CN103120718B - A kind of pharmaceutical composition for the treatment of chronic hepatitis, liver cirrhosis - Google Patents

Abstract

The present invention discloses a kind of pharmaceutical composition and preparation method thereof for the treatment of chronic hepatitis, liver cirrhosis.Said composition is made primarily of Fructus Schisandrae Sphenantherae, Fructus Ligustri Lucidi, Fructus Forsythiae, Sonchus brachyotus DC. four taste crude drug.Preparation method is: above a few taste, is ground into fine powder, sieves, and mixing, adds refined honey, add water pill, dry, makes water-honeyed pill; Add customary adjuvant, conveniently technique, make clinical acceptable any dosage form.Present invention also offers the method for quality control of this pharmaceutical composition and the purposes in preparation treatment chronic hepatitis, liver cirrhosis medicine thereof.

Description

A kind of pharmaceutical composition for treating chronic hepatitis and liver cirrhosis

technical field

The invention relates to a pharmaceutical composition, a preparation method and a quality control method thereof, in particular to a pharmaceutical composition for treating chronic hepatitis and liver cirrhosis, a preparation method and a quality control method thereof.

Background technique

Chronic hepatitis refers to chronic inflammatory changes in the liver caused by various etiologies, and the course of the disease is at least half a year. Visible jaundice, hypochondriac pain, swelling, accumulation and other manifestations. Usually, the activity of cell enzymes is abnormal, and the enzyme activity in normal human body is relatively stable. Changes in enzyme activity indicate that some organs and tissues of the human body are damaged or some enzymes are released into blood, urine or body fluids after disease occurs. Therefore, the occurrence and development of certain diseases can be understood or judged by the determination of enzyme activity in blood, urine or body fluids. In traditional Chinese medicine, it is mostly manifested as deficiency of liver qi, loss of both yin and blood, liver qi invading the spleen, stagnation of qi stagnation due to liver depression, and residual toxins are not cleared. The course of chronic hepatitis is protracted, even repeated for several years or more than ten years, and some may develop into liver cirrhosis.

Liver cirrhosis (liver cirrhosis) is a common chronic liver disease, liver damage can be caused by one or more reasons, and the liver is progressive, diffuse, and fibrous. The specific manifestation is diffuse degeneration and necrosis of liver cells, followed by fibrous tissue hyperplasia and nodular regeneration of liver cells. These three changes are repeated and interlaced. As a result, the structure of liver lobules and blood circulation pathways are gradually remodeled, causing the liver to deform, harden and rot. lead to cirrhosis of the liver. The disease had no obvious symptoms in the early stage, but in the later stage, a series of portal hypertension and liver dysfunction occurred in different degrees, until the upper gastrointestinal bleeding, hepatic encephalopathy and other complications occurred and died. Liver shrinkage, "liver atrophy" refers to B-ultrasound or CT detection, the liver volume shrinks, which belongs to the "swelling" of traditional medicine, and modern medicine is characterized as "cirrhosis of the liver". treatment method.

The development of modern medicine has deeply studied the etiology, pathogenesis, occurrence and development of chronic hepatitis from basic to clinical. Currently, there is no effective treatment for chronic viral hepatitis. No drug has ever shown a clear and definitive effect on chronic viral hepatitis, so various experimental treatments are still underway. TCM treatment of chronic hepatitis based on syndrome differentiation has incomparable advantages in the prevention and treatment of chronic hepatitis and liver cirrhosis due to its simple, convenient, practical and inexpensive features.

Contents of the invention

The object of the present invention is to provide a pharmaceutical composition and a preparation method thereof, and another object of the present invention is to provide a quality control method and application of the pharmaceutical composition.

The purpose of the invention is to realize through the following technical solutions

The bulk drug of the pharmaceutical composition of the present invention consists of:

Southern Schisandra 1000-2000 parts by weight Ligustrum lucidum 300-700 parts by weight

Forsythia 300-700 parts by weight Beibaijiang 200-450 parts by weight

The preferred weight ratio of the above-mentioned raw materials is as follows:

Southern Schisandra 1667 parts by weight, Ligustrum lucidum 500 parts by weight, Forsythia 500 parts by weight, Beibaijiang 333 parts by weight

The preferred weight ratio of the above-mentioned raw materials is as follows:

1950 parts by weight of Schisandra chinensis, 350 parts by weight of privet fruit, 650 parts by weight of forsythia, 250 parts by weight of Beibaijiang

The preferred weight ratio of the above-mentioned raw materials is as follows:

1100 parts by weight of Schisandra chinensis, 600 parts by weight of privet fruit, 350 parts by weight of forsythia, 400 parts by weight of Beibaijiang

The crude drug of the pharmaceutical composition of the present invention is prepared by adding conventional auxiliary materials and following conventional processes into clinically acceptable large honeyed pills, honeyed water pills, watered pills, capsules, granules, and tablets.

The preparation technology of pharmaceutical composition water honey pill of the present invention is as follows:

Take the above-mentioned four raw materials, grind them into fine powder, pass through a 100-mesh sieve, mix evenly, add 20 to 25 parts by weight of refined honey for every 100 parts by weight of raw drug powder, add water to make pills, and dry to make honeyed pills.

The quality detection method of pharmaceutical composition of the present invention comprises following identification and/or assay:

Identification includes one or more of the following methods:

A. Take 2g of honeyed pills of the pharmaceutical composition of the present invention, grind finely, add 10-30ml of petroleum ether at 30-60°C, ultrasonically treat for 20 minutes, filter, evaporate the filtrate to dryness, add 0.5-2ml of petroleum ether at 30-60°C to dissolve the residue , as the test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and make the reference medicinal solution in the same way; then take the reference substance of Schisandrin A, add 30-60°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference solution According to the thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition) test, draw each 5 μ l of the above three solutions and place them on the same silica gel GF254 thin-layer plate respectively, and use 30-60 ℃ of petroleum ether-ethyl formate-formic acid The upper solution of 10～20:3～7:0.8～1.2 is the developing agent, develop, take out, dry, and put it under 254nm ultraviolet lamp for inspection, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference substance, Spots of the same color;

B. Get 6g of honeyed pills of the pharmaceutical composition of the present invention, grind them finely, add 10-30ml of petroleum ether at 30-60°C, seal them up, ultrasonically treat them for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add Methanol 10-30ml, plugged, sonicated for 20 minutes, filtered, the filtrate was evaporated to dryness, the residue was dissolved in 5ml of methanol, and used as the test solution; another 1g of forsythia reference drug was taken, and the reference drug solution was prepared in the same way; Forsythin reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution; test according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B), draw each 5 μ l of the above three solutions, respectively Spot on the same silica gel G thin-layer plate, use water-saturated chloroform-methanol-ethyl acetate-formic acid=8～12:1.5～2.5:0.8～1.2:0.3～0.7 as developing agent, develop, take out, dry in the air, spray With 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and in the chromatogram of the test product, the spots of the same color appeared at the positions corresponding to the chromatogram of the reference substance;

C. get the pharmaceutical composition water honey pill of the present invention, put and observe under the microscope: pericarp epidermal cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; Endocarp fiber bundle upper and lower layers are arranged obliquely or vertically;

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 Edition), chromatographic conditions and system suitability test: use octadecylsilane bonded silica gel as filler; acetonitrile-water-acetic acid=60～80: 20～40: 0.8～1.2 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should not be less than 4000 according to the calculation of schisandrin; the preparation of the reference solution: accurately weigh the reference substance of schisandrin , put in a measuring bottle, add methanol to make a solution containing 40 μg per 1ml; preparation of the test solution: take the pharmaceutical composition water honey pill of the present invention, grind it finely, take 1.0g, accurately weigh, and place a stoppered conical flask Add 20-30ml of methanol precisely, seal it tightly, weigh it, ultrasonically treat it with a power of 120W, 40kHz, for 25-35 minutes, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake well, Filter and get the filtrate to get the product; Assay method: respectively accurately draw 10 μl of the reference substance solution and the solution of the test product, inject it into a liquid chromatograph, and measure to get the product; the pharmaceutical composition water honey pill of the present invention contains Schisandra chinensis per 1g Based on Schizandrin A C ₂₄ H ₃₂ O ₆ , it should not be less than 0.9mg.

