CN103120718B - A kind of pharmaceutical composition for the treatment of chronic hepatitis, liver cirrhosis - Google Patents
A kind of pharmaceutical composition for the treatment of chronic hepatitis, liver cirrhosis Download PDFInfo
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- CN103120718B CN103120718B CN201110367498.1A CN201110367498A CN103120718B CN 103120718 B CN103120718 B CN 103120718B CN 201110367498 A CN201110367498 A CN 201110367498A CN 103120718 B CN103120718 B CN 103120718B
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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
Technical field
The present invention relates to a kind of pharmaceutical composition and preparation method thereof and method of quality control, particularly a kind ofly treat chronic hepatitis, the pharmaceutical composition and preparation method thereof of liver cirrhosis and method of quality control.
Background technology
Chronic hepatitis refers to that causing liver generation chronic inflammatory disease by the various cause of disease changes, and the course of disease is at least more than half a year.Visible jaundice, the side of body costalgia, tympanites, the performance such as to gather.Usual cell enzyme activity is abnormal, and in normal human, enzymatic activity is more stable, and enzyme activity change imply that some Organ and tissue of human body is impaired or after there is disease, some enzyme is released in blood, urine or body fluid.Therefore, by the determination of activity of blood, urine or body fluid endoenzyme, can understand or judge generation and the development of some disease.The diseases such as the traditional Chinese medical science shows as deficiency of liver-QI more, cloudy blood two loses, hyperactive liver-QI attacking the spleen, stagnation of QI due to depression of the liver, residual poison are unclear.The chronic hepatitis course of disease is delayed, several years even repeatedly, more than ten years, the developed into liver cirrhosis had.
Liver cirrhosis (liver cirrhosis) is a kind of common chronic hepatopathy, can cause liver injury by one or more reasons, and liver is Progressive symmetric erythrokeratodermia, diffusivity, fibrous lesions.Be embodied in hepatocyte diffusivity degeneration necrosis, then there is proliferation of fibrous tissue and the regeneration of liver cell nodules shape, these three kinds change repeatedly staggered carrying out, and result lobules of liver structure and blood circulation path are reconstructed gradually, liver are out of shape, hardening and cause liver cirrhosis.The early stage non-evident sympton of this disease, there is a series of portal hypertension in various degree and hepatic insufficiency in stage, until occur that the complication such as upper gastrointestinal hemorrhage, hepatic encephalopathy is dead.Liver reduces, " hepatic atrophy " refers to that B ultrasonic or CT detect, and liver volume reduces, and belongs to " tympanites " of traditional medicine, and modern medicine is qualitative is " liver cirrhosis ", and liver reduces and except liver transplantation, there is no other effective Therapeutic Method both at home and abroad at present.
The etiology and pathogenesis of chronic hepatitis, regularity of occurrence and development etc. are studied very deep from basis to clinical by the development of modern medicine.Chronic viral hepatitis still lacks the Therapeutic Method of specially good effect at present.Any medicine all never shows its definite curative effect to chronic viral hepatitis, is therefore still carrying out various test of cure at present.Tcm syndrome differentiation and treatment chronic hepatitis, with its letter, just, test, honest and clean characteristic, preventing and treating chronic hepatitis, liver cirrhosis has incomparable advantage.
Summary of the invention
The object of the invention is to provide a kind of pharmaceutical composition and preparation method thereof, and another object of the present invention is the method for quality control and the purposes that provide this pharmaceutical composition.
The present invention seeks to be achieved through the following technical solutions
The crude drug of pharmaceutical composition of the present invention consists of:
Fructus Schisandrae Sphenantherae 1000-2000 weight portion Fructus Ligustri Lucidi 300-700 weight portion
Fructus Forsythiae 300-700 weight portion Sonchus brachyotus DC. 200-450 weight portion
Above-mentioned raw materials medicine preferred weight proportioning is as follows:
Fructus Schisandrae Sphenantherae 1667 weight portion Fructus Ligustri Lucidi 500 weight portion Fructus Forsythiae 500 weight portion Sonchus brachyotus DC. 333 weight portion
Above-mentioned raw materials medicine preferred weight proportioning is as follows:
Fructus Schisandrae Sphenantherae 1950 weight portion Fructus Ligustri Lucidi 350 weight portion Fructus Forsythiae 650 weight portion Sonchus brachyotus DC. 250 weight portion
Above-mentioned raw materials medicine preferred weight proportioning is as follows:
Fructus Schisandrae Sphenantherae 1100 weight portion Fructus Ligustri Lucidi 600 weight portion Fructus Forsythiae 350 weight portion Sonchus brachyotus DC. 400 weight portion
Pharmaceutical composition crude drug of the present invention, adds customary adjuvant, conveniently technique, makes clinical acceptable big honeyed pills, water-honeyed pill, the watered pill, capsule, granule, tablet.
The preparation technology of pharmaceutical composition water-honeyed pill of the present invention is as follows:
Get above-mentioned four taste crude drug, be ground into fine powder, cross 100 mesh sieves, mixing, every 100 raw material medicated powder ends add 20 ~ 25 weight portion refined honeys, add water pill, dry, make water-honeyed pill.
