CN1323700C - Traditional Chinese medicine composition for treating asthenic disease, prepn. method and quality-standard thereof - Google Patents
Traditional Chinese medicine composition for treating asthenic disease, prepn. method and quality-standard thereof Download PDFInfo
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Abstract
The present invention discloses a medicine composition for treating consumptive diseases, and simultaneously discloses a preparation method and a quality standard of the medicine composition. The medicine composition is mainly composed of dodder seed, epimedium herb, himalayan teasel root, songaria cynomorium herb, cibot rhizome, spine date seed, fleece flower root, prepared liauorice root, prepared liauorice root, dried tangerine peel, deerhorn glue, prepared rehmannia root, glue of tortoise plastron, cherokee rose fruit, prepared astragalua root, common yan rhizome and palmleaf raspberry fruit. The preparation method comprises: the dried tangerine peel, the common yan rhizome, the prepared liauorice root, the deerhorn glue and the glue powder of tortoise plastron are pulverized into be in a micro-powder state for standby. The rest is decocted, and the decocted liquid is merged, filtered, concentrated and refrigerated for standing; the supernatant fluid is filtered, and is concentrated and dried to be pulverized. The supernatant fluid is mixed with the micro-powder of the materials to be pulverized; auxiliary materials are added to the materials to be prepared into tablets or granules. The method for controlling the quality of the prepared medicine preparation of the medicine composition comprises identification and content detection. The identifying method comprises the thin layer chromatography test to the fleece flower root, the radix glycyrrhiza and the milk vetch; the content detection method comprises an HPLC method for detecting the contents of the epimedium herb. The present invention is mainly used for treating consumptive diseases, and specially aims at kidney-yang deficiency and kidney qi deficiency.
Description
Invention field
The present invention relates to the preparation method and the quality standard of a kind of pharmaceutical composition and said composition, particularly a kind of Chinese medicine composition that is used for the treatment of deficiency of five viscera relates to the preparation method and the quality standard of said composition simultaneously.
Background technology
Deficiency of five viscera claims asthenia again.Due to multiple reason, with the internal organs loss, the negative and positive of qi and blood deficiency is the general name of the multiple chronic weak syndrome of main pathogenesis.
The deficiency of five viscera pathogenic factor is a lot of, and common reason has the natural endowment weakness, and body constitution is not strong; Bother excessively, undermine the five internal organs; Eating and drinking without temperance, impairing the spleen and stomach; Serious disease prolonged illness, lose in the conditioning etc.Deficient treatment is a basic principle with the tonification, and " element is asked a three body parts and nine pulse-taking sites opinion piece of writing " said: " deficiency syndrome should be treated by tonifying method." " element asks YIN YANG classification of natural phenomena a big opinion piece of writing " point out: " weakened configuration person, warming with drugs of thick nature; The insufficiency of essence person, tonifying with drugs of thick flavor ", in addition, the spleen being the foundation of acquired constitution is the source of water paddy, QI and bloodization life; The kidney being the origin of congenital constitution, and the residence nephroyin and nephroyang is this yuan of life, so invigorating spleen and kidney is significant in " deficiency of five viscera " treatment.
The present invention focuses on the Yuanyang with Semen Cuscutae, Herba Cynomorii, Herba Epimedii the kidney invigorating, with the Kidney-Yin of Radix Polygoni Multiflori, Colla cornus cervi, Colla Plastri Testudinis, Radix Rehmanniae Preparata the kidney invigorating; With Rhizoma Cibotii, the strong waist knee joint of Radix Dipsaci; Fructus Rosae Laevigatae, the astringent or styptic treatment for spontaneous sweating vital essence of Fructus Rubi; Radix Glycyrrhizae Preparata, Radix Astragali Preparata, the Rhizoma Dioscoreae tonification temper day after tomorrow; Assistant is with the Semen Ziziphi Spinosae mind tranquilizing and the heart calming.Full side is the merit of strengthening bone and muscle, replenishing QI and blood, kidney invigorating and YANG supporting altogether, for deficiency of five viscera show as physical weakness, soreness of the waist and knees, have a dizzy spell, deficiency of the kidney cold sperm, libido lower, urinate night more, forgetful insomniac, with more than obtain prompt effect.
Summary of the invention
One object of the present invention is to disclose a kind of Chinese medicine composition; Another object of the present invention is to disclose a kind of Chinese medicine composition and this Chinese medicine composition preparation method for the treatment of deficiency of five viscera; The 3rd purpose of the present invention is to disclose the method for quality control of this Chinese medicine composition.