The quality control method of pharmaceutical composition of the present invention is preferably following identification and/or assay:

A. Take 2 g of the pharmaceutical composition water honey pill of the present invention, grind it finely, add 20 ml of petroleum ether at 30 to 60 ° C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add 1 ml of petroleum ether at 30 to 60 ° C to dissolve the residue, and use it as a test Take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution by the same method; then take the reference substance of Schizandrin A, add 30-60°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; Chromatography (Appendix VI B, Chinese Pharmacopoeia 2010 Edition) test, draw 5 μl of each of the above three solutions and place them on the same silica gel GF254 thin-layer plate, and use 30-60°C petroleum ether-ethyl formate-formic acid as 15:5 : The upper layer solution of 1 is developing agent, develops, takes out, dries, puts inspection under the 254nm ultraviolet light lamp, in the test sample chromatogram, on the corresponding position with reference substance chromatogram, show the spot of same color;

B. Take 6g of honeyed pills of the pharmaceutical composition of the present invention, grind them finely, add 20ml of petroleum ether at 30-60°C, seal them tightly, treat with ultrasound for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, and add 20ml of methanol , dense plug, sonicated for 20 minutes, filtered, the filtrate was evaporated to dryness, the residue was dissolved in 5ml of methanol, and used as the test solution; another 1g of forsythia reference drug was taken, and the reference drug solution was prepared in the same way; Add methanol to make a solution containing 0.25mg per 1ml, as a reference solution; test according to thin-layer chromatography (Appendix VI B, Chinese Pharmacopoeia, 2010 edition), draw 5 μl of each of the above three solutions, and spot on the same silica gel respectively On G thin-layer board, use water-saturated chloroform-methanol-ethyl acetate-formic acid=10:2:1:0.5 as developing solvent, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, and heat at 105°C to The color of the spots is clear, and in the chromatogram of the test product, the spots of the same color appear on the position corresponding to the chromatogram of the reference product;

C. get the pharmaceutical composition water honey pill of the present invention, put and observe under the microscope: pericarp epidermal cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; Endocarp fiber bundle upper and lower layers are arranged obliquely or vertically;

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: use octadecylsilane bonded silica gel as filler; acetonitrile-water-acetic acid=64:35: 1 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should be no less than 4000 according to the calculation of schisandrin A; the preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, Add methanol to make a solution containing 40 μg per 1ml; the preparation of the test solution: take the content of the pharmaceutical composition water honey pill of the present invention under the difference in loading amount, grind it finely, get 1.0g, accurately weigh, and put a stopper cone Add 25ml of methanol to the bottle, seal it tightly, weigh it, ultrasonically treat it at 120W, 40kHz for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake well, and filter , get the filtrate, to get it; Assay method: respectively accurately draw 10 μl of the reference substance solution and the solution of the test sample, inject it into a liquid chromatograph, and measure it to get it; Based on C ₂₄ H ₃₂ O ₆ , it should not be less than 0.9mg.

The quality control method of pharmaceutical composition of the present invention is preferably following identification and/or assay:

A. Take 2 g of honeyed pills of the pharmaceutical composition of the present invention, grind finely, add 28 ml of petroleum ether at 35 ° C, ultrasonically treat for 20 minutes, filter, evaporate the filtrate to dryness, add 0.6 ml of petroleum ether at 35 ° C to dissolve the residue, and use it as the test solution; Take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution in the same way; then take the reference substance of Schisandrin A, add 35°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B) test, draw 5 μl of each of the above three solutions and spot on the same silica gel GF254 thin-layer plate, and use the upper layer solution of 35 ℃ petroleum ether-ethyl formate-formic acid as 11:6:0.9 as the development agent, unfolded, taken out, dried, and inspected under a 254nm ultraviolet lamp, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference substance, spots of the same color are displayed;

B. Take 6g of honeyed pills of the pharmaceutical composition of the present invention, grind them finely, add 12ml of petroleum ether at 55°C, seal them tightly, ultrasonically treat them for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add 28ml of methanol, seal them tightly stopper, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take another 1 g of forsythia reference drug, and make the reference drug solution in the same way; then take the forsythin reference substance, Add methanol to make a solution containing 0.25 mg per 1 ml, as a reference solution; test according to thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition), draw 5 μl of each of the above three solutions, and spot them on the same silica gel G thin film respectively. On the layer board, use water-saturated chloroform-methanol-ethyl acetate-formic acid=9:2.4:0.9:0.6 as the developing agent, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, and heat at 105°C until the spots appear. The color is clear, and in the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product;

C. get the pharmaceutical composition water honey pill of the present invention, put and observe under the microscope: pericarp epidermal cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; Endocarp fiber bundle upper and lower layers are arranged obliquely or vertically;

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=62:38: 0.9 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should not be less than 4000 according to the calculation of schisandrin; Add methanol to make a solution containing 40 μg per 1ml; the preparation of the test solution: take the pharmaceutical composition water honey pill of the present invention, grind it finely, get 1.0g, accurately weigh, put in a stoppered Erlenmeyer flask, and accurately add 22ml of methanol , dense plug, weighed, ultrasonic treatment, power 120W, 40kHz, 34 minutes, take out, let cool, weigh again, make up the lost weight with methanol, shake well, filter, take the filtrate to get final product; Measuring method: accurately draw 10 _μl each of the reference substance solution and the test solution, _inject it into a liquid chromatograph, measure it, and get it _; It should not be less than 0.9mg.

The quality control method of pharmaceutical composition of the present invention is preferably following identification and/or assay:

A. Take 2 g of honeyed pills of the pharmaceutical composition of the present invention, grind finely, add 12 ml of petroleum ether at 55 ° C, ultrasonically treat for 20 minutes, filter, evaporate the filtrate to dryness, add 1.8 ml of petroleum ether at 55 ° C to dissolve the residue, and use it as the test solution; Take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution in the same way; then take the reference substance of Schizandrin A, add 55°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B) test, draw 5 μl of each of the above three solutions and spot on the same silica gel GF254 thin-layer plate, and use the upper layer solution of 55 ℃ petroleum ether-ethyl formate-formic acid as 18:4:1.1 as the development agent, unfolded, taken out, dried, and inspected under a 254nm ultraviolet lamp, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference substance, spots of the same color are displayed;

B. Take 6g of honeyed pills of the pharmaceutical composition of the present invention, grind them finely, add 28ml of petroleum ether at 35°C, seal them tightly, treat them ultrasonically for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add 12ml of methanol, seal them tightly stopper, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take another 1 g of forsythia reference drug, and make the reference drug solution in the same way; then take the forsythin reference substance, Add methanol to make a solution containing 0.25 mg per 1 ml, as a reference solution; test according to thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition), draw 5 μl of each of the above three solutions, and spot them on the same silica gel G thin film respectively. On the layer board, use water-saturated chloroform-methanol-ethyl acetate-formic acid=11:1.6:1.1:0.4 as the developer, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, and heat at 105°C until the spots appear. The color is clear, and in the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product;

C. get the pharmaceutical composition water honey pill of the present invention, put and observe under the microscope: pericarp epidermis cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; Endocarp fiber bundle upper and lower layers are arranged obliquely or vertically.

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=78:22: 1.1 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should not be less than 4000 according to the calculation of schisandrin; the preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, Add methanol to make a solution containing 40 μg per 1ml; preparation of the test solution: take the pharmaceutical composition water honey pill of the present invention, grind finely, get 1.0g, accurately weigh, put in a stoppered Erlenmeyer flask, and accurately add 28ml of methanol , dense plug, weighed, ultrasonic treatment, power 120W, 40kHz, 26 minutes, take out, let cool, weigh again, make up the lost weight with methanol, shake well, filter, take the filtrate to get final product; Measuring method: accurately draw 10 _μl each of the reference substance solution and the test solution, _inject it into a liquid chromatograph, measure it, and get it _; It should not be less than 0.9mg.

The pharmaceutical composition of the present invention has the effects of nourishing yin and nourishing the liver, and clearing away residual toxins. It is mainly used to treat chronic hepatitis and liver cirrhosis, which belong to the syndrome of TCM differentiation of both deficiency of yin and blood, and residual toxins. The clinical curative effect is very definite. It has a good curative effect on promoting blood circulation and promoting qi, nourishing yin and clearing away heat, reducing jaundice and improving protein metabolism. The preparation method of the pharmaceutical composition of the invention is advanced, the quality is controllable, and it is safe and effective. The water honeyed pill of the pharmaceutical composition of the present invention has the characteristics of being convenient to take, convenient to carry, remarkable economic benefit and the like. The separation and reproducibility of the identification method are good; the negative control sample has no interference; the specificity is strong. The method of content determination has good instrument precision and operation precision, good linear relationship, good experimental reproducibility, and this method has good recovery rate.