The quality determining method of pharmaceutical composition of the present invention comprises following discriminating and/or assay:
Differentiate comprise in following method one or more:
A. get pharmaceutical composition water-honeyed pill 2g of the present invention, porphyrize, add 30 ~ 60 DEG C of petroleum ether 10 ~ 30ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 0.5 ~ 2ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 10 ~ 20: 3 ~ 7: 0.8 ~ 1.2 is for developing solvent with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get pharmaceutical composition water-honeyed pill 6g of the present invention, porphyrize, add 30 ~ 60 DEG C of petroleum ether 10 ~ 30ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 10 ~ 30ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=8 ~ 12: 1.5 ~ 2.5: 0.8 ~ 1.2: 0.3 ~ 0.7 is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. get pharmaceutical composition water-honeyed pill of the present invention, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=60 ~ 80: 20 ~ 40: 0.8 ~ 1.2 is mobile phase, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: get pharmaceutical composition water-honeyed pill of the present invention, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 20 ~ 30ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 25 ~ 35 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
The method of quality control of pharmaceutical composition of the present invention is preferably as follows discriminating and/or assay:
A. get pharmaceutical composition water-honeyed pill 2g of the present invention, porphyrize, add 30 ~ 60 DEG C of petroleum ether 20ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 1ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 15: 5: 1 is for developing solvent with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get pharmaceutical composition water-honeyed pill 6g of the present invention, porphyrize, add 30 ~ 60 DEG C of petroleum ether 20ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 20ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=10: be developing solvent at 2: 1: 0.5, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. get pharmaceutical composition water-honeyed pill of the present invention, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=64: be mobile phase at 35: 1, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: get the pharmaceutical composition water-honeyed pill content of the present invention under content uniformity item, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 25ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 30 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
The method of quality control of pharmaceutical composition of the present invention is preferably as follows discriminating and/or assay:
A. get pharmaceutical composition water-honeyed pill 2g of the present invention, porphyrize, add 35 DEG C of petroleum ether 28ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds 35 DEG C of petroleum ether 0.6ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 35 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 11: 6: 0.9 is for developing solvent with 35 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get pharmaceutical composition water-honeyed pill 6g of the present invention, porphyrize, add 55 DEG C of petroleum ether 12ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 28ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=9: be developing solvent at 2.4: 0.9: 0.6, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. get pharmaceutical composition water-honeyed pill of the present invention, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=62: be mobile phase at 38: 0.9, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: get pharmaceutical composition water-honeyed pill of the present invention, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 22ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 34 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
The method of quality control of pharmaceutical composition of the present invention is preferably as follows discriminating and/or assay:
A. get pharmaceutical composition water-honeyed pill 2g of the present invention, porphyrize, add 55 DEG C of petroleum ether 12ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds 55 DEG C of petroleum ether 1.8ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 55 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 18: 4: 1.1 is for developing solvent with 55 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get pharmaceutical composition water-honeyed pill 6g of the present invention, porphyrize, add 35 DEG C of petroleum ether 28ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 12ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=11: be developing solvent at 1.6: 1.1: 0.4, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. get pharmaceutical composition water-honeyed pill of the present invention, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered.
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=78: be mobile phase at 22: 1.1, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: get pharmaceutical composition water-honeyed pill of the present invention, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 28ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 26 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
Pharmaceutical composition of the present invention has YIN nourishing nourishing the liver, effect of expelling remnant toxicity, cure mainly chronic hepatitis, hepatitis interstitialis chronica belongs to that the cloudy blood two of Chinese medical discrimination loses, the clear person of residual poison, clinical efficacy is very definite, the liver protecting and ALT lowering good medicine, and at blood-activating and qi-promoting, replenishing YIN and removing heat, jaundice eliminating and improve all aobvious good therapeutic effect in the aspects such as Proteometabolism.Pharmaceutical composition preparation method of the present invention is advanced, quality controllable, safe and effective.The features such as pharmaceutical composition water-honeyed pill of the present invention has taking convenience, it is convenient to carry, remarkable in economical benefits.The separating degree of discrimination method and repeatability are all good; Negative control sample is noiseless; Specificity is stronger.Better, linear relationship is good for content assaying method instrument precision and operation precision, and experiment repeatability is good, and this law has good recovery.
Accompanying drawing illustrates:
The indentification by TLC Test Drawing of Fig. 1 Fructus Schisandrae Chinensis
(1-6 is respectively negative control, test sample three batches (080220,08022023,080226), reference substance deoxyschizandrin, control medicinal material Fructus Schisandrae Sphenantherae)
The indentification by TLC Test Drawing of Fig. 2 Fructus Forsythiae
(1-6 is respectively negative control, test sample three batches (080220,08022023,080226), reference substance phillyrin, control medicinal material Fructus Forsythiae)
Fig. 3 test sample, reference substance, negative control solution are separated HPLC chromatogram
(A negative control solution; B reference substance solution; C need testing solution; 1 deoxyschizandrin)
The ultraviolet spectrogram of deoxyschizandrin in Fig. 4 test sample, reference substance solution chromatograph
Fig. 5 deoxyschizandrin concentration-peak area linear relationship chart
Fig. 6 assay chromatogram-reference substance (1)
Fig. 7 assay chromatogram-reference substance (2)
Fig. 8 assay chromatogram-lot number 080220 sample (1)
Fig. 9 assay chromatogram-lot number 080220 sample (2)
Figure 10 assay chromatogram-lot number 080220 sample (3)
Figure 11 assay chromatogram-lot number 080223 sample (1)
Figure 12 assay chromatogram-lot number 080223 sample (2)
Figure 13 assay chromatogram-lot number 080223 sample (3)
Figure 14 assay chromatogram-lot number 080223 sample (4)
Figure 15 assay chromatogram-lot number 080226 sample (1)
Figure 16 assay chromatogram-lot number 080226 sample (2)
Figure 17 assay chromatogram-lot number 080226 sample (3)
Figure 18 assay chromatogram-lot number 080226 sample (4)
Following experimental example and embodiment are used for further illustrating but are not limited to the present invention.
Experimental example 1 pharmaceutical composition water-honeyed pill of the present invention preparation method is tested
1. disintegrating process is investigated
1.1 pulverization conditions
Instrument and equipment: pulverizer, Shandong Yantai City Zhi Gang pharmaceutical machine factory produces, and is pulverized three major parts such as unit control station, WZS3-470 type Horizontal revolving screen and hF3-200 type pulverizer form by the automatic control of ZKF-3 type; Medical material: above four Chinese medicine material is all dried to bone dry in drying room or baking oven.
1.2 disintegrating process
Crushed work efficiency is pulverizing 20 ~ 200kg medical material per hour, and granularity of sieving can adjust arbitrarily between 20 ~ 400 orders, actually selects No. 8 sieves (80 order) to pulverize, and owing to being automatic circulation type grinding mode, its flour extraction can reach more than 95%.