The present invention seeks to be achieved through the following technical solutions:
The crude drug composition and the proportioning of pharmaceutical composition of the present invention are as follows:
Semen Cuscutae 120-140 weight portion Herba Epimedii 100-110 weight portion Radix Dipsaci 100-110 weight portion
Herba Cynomorii 120-140 weight portion Rhizoma Cibotii 150-170 weight portion Semen Ziziphi Spinosae 100-110 weight portion
Radix Polygoni Multiflori 150-170 weight portion Radix Glycyrrhizae Preparata 50-60 weight portion Pericarpium Citri Reticulatae 50-60 weight portion
Colla cornus cervi 20-30 weight portion Radix Rehmanniae Preparata 150-170 weight portion Colla Plastri Testudinis 30-40 weight portion
Fructus Rosae Laevigatae 120-140 weight portion Radix Astragali Preparata 100-110 weight portion Rhizoma Dioscoreae 100-110 weight portion
Fructus Rubi 210-230 weight portion
The above-mentioned raw materials optimum ratio is:
Semen Cuscutae (stir-fry) 128 weight portion Herba Epimedii (steaming) 106 weight portion Radix Dipsacis (steaming) 106 weight portions
Herba Cynomorii (steaming) 128 weight portion Rhizoma Cibotii (steaming) 160 weight portion Semen Ziziphi Spinosaes (stir-fry) 106 weight portions
Radix Polygoni Multiflori Preparata 160 weight portion Radix Glycyrrhizae Preparatas 53 weight portion Pericarpium Citri Reticulataes (steaming) 53 weight portions
Colla cornus cervi (stir-fry) 23 weight portion Radix Rehmanniae Preparata 160 weight portion Colla Plastri Testudiniss (stir-fry) 34 weight portions
Fructus Rosae Laevigatae (steaming) 128 weight portion Radix Astragali Preparatas 106 weight portion Rhizoma Dioscoreaes (stir-fry) 106 weight portions
Fructus Rubi (steaming) 213 weight portions
Preparation method is: get broken 10～75 microns of Pericarpium Citri Reticulatae, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micropowder, and standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2-3 hour for the first time, add 10-14 times of water gaging; 1-2 hour for the second time, add 8-12 times of water gaging; Collecting decoction filters, and concentrates, and cold preservation was left standstill 12-24 hour, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders mix and grind, and add 10-20%PVK30 ethanol liquid and granulate 70-75 ℃ of drying; Add adjuvant, mixing, direct compression or make granule.
The method of quality control that this compositions is made medicament comprises discriminating and/or assay.
Discrimination method comprises a kind of and/or several in the following method:
A. get 10 in tablet of the present invention, remove coating, porphyrize, add methanol 40-60ml, supersound process 10-20 minute, filter, the filtrate evaporate to dryness, residue adds water 20-40ml makes dissolving, uses ethyl acetate 20-40ml at every turn, and jolting is extracted 2-4 time, merge ethyl acetate liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Radix Polygoni Multiflori control medicinal material 0.5g, adds ethanol 40-60ml reflux 1-1.5 hour, filters, and filtrate is concentrated into 3ml, in contrast medical material solution; Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, control medicinal material and reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of binding agent, to make into ribbon with the carboxymethylcellulose sodium solution, with 10-20: 2-4: 1-2 toluene-ethyl acetate-formic acid is placed and was treated that stratified upper solution was developing solvent in 20-40 minute, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with control medicinal material and the corresponding position of contrast mouthful chromatograph on, show the fluorescence streak of same color; Behind the ammonia cure, become redness under the streak daylight.
B. get the aqueous solution behind the A item ethyl acetate extraction, use water saturated n-butyl alcohol 20-40ml at every turn, extract 2-4 time, merge n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, add the 0.5-1g neutral alumina and mix thoroughly, drying is added in 100～120 orders, 2g is on the neutral alumina post of internal diameter 10mm, with 30-50% methanol 20-40ml eluting, collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds water 30-40ml in addition, and reflux 1-1.5 hour, filter, the n-butyl alcohol that filtrate water is saturated shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, ethyl acetate-formic acid-glacial acetic acid-water with 15-20: 1-2: 1-2: 2-4 is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 100 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, on the corresponding position of control medicinal material chromatograph, daylight shows identical yellow principal spot down; Under the ultra-violet lamp, show identical yellow fluorescence principal spot.
C. get 10 in tablet of the present invention, remove coating, porphyrize, add methanol 30-50ml, reflux 1-1.5 hour, filter, filtrate evaporate to dryness, residue add water 30-40ml makes dissolving, uses ethyl acetate 20-40ml at every turn, extract 2-4 time, discard ethyl acetate liquid, water layer extracts 2-4 time with water saturated n-butyl alcohol 20-40ml at every turn, merge n-butanol extracting liquid, use 1% sodium hydroxide solution 20-40ml at every turn, wash 2-4 time, discard alkali wash water, each water 20-30ml washs 2-4 time, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, adding the 0.5-1g neutral alumina mixes thoroughly, drying is added in 100～120 orders, 2g, on the neutral alumina post of internal diameter 10mm, with 30-50% methanol 40-60ml eluting, collect eluent, evaporate to dryness, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 10 μ l, reference substance solution 5 μ l, put respectively on the same silica gel g thin-layer plate of thick 0.5mm, with 10-15: 20-25: 10-15: 5-10 chloroform-ethyl acetate-methanol-water, at lower floor's solution of placing 10-20 hour below 10 ℃ is developing solvent, launches; Take out, dry, spray is with the 10-15% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105-110 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, daylight shows down identical sepia speckle, ultra-violet lamp shows identical orange-yellow fluorescence speckle down.