Description of drawings:

Figure 1 TLC identification test diagram of Schisandra chinensis

(1-6 are the negative control, three batches of the test product (080220, 08022023, 080226), the reference substance Schisandrin A, and the reference medicinal material Southern Schisandra)

Figure 2 TLC identification test diagram of forsythia

(1-6 are the negative control, three batches of the test product (080220, 08022023, 080226), the reference substance forsythin, and the reference drug forsythia)

Fig. 3 separation HPLC chromatogram of test sample, reference substance, negative control solution

(A negative control solution; B reference substance solution; C need testing solution; 1 Schizandrin A)

The ultraviolet spectrogram of Schizandrin A in Fig. 4 test sample, reference substance solution chromatography

Fig. 5 Schizandrin A concentration-peak area linear relationship diagram

Figure 6 content determination chromatogram - reference substance (1)

Figure 7 content determination chromatogram - reference substance (2)

Figure 8 Content determination chromatogram - batch number 080220 sample (1)

Figure 9 content determination chromatogram - batch number 080220 sample (2)

Figure 10 content determination chromatogram - batch number 080220 sample (3)

Figure 11 Content determination chromatogram - batch number 080223 sample (1)

Figure 12 Content determination chromatogram - batch number 080223 sample (2)

Figure 13 content determination chromatogram - batch number 080223 sample (3)

Figure 14 content determination chromatogram - batch number 080223 sample (4)

Figure 15 Content determination chromatogram - batch number 080226 sample (1)

Figure 16 content determination chromatogram - batch number 080226 sample (2)

Figure 17 content determination chromatogram - batch number 080226 sample (3)

Figure 18 content determination chromatogram - batch number 080226 sample (4)

The following experimental examples and examples are used to further illustrate but not limit the present invention.

Experimental example 1 preparation method experiment of pharmaceutical composition water honey pill of the present invention

1. Inspection of crushing process

1.1 Crushing conditions

Instruments and equipment: Pulverizer, produced by Zhigang Pharmaceutical Machinery Factory in Yantai City, Shandong Province, consists of three main parts: ZKF-3 self-controlled pulverizer console, WZS3-470 horizontal rotary sieve and hF3-200 pulverizer; medicinal materials : The above four medicinal materials are all dried in a drying room or an oven until completely dry.

1.2 Crushing process

The crushing efficiency is to crush 20-200kg of medicinal materials per hour, and the sieving particle size can be adjusted arbitrarily between 20-400 mesh. Actually, No. 8 sieve (80 mesh) is used for crushing. Since it is an automatic circulation crushing method, its powder yield It can reach more than 95%.

2 Soft material pelletizing process conditions

On the basis of the original large honeyed pills, the specifications of watery honeyed pills are added; watery honeyed pills: refer to pills made of fine powder of medicinal materials with honey and water as a binder. The prepared pellets should be round and uniform, with consistent color.

The specification is 12g per 100 capsules.

2.1 Soft materials

The degree of honey and the amount of honey added are two more important factors in the production of soft wood, and the L9(34) orthogonal table is used to test and investigate.

Honey can be divided into three specifications according to the degree of refining: tender honey, medium honey, and old honey.

Tender honey: Honey is heated to 105°C-115°C, the water content is 14%-20%, the density is about 1.35, the color has no obvious change, and there is little stickiness.

Medium honey: the tender honey continues to be heated, the temperature reaches 116-118°C, the water content is 14%-16%, the density is about 1.37, and there are light yellow, glossy, churning uniform bubbles, which are sticky when twisted by hand. When the two hands are separated No white threads appear.

Old honey: continue to heat the medium honey, the temperature reaches 119-122°C, the water content is below 10%, the density is about 1.4, and reddish-brown glossy bubbles appear, which is very sticky when the hands are twisted. When the two fingers are separated, long white threads appear. Drop into water to form beads.

2.1.1 Factor level table, see Table 1

Table 1 Orthogonal test factor level table

2.1.2 Sample preparation: Weigh 0.5 kg of medicinal powder, and prepare samples according to the L ₉ (3 ⁴ ) orthogonal table condition.

2.1.3 Evaluation indicators: The pellets should be able to be shaped freely without cracking, rubbing with hands without sticking to hands, and not adhering to the wall of the device.

The experimental results are shown in Table 2:

Table 2 Orthogonal experiment results

The results show that both experiments 8 and 9 can be molded, but experiment 9 consumes a lot of honey, so in order to save costs, experiment 8 is selected, that is, tender honey is selected, and the amount of honey is 25% of the medicinal powder.

2.2 Baked noodles

The prepared soft material is refined by a refining machine and put into pellets in a pill making machine. After the pellets are made, they need to be powdered and rounded to make the pellets more round and smooth.

In the process of powder cultivation and rounding, it is necessary to investigate the honey water (the ratio of honey to water), and the smooth and rounded pellets should be used as the evaluation index.

experiment procedure:

Experiment 1: Use honey water (honey: water = 1: 4) to make powder and round,

The resulting pellets were smooth, but not round and had a poor appearance.

Experiment 2: Use honey water (honey: water = 1: 5) to make powder and round,

The obtained pellets are smooth, round, uniform and have a good appearance.

Experiment 3: Use honey water (honey: water = 1: 6) to make powder and round,

The obtained pellets are round and uniform, but rough and have a bad appearance.

Through the above experiments, it can be seen that the pellets can be rounded with honey: water of 1:5 and 1:6, but the pellets obtained with honey water of 1:5 are smooth, round, uniform and have a better appearance.

2.3 drying

When drying the samples, it is necessary to examine the drying temperature and time, and control the moisture content within 12%.

Under Chinese Pharmacopoeia Appendix IA Pills:

Unless otherwise specified, honeyed pills, watered pills, concentrated honeyed pills and concentrated watered pills should be dried below 80°C.

【Moisture】Water honey pills must not exceed 12.0%

Therefore, the water honey pill of the pharmaceutical composition of the present invention is planned to be dried at 80° C., and it is inspected, taking the water content of the dried pill and whether the pill has cracks as the inspection indexes. Samples were taken at different drying times to measure their moisture content. The results are shown in Table 3:

Table 3 The moisture detection results of sampling in different drying time periods

The results show that the pellets meet the requirements when they are dried for 120 minutes at 80°C.

Conclusion: comprehensive above experimental data, the production technology parameter of pharmaceutical composition water honey pill of the present invention is:

Honey type: tender honey; amount of honey added: 25% of medicinal powder; ratio of powdered and round honey to water: 1:5; drying temperature: 80°C; drying time: 120min

3. Sources and standards of excipients

Honey: registration number 01707Q10265ROS (food additive); produced by Beijing Baiji Jianle Bee Products Co., Ltd.; production batch number: 20080228.

Experimental Example 2 Differentiation Experiment of Pharmaceutical Composition Water Honey Pills of the present invention

Identification (1) [Prescription] Southern Schisandra 1667g, Ligustrum lucidum 500g, Forsythia 500g, Beibaijiang 333g

[Preparation method] The above four flavors are crushed into fine powder, sieved, and mixed. For every 100g of powder, use 20-30g of refined honey, add appropriate amount of water pan pills, and dry them to make water honey pills.

[Properties] The honeyed pills of the pharmaceutical composition of the present invention are brown-black honeyed pills; the smell is slight and the taste is slightly sour.

[identification] (1) get the pharmaceutical composition water honey pill of the present invention, put and observe under the microscope: pericarp epidermis cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; Staggered vertically.

(2) Take 2 g of honeyed pills of the pharmaceutical composition of the present invention, grind them finely, add 20 ml of petroleum ether (30-60° C.), sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 1 ml of petroleum ether (30-60° C.) to the residue Dissolved as the test solution. Take another 1 g of Schisandra chinensis reference medicinal material, and make a reference medicinal material solution in the same way. Then take Schizandrin A reference substance, add petroleum ether (30-60°C) to make a solution containing 2mg per 1ml, and use it as the reference substance solution. According to the test of thin-layer chromatography (Appendix VI B, Chinese Pharmacopoeia 2010 Edition), draw 5 μl of each of the above three solutions and spot on the same silica gel GF ₂₅₄ thin-layer plate, and use petroleum ether (30-60 ° C)-ethyl formate -The upper layer solution of formic acid (15:5:1) is used as developing agent, developed, taken out, dried in the air, and inspected under ultraviolet light (254nm). In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product.

(3) Take 6 g of the pharmaceutical composition water honey pill of the present invention, grind finely, add petroleum ether (30～60 ℃) 20ml, close up, ultrasonic treatment for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, Add 20ml of methanol, plug it tightly, treat it with ultrasound for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve it, and use it as the test solution. Another 1 g of forsythia reference medicinal material was taken, and the reference medicinal material solution was prepared in the same way. Then take the forsythin reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution. Test according to thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 edition), draw each 5 μ l of the above-mentioned three kinds of solutions, spot on the same silica gel G thin-layer plate respectively, and use chloroform (water saturated)-methanol-ethyl acetate- Formic acid (10:2:1:0.5) was used as the developer, developed, taken out, dried in the air, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product.