2 soft material pill making craft conditions
Former big honeyed pills basis increases water-honeyed pill specification; Water-honeyed pill: refer to the pill that fine medicinal material powder is made for adhesive with Mel and water.Made pill shall be rounded up evenly, color and luster is consistent.
Specification is every 100 12g.
2.1 soft materials processed
Honey degree and add two factors of amount outbalance when being soft material processed of honey, employing L9 (34) orthogonal table is tested, and investigates.
Mel can be divided into three kinds of specifications according to the degree of refining: tender honey, middle honey, always honey.
Tender honey: Mel is heated to 105 DEG C ~ 115 DEG C, water content is 14% ~ 20%, and density is about 1.35, color and luster without significant change, rare viscosity.
Middle honey: tender honey continues heating, and temperature reaches 116 ~ 118 DEG C, and water content is 14% ~ 16%, and density is about 1.37, occurs the light yellow glossiness uniform bubble seethed, twists with the fingers toughness, occur when two handss separate without white silk with hands.
Old honey: middle honey continues heating, and temperature reaches 119 ~ 122 DEG C, and water content is below 10%, and density is about 1.4, occurs the bubble that rufous gloss is larger, hands sth.s made by twisting very sticky, when long white silk separately appears in two fingers, instills beading in water.
2.1.1 factor level table, in table 1
Table 1 orthogonal test factor level table
2.1.2 sample preparation: take medicated powder 0.5kg, by L
9(3
4) orthogonal table condition prepares sample.
2.1.3 evaluation index: ball block should be able to be arbitrarily moulding and do not ftracture, hand rubbing alunite and tack-free, does not adhere to wall.
Experimental result is in table 2:
Table 2 Orthogonal experiment results
Result shows: experiment 8,9 is all plastic, but experiment 9 consumption honey is many, is save cost, selects experiment 8, namely select tender honey, and honey amount is 25% of medicated powder.
2.2 training powder draw circle
The soft material made is put into pellet processing machine pill after medicine refining machine refining, and need training powder to draw circle after obtained ball, rounding is smooth more to make pill.
At training powder, draw in round process, need to investigate, using the smooth rounding of pill as well as evaluation index mulse (ratio of honey and water).
Experimentation:
Experiment one: with mulse (sweet: water=1: 4) carry out training powder, draw circle,
Make pill smooth, but not rounding, outward appearance is poor.
Experiment two: with mulse (sweet: water=1: 5) carry out training powder, draw circle,
Gained pill is smooth, rounding, evenly, outward appearance is better.
Experiment three: with mulse (sweet: water=1: 6) carry out training powder, draw circle,
Gained pill rounding, evenly, but more coarse, and outward appearance is bad.
Known by testing above: with honey: water be 1: 5 and 1: 6 mulse pill can be drawn circle, but, rounding smooth with the mulse gained pill of 1: 5, even, outward appearance is better.
2.3 dry
Sample need investigate dry temperature and time when drying, and makes its moisture Control within 12%.
Under Chinese Pharmacopoeia annex IA pill item:
Unless otherwise specified, water-honeyed pill, the watered pill, condensed water honeyed pill and concentrated watered pill all should be dry below 80 DEG C.
[moisture] water-honeyed pill must not cross 12.0%
So pharmaceutical composition water-honeyed pill of the present invention is drafted carry out drying at 80 DEG C, it is investigated, have flawless for inspection target with the dried moisture of pill, pill.In different section samplings drying time, survey its moisture.The results are shown in Table 3:
The section sampling drying time water content detection result that table 3 is different
Result shows: pill is at 80 DEG C, and dry 120min, namely conforms with the regulations.
Conclusion: comprehensive above experimental data, the processing parameter of pharmaceutical composition water-honeyed pill of the present invention is:
Honey kind: tender honey; Add honey amount: 25% of medicated powder; Training powder draws round mulse to compare: 1: 5; Baking temperature: 80 DEG C; Drying time: 120min
3. adjuvant source and standard
Mel: number of registration food 01707Q10265ROS (food additive); Beijing Bai Ji Jian Le bee product company limited produces; Product batch number: 20080228.
Experimental example 2 pharmaceutical composition water-honeyed pill of the present invention identification experiment
Differentiate (1) [prescription] Fructus Schisandrae Sphenantherae 1667g Fructus Ligustri Lucidi 500g Fructus Forsythiae 500g Sonchus brachyotus DC. 333g
More than [method for making], four tastes, are ground into fine powder, sieve, mixing.Every 100g powder refined honey 20 ~ 30g adds appropriate water pill, dry, makes water-honeyed pill.
[character] pharmaceutical composition water-honeyed pill of the present invention is the water-honeyed pill of brownish black; Feeble QI, the micro-acid of taste.
Pharmaceutical composition water-honeyed pill of the present invention is got in [discriminating] (1), puts basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, and cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered.
(2) get pharmaceutical composition water-honeyed pill 2g of the present invention, porphyrize, add petroleum ether (30 ~ 60 DEG C) 20ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds petroleum ether (30 ~ 60 DEG C) 1ml and dissolves, as need testing solution.Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution.Get deoxyschizandrin reference substance again, add petroleum ether (30 ~ 60 DEG C) and make the solution of every 1ml containing 2mg, product solution in contrast.According to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B) test, draw each 5 μ l of above-mentioned three kinds of solution and put respectively in same silica gel G F
254on lamellae, with the upper solution of petroleum ether (30 ~ 60 DEG C)-Ethyl formate-formic acid (15: 5: 1) for developing solvent, launch, take out, dry, inspect under putting ultra-violet lamp (254nm).In test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color.
(3) get pharmaceutical composition water-honeyed pill 6g of the present invention, porphyrize, add petroleum ether (30 ~ 60 DEG C) 20ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 20ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution.Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution.Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast.Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform (water saturation)-methanol-ethyl acetate-formic acid (10: 2: 1: 0.5) for developing solvent, launch, take out, dry, spray, with 10% ethanol solution of sulfuric acid, is heated to spot development at 105 DEG C clear.In test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color.