Content assaying method comprises following assay method:
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Water-acetonitrile-oxolane-glacial acetic acid with 750-760: 230-240: 10-15: 5-10 is a mobile phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the icariin peak should be not less than 3500;
The preparation of reference substance solution: take by weighing the icariin reference substance, add methanol and make the solution that 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get 10 in tablet of the present invention, remove coating, it is fixed to claim, get the single taking dose, it is fixed to claim, puts in the tool plug conical flask, adding concentration is 50% ethanol 50ml, and close plug claims to decide weight, supersound process 1-1.5 hour, put coldly, claim to decide weight again, with concentration is that 50% ethanol is supplied the weight that subtracts mistake, shakes up, and filters, draw subsequent filtrate 25ml, put in the evaporating dish evaporate to dryness, residue adds water 5ml makes dissolving, is added in 13～80 orders of anticipating, 5g, internal diameter 1.5cm, on the polyamide column of dry column-packing, water 50-60ml eluting, discard eluent, continue, collect eluent with ethanol 80-100ml eluting, evaporate to dryness, residue adds dissolve with methanol and is transferred in the 5ml measuring bottle, adds methanol to scale, shakes up, filter, promptly;
Algoscopy: draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly;
Drug combination preparation per unit preparation (being equivalent to conventional tablet) per unit preparation contains Herba Epimedii by icariin C
33H
40O
15Meter must not be less than 0.05mg.
The present invention is mainly used in the treatment spleen fatiguing disease, and especially at insufficiency of kidney-YANG and syndrome of deficiency of kidney-QI, following experimental example further specifies the present invention.
Experimental example 1: the thin layer chromatography of Radix Polygoni Multiflori is differentiated comparative experiments
Get 10 in tablet of the present invention, remove coating, porphyrize adds methanol 50ml, supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 30ml makes dissolving, adds the ethyl acetate jolting and extracts 2 times, each 30ml, merge ethyl acetate liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Radix Polygoni Multiflori control medicinal material 0.5g, adds ethanol 50ml reflux 1 hour, filters, and filtrate is concentrated into 3ml, in contrast medical material solution; Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, control medicinal material and reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of binding agent, to make into ribbon with the carboxymethylcellulose sodium solution, with 15: 2: 1 toluene-ethyl acetate-formic acid, place and treated that stratified upper solution was developing solvent in about 30 minutes, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with control medicinal material and the corresponding position of contrast mouthful chromatograph on, show the fluorescence streak of same color; Behind the ammonia cure, become redness under the streak daylight;
Other gets need testing solution, with reference to " " Radix Polygoni Multiflori " thin layer chromatography of Chinese pharmacopoeia version in 2000 discrimination method launches as the developing solvent secondary with benzene-ethanol, sprays with the phosphomolybdic acid test solution again, with stilbene glucoside category feature composition in the discriminating Radix Polygoni Multiflori, but negative control has interference.
Get tablet of the present invention, with reference to " the thin layer chromatography discrimination method under " Radix Polygoni Multiflori " item of Chinese pharmacopoeia version in 2000 gets the Radix Polygoni Multiflori thin layer and differentiates collection of illustrative plates.From thin layer chromatography as seen, in the test sample chromatograph, with Radix Polygoni Multiflori control medicinal material chromatograph relevant position on, streak is difficult to be observed, this may prepare relevantly with need testing solution, and the need testing solution viscosity is big, influence launches effect.
Experimental example 2: the thin layer chromatography of Radix Glycyrrhizae is differentiated comparative experiments
Get the aqueous solution after Radix Polygoni Multiflori thin layer in the experimental example 1 is differentiated an ethyl acetate extraction, with water saturated n-butanol extraction 2 times, 30ml at every turn, merge n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, add an amount of neutral alumina and mix thoroughly, drying is added in 100～120 orders, 2g is on the internal diameter 10mm neutral alumina post, with 40% methanol 30ml eluting, collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds water 30ml in addition, and reflux 1 hour filters, and the n-butyl alcohol that filtrate water is saturated shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 15: 1: 1: 2 ethyl acetates-formic acid-glacial acetic acid-water was developing solvent, launched, and took out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 100 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, on the corresponding position of control medicinal material chromatograph, daylight shows identical yellow principal spot down; Under the ultra-violet lamp, show identical yellow fluorescence principal spot;
Radix Glycyrrhizae in the prescription is adopted the microscopical identification method, because among this product preparation technology, Radix Glycyrrhizae powder is broken into impalpable powder (10～75 μ m), the microscopic features difficulty is examined and seen, so adopt the thin layer discrimination method.
In addition, get 10 in this pharmaceutical composition tablet, with reference to " the thin layer chromatography discrimination method under Chinese pharmacopoeia version " Radix Glycyrrhizae " in 2000 item, with the ammonium glycyrrhizinate reference substance in contrast, experimental result test sample color vinegar with reference substance chromatograph relevant position on, speckle is unintelligible, inferior separating effect.
Experimental example 3: the thin layer chromatography of the Radix Astragali is differentiated comparative experiments
Get 10 in this pharmaceutical composition tablet, remove coating, porphyrize, add methanol 40ml, reflux 1 hour filters, filtrate evaporate to dryness, residue add water 30ml makes dissolving, uses ethyl acetate extraction 2 times, each 30ml discards ethyl acetate liquid, and water layer is with water saturated n-butanol extraction 2 times, each 30ml merges n-butanol extracting liquid, with 1% sodium hydroxide solution washing 2 times, each 30ml discards alkali wash water, washes with water 2 times, each 25ml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, adds an amount of neutral alumina and mixes drying thoroughly, be added in 100～120 orders, 2g is on the internal diameter 10mm neutral alumina post, with 40% methanol 50ml eluting, collect eluent, evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 10 μ l, reference substance solution 5 μ l put respectively on the same silica gel g thin-layer plate of thick 0.5mm, with 10: 20: 11: lower floor's solution that 5 chloroforms-ethyl acetate-methanol-water was placed 12 hours below 10 ℃ was developing solvent, launched; Take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, composing on the corresponding position with the reference substance bag, daylight shows identical sepia speckle down, and ultra-violet lamp shows identical orange-yellow fluorescence speckle down.