Identification (2) is the TLC identification of Schisandra chinensis in the pharmaceutical composition Shuimi Pill of the present invention: Schisandra chinensis is the monarch drug in the prescription. It is known that the lignans represented by schisandrin are the main active ingredients and have strong specificity. In this identification test, the treatment method and chromatographic conditions of the test solution refer to the quality standard of the original preparation of "Gandening Pills" and the method (P.169) under the identification item of "Schisandra chinensis" in the 2005 edition of the Chinese Pharmacopoeia, and the obtained spots The R _f value is more appropriate, and the spots are clear. The separation degree and reproducibility of the method are all good; the negative control sample (lack of Schisandra chinensis, negative sample is prepared according to the preparation process, and then the negative control solution is prepared according to the preparation method of the test solution) has no interference; The medicinal materials are also used as the control, and the specificity is strong. (see accompanying drawing 1).

Identification (3) is the identification of Forsythia in the pharmaceutical composition water honey pill of the present invention: in this identification test, the processing method and the chromatographic conditions reference method of the test solution are improved, and the gained spot _Rf value is more appropriate, and the spot is clear . The separation degree and reproducibility of the method are all good, and the negative control sample (lack of forsythia, negative sample is prepared according to the preparation process, and then the negative control solution is prepared according to the preparation method of the test solution) has no interference; The medicinal material is the control, which has strong specificity. Through many batches of samples and negative control test observation (seeing accompanying drawing 2), confirm that this law is established.

【Inspection】According to the regulations under the pill item in the first appendix of the Chinese Pharmacopoeia 2010 edition, the results were all in compliance with the regulations.

1. Moisture Get the pharmaceutical composition water honeyed pill of the present invention, measure according to moisture determination method (Chinese Pharmacopoeia version one appendix IX H in 2010). Should not exceed 12.0%. The results are shown in Table 4.

Table 4 moisture measurement results

Conclusion: The moisture determination results of the three batches of samples are all in compliance with the regulations.

2. Weight difference Get the pharmaceutical composition water honeyed pill of the present invention, measure according to law (Chinese Pharmacopoeia 2010 edition an appendix IA item weight difference inspection method).

Table 5 Weight Difference Check

Conclusion: The weight differences of the three batches of samples are all within the limit (±10%), and all meet the regulations.

3. Difference in loading: Take 10 bags of honeyed pills of the pharmaceutical composition of the present invention, and check according to the law. The results are shown in Table 6.

Table 6 Loading difference check

Conclusion: The differences in loading of the three batches of samples are all within the limit (±5%), and all meet the regulations.

4. Time limit for dissolving and dispersing Take 6 pills of the pharmaceutical composition of the present invention and check according to the law. The results are shown in Table 7.

Table 7 Dissolution Time Limit Determination Result

Conclusion: The determination results of dissolution time of three batches of samples are all in compliance with the regulations.

5. Microbial limit

Check the microbial limit according to Appendix XIII C of Chinese Pharmacopoeia 2010 Edition. Get the pharmaceutical composition water honey pill 10g of the present invention, add pH 7.0 sterile sodium chloride-peptone buffer to 100ml, make the test solution with the insulation shaking method, check with the plate method. The results are shown in Table 8.

Table 8 Microbiological Limit Checklist

Conclusion: The microbial limit results of the three batches of samples all meet the regulations.

6. Heavy metal inspection

Measured according to the second method of heavy metal inspection in Appendix IX E of Chinese Pharmacopoeia 2010 edition. The results are shown in Table 9.

Table 9 Heavy metal determination table

The results showed that the heavy metal contents of the three batches of samples were all less than 20PPM.

7. Arsenic salt inspection

Determination according to the first method of "Chinese Pharmacopoeia" 2010 edition of Appendix IX F arsenic salt inspection method (ancient Cai's method). The results are shown in Table 10:

Table 10 Determination table of arsenic salt

The results showed that the content of arsenic salt in the three batches of samples was all less than 2PPM.

Experimental example 3 pharmaceutical composition of the present invention (preparation is the same as embodiment 1) honeyed water pill content determination experiment

1. Selection of index components for content determination

One of the main medicinal flavors of the preparation, Schisandra chinensis, is listed as a monarch drug in the prescription. The content limit of schisandrin is determined according to the determination results of three batches of samples. It is known that the lignans represented by schisandrin are its main active ingredients, so it is selected as the content determination object of the pharmaceutical composition water honey pill of the present invention. This quality standard mainly refers to the content determination method of schisandrin A under the item of Shenqi Wuweizi Tablets (Chinese Pharmacopoeia 2005, p.508), and the content determination of schisandrin A, one of the index components of Schisandra chinensis, was carried out by HPLC method. The method is verified to meet the content determination requirements, and can be used as a method for determining the content of schisandra in the pharmaceutical composition of the present invention.

2. Instruments and reagents:

High-performance liquid chromatography: Agilent 1100 quaternary gradient pump; Agilent 1100 autosampler; Agilent 1100DAD detector; Agilent ChemStation; HP DeskJet Laser P1015 printer.

Reagents: Methanol and acetonitrile are chromatographically pure; water is ultrapure water, and other reagents are analytically pure.

Chromatographic conditions: analytical column is phenomenex Luna C-18 (5μm, 250×4.6mm) (Phenomenex, USA); mobile phase is acetonitrile-water-acetic acid (64:35:1), filtered through a 0.45μm filter membrane before use ; Flow rate 1.0ml/min, detection wavelength 249nm; Column temperature 25°C.

Under the proposed chromatographic conditions, the retention time of schisandrin is about 22 minutes, and the theoretical plate number of schisandrin is not less than 4000, so the tentative theoretical plate number is not less than 4000 based on schisandrin.

3. Methodological investigation

Selection of extraction method According to the properties of schisandrin, refer to the preparation method of the test product solution of Shenqi schisandra (Chinese Pharmacopoeia 2005 edition, P.508), consider using methanol as a solvent, ultrasonic extraction, and investigate the extraction effect of different ultrasonic extraction time . The method is as follows: get the content of the pharmaceutical composition water honeyed pill of the present invention, grind it finely, get about 1 g of the fine powder, accurately weigh it, put it in a stoppered Erlenmeyer flask, accurately add 25 ml of methanol, seal it tightly, weigh it, and perform ultrasonic treatment ( Power 120W, 40kHz), starting from 10 minutes, take it out every 10 minutes, let it cool, then weigh it, make up the lost weight with methanol, shake well, take about 1ml of the solution, filter, take 10μl of the filtrate, and inject A liquid chromatograph, record the chromatogram, and measure the area of schisandrin A; the remaining solution is repacked, weighed, and ultrasonically treated. The results are shown in Table 11.

Table 11 is the extraction result of different ultrasonic extraction time with methanol as solvent

It can be seen from the above table that after ultrasonic treatment for 30 minutes, the peak area of schisandrin A no longer increases, and the ultrasonic treatment time is determined to be 30 minutes.

blank test

Self-prepared negative samples that do not contain Schisandra chinensis according to the ratio of herbal flavor in the prescription, made a blank solution according to the method in the text, and measured according to the law. As a result, no obvious chromatographic peaks were seen in the blank solution at the same retention time as the Schisandrin A reference substance, and it was considered that there was no interference. The peak shape of schisandrin A, the degree of separation with adjacent peaks, etc. all meet the test requirements (Fig. 3); the reference substance and the test sample are scanned by using a diode array detector and a chromatographic workstation, and the results are shown in the ultraviolet spectrum scanning diagram (Fig. 4).

Stability test Take the test solution and measure it at 0, 1, 2, 4, 6, and 8 hours after preparation. The result shows that need testing solution is basically stable (table 12) in 8 hours.

Table 12 test product solution stability test result

Precision experiment Precisely draw 10 μl of Schizandrin A reference substance solution, inject 6 times continuously, measure the peak area, the results are shown in Table 13. It shows that the instrument precision and operation precision are good.

Table 13 precision test results

Examination of linear relationship Accurately measure 4.0, 5.0, 6.0, 7.0, 8.0ml of the stock solution of Schizandrin A reference substance (164.0μg/ml), respectively, put them in different 25ml brown measuring bottles, add methanol to the mark, shake well, Precisely draw 10 μl into the high-performance liquid chromatograph in turn, measure the peak area according to the above chromatographic conditions, take the peak area integral value (A) as the ordinate, and the concentration of schisandrin A (C, μg/ml) as the abscissa, draw the standard Curve (see Figure 5), regression equation is A=23.4071C+1.1763, correlation coefficient r=0.9999 (n=5).

The results showed that schisandrin had a good linear relationship in the concentration range of 26.24～52.48μg/ml.

Repeatability test Take the same batch of samples (080220), prepare 6 copies of the test solution in parallel, measure them, and calculate the content. The results are shown in Table 14. It shows that the experimental method has good reproducibility.