Differentiate the TLC distinguish that (2) are Fructus Schisandrae Sphenantherae in pharmaceutical composition water-honeyed pill of the present invention: monarch drug in the Fructus Schisandrae Sphenantherae side of being.Known be wherein the lignan component of representative with deoxyschizandrin is its main active, and specificity is stronger.In this discrimination test, the processing method of need testing solution and chromatographic condition differentiate the method (P.169) under item with reference to " liver obtains peaceful ball " former preparation quality standard and China's coastal port one " Fructus Schisandrae Sphenantherae ", gained speckle R
fvalue is comparatively suitable, clear spot.The separating degree of method and repeatability are all good; Negative control sample (lack Fructus Schisandrae Sphenantherae, prepare negative sample, then prepare negative control solution by need testing solution preparation method by preparation technology) is noiseless; With deoxyschizandrin and Fructus Schisandrae Sphenantherae control medicinal material simultaneously for contrast, specificity is stronger.(see accompanying drawing 1).
Differentiate (3) discriminating that is Fructus Forsythiae in pharmaceutical composition water-honeyed pill of the present invention: in this discrimination test, the processing method of need testing solution and chromatographic condition list of references method are also improved, gained speckle R
fvalue is comparatively suitable, clear spot.The separating degree of method and repeatability are all good, and negative control sample (lack Fructus Forsythiae, prepare negative sample, then prepare negative control solution by need testing solution preparation method by preparation technology) is noiseless; With phillyrin and Fructus Forsythiae control medicinal material for contrast, specificity is strong.Observe (see accompanying drawing 2) through too much batch sample and negative control experiments, confirm that this law is set up.
[inspection], by every regulation under Chinese Pharmacopoeia version in 2010 annex pill item, result all conforms with the regulations.
1. moisture gets pharmaceutical composition water-honeyed pill of the present invention, measures according to aquametry (Chinese Pharmacopoeia version in 2010 annex IX H).Should 12.0% be crossed.The results are shown in Table 4.
Table 4 determination of water result
Conclusion: three batch sample determination of water results all conform with the regulations.
2. weight differential gets pharmaceutical composition water-honeyed pill of the present invention, and (under Chinese Pharmacopoeia version in 2010 annex I A item weight differential inspection technique) measures in accordance with the law.
The inspection of table 5 weight differential
Conclusion: three batch sample weight differentials all, within limit (± 10%), all conform with the regulations.
3. content uniformity gets pharmaceutical composition water-honeyed pill of the present invention 10 bags, checks in accordance with the law.The results are shown in Table 6.
The inspection of table 6 content uniformity
Conclusion: three batch sample content uniformity all, within limit (± 5%), all conform with the regulations.
4. dissolve scattered time limit gets pharmaceutical composition water-honeyed pill 6 ball of the present invention, checks in accordance with the law, the results are shown in Table 7.
Table 7 dissolve scattered time limit measurement result
Conclusion: three batch sample dissolve scattered time limit measurement results all conform with the regulations.
5. microbial limit
Microbial limit is checked according to Chinese Pharmacopoeia version in 2010 annex XIII C.Get pharmaceutical composition water-honeyed pill 10g of the present invention, add pH 7.0 sterile NaCl-peptone buffer agent to 100ml, make test liquid by insulation jolting method, check with Plating.The results are shown in Table 8.
Table 8 limit test of microbe table
Conclusion: three batch sample microbial limit results all conform with the regulations.
6. heavy metal inspection
Check that the second method measures according to Chinese Pharmacopoeia version in 2010 annex IX E heavy metal.The results are shown in Table 9.
Table 9 determining heavy metals table
Result shows, three batch sample content of beary metal are all less than 20PPM.
7. arsenic salt checks
Measure according to " Chinese Pharmacopoeia " version in 2010 annex IX F arsenic salt inspection technique first method (ancient Cai Shi method).The results are shown in Table 10:
Table 10 arsenic salt chart
Result shows, three batch sample arsenic salt contents are all less than 2PPM.
Experimental example 3 pharmaceutical composition of the present invention (preparing same embodiment 1) water-honeyed pill assay is tested
1. the selection of assay index composition
One of the main flavour of a drug of said preparation Fructus Schisandrae Sphenantherae, is classified as monarch drug in prescription.Deoxyschizandrin content limit is determined according to the measurement result of three batch samples.Known be wherein the lignan component of representative with deoxyschizandrin is its main active, thus selected its be pharmaceutical composition water-honeyed pill assay object of the present invention.This quality standard is main, and with reference to SHENQI WUWEIZI PIAN, (Chinese Pharmacopoeia 2005 editions, p.508) content assaying method of deoxyschizandrin under item, adopts HPLC method to carry out assay to one of Fructus Schisandrae Sphenantherae index components deoxyschizandrin.Meet assay requirement through Method validation, can be used as the method for pharmaceutical composition water-honeyed pill deoxyschizandrin assay of the present invention.
2. instrument and reagent:
High performance liquid chromatograph: Agilent 1100 quaternary gradient pump; Agilent 1100 automatic sampler; Agilent1100DAD detector; Agilent chem workstation; HP DeskJet Laser P1015 printer.
Reagent: methanol and acetonitrile are chromatographically pure; Water is ultra-pure water, and other reagent is analytical pure.
Chromatographic condition: analytical column is phenomenex Luna C-18 (5 μm, 250 × 4.6mm) (phenomenex company of the U.S.); Mobile phase is acetonitrile-water-acetic acid (64: 35: 1), filters through 0.45 μm of filter membrane with front; Flow velocity 1.0ml/min, determined wavelength 249nm; Column temperature 25 DEG C.
Under institute's mimic colouration spectral condition, the retention time of deoxyschizandrin is about 22 minutes, and the theoretical cam curve of deoxyschizandrin is for being not less than 4000, therefore tentative theoretical cam curve is not less than 4000 by deoxyschizandrin.