In addition, with reference to " the thin layer chromatography discrimination method under Chinese pharmacopoeia version " Radix Astragali " in 2000 item, with chloroform-methanol-water (13: 7: 2) as developing solvent, speckle can suitably separate than former method as a result, but speckle comparatively spreads relatively, particularly the hands bed board because thin layer collection of illustrative plates speckle is more, influences result of the test and observes.
Experimental example 4: the assay condition is selected test
This product prescription is made up of 16 flavor Chinese medicines, and wherein Herba Epimedii is a principal agent, and icariin is its main effective ingredient, so be elected to be the assay index.
The selection of extraction solvent amount: in the official method, the 0.2g crude drug adds 50% ethanol 20ml and extracts.Press recipe quantity and calculate, the every capsules of this product contains Herba Epimedii and is equivalent to crude drug 0.212g, except that the floating sheep leaves of pulse plants, also contains other Chinese crude drug in considering to write out a prescription, and may influence the extraction of icariin, is that 50ml is as extracting solvent so select 50% consumption of ethanol.
The selection of extracting method: pharmacopeia adopts 1 hour method of supersound process to extract.For investigating extracting method, get 2 parts of each 1g of same test sample, it is fixed to claim, respectively adds 50% ethanol 50ml, adopts supersound process respectively 1 hour and after reflux extracted in 1 hour, measures content Determination of Icariin by the assay condition of drafting, and the results are shown in Table 1.
Table 1:
| Extracting method | Supersound process 1 hour | Reflux 1 hour |
| Content (mg/ grain) | 0.0875 | 0.0869 |
By The above results as can be known, the result of two kinds of methods is basic identical, selects 1 hour extracting method as assay of supersound process for use.
Need testing solution is removed the method for interference component: get the extracting solution after the supersound process, filter, get subsequent filtrate 25ml, water bath method, residue add dissolve with methanol and are settled to 5ml, improve the need testing solution content Determination of Icariin, but other interference component concentration improves relatively, after the filtration, directly get filtrate and inject chromatograph of liquid, influence separating effect.Because this product content Determination of Icariin is lower, Herba Epimedii contains flavonoid chemical constituents such as icariin, select for use to flavones ingredient absorption, separating effect preferably polyamide reduce the interference of need testing solution impurity as purifying pillar.
The selection of eluant: get 3 parts of each 1g of same test sample, it is fixed to claim,, measures by the content assaying method of drafting as eluant with 50% ethanol, 70% ethanol, each 100ml of 100% ethanol, the results are shown in Table 2.
Table 2:
| Extract solvent | 50% ethanol | 70% ethanol | 100% ethanol |
| Content (mg/ grain) | 0.0769 | 0.0826 | 0.0860 |
The above results shows that ethanol is good than other solvent to the eluting power of icariin, and as eluant.
The consumption of eluant: get 2 parts of each 1g of same test sample, it is fixed to claim, by drafting the content assaying method operation, add ethanol 60ml, 80ml respectively and carry out eluting, and the 81～100ml eluent behind the collection 80ml ethanol elution, measure content Determination of Icariin behind the evaporate to dryness, result of the test such as table 3:
Table 3:
| Eluant | 60ml | 80ml | 81～100ml |
| Content (mg/ grain) | 0.0876 | 0.0811 | 0 |
The above results shows, and is that the icariin eluting is complete with 60ml ethanol, considers that every batch sample content Determination of Icariin has difference, for guaranteeing the eluting of tested composition, and the usefulness of the determining eluant 80ml that measures one's own ability.
Measure the selection of wavelength: get excessive sheep flower bud glycosides reference substance solution, in the interscan of 200～320nm wave-length coverage, experimental result must have absorption maximum at 270nm wavelength place.
The selection of mobile phase: the mobile phase that pharmacopeia is recorded under " Herba Epimedii " assay item is acetonitrile-water (30: 70), through test, test sample is because the component content complexity, tested component fails effectively to separate with impurity, find that with the different proportionings of water acetonitrile-water (23: 77) is all better to the separation of icariin than other proportioning by changing acetonitrile, but still it is undesirable, selected for use afterwards flavone compound separation selectivity oxolane preferably, and add a certain amount of glacial acetic acid, make faintly acid compound separation effect better, through adjusting different proportion, test of many times, determined water-acetonitrile-oxolane-glacial acetic acid at last (755: 230: 10: 5) be best proportioning, behind the tested peak and do not have Interference Peaks substantially, can reach baseline separation.The methanol-water with different proportion, methanol-glacial acetic acid-water, methanol-0.2mol/L NaH have also been attempted in this test
2PO
4Make mobile phase, but separating effect is all not good enough, therefore selected water-acetonitrile-oxolane-glacial acetic acid for use (755: 230: 10: 5) be the optimal flow phase of " Herba Epimedii " assay.
