starch, mix, and drying, pulverizing gets medicinal powder; It is an amount of to add 65% ethanol, ball processed, and packing, packing, and get final product.

Embodiment 3

1. will pull root vegetables 8300g, boiling 3 times, 2 hours for the first time, 3 hours for the second time, 1 hour for the third time, merge decocting liquid, filter, filtrate is condensed under 60-65 ℃ of condition and surveys relative density is 1.10 clear cream, and cooling adds ethanol and makes and contain the alcohol amount and reach 55%, stir, leave standstill, filter, get filter residue and filtrate A;

2. filter residue merges washing lotion and filtrate A with 90% ethanol washing 3 times, and Recycled ethanol and being condensed into is surveyed relative density under 40 ℃ of conditions be 1.30 thick paste, adds appropriate amount of starch, mix, and drying, pulverizing gets medicinal powder; It is an amount of to add 85% ethanol, ball processed, and packing, packing, and get final product.

Embodiment 4

1. will pull root vegetables 8320g, boiling 3 times, 3 hours for the first time, 2 hours for the second time, 1 hour for the third time, merge decocting liquid, filter, filtrate is condensed under 60-65 ℃ of condition and surveys relative density is 1.18 clear cream, and cooling adds ethanol and makes and contain the alcohol amount and reach 65%, stir, leave standstill, filter, get filter residue and filtrate A;

2. filter residue merges washing lotion and filtrate A with 80% ethanol washing 3 times, and Recycled ethanol and being condensed into is surveyed relative density under 40 ℃ of conditions be 1.30 thick paste, adds appropriate amount of starch, mix, and drying, pulverizing gets medicinal powder; It is an amount of to add 70% ethanol, ball processed, and packing, packing, and get final product.

Claims (5)

1, a kind of preparation method who is used for the treatment of the medicine pill of hepatitis is characterized in that described method comprises following steps:

1. will pull root vegetables boiling 2-3 time, each 1-3 hour, merge decocting liquid, filter, filtrate is condensed under 60-65 ℃ of condition and surveys relative density is the clear cream of 1.10-1.20, and cooling adds ethanol and makes and contain the alcohol amount and reach 55-70%, stirs, leave standstill, filter, get filter residue and filtrate A;

2. filter residue merges washing lotion and filtrate A with the ethanol of 50-90% washing 2-3 time, and Recycled ethanol also is condensed into that to survey relative density under 40 ℃ of conditions be the thick paste of 1.30-1.35, adds appropriate amount of starch, mixing, and drying, pulverizing gets medicinal powder; It is an amount of to add 65-85% ethanol, ball processed, and packing, packing, and get final product.

2, according to the preparation method of medicine pill claimed in claim 1, it is characterized in that 1. described step is to pull the root vegetables boiling 3 times, each 2 hours, merge decocting liquid, filter, filtrate is condensed into and surveys relative density under 60-65 ℃ of condition is the clear cream of 1.15-1.18.

3, according to the preparation method of medicine pill claimed in claim 2, it is characterized in that described step amount of water 1. is respectively: for the first time 12 times of medicinal material amounts, for the second time 10 times of medicinal material amounts, for the third time 8 times of medicinal material amounts.

4, according to the preparation method of claim 1 or 2 described medicine pills, it is characterized in that 1. described step adds ethanol and make and contain alcohol amount and reach 60%.

5, according to the preparation method of medicine pill claimed in claim 1, it is characterized in that described step 2. filter residue with 60% ethanol washing 3 times, merge washing lotion and filtrate A, Recycled ethanol and to be condensed into 40 ℃ of condition downside relative densities be the thick paste of 1.30-1.32.