3. methodological study
The selection of extracting method is according to the character of deoxyschizandrin, with reference to the preparation method of joining stilbene Fructus Schisandrae Chinensis need testing solution, (Chinese Pharmacopoeia 2005 editions, P.508), considers to take methanol as solvent, supersound extraction, investigates the extraction effect of different supersound extraction time.Method is: get pharmaceutical composition water-honeyed pill content of the present invention, porphyrize, gets fine powder and be about 1g, accurately weighed, put in tool plug conical flask, precision adds methanol 25ml, close plug, weighed weight, supersound process (power 120W, 40kHz), from 10 minutes, took out every 10 minutes, let cool, more weighed weight, supply the weight of less loss with methanol, shake up, get solution and be about 1ml, filter, get subsequent filtrate 10 μ l, injection liquid chromatography, record chromatogram, measures deoxyschizandrin area; Surplus solution is close plug again, weighed weight, supersound process.The results are shown in Table 11.
Table 11 take methanol as the extraction result of solvent different supersound extraction time
From upper table, supersound process is after 30 minutes, and deoxyschizandrin peak area no longer increases, and determines that sonication treatment time is 30 minutes.
Blank assay
In the autogamy of square taste of Chinese medicine ratio not containing the negative sample of Fructus Schisandrae Sphenantherae, blank solution is made by text method, measure in accordance with the law, result blank solution has no obvious chromatographic peak at retention time place identical with deoxyschizandrin reference substance, think noiseless, deoxyschizandrin peak shape, all meet test request (Fig. 3) with the separating degree etc. of adjacent peak; Application diode array detector and chromatographic work station, scan reference substance and test sample, the results are shown in uv scan figure (Fig. 4).
Stability experiment gets need testing solution, respectively at preparation rear 0,1,2,4, within 6,8 hours, measures.Result shows, need testing solution is basicly stable (table 12) in 8 hours.
Table 12 need testing solution stability test result
Precision Experiment precision draws deoxyschizandrin reference substance solution 10 μ l, and continuous sample introduction 6 times, measures peak area, the results are shown in Table 13.Show instrument precision and operate precision better.
Table 13 Precision Experiment result
The investigation of linear relationship respectively precision measures deoxyschizandrin reference substance stock solution (164.0 μ g/ml) 4.0, 5.0, 6.0, 7.0, 8.0ml, put in the brown measuring bottle of different 25ml respectively, add methanol to scale, shake up, the accurate 10 μ l that draw inject high performance liquid chromatograph successively respectively, peak area is measured by above-mentioned chromatographic condition, with integrating peak areas value (A) for vertical coordinate, concentration (the C of deoxyschizandrin, μ g/ml) be abscissa, drawing standard curve (see Fig. 5), regression equation is A=23.4071C+1.1763, correlation coefficient r=0.9999 (n=5).
Result shows deoxyschizandrin within the scope of concentration 26.24 ~ 52.48 μ g/ml in good linear relationship.
Replica test gets same batch sample (080220), and parallel preparation 6 parts of need testing solutions also measure, and calculates content.The results are shown in Table 14.Show that experimental technique repeatability is good.
Table 14 repeated experiment result (n=6)
Average recovery is tested precision respectively and is taken known content (0.9999mg/g) same batch sample (080220) about 0.5g totally 6 parts, and precision adds each 0.5248mg of deoxyschizandrin reference substance (104.96 μ g/ml × 5.0ml) respectively.Preparation method and the above-mentioned chromatographic condition of pressing text need testing solution measure, and calculate the response rate, the results are shown in Table 15 with following formula.
Table 15 average recovery result of the test (n=6)
Above measurement result shows, this law has good recovery.
Sample determination
Get three batch samples, record by text the content that method measures deoxyschizandrin, the results are shown in Table 16, chromatogram is shown in Fig. 6 ~ 18.
The assay result of deoxyschizandrin in table 16 three batch sample
According to the assay result of above three batch samples, the content limit of pharmaceutical composition water-honeyed pill of the present invention is fixed tentatively and is: every 1g contains Fructus Schisandrae Sphenantherae with deoxyschizandrin (C
16h
18o
9) must not count and be less than 0.90mg (being about equivalent to 70% of 080220 batch sample content).
The clinical experiment of experimental example 4 pharmaceutical composition water-honeyed pill of the present invention (prepared by embodiment 1) treatment chronic type B viral hepatitis
1. diagnostic criteria
1.1 Western medicine diagnose standards:
With reference to Chinese Medical Association's hepatopathy credit meeting in 2005, infect disease credit and can formulate " the guideline " diagnostic criteria about chronic hepatitis B.
1.2 tcm diagnosis standards: reference virus hepatitis Standards of Chinese Medical Syndrome Differentiation (trying)
1.2.1 TCM syndrome: cloudy blood two loses, syndrome of remained toxicity person
1.2.2 CM syndrome differentiation criterion
(1) pharyngoxerosis, (2) dysphoria with feverish sensation in the chest palms and soles, (3) two dryness and uneasy feeling of the eye, (4) weak, (5) poor appetite, (6) yellowish urine, (7) dimly red tongue, greasy and thin fur, (8) thready,rolling and rapid pulse.
2. inclusive criteria
In 2.1 age 18-65 years, men and women does not limit.
2.2 clinical diagnosis is chronic viral hepatitis B.
2.3 Chinese medical discriminations meet cloudy blood two and lose, syndrome of remained toxicity.
2.4ALT > upper limits of normal more than 2 times, BIL < upper limits of normal 2 times.
3. exclusion standard
Other clinical drug research is accepted in 3.1 or 30 days.
3.2 patients that can not take medicine on request or be checked.
3.3 turn out to be the hepatitis (as drug intoxication, Alcoholic, autoimmunity and other non-hepatitis B etc.) of other type through inspection.
The whole latter stage of 3.4 serious anaphylaxiss and chronic disease or disease.
3.5 gestation and the women of age of sucking.
4. experimental technique and grouping
4.1 methods: adopt random, open, positive drug controlled trial method.