Experimental example 5: sample size determination test
Get it filled three batches in compositions tablet is measured by content assaying method in the technical scheme, with content Determination of Icariin in the external standard method calculation sample.The results are shown in Table 4.
Table 4:
| Lot number | 1 | 2 | On average (mg/ sheet) |
| 020301 020302 020303 | 0.0720 0.0843 0.0884 | 0.0751 0.0888 0.0915 | 0.074 0.086 0.090 |
Because the place of production and the difference in season, the quantity of the contained blade of crude drug are difficult to control, carry out an acceptance inspection by standard test, crude drug content acceptable ranges is bigger, so according to above experimental result, be to guarantee product quality, just decide this product and contain icariin and must not be less than the 0.05mg/ unit formulation.
Experimental example 6: assay test
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With 755: 230: 10: 5 water-acetonitrile-oxolane-glacial acetic acid was a mobile phase; The detection wavelength is 270nm; Theory is pulled number should be not less than 3500 by the calculating of icariin peak;
The preparation of reference substance solution: take by weighing the icariin reference substance, add methanol and make the solution that 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get 10 in tablet of the present invention, remove coating, it is fixed to claim, get about 2 amount, it is fixed to claim, puts in the tool plug conical flask, adding concentration is 50% ethanol 50ml, and close plug claims to decide weight, supersound process 1 hour is put coldly, claims to decide weight again, with concentration is that 50% ethanol is supplied the weight that subtracts mistake, shakes up, and filters, draw subsequent filtrate 25ml, put in the evaporating dish evaporate to dryness, residue adds water 5ml makes dissolving, is added in 13～80 orders of anticipating, 5g, internal diameter 1.5cm, on the polyamide column of dry column-packing, water 50ml eluting, discard eluent, continue, collect eluent with ethanol 80ml eluting, evaporate to dryness, residue adds dissolve with methanol and is transferred in the 5ml measuring bottle, adds methanol to scale, shakes up, filter, promptly;
Algoscopy: draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly;
Every in tablet of the present invention contains Herba Epimedii by icariin C
33H
40O
15Meter must not be less than 0.05mg;
Measure and the test of medical material assay through blank assay, precision test, replica test, stability test, application of sample recovery test, sample size, prove that above-mentioned assay test method can improve the control drug quality.
Experimental example 7: clinical drug trial
With reference to " syndrome of deficiency of kidney-YANG " standard of carrying the whole nation the 3rd traditional Chinese medical science combination of Chinese and Western medicine deficiency syndrome meeting formulation of record in national high traditional Chinese medical science universities and colleges' teaching materials " Chinese Internal Medicine " spleen fatiguing diseases " syndrome of deficiency of kidney-YANG " and the Ministry of Public Health " the clinical research guideline of new Chinese medicine treatment syndrome of deficiency of kidney-YANG ".30 routine patients, male 14 examples, women 16 examples, the oldest 68 years old, minimum 40 years old.Primary symptom: soreness of waist and knee joint, aversion to cold and cold limbs, hyposexuality, body of the tongue is fat big, and tongue is white, deep pulse.Inferior disease: lassitude, nocturia, big loose stool.Take tablet of the present invention, each 4,2 times on the one, 4 weeks were 1 course of treatment.
Table 4: mild symptoms weight classification
| Symptom | Gently (+) | In (++) | Heavy (+++) |
| Soreness of waist and knee joint sexual disorder lassitude nocturia | Idol has the outbreak libido to reduce the lassitude nocturia 1-2 time | Even sexual lethargy nocturia 3-4 time that require of outbreak repeatedly | Continue outbreak, be difficult for alleviating sexual impotence or infertility lethargy, bradykinesia nocturia frequency, number of times is a lot |
Curative effect judging standard
1, clinical recovery: deficiency of kidney-QI or insufficiency of kidney-YANG symptom disappear.
2, produce effects: deficiency of kidney-QI or insufficiency of kidney-YANG symptom obviously improve more than 2 grades (by +++become+): or indivedual symptom changes at 1 grade, and other symptoms all disappear.
3, effective: deficiency of kidney-QI or deficiency of kidney-YANG syndrome improve more than 1 grade, or indivedual symptom be significantly improved (by ++ become+).
4, invalid: deficiency of kidney-QI or insufficiency of kidney-YANG symptom do not have improvement.
Result of the test:
1, total effects: clinical recovery 8 examples (accounting for 26.67%), produce effects 14 examples (accounting for 46.67%), effective 7 examples (accounting for 23.33%), invalid 1 example (accounting for 3.33%).
2, symptom curative effect
Soreness of waist and knee joint effective percentage 96.67% (29/30)
Sexual disorder effective percentage 50% (6/12)
Lassitude effective percentage 71.43% (20/28)
Nocturia effective percentage 72% (18/25)
This drug combination preparation treatment deficiency of five viscera syndrome of deficiency of kidney-QI and syndrome of deficiency of kidney-YANG, effective percentage is remarkable, and determined curative effect is safe in utilization.