Table 14 Repeatability Experiment Results (n=6)

Sample addition recovery rate test Precisely weigh 0.5g of the same batch of samples (080220) with known content (0.9999mg/g) and 6 parts in total, and add 0.5248mg (104.96μg/ml×5.0ml) of Schizandrin A reference substance respectively ). According to the preparation method of the test solution in the text and the above-mentioned chromatographic conditions, the recovery is calculated by the following formula, and the results are shown in Table 15.

Table 15 Sample addition recovery test results (n=6)

The above determination results show that this method has a good recovery rate.

Sample determination

Three batches of samples were taken, and the content of schisandrin A was determined according to the method stated in the main text. The results are shown in Table 16, and the chromatograms are shown in Figures 6-18.

The content determination result of Schizandrin A in three batches of samples of table 16

According to the content measurement results of the above three batches of samples, the content limit of _the pharmaceutical composition water honeyed pill of the present invention is tentatively determined as: every 1g containing Schisandra _chinensis shall not be less than _0.90mg (about equivalent 70% of the sample content in batch 080220).

Experimental Example 4 The clinical experiment of treating chronic viral hepatitis B with pharmaceutical composition Shuimi Wan (prepared in Example 1) of the present invention

1. Diagnostic criteria

1.1 Western medicine diagnostic criteria:

Refer to the diagnostic criteria for chronic hepatitis B in the "Guidelines for the Prevention and Treatment of Chronic Hepatitis B" formulated by the Society of Hepatology and Infectious Diseases of the Chinese Medical Association in 2005.

1.2 TCM diagnostic criteria: refer to TCM syndrome differentiation criteria for viral hepatitis (trial implementation)

1.2.1 Symptoms of TCM: Deficiency of both yin and blood, residual toxins are not cleared

1.2.2 Differential criteria

(1) dry mouth and throat, (2) dysphoria, (3) dry eyes, (4) fatigue, (5) poor appetite, (6) yellow urine, (7) dark red tongue, thin greasy fur, (8) thready and slippery pulse.

2. Inclusion criteria

2.1 Age 18-65 years old, male or female.

2.2 The clinical diagnosis is chronic hepatitis B.

2.3 TCM syndrome differentiation conforms to Yin and blood deficiency, and residual poison is not cleared.

2.4 ALT > more than 2 times the upper limit of normal, BIL < 2 times the upper limit of normal.

3. Exclusion criteria

3.1 Accepting other clinical drug studies within or within 30 days.

3.2 Patients who cannot take medicine or receive examinations as required.

3.3 Other types of hepatitis (such as drug poisoning, alcoholism, autoimmunity and other non-B virus hepatitis, etc.) have been confirmed by inspection.

3.4 Severe allergic reactions and chronic diseases or end-stage diseases.

3.5 Pregnant and lactating women.

4. Experimental method and grouping

4.1 Method: A randomized, open, positive drug-controlled trial method was adopted.

4.2 Source and grouping of cases: The cases in this group are all inpatients in the 302 Hospital of the People's Liberation Army, including 60 cases in the treatment group and 30 cases in the control group. In the treatment group, there were 56 males and 4 females, with an average age of 30.6±9.3 years (17-57 years old), and in the control group, there were 28 males and 2 females, with an average age of 32.8±12.5 years old (19-59 years old). The two groups were comparable before treatment.

5. Selection of control drugs

The commonly used clinical enzyme-lowering chemical drug "Bifendate Dropping Pills" (National Drug Accreditation H11020980, produced by Peking Union Medical College Pharmaceutical Factory) was selected.

6. Test drug:

6.1 Observation of drugs: the pharmaceutical composition water honey pill of the present invention

Source: Provided by PLA 302 Hospital

Specifications: 7.2g/bag, 60 pills/bag

Batch number: 070919

Storage conditions: room temperature, away from light, in a cool place.

6.2 Control drug: bifendate dropping pills (positive drug)

Source: Peking Union Medical College

Specification: 1.5mg/pill

Batch number: 07100104

Usage and dosage: Oral: 7.5mg (5 pills) once, 3 times a day. 3 months a course of treatment.

Storage: Sealed and stored in a dry place.

7. Random design: use random number table method.

8. Medication method

Dosage: treatment group: 1 bag/time of the pharmaceutical composition of the present invention, 2 times/day

Control group: bifendate dropping pills 5 pills/time, 3 times/day

Course of treatment: 3 months

Administration route: Oral.

If the ALT still rises after 4 weeks of medication, the treatment can be stopped and changed.

9. Observation items and indicators

9.1 Safety indicators: Check renal function (BUN, Cr), ECG, blood, urine and stool routine before treatment and at the end of treatment.

9.2 Liver and spleen color Doppler B-ultrasound examination before treatment and at the end of treatment.

9.3 Liver function (ALT, AST, TBIL, DBIL, A/G, PT/PA) and other laboratory indicators should be checked at least once before and at the end of treatment, but liver function can be rechecked regularly according to the condition during treatment.

10. Efficacy Judgment Criteria

Referring to the "Guiding Principles for Clinical Research of New Chinese Medicines for the Treatment of Viral Hepatitis", the criteria for judging the curative effect are basically cured, effective, and ineffective.

●Basic cure: ALT returns to normal

●Effective: ALT decreased by more than 50% compared with before treatment

●Invalid: Those who do not meet the above indicators

11. Curative effect analysis

The comparative analysis of the total curative effect is shown in Table 17

Table 17 The comparative analysis of the total curative effect between the two groups

Through X2 analysis, the total curative effect of the ^two groups is compared, P=0.6607, and the difference between the two groups is not statistically significant, showing that in the treatment of chronic viral hepatitis B, there is no difference in the curative effect of the pharmaceutical composition Shuimiwan and bifendate . No adverse reactions occurred during the whole treatment process.

The following embodiments can all achieve the effects described in the above experimental examples.

The preparation of embodiment 1 water honey pill

Southern Schisandra 1667g Ligustrum lucidum 500g Forsythia 500g Beibaijiang 333g

Take the above four raw materials, grind them into fine powder, pass through a 100-mesh sieve, mix evenly, add 20 g of refined honey for every 100 g of raw drug powder, add appropriate amount of water pan pills, and dry to make honeyed water pills. Specification 7.2g/bag (60 pills/bag), 120mg/pill.

The preparation of embodiment 2 water honey pills

Southern Schisandra 1950g Ligustrum lucidum 350g Forsythia 650g Beibaijiang 250g

Take the above four raw materials, grind them into fine powder, pass through a 100-mesh sieve, mix evenly, add 25g of refined honey for every 100g of raw material powder, add appropriate amount of water pan pills, dry to make honeyed water pills. Specification 7.2g/bag (60 pills/bag), 120mg/pill.

The preparation of embodiment 3 water honey pills

Southern Schisandra 1100g Ligustrum lucidum 600g Forsythia 350g Beibaijiang 400g

Take the above four raw materials, grind them into fine powder, pass through a 100-mesh sieve, mix evenly, add 23g of refined honey for every 100g of raw drug powder, add appropriate amount of water pan pills, and dry to make honeyed water pills. Specification 7.2g/bag (60 pills/bag), 120mg/pill.

The preparation of embodiment 4 capsules

Southern Schisandra 1667g Ligustrum lucidum 500g Forsythia 500g Beibaijiang 333g

The above-mentioned raw materials are taken, added with conventional auxiliary materials, boiled in water, alcoholized, dried and pulverized according to the conventional process to make powder into capsules.

The preparation of embodiment 5 tablet

Southern Schisandra 1950g Ligustrum lucidum 350g Forsythia 650g Beibaijiang 250g

Take the above-mentioned raw materials, add conventional auxiliary materials, and make tablets according to the conventional process of alcohol extraction and water decoction.

The preparation of embodiment 6 big honeyed pills

Southern Schisandra 1100g Ligustrum lucidum 600g Forsythia 350g Beibaijiang 400g

Take the above-mentioned four raw materials, dry and pulverize, and make large honeyed pills according to the conventional process.

The preparation of embodiment 7 big honeyed pills

Southern Schisandra 1667g Ligustrum lucidum 500g Forsythia 500g Beibaijiang 333g

Take Schisandra chinensis, Ligustrum lucidum, and forsythia three flavors and decoct in alcohol, and Beibaijiang in water to decoct in alcohol to make big honey pills.

The preparation of embodiment 8 water pills

Southern Schisandra 1950g Ligustrum lucidum 350g Forsythia 650g Beibaijiang 250g

Take the above-mentioned four raw materials, dry and pulverize, and make large honeyed pills according to the conventional process.