4.2 case source and groupings: this group case is PLA 302 hospital case, wherein treatment group 60 example, matched group 30 example.Male 56 example in treatment group, women 4 example, 30.6 ± 9.3 years old mean age (17-57 year), matched group male 28 example, female 2 example, 32.8 ± 12.5 years old mean age (19-59 year).Treat first two groups and there is comparability.
5. control drug is selected
Select clinical commonly using to fall enzymology medicine " bifendate drop pill " (the accurate word H11020980 of traditional Chinese medicines, Beijing XieHe medicine Factory produces), this medicine records in China's coastal port two.
6. test medication:
6.1 observe medicine: pharmaceutical composition water-honeyed pill of the present invention
Source: hospital of PLA 302 provides
Specification: 7.2g/ bag, 60 balls/bag
Lot number: 070919
Preservation condition: room temperature, lucifuge, is placed in sombre place.
6.2 control drug: bifendate drop pill (positive drug)
Source: Beijing XieHe medicine Factory
Specification: 1.5mg/ ball
Lot number: 07100104
Usage and dosage: oral a: 7.5mg (5 ball), 3 times on the one.3 months courses for the treatment of.
Storage: sealing, preserves in drying place.
7. Random Design: adopt random digits table.
8. administrated method
Dosage: treatment group: pharmaceutical composition water-honeyed pill of the present invention 1 bag/time, 2 times/day
Matched group: bifendate drop pill 5 ball/time, 3 times/day
The course for the treatment of: 3 months
Route of administration: oral.
Medication 4 weeks ALT still ascensionist can stop to change treatment.
9. observation item and index
9.1 safety indexes: respectively look into kidney merit (BUN, Cr), ECG and blood, a urine, feces routine before treatment and at the end for the treatment of.
The ultrasound diagnosis of liver spleen color Doppler is looked into before 9.2 treatments and at the end for the treatment of.
The lab index such as 9.3 liver functions (ALT, AST, TBIL, DBIL, A/G, PT/PA) before treatment and at the end of at least respectively look into once, but liver function over the course for the treatment of can according to state of an illness periodic review.
10. curative effect determinate standard
With reference to " guideline of clinical investigations of new Chinese medicine treatment viral hepatitis ", curative effect determinate standard, point basic healing is effectively, invalid.
● substantially cure: ALT recovers normal
● effectively: before ALT comparatively treats, decline more than 50%
● invalid: not reach These parameters person
11. efficacy analysis
Total effects relative analysis is in table 17
Total effects relative analysis situation between table 17 liang group
Through X
2analyze, two groups of total effectses compare, P=0.6607, two groups of no significant difference, show to fall in enzyme at treatment chronic type B viral hepatitis, pharmaceutical composition water-honeyed pill of the present invention and bifendate curative effect zero difference.Whole therapeutic process occurs there are no untoward reaction.
Following embodiment all can realize the effect described in above-mentioned experimental example.
The preparation of embodiment 1 water-honeyed pill
Fructus Schisandrae Sphenantherae 1667g Fructus Ligustri Lucidi 500g Fructus Forsythiae 500g Sonchus brachyotus DC. 333g
Get above-mentioned four taste crude drug, be ground into fine powder, cross 100 mesh sieves, mixing, every 100g crude drug powder adds 20g refined honey, adds appropriate water pill, dry, makes water-honeyed pill.Specification 7.2g/ bag (60 balls/bag), 120mg/ ball.
The preparation of embodiment 2 water-honeyed pill
Fructus Schisandrae Sphenantherae 1950g Fructus Ligustri Lucidi 350g Fructus Forsythiae 650g Sonchus brachyotus DC. 250g
Get above-mentioned four taste crude drug, be ground into fine powder, cross 100 mesh sieves, mixing, every 100g crude drug powder adds 25g refined honey, adds appropriate water pill, dry, makes water-honeyed pill.Specification 7.2g/ bag (60 balls/bag), 120mg/ ball.
The preparation of embodiment 3 water-honeyed pill
Fructus Schisandrae Sphenantherae 1100g Fructus Ligustri Lucidi 600g Fructus Forsythiae 350g Sonchus brachyotus DC. 400g
Get above-mentioned four taste crude drug, be ground into fine powder, cross 100 mesh sieves, mixing, every 100g crude drug powder adds 23g refined honey, adds appropriate water pill, dry, makes water-honeyed pill.Specification 7.2g/ bag (60 balls/bag), 120mg/ ball.
The preparation of embodiment 4 capsule
Fructus Schisandrae Sphenantherae 1667g Fructus Ligustri Lucidi 500g Fructus Forsythiae 500g Sonchus brachyotus DC. 333g
Get above-mentioned raw materials medicine, add customary adjuvant, conveniently technique decocting in water, precipitate with ethanol, oven dry pulverizing are made powder and are entered capsule.
The preparation of embodiment 5 tablet
Fructus Schisandrae Sphenantherae 1950g Fructus Ligustri Lucidi 350g Fructus Forsythiae 650g Sonchus brachyotus DC. 250g
Get above-mentioned raw materials medicine, add customary adjuvant, conveniently process alcohols water lift is fried into tablet.
The preparation of embodiment 6 big honeyed pills
Fructus Schisandrae Sphenantherae 1100g Fructus Ligustri Lucidi 600g Fructus Forsythiae 350g Sonchus brachyotus DC. 400g
Get above-mentioned four taste crude drug, dry, pulverizing, conveniently technique makes big honeyed pills.
The preparation of embodiment 7 big honeyed pills
Fructus Schisandrae Sphenantherae 1667g Fructus Ligustri Lucidi 500g Fructus Forsythiae 500g Sonchus brachyotus DC. 333g
Get Fructus Schisandrae Sphenantherae, Fructus Ligustri Lucidi, Fructus Forsythiae three taste alcohol decocts, Sonchus brachyotus DC. water decoction-alcohol sedimentation, makes big honeyed pills.
The preparation of embodiment 8 watered pill
Fructus Schisandrae Sphenantherae 1950g Fructus Ligustri Lucidi 350g Fructus Forsythiae 650g Sonchus brachyotus DC. 250g
Get above-mentioned four taste crude drug, dry, pulverizing, conveniently technique makes big honeyed pills.