Specific embodiments of the invention are as follows
Embodiment 1 preparation tablet
Semen Cuscutae (stir-fry) 128g Herba Epimedii (steaming) 106g Radix Dipsaci (steaming) 106g
Herba Cynomorii (steaming) 128g Rhizoma Cibotii (steaming) 160g Semen Ziziphi Spinosae (stir-fry) 106g
Radix Polygoni Multiflori Preparata 160g Radix Glycyrrhizae Preparata 53g Pericarpium Citri Reticulatae (steaming) 53g
Colla cornus cervi (stir-fry) 23g Radix Rehmanniae Preparata 160g Colla Plastri Testudinis (stir-fry) 34g
Fructus Rosae Laevigatae (steaming) 128g Radix Astragali Preparata 106g Rhizoma Dioscoreae (stir-fry) 106g
Fructus Rubi (steaming) 213g
Get 75 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2 hours for the first time, add 12 times of water gagings; 1 hour for the second time, add 10 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 16 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders mix and grind, and add 10%PVK30 ethanol liquid and granulate 75 ℃ of dryings, granulate; Dry granular adds carboxymethyl starch sodium 14g again, and is above-mentioned by the medicated powder mixing, adds 50% alcohol granulation, 75 ℃ of dryings, and granulate adds carboxymethyl starch sodium 13g, silica 1 0g and magnesium stearate 5g, and mixing is pressed special-shaped tablets, wraps pink film-coat, promptly.
Embodiment 2 preparation tablets
Semen Cuscutae (stir-fry) 120g Herba Epimedii (steaming) 100g Radix Dipsaci (steaming) 100g
Herba Cynomorii (steaming) 120g Rhizoma Cibotii (steaming) 150g Semen Ziziphi Spinosae (stir-fry) 100g
Radix Polygoni Multiflori Preparata 150g Radix Glycyrrhizae Preparata 50g Pericarpium Citri Reticulatae (steaming) 50g
Colla cornus cervi (stir-fry) 20g Radix Rehmanniae Preparata 150g Colla Plastri Testudinis (stir-fry) 30g
Fructus Rosae Laevigatae (steaming) 120g Radix Astragali Preparata 100g Rhizoma Dioscoreae (stir-fry) 100g
Fructus Rubi (steaming) 210g
Get 20 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2 hours for the first time, add 11 times of water gagings; 1 hour for the second time, add 8 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 15 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders mix and grind, and add 10%PVK30 ethanol liquid and granulate 70 ℃ of dryings, granulate; Dry granular adds carboxymethyl starch sodium 10g again, and is above-mentioned by the medicated powder mixing, adds 40% alcohol granulation, 70 ℃ of dryings, and granulate adds carboxymethyl starch sodium 10g, silicon dioxide 8g and magnesium stearate 4g, and mixing, tabletting are promptly.
Embodiment 3 preparation tablets
Semen Cuscutae (stir-fry) 140g Herba Epimedii (steaming) 110g Radix Dipsaci (steaming) 110g
Herba Cynomorii (steaming) 140g Rhizoma Cibotii (steaming) 170g Semen Ziziphi Spinosae (stir-fry) 110g
Radix Polygoni Multiflori Preparata 170g Radix Glycyrrhizae Preparata 60g Pericarpium Citri Reticulatae (steaming) 60g
Colla cornus cervi (stir-fry) 30g Radix Rehmanniae Preparata 170g Colla Plastri Testudinis (stir-fry) 40g
Fructus Rosae Laevigatae (steaming) 140g Radix Astragali Preparata 110g Rhizoma Dioscoreae (stir-fry) 110g
Fructus Rubi (steaming) 230g
Get 40 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2.5 hours for the first time, add 12 times of water gagings; 1.5 hours for the second time, add 10 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 20 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders mix and grind, and add 15%PVK30 ethanol liquid and granulate 75 ℃ of dryings, granulate; Dry granular adds carboxymethyl starch sodium 12g again, and is above-mentioned by the medicated powder mixing, adds 50% alcohol granulation, 75 ℃ of dryings, and granulate adds carboxymethyl starch sodium 12g, silica 1 0g and magnesium stearate 6g, mixing, direct compression.
Embodiment 4 preparation tablets
Semen Cuscutae (stir-fry) 120g Herba Epimedii (steaming) 110g Radix Dipsaci (steaming) 100g
Herba Cynomorii (steaming) 140g Rhizoma Cibotii (steaming) 150g Semen Ziziphi Spinosae (stir-fry) 110g
Radix Polygoni Multiflori Preparata 150g Radix Glycyrrhizae Preparata 60g Pericarpium Citri Reticulatae (steaming) 50g
Colla cornus cervi (stir-fry) 30g Radix Rehmanniae Preparata 150g Colla Plastri Testudinis (stir-fry) 40g
Fructus Rosae Laevigatae (steaming) 120g Radix Astragali Preparata 110g Rhizoma Dioscoreae (stir-fry) 100g
Fructus Rubi (steaming) 230g
Get 60 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 3 hours for the first time, add 13 times of water gagings; 2 hours for the second time, add 12 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 4 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders mix and grind, and add 20%PVK30 ethanol liquid, 75 ℃ of dryings, granulate; Dry granular adds carboxymethyl starch sodium 15g again, and is above-mentioned by the medicated powder mixing, adds 60% alcohol granulation, 75 ℃ of dryings, and granulate adds carboxymethyl starch sodium 15g, silica 1 2g and magnesium stearate 8g, mixing, direct compression.