The preparation of embodiment 9 water pills

Southern Schisandra 1100g Ligustrum lucidum 600g Forsythia 350g Beibaijiang 400g

Take Schisandra chinensis, Ligustrum lucidum, and forsythia three flavors and decoct in alcohol, and Beibaijiang in water to decoct in alcohol to make big honey pills.

Embodiment 10 quality detection method

A. Take 2 g of the honeyed pills of the pharmaceutical composition of the present invention made in Example 1, grind finely, add 20 ml of petroleum ether at 30 to 60 ° C, ultrasonically treat for 20 minutes, filter, evaporate the filtrate to dryness, add petroleum ether at 30 to 60 ° C to the residue Dissolve 1ml and use it as the test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and make the reference medicinal solution in the same way; then take the reference substance of Schisandrin A, add petroleum ether at 30-60°C to make a solution containing 2mg per 1ml, and use it as a control product solution; test according to thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition), draw 5 μl of each of the above three solutions and place them on the same silica gel GF254 thin-layer plate, and use 30-60 ℃ petroleum ether-ethyl formate - formic acid is 15: 5: 1 upper layer solution is developing agent, develops, takes out, dries, puts inspection under 254nm ultraviolet lamp, in the test sample chromatogram, on the position corresponding to reference substance chromatogram, show the same color spot;

B. Take 6 g of the pharmaceutical composition water honey pill of the present invention made in Example 1, grind it finely, add 20 ml of petroleum ether at 30 to 60 ° C, seal it up, ultrasonically treat it for 15 minutes, filter, discard the petroleum ether liquid, and evaporate the residue to dryness Petroleum ether, add methanol 20ml, plug, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take 1g of forsythia reference medicinal material, and prepare the reference medicinal material solution in the same way; Then take the forsythin reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution; test according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B), draw each of the above three solutions 5 μl , respectively spot on the same silica gel G thin-layer plate, use water-saturated chloroform-methanol-ethyl acetate-formic acid=10:2:1:0.5 as the developer, develop, take out, dry in the air, and spray with 10% sulfuric acid ethanol solution , heated at 105°C until the color of the spots is clear, and in the chromatogram of the test product, the spots of the same color appear at the position corresponding to the chromatogram of the reference product;

C. get the pharmaceutical composition honeyed pill of the present invention that embodiment 1 makes, put and observe under the microscope: pericarp epidermis cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; staggered vertically or vertically;

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: use octadecylsilane bonded silica gel as filler; acetonitrile-water-acetic acid=64:35: 1 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should be no less than 4000 according to the calculation of schisandrin A; the preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, Add methyl alcohol and make every 1ml contain the solution of 40 μ g; The preparation of need testing solution: take the content of the pharmaceutical composition water honey pill of the present invention that the embodiment 1 under the item of loading difference is made, grind finely, get 1.0g, accurately weigh Set, put in a stoppered Erlenmeyer flask, add 25ml of methanol precisely, seal the plug, weigh, ultrasonically treat with 120W power, 40kHz, 30 minutes, take it out, let it cool, weigh again, make up the lost weight with methanol , shake up, filter, and get the filtrate to get final product; Assay method: respectively accurately draw 10 μl of reference substance solution and need testing solution, inject liquid chromatograph, measure, and get final product; 1g contains Schisandra chinensis, calculated as schisandrin A C ₂₄ H ₃₂ O ₆ , should not be less than 0.9mg.

Embodiment 11 quality detection method

A. Take 2 g of the pharmaceutical composition water honey pill of the present invention made in Example 2, grind it finely, add 28 ml of 35° C. petroleum ether, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add 0.6 ml of 35° C. petroleum ether to dissolve the residue, As the test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution by the same method; then take the reference substance of Schizandrin A, add 35 ℃ petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; Layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition) test, draw 5 μ l of each of the above three solutions and place them on the same silica gel GF254 thin-layer plate, and use 35 ℃ petroleum ether-ethyl formate-formic acid as 11:6: The upper layer solution of 0.9 is a developing agent, developed, taken out, dried, and inspected under a 254nm ultraviolet light, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference substance, spots of the same color are shown;

B. Take 6g of the pharmaceutical composition water honey pill of the present invention made in Example 2, grind it finely, add 12ml of petroleum ether at 55°C, seal it tightly, treat it ultrasonically for 15 minutes, filter, discard the petroleum ether liquid, and evaporate the residue to dry petroleum ether , add 28ml of methanol, close plug, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take 1g of Forsythia reference medicinal material, and prepare the reference medicinal material solution in the same way; Forsythin reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution; test according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B), draw each 5 μ l of the above three solutions, respectively Spot on the same silica gel G thin-layer plate, use water-saturated chloroform-methanol-ethyl acetate-formic acid=9:2.4:0.9:0.6 as developing solvent, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, in Heat at 105°C until the color of the spots is clear, and in the chromatogram of the test product, the spots of the same color appear at the position corresponding to the chromatogram of the reference product;

C. get the pharmaceutical composition honeyed pill of the present invention that embodiment 2 makes, put and observe under the microscope: pericarp epidermis cell surface view class polygon, anticlinal wall is uneven in thickness, and cell cavity contains light-colored matter; staggered vertically or vertically;

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=62:38: 0.9 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should not be less than 4000 according to the calculation of schisandrin; Add methyl alcohol and make every 1ml contain the solution of 40 μ g; The preparation of need testing solution: get the pharmaceutical composition water honey pill of the present invention that embodiment 2 makes, grind finely, get 1.0g, accurately weigh, put the conical flask of tool stopper Add 22ml of methanol precisely, plug it tightly, weigh it, ultrasonically treat it, power 120W, 40kHz, 34 minutes, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake well, filter, take Continue the filtrate to get it; Assay method: accurately draw 10 μl each of the reference substance solution and the test solution, inject it into a liquid chromatograph, and measure it to get it; Based on C ₂₄ H ₃₂ O ₆ , it should not be less than 0.9mg.

Embodiment 12 quality detection method

A. Take 2 g of the pharmaceutical composition water honey pill of the present invention made in Example 3, grind it finely, add 12 ml of petroleum ether at 55 ° C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add 1.8 ml of petroleum ether at 55 ° C to the residue to dissolve, As the test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution by the same method; then take the reference substance of Schizandrin A, add 55°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; Layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition) test, draw 5 μ l of each of the above three solutions and place them on the same silica gel GF254 thin-layer plate, and use 55 ℃ petroleum ether-ethyl formate-formic acid as 18:4: The upper layer solution of 1.1 is a developer, developed, taken out, dried, and inspected under a 254nm ultraviolet light. In the chromatogram of the test product, on the position corresponding to the chromatogram of the reference product, spots of the same color appear;

B. Take 6g of the pharmaceutical composition water honey pill of the present invention made in Example 3, grind it finely, add 28ml of petroleum ether at 35°C, plug it tightly, treat it ultrasonically for 15 minutes, filter, discard the petroleum ether liquid, and evaporate the residue to dry petroleum ether , add 12ml of methanol, close plug, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take 1g of Forsythia reference drug, and prepare the reference drug solution in the same way; Forsythin reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution; test according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B), draw each 5 μ l of the above three solutions, respectively Spot on the same silica gel G thin-layer plate, use water-saturated chloroform-methanol-ethyl acetate-formic acid=11:1.6:1.1:0.4 as developing solvent, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, in Heat at 105°C until the color of the spots is clear, and in the chromatogram of the test product, the spots of the same color appear at the position corresponding to the chromatogram of the reference product;

C. get the pharmaceutical composition honeyed pill of the present invention that embodiment 3 makes, put and observe under the microscope: pericarp epidermis cell surface view class polygon, anticlinal wall thickness is uneven, and cell cavity contains light-colored matter; staggered vertically or vertically;

The content determination in the quality inspection method is as follows:

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=78:22: 1.1 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should not be less than 4000 according to the calculation of schisandrin; the preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, Add methyl alcohol and make every 1ml contain the solution of 40 μ g; The preparation of need testing solution: get the pharmaceutical composition water honey pill of the present invention that embodiment 3 makes, grind finely, get 1.0g, accurately weighed, put the conical flask with stopper Add 28ml of methanol precisely, plug it tightly, weigh it, ultrasonically treat it, power 120W, 40kHz, 26 minutes, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake well, filter, take Continue the filtrate to get it; Assay method: accurately draw 10 μl each of the reference substance solution and the test solution, inject it into a liquid chromatograph, and measure it to get it; Based on C ₂₄ H ₃₂ O ₆ , it should not be less than 0.9mg.