The preparation of embodiment 9 watered pill
Fructus Schisandrae Sphenantherae 1100g Fructus Ligustri Lucidi 600g Fructus Forsythiae 350g Sonchus brachyotus DC. 400g
Get Fructus Schisandrae Sphenantherae, Fructus Ligustri Lucidi, Fructus Forsythiae three taste alcohol decocts, Sonchus brachyotus DC. water decoction-alcohol sedimentation, makes big honeyed pills.
Embodiment 10 quality determining method
A. the pharmaceutical composition water-honeyed pill 2g of the present invention that makes of Example 1, porphyrize, adds 30 ~ 60 DEG C of petroleum ether 20ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 1ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 15: 5: 1 is for developing solvent with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. the pharmaceutical composition water-honeyed pill 6g of the present invention that makes of Example 1, porphyrize, adds 30 ~ 60 DEG C of petroleum ether 20ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 20ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=10: be developing solvent at 2: 1: 0.5, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. the pharmaceutical composition water-honeyed pill of the present invention made of Example 1, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=64: be mobile phase at 35: 1, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: the pharmaceutical composition water-honeyed pill content of the present invention that the embodiment 1 of getting under content uniformity item is made, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 25ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 30 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
Embodiment 11 quality determining method
A. the pharmaceutical composition water-honeyed pill 2g of the present invention that makes of Example 2, porphyrize, adds 35 DEG C of petroleum ether 28ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 35 DEG C of petroleum ether 0.6ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 35 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 11: 6: 0.9 is for developing solvent with 35 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. the pharmaceutical composition water-honeyed pill 6g of the present invention that makes of Example 2, porphyrize, adds 55 DEG C of petroleum ether 12ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 28ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=9: be developing solvent at 2.4: 0.9: 0.6, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. the pharmaceutical composition water-honeyed pill of the present invention made of Example 2, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=62: be mobile phase at 38: 0.9, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: the pharmaceutical composition water-honeyed pill of the present invention that Example 2 is made, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 22ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 34 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
Embodiment 12 quality determining method
A. the pharmaceutical composition water-honeyed pill 2g of the present invention that makes of Example 3, porphyrize, adds 55 DEG C of petroleum ether 12ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 55 DEG C of petroleum ether 1.8ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 55 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 18: 4: 1.1 is for developing solvent with 55 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. the pharmaceutical composition water-honeyed pill 6g of the present invention that makes of Example 3, porphyrize, adds 35 DEG C of petroleum ether 28ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 12ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=11: be developing solvent at 1.6: 1.1: 0.4, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. the pharmaceutical composition water-honeyed pill of the present invention made of Example 3, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=78: be mobile phase at 22: 1.1, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: the pharmaceutical composition water-honeyed pill of the present invention that Example 3 is made, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 28ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 26 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
Embodiment 13 discrimination method
A. the pharmaceutical composition water-honeyed pill 2g of the present invention that makes of Example 2, porphyrize, adds 30 ~ 60 DEG C of petroleum ether 20ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 1ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 15: 5: 1 is for developing solvent with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. the pharmaceutical composition water-honeyed pill 6g of the present invention that makes of Example 1, porphyrize, adds 30 ~ 60 DEG C of petroleum ether 20ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 20ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=10: be developing solvent at 2: 1: 0.5, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color.
Embodiment 14 content assaying method
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D), chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=64: be mobile phase at 35: 1, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: get the pharmaceutical composition water-honeyed pill content of the present invention under content uniformity item, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 25ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 30 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill of the present invention contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
Embodiment 15 discrimination method
The pharmaceutical composition water-honeyed pill 2g of the present invention that Example 3 is made, porphyrize, adds 30 ~ 60 DEG C of petroleum ether 20ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 1ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; Test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 15: 5: 1 is for developing solvent with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color.
Claims (6)
1. treat the pharmaceutical composition water-honeyed pill of chronic hepatitis, liver cirrhosis for one kind, it is characterized in that described water-honeyed pill is made by the following method: get crude drug Fructus Schisandrae Sphenantherae 1667g, Fructus Ligustri Lucidi 500g, Fructus Forsythiae 500g, Sonchus brachyotus DC. 333g, be ground into fine powder, cross 100 mesh sieves, mixing, every 100g crude drug powder adds 20g refined honey, adds appropriate water pill, drying, makes water-honeyed pill.