Embodiment 5 preparation granules
Semen Cuscutae (stir-fry) 128g Herba Epimedii (steaming) 106g Radix Dipsaci (steaming) 106g
Herba Cynomorii (steaming) 128g Rhizoma Cibotii (steaming) 160g Semen Ziziphi Spinosae (stir-fry) 106g
Radix Polygoni Multiflori Preparata 160g Radix Glycyrrhizae Preparata 53g Pericarpium Citri Reticulatae (steaming) 53g
Colla cornus cervi (stir-fry) 23g Radix Rehmanniae Preparata 160g plate glue (stir-fry) 34g
Fructus Rosae Laevigatae (steaming) 128g Radix Astragali Preparata 106g Rhizoma Dioscoreae (stir-fry) 106g
Fructus Rubi (steaming) 213g
Get 75 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2 hours for the first time, add 12 times of water gagings; 1 hour for the second time, add 10 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 16 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders add cane sugar powder 500g again, and dextrin 72g mixes and grinds, and add 10%PVK30 ethanol liquid and granulate, 75 ℃ of dryings, granulate is promptly.
Embodiment 6 preparation granules
Semen Cuscutae (stir-fry) 140g Herba Epimedii (steaming) 100g Radix Dipsaci (steaming) 110g
Herba Cynomorii (steaming) 120g Rhizoma Cibotii (steaming) 170g Semen Ziziphi Spinosae (stir-fry) 100g
Radix Polygoni Multiflori Preparata 170g Radix Glycyrrhizae Preparata 50g Pericarpium Citri Reticulatae (steaming) 60g
Colla cornus cervi (stir-fry) 20g Radix Rehmanniae Preparata 170g Colla Plastri Testudinis (stir-fry) 30g
Fructus Rosae Laevigatae (steaming) 140g Radix Astragali Preparata 100g Rhizoma Dioscoreae (stir-fry) 110g
Fructus Rubi (steaming) 210g
Get 20 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2 hours for the first time, add 11 times of water gagings; 1 hour for the second time, add 8 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 15 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders add cane sugar powder 500g again, and dextrin 38g mixes and grinds, and add 10%PVK30 ethanol liquid and granulate, 70 ℃ of dryings, granulate is promptly.
Embodiment 7 preparation granules
Semen Cuscutae (stir-fry) 130g Herba Epimedii (steaming) 105g Radix Dipsaci (steaming) 105g
Herba Cynomorii (steaming) 130g Rhizoma Cibotii (steaming) 160g Semen Ziziphi Spinosae (stir-fry) 105g
Radix Polygoni Multiflori Preparata 160g Radix Glycyrrhizae Preparata 55g Pericarpium Citri Reticulatae (steaming) 55g
Colla cornus cervi (stir-fry) 25g Radix Rehmanniae Preparata 160g Colla Plastri Testudinis (stir-fry) 35g
Fructus Rosae Laevigatae (steaming) 130g Radix Astragali Preparata 105g Rhizoma Dioscoreae (stir-fry) 105g
Fructus Rubi (steaming) 220g
Get 40 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 2.5 hours for the first time, add 12 times of water gagings; 1.5 hours for the second time, add 10 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 20 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders add cane sugar powder 500g again, and dextrin 35g mixes and grinds, and add 15%PVK30 ethanol liquid and granulate, 75 ℃ of dryings, granulate is promptly.
Embodiment 8 preparation granules
Semen Cuscutae (stir-fry) 140g Herba Epimedii (steaming) 110g Radix Dipsaci (steaming) 110g
Herba Cynomorii (steaming) 140g Rhizoma Cibotii (steaming) 170g Semen Ziziphi Spinosae (stir-fry) 110g
Radix Polygoni Multiflori Preparata 170g Radix Glycyrrhizae Preparata 60g Pericarpium Citri Reticulatae (steaming) 60g
Colla cornus cervi (stir-fry) 30g Radix Rehmanniae Preparata 170g Colla Plastri Testudinis (stir-fry) 40g
Fructus Rosae Laevigatae (steaming) 140g Radix Astragali Preparata 110g Rhizoma Dioscoreae (stir-fry) 110g
Fructus Rubi (steaming) 230g
Get 60 microns of Pericarpium Citri Reticulataes, Rhizoma Dioscoreae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Carapax et Plastrum Testudinis rubber powder micronizing, standby; All the other Semen Cuscutae etc. ten decoct with water secondary simply in the prescription, 3 hours for the first time, add 13 times of water gagings; 2 hours for the second time, add 12 times of water gagings; Collecting decoction filters, and concentrates, and cold preservation was left standstill 4 hours, gets supernatant and filters, and is concentrated into the thick paste shape, and 75 ℃ of dryings get dry extract, and are ground into fine powder, and are standby; Above-mentioned dried cream fine powder and Pericarpium Citri Reticulatae, Radix Glycyrrhizae Preparata, Colla cornus cervi, Colla Plastri Testudinis four flavor fine powders add cane sugar powder 470g again, and dextrin 29g mixes and grinds, and adds 20%PVK30 ethanol liquid, 75 ℃ of dryings, and granulate is promptly.