Embodiment 13 identification method

A. Take 2 g of the pharmaceutical composition water honey pill of the present invention made in Example 2, grind it finely, add 20 ml of petroleum ether at 30 to 60 ° C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add petroleum ether at 30 to 60 ° C to the residue Dissolve 1ml as the test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and make the reference medicinal solution in the same way; then take the reference substance of Schisandrin A, add petroleum ether at 30-60°C to make a solution containing 2mg per 1ml, as a control product solution; test according to thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition), draw 5 μl of each of the above three solutions and place them on the same silica gel GF254 thin-layer plate respectively, and use petroleum ether-ethyl formate at 30-60°C - Formic acid is 15: 5: 1 upper layer solution is developing agent, develops, takes out, dries, puts inspection under 254nm ultraviolet lamp, in the chromatogram of test product, on the position corresponding to chromatogram of reference substance, show the same color spot;

B. Take 6 g of the pharmaceutical composition water honey pill of the present invention made in Example 1, grind it finely, add 20 ml of petroleum ether at 30 to 60 ° C, seal it up, ultrasonically treat it for 15 minutes, filter, discard the petroleum ether liquid, and evaporate the residue to dryness Petroleum ether, add methanol 20ml, plug, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take 1g of forsythia reference medicinal material, and prepare the reference medicinal material solution in the same way; Then take the forsythin reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution; test according to thin-layer chromatography (Chinese Pharmacopoeia 2010 edition one appendix VI B), draw each of the above three solutions 5 μl , respectively spot on the same silica gel G thin-layer plate, use water-saturated chloroform-methanol-ethyl acetate-formic acid=10:2:1:0.5 as the developer, develop, take out, dry in the air, and spray with 10% sulfuric acid ethanol solution , heated at 105°C until the spots are clearly colored, and in the chromatogram of the test product, spots of the same color appear at the positions corresponding to the chromatogram of the reference product.

Embodiment 14 Assay method

Measure according to high performance liquid chromatography (Appendix VI D of Chinese Pharmacopoeia 2010 edition), chromatographic conditions and system suitability test: use octadecylsilane bonded silica gel as filler; acetonitrile-water-acetic acid=64:35: 1 is the mobile phase, flow rate: 1.0ml/min; the detection wavelength is 249nm, and the number of theoretical plates should be no less than 4000 according to the calculation of schisandrin A; the preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, Add methanol to make a solution containing 40 μg per 1ml; the preparation of the test solution: take the content of the pharmaceutical composition water honey pill of the present invention under the difference in loading amount, grind it finely, get 1.0g, accurately weigh, and put a stopper cone Add 25ml of methanol to the bottle, seal it tightly, weigh it, ultrasonically treat it at 120W, 40kHz for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake well, and filter , get the filtrate, to get it; Assay method: respectively accurately draw 10 μl of the reference substance solution and the solution of the test sample, inject it into a liquid chromatograph, and measure it to get it; Based on C ₂₄ H ₃₂ O ₆ , it should not be less than 0.9mg.

Embodiment 15 identification method

Take 2 g of the pharmaceutical composition water honey pill of the present invention prepared in Example 3, grind it finely, add 20 ml of petroleum ether at 30 to 60 ° C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add 1 ml of petroleum ether at 30 to 60 ° C to dissolve the residue , as the test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and make the reference medicinal solution in the same way; then take the reference substance of Schisandrin A, add 30-60°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference solution According to the thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia 2010 Edition) test, draw each 5 μ l of the above three solutions and place them on the same silica gel GF254 thin-layer plate respectively, and use 30-60 ℃ of petroleum ether-ethyl formate-formic acid The upper layer solution of 15:5:1 is the developer, developed, taken out, dried, and inspected under a 254nm ultraviolet light. In the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product.

Claims (6)