2. the detection method of pharmaceutical composition water-honeyed pill as claimed in claim 1, is characterized in that the method comprises following discriminating and/or one or more in assay:
A. get it filled compositions water-honeyed pill 2g, and porphyrize adds 30 ~ 60 DEG C of petroleum ether 10 ~ 30ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 0.5 ~ 2ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, be that the upper solution of 10 ~ 20:3 ~ 7:0.8 ~ 1.2 is for developing solvent with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get it filled compositions water-honeyed pill 6g, and porphyrize adds 30 ~ 60 DEG C of petroleum ether 10 ~ 30ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 10 ~ 30ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=8 ~ 12:1.5 ~ 2.5:0.8 ~ 1.2:0.3 ~ 0.7 for developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. to get it filled compositions water-honeyed pill, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
Assay in quality determining method is as follows:
According to Chinese Pharmacopoeia version in 2010 annex VI D high effective liquid chromatography for measuring, chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=60 ~ 80:20 ~ 40:0.8 ~ 1.2 are mobile phase, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: compositions water-honeyed pill of getting it filled, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 20 ~ 30ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 25 ~ 35 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should 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 following discriminating and/or one or more in assay:
A. get it filled compositions water-honeyed pill 2g, and porphyrize adds 30 ~ 60 DEG C of petroleum ether 20ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 30 ~ 60 DEG C of petroleum ether 1ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 30 ~ 60 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, the upper solution being 15:5:1 with 30 ~ 60 DEG C of petroleum ether-Ethyl formate-formic acid is for developing solvent, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get it filled compositions water-honeyed pill 6g, and porphyrize adds 30 ~ 60 DEG C of petroleum ether 20ml, close plug, supersound process 15 minutes, filters, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 20ml, close plug, supersound process 20 minutes, filters, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=10:2:1:0.5 for developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. to get it filled compositions water-honeyed pill, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
According to Chinese Pharmacopoeia version in 2010 annex VI D high effective liquid chromatography for measuring, chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=64:35:1 is mobile phase, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: get the pharmaceutical composition water-honeyed pill content under content uniformity item, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 25ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 30 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
4. the detection method of pharmaceutical composition water-honeyed pill as claimed in claim 2, is characterized in that the method comprises following discriminating and/or one or more in assay:
A. get it filled compositions water-honeyed pill 2g, and porphyrize adds 35 DEG C of petroleum ether 28ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 35 DEG C of petroleum ether 0.6ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 35 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, the upper solution being 11:6:0.9 with 35 DEG C of petroleum ether-Ethyl formate-formic acid is for developing solvent, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get it filled compositions water-honeyed pill 6g, and porphyrize adds 55 DEG C of petroleum ether 12ml, close plug, supersound process 15 minutes, filter, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 28ml, close plug, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=9:2.4:0.9:0.6 for developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. to get it filled compositions water-honeyed pill, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
According to Chinese Pharmacopoeia version in 2010 annex VI D high effective liquid chromatography for measuring, chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=62:38:0.9 is mobile phase, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: compositions water-honeyed pill of getting it filled, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 22ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 34 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
5. the detection method of pharmaceutical composition water-honeyed pill as claimed in claim 2, is characterized in that the method comprises following discriminating and/or one or more in assay:
A. get it filled compositions water-honeyed pill 2g, and porphyrize adds 55 DEG C of petroleum ether 12ml, supersound process 20 minutes, and filter, filtrate evaporate to dryness, residue adds 55 DEG C of petroleum ether 1.8ml and dissolves, as need testing solution; Separately get Fructus Schisandrae Sphenantherae control medicinal material 1g, be made in the same way of control medicinal material solution; Get deoxyschizandrin reference substance again, add 55 DEG C of petroleum ether and make the solution of every 1ml containing 2mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, drawing each 5 μ l of above-mentioned three kinds of solution puts respectively on same silica GF254 lamellae, the upper solution being 18:4:1.1 with 55 DEG C of petroleum ether-Ethyl formate-formic acid is for developing solvent, launch, take out, dry, inspect under putting 254nm ultra-violet lamp, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
B. get it filled compositions water-honeyed pill 6g, and porphyrize adds 35 DEG C of petroleum ether 28ml, close plug, supersound process 15 minutes, filter, discard petroleum ether liquid, residue volatilizes petroleum ether, adds methanol 12ml, close plug, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue adds methanol 5ml makes dissolving, as need testing solution; Separately get Fructus Forsythiae control medicinal material 1g, be made in the same way of control medicinal material solution; Get phillyrin reference substance again, add methanol and make the solution of every 1ml containing 0.25mg, product solution in contrast; According to the annex VI B thin layer chromatography test of Chinese Pharmacopoeia version in 2010, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with water saturation chloroform-methanol-acetic ether-methanoic acid=11:1.6:1.1:0.4 for developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color;
C. to get it filled compositions water-honeyed pill, put basis of microscopic observation: peel epidermal surface sees class polygon, anticlinal wall gage distortion, cell is containing light color thing; Endocarp fibre bundle levels is oblique or be vertically staggered;
According to Chinese Pharmacopoeia version in 2010 annex VI D high effective liquid chromatography for measuring, chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Acetonitrile-water-acetic acid=78:22:1.1 is mobile phase, flow velocity: 1.0ml/min; Determined wavelength 249nm, number of theoretical plate calculates should be not less than 4000 by deoxyschizandrin; The preparation of reference substance solution: precision takes deoxyschizandrin reference substance, puts in measuring bottle, adds methanol and makes the solution of every 1ml containing 40 μ g; The preparation of need testing solution: compositions water-honeyed pill of getting it filled, porphyrize, gets 1.0g, accurately weighed, put in tool plug conical flask, precision adds methanol 28ml, close plug, weighed weight, supersound process, power 120W, 40kHz, 26 minutes, take out, let cool, weighed weight again, supplies the weight of less loss, shakes up with methanol, filter, get subsequent filtrate, to obtain final product; Algoscopy: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product; The every 1g of pharmaceutical composition water-honeyed pill contains Fructus Schisandrae Sphenantherae with deoxyschizandrin C
24h
32o
6meter, should be less than 0.9mg.
6. the application of pharmaceutical composition water-honeyed pill as claimed in claim 1 in preparation treatment chronic hepatitis, liver cirrhosis medicine.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1759884A (en) * | 2005-09-08 | 2006-04-19 | 闫敬武 | Chinese traditional medicine prescription for treating hepatitis B and preparation technics |
| CN101417007A (en) * | 2008-12-11 | 2009-04-29 | 浙江省中药研究所有限公司 | Traditional Chinese medicine formulation for treating non-insulin dependent diabetes mellitus and preparation method and use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1759884A (en) * | 2005-09-08 | 2006-04-19 | 闫敬武 | Chinese traditional medicine prescription for treating hepatitis B and preparation technics |
| CN101417007A (en) * | 2008-12-11 | 2009-04-29 | 浙江省中药研究所有限公司 | Traditional Chinese medicine formulation for treating non-insulin dependent diabetes mellitus and preparation method and use thereof |
Non-Patent Citations (3)
| Title |
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| 刘峰群等,.高效液相色谱法同时测定肝得宁-Ⅱ号丸中五味子甲素和五味子乙素的含量.《中国中药杂志》.2000,第25卷(第3期),第157-158页,特别是第157页左栏第1段. * |
| 沈连生.败酱草 北败酱.《彩色图解中药饮片鉴别手册》.华夏出版社,2002,(第1版),第36页. * |
| 潘五九等,.肝得宁片中五味子、女贞子提取工艺研究.《中国中药杂志》.2004,第29卷(第8期),第743-745页. * |
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