1. A medicinal composition honeyed water pill for the treatment of chronic hepatitis and liver cirrhosis, characterized in that said honeyed water pill is made by the following method: get raw material medicine Schisandra chinensis 1667g, fruit of privet lucidum 500g, forsythia 500g, beipaijiang 333g , crushed into fine powder, passed through a 100-mesh sieve, mixed evenly, added 20g of refined honey for every 100g of crude drug powder, added appropriate amount of water pan pills, dried, and made into water honey pills. 2. the detection method of pharmaceutical composition water honeyed pill as claimed in claim 1 is characterized in that the method comprises one or more in following identification and/or content determination: A. Take 2g of the medicinal composition water honey pill, grind it finely, add 10-30ml of petroleum ether at 30-60°C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add 0.5-2ml of petroleum ether at 30-60°C to dissolve the residue, and use it as The test solution; take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution in the same way; then take the reference substance of Schisandrin A, add 30-60°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; Chinese Pharmacopoeia 2010 edition one appendix VIB thin-layer chromatography test, draw each 5 μ l of above-mentioned three kinds of solutions and point respectively on the same silica gel GF254 thin-layer plate, with 30～60 ℃ petroleum ether-ethyl formate-formic acid as 10～20: 3～7: The upper layer solution of 0.8～1.2 is the developing agent, develop, take out, dry, put it under 254nm ultraviolet light for inspection, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference product, spots of the same color appear ; B. Take 6g of the medicinal composition water honey pill, grind it finely, add 10-30ml of petroleum ether at 30-60°C, plug it tightly, treat it with ultrasound for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add methanol 10 ～30ml, plugged, sonicated for 20 minutes, filtered, the filtrate was evaporated to dryness, the residue was dissolved in 5ml of methanol, and used as the test solution; another 1g of Forsythia reference drug was taken, and the reference drug solution was prepared in the same way; Glycoside reference substance, add methanol to make a solution containing 0.25mg per 1ml, as the reference substance solution; according to the Chinese Pharmacopoeia 2010 edition, appendix VIB thin-layer chromatography test, draw 5μl of each of the above three solutions, and spot on the same silica gel G On a thin-layer board, use water-saturated chloroform-methanol-ethyl acetate-formic acid=8～12:1.5～2.5:0.8～1.2:0.3～0.7 as the developer, develop, take out, dry in the air, and spray with 10% sulfuric acid ethanol Solution, heated at 105°C until the color of the spots is clear, and in the chromatogram of the test product, the spots of the same color appear at the position corresponding to the chromatogram of the reference substance; C. Take the medicinal composition water honey pill, put it under a microscope and observe: pericarp epidermis cells are polygonal in surface view, anticlinal wall thickness is uneven, and the cell cavity contains light-colored matter; the upper and lower layers of endocarp fiber bundles are arranged obliquely or vertically; The content determination in the quality inspection method is as follows: Measure according to Chinese Pharmacopoeia 2010 edition one appendix VID high performance liquid chromatography, chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=60～80:20～40 : 0.8～1.2 is the mobile phase, flow rate: 1.0ml/min; detection wavelength is 249nm, the number of theoretical plates should not be less than 4000 according to the calculation of schisandrin A; preparation of reference solution: accurately weigh the schisandrin reference substance, put In the bottle, add methanol to make a solution containing 40 μg per 1ml; preparation of the test solution: take the medicinal composition water honey pill, grind it finely, take 1.0g, accurately weigh it, put it in a stoppered conical flask, and add methanol precisely 20 ~ 30ml, sealed, weighed, ultrasonic treatment, power 120W, 40kHz, 25 ~ 35 minutes, take out, let cool, weigh again, make up the lost weight with methanol, shake well, filter, take continued The filtrate is ready to be obtained; the determination method is to accurately draw 10 μl of each of the reference substance solution and the test solution, inject it into a liquid chromatograph, and measure to obtain the product; the pharmaceutical composition water honey pill contains Schisandra chinensis and Schizandrin A C ₂₄ H per 1 g ₃₂ O ₆ should not be less than 0.9mg. 3. the detection method of pharmaceutical composition water honeyed pill as claimed in claim 2 is characterized in that the method comprises one or more in following identification and/or assay: A. Take 2g of the medicinal composition water honey pill, grind it finely, add 20ml of petroleum ether at 30-60°C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, add 1ml of petroleum ether at 30-60°C to dissolve the residue, and use it as the test solution ; Take another 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution in the same way; then take the reference substance of Schizandrin A, add 30-60°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; according to the Chinese Pharmacopoeia 2010 edition An appendix VIB thin-layer chromatography test, draw 5 μl of each of the above three solutions and spot them on the same silica gel GF254 thin-layer plate, and use 30-60°C petroleum ether-ethyl formate-formic acid as the upper layer solution of 15:5:1 It is a developing agent, developed, taken out, dried, and inspected under a 254nm ultraviolet lamp. In the chromatogram of the test product, spots of the same color appear on the position corresponding to the chromatogram of the reference product; B. Take 6g of the medicinal composition water honey pill, grind it finely, add 20ml of petroleum ether at 30-60°C, seal it tightly, treat it with ultrasound for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add 20ml of methanol, seal it tightly stopper, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take another 1 g of forsythia reference drug, and make the reference drug solution in the same way; then take the forsythin reference substance, Add methanol to make a solution containing 0.25mg per 1ml, as a reference solution; according to the Chinese Pharmacopoeia 2010 Edition, Appendix VIB thin-layer chromatography test, draw 5μl of each of the above three solutions, and spot them on the same silica gel G thin-layer plate , use water-saturated chloroform-methanol-ethyl acetate-formic acid=10:2:1:0.5 as the developer, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clear in color, In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product; C. Take the medicinal composition water honey pill, put it under a microscope and observe: pericarp epidermis cells are polygonal in surface view, anticlinal wall thickness is uneven, and the cell cavity contains light-colored matter; the upper and lower layers of endocarp fiber bundles are arranged obliquely or vertically; Measure according to Chinese Pharmacopoeia 2010 edition one appendix VID high-performance liquid chromatography, chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=64:35:1 is flow Phase, flow rate: 1.0ml/min; detection wavelength 249nm, the number of theoretical plates should not be less than 4000 based on the calculation of schisandrin A; preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, add methanol to prepare into a solution containing 40 μg per 1ml; the preparation of the test solution: take the contents of the pharmaceutical composition water honey pill under the item of difference in loading amount, grind it finely, get 1.0g, accurately weigh it, put it in a stoppered conical flask, and accurately weigh it. Add 25ml of methanol, plug it tightly, weigh it, ultrasonically treat it, power 120W, 40kHz, 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake well, filter, and take the filtrate. Determination method: each 10 μ l of the reference substance solution and the test solution are precisely drawn, injected into a liquid chromatograph, and measured, and the product is obtained; the pharmaceutical composition water honey pill contains Schisandra chinensis and Schizandrin A C ₂₄ H ₃₂ O per 1 g ₆ , should not be less than 0.9mg. 4. the detection method of pharmaceutical composition honeyed pill as claimed in claim 2 is characterized in that the method comprises one or more of following identification and/or content determination: A. Take 2g of the medicinal composition water honey pill, grind it finely, add 28ml of petroleum ether at 35°C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue with 0.6ml of petroleum ether at 35°C, and use it as the test solution; Take 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution in the same way; then take the reference substance of Schizandrin A, add 35°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; according to the Chinese Pharmacopoeia 2010 Edition, Appendix VIB For layer chromatography test, absorb 5 μl of each of the above three solutions and spot them on the same silica gel GF254 thin-layer plate, and use the upper layer solution of 35°C petroleum ether-ethyl formate-formic acid as the developing agent at 11:6:0.9 to develop and take out , dried, and inspected under a 254nm ultraviolet lamp, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference substance, spots of the same color are displayed; B. Take 6g of the medicinal composition water honey pill, grind it finely, add 12ml of petroleum ether at 55°C, seal it up, treat it with ultrasound for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add 28ml of methanol, seal it up, Sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take another 1g of forsythia reference drug, and make a reference drug solution in the same way; then take forsythin reference substance, add methanol Make a solution containing 0.25 mg per 1 ml as the reference substance solution; according to the Chinese Pharmacopoeia 2010 edition, an appendix VIB thin-layer chromatography test, draw 5 μl of each of the above three solutions, and spot them on the same silica gel G thin-layer plate respectively to Water-saturated chloroform-methanol-ethyl acetate-formic acid=9:2.4:0.9:0.6 as developing agent, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots develop clear color, for testing In the chromatogram of the product, spots of the same color appear at the position corresponding to the chromatogram of the reference product; C. Take the medicinal composition water honey pill, put it under a microscope and observe: pericarp epidermis cells are polygonal in surface view, anticlinal wall thickness is uneven, and the cell cavity contains light-colored matter; the upper and lower layers of endocarp fiber bundles are arranged obliquely or vertically; Measure according to Chinese Pharmacopoeia 2010 edition one appendix VID high-performance liquid chromatography, chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=62:38:0.9 is flow Phase, flow rate: 1.0ml/min; detection wavelength 249nm, the number of theoretical plates should not be less than 4000 based on the calculation of schisandrin A; preparation of reference solution: accurately weigh the reference substance of schisandrin A, put it in a volumetric bottle, add methanol to prepare into a solution containing 40 μg per 1 ml; the preparation of the test solution: take the pharmaceutical composition honeyed pills, grind finely, get 1.0 g, accurately weigh, put in a stoppered Erlenmeyer flask, add 22 ml of methanol precisely, seal it tightly, weigh Determine the weight, ultrasonic treatment, power 120W, 40kHz, 34 minutes, take it out, let it cool, weigh again, make up the lost weight with methanol, shake well, filter, and take the filtrate to get it; determination method: separate precision Take 10 μl each of the reference substance solution and the test solution, inject it into a liquid chromatograph, measure it, and get it; every 1 g of the pharmaceutical composition Shuimiwan contains Schisandra chinensis, calculated as Schizandrin A C ₂₄ H ₃₂ O ₆ , should not be less than 0.9 mg. 5. the detection method of pharmaceutical composition honeyed pill as claimed in claim 2 is characterized in that the method comprises one or more of following identification and/or content determination: A. Take 2g of the medicinal composition water honey pill, grind it finely, add 12ml of petroleum ether at 55°C, ultrasonically treat it for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue with 1.8ml of petroleum ether at 55°C, and use it as the test solution; Take 1g of the reference medicinal material of Schisandra chinensis, and prepare the reference medicinal material solution in the same way; then take the reference substance of Schizandrin A, add 55°C petroleum ether to make a solution containing 2mg per 1ml, and use it as the reference substance solution; according to the Chinese Pharmacopoeia 2010 edition, appendix VIB thin For layer chromatography test, draw 5 μl of each of the above three solutions and spot them on the same silica gel GF254 thin-layer plate, and use the upper layer solution of 55°C petroleum ether-ethyl formate-formic acid as the developing agent at 18:4:1.1 to develop and take out , dried, and inspected under a 254nm ultraviolet lamp, in the chromatogram of the test product, on the position corresponding to the chromatogram of the reference substance, spots of the same color are displayed; B. Take 6g of the medicinal composition water honey pill, grind it finely, add 28ml of petroleum ether at 35°C, seal it up, treat it with ultrasound for 15 minutes, filter, discard the petroleum ether liquid, evaporate the residue to dry petroleum ether, add 12ml of methanol, seal it up, Sonicate for 20 minutes, filter, evaporate the filtrate to dryness, add 5ml of methanol to the residue to dissolve, and use it as the test solution; take another 1g of forsythia reference drug, and make a reference drug solution in the same way; then take forsythin reference substance, add methanol Make a solution containing 0.25 mg per 1 ml as the reference substance solution; according to the Chinese Pharmacopoeia 2010 edition, an appendix VIB thin-layer chromatography test, draw 5 μl of each of the above three solutions, and spot them on the same silica gel G thin-layer plate respectively to Water-saturated chloroform-methanol-ethyl acetate-formic acid=11:1.6:1.1:0.4 is used as developer, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots develop clear color, for testing In the chromatogram of the product, spots of the same color appear at the position corresponding to the chromatogram of the reference product; C. Take the medicinal composition water honey pill, put it under a microscope and observe: pericarp epidermis cells are polygonal in surface view, anticlinal wall thickness is uneven, and the cell cavity contains light-colored matter; the upper and lower layers of endocarp fiber bundles are arranged obliquely or vertically; Measure according to Chinese Pharmacopoeia 2010 edition one appendix VID high-performance liquid chromatography, chromatographic conditions and system suitability test: be filler with octadecylsilane bonded silica gel; Acetonitrile-water-acetic acid=78:22:1.1 is flow Phase, flow rate: 1.0ml/min; detection wavelength 249nm, the number of theoretical plates should not be less than 4000 based on the calculation of schisandrin A; preparation of the reference solution: accurately weigh the schisandrin reference substance, put it in a volumetric bottle, add methanol to prepare into a solution containing 40 μg per 1 ml; the preparation of the test solution: take the pharmaceutical composition honeyed pills, grind finely, get 1.0 g, accurately weigh, put in a stoppered Erlenmeyer flask, accurately add 28 ml of methanol, seal it tightly, weigh Determine the weight, ultrasonic treatment, power 120W, 40kHz, 26 minutes, take it out, let it cool, weigh again, make up the lost weight with methanol, shake well, filter, and take the filtrate to get it; determination method: separate precision Take 10 μl each of the reference substance solution and the test solution, inject it into a liquid chromatograph, measure it, and get it; every 1 g of the pharmaceutical composition Shuimiwan contains Schisandra chinensis in terms of Schizandrin A C ₂₄ H ₃₂ O ₆ , which should not be less than 0.9 mg. 6. the application of pharmaceutical composition honeyed water pill as claimed in claim 1 in the preparation treatment chronic hepatitis, liver cirrhosis medicine.