The test of embodiment 9 assays
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; With 755: 230: 10: 5 water-acetonitrile-oxolane-glacial acetic acid was a mobile phase; The detection wavelength is 270nm; Theory is pulled number should be not less than 3500 by the calculating of icariin peak;
The preparation of reference substance solution: take by weighing the icariin reference substance, add methanol and make the solution that 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get 10 in tablet of the present invention, remove coating, the accurate title, decide, get about 2 amount, it is fixed to claim, puts in the tool plug conical flask, adding concentration is 50% ethanol 50ml, and close plug claims to decide weight, supersound process 1 hour is put coldly, claims to decide weight again, with concentration is that 50% ethanol is supplied the weight that subtracts mistake, shakes up, and filters, draw subsequent filtrate 25ml, put in the evaporating dish evaporate to dryness, residue adds water 5ml makes dissolving, is added in 13～80 orders of anticipating, 5g, internal diameter 1.5cm, on the polyamide column of dry column-packing, water 50ml eluting, discard eluent, continue, collect eluent with ethanol 80ml eluting, evaporate to dryness, residue adds dissolve with methanol and is transferred in the 5ml measuring bottle, adds methanol to scale, shakes up, filter, promptly;
Algoscopy: draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly;
Every in tablet of the present invention contains Herba Epimedii by icariin C
33H
40O
15Meter must not be less than 0.05mg;
Embodiment 10 preparation tablets
Semen Cuscutae (stir-fry) 140g Herba Epimedii (steaming) 110g Radix Dipsaci (steaming) 110g
Herba Cynomorii (steaming) 140g Rhizoma Cibotii (steaming) 170g Semen Ziziphi Spinosae (stir-fry) 110g
Radix Polygoni Multiflori Preparata 170g Radix Glycyrrhizae Preparata 60g Pericarpium Citri Reticulatae (steaming) 60g
Colla cornus cervi (stir-fry) 30g Radix Rehmanniae Preparata 170g Colla Plastri Testudinis (stir-fry) 40g
Fructus Rosae Laevigatae (steaming) 140g Radix Astragali Preparata 110g Rhizoma Dioscoreae (stir-fry) 110g
Fructus Rubi (steaming) 230g
Get the above-mentioned raw materials medicine and make tablet, every heavy 0.5g, the deficiency of five viscera patient is oral, each 2-4 sheet, every day 2 times.
Embodiment 11 preparation granules
Semen Cuscutae (stir-fry) 120g Herba Epimedii (steaming) 100g Radix Dipsaci (steaming) 100g
Herba Cynomorii (steaming) 120g Rhizoma Cibotii (steaming) 150g Semen Ziziphi Spinosae (stir-fry) 100g
Radix Polygoni Multiflori Preparata 150g Radix Glycyrrhizae Preparata 50g Pericarpium Citri Reticulatae (steaming) 50g
Colla cornus cervi (stir-fry) 20g Radix Rehmanniae Preparata 150g Colla Plastri Testudinis (stir-fry) 30g
Fructus Rosae Laevigatae (steaming) 120g Radix Astragali Preparata 100g Rhizoma Dioscoreae (stir-fry) 100g
Fructus Rubi (steaming) 210g
Get the above-mentioned raw materials medicine and make granule, every bag heavy 2g, the deficiency of five viscera patient is oral, each 1-2 bag, every day 2 times.
Claims (5)
1, a kind of method of quality control of Chinese medicine composition is characterized in that the assay in this method is:
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Water-acetonitrile-oxolane-glacial acetic acid with 750-760: 230-240: 10-15: 5-10 is a mobile phase; The detection wavelength is 270nm; Theory is pulled number should be not less than 3500 by the calculating of icariin peak; The preparation of reference substance solution: take by weighing the icariin reference substance, add methanol and make the solution that 1ml contains 20 μ g, promptly; The preparation of need testing solution: get 10 in this pharmaceutical composition tablet, remove coating, it is fixed to claim, get the single taking dose, it is fixed to claim, puts in the tool plug conical flask, adding concentration is 50% ethanol 50ml, and close plug claims to decide weight, supersound process 1-1.5 hour, put coldly, claim to decide weight again, with concentration is that 50% ethanol is supplied the weight that subtracts mistake, shakes up, and filters, draw subsequent filtrate 25ml, put in the evaporating dish evaporate to dryness, residue adds water 5ml makes dissolving, is added in 13～80 orders of anticipating, 5g, internal diameter 1.5cm, on the polyamide column of dry column-packing, water 50-60ml eluting, discard eluent, continue, collect eluent with ethanol 80-100ml eluting, evaporate to dryness, residue adds dissolve with methanol and is transferred in the 5ml measuring bottle, adds methanol to scale, shakes up, filter, promptly; Algoscopy: draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly; This drug combination preparation per unit preparation contains Herba Epimedii by icariin C
33H
40O
15Meter must not be less than 0.05mg;
Wherein the crude drug of this Chinese medicine composition consists of:
Semen Cuscutae 120-140 weight portion Herba Epimedii 100-110 weight portion
Radix Dipsaci 100-110 weight portion Herba Cynomorii 120-140 weight portion
Rhizoma Cibotii 50-170 weight portion Semen Ziziphi Spinosae 100-110 weight portion
Radix Polygoni Multiflori 150-170 weight portion Radix Glycyrrhizae Preparata 50-60 weight portion
Pericarpium Citri Reticulatae 50-60 weight portion Colla cornus cervi 20-30 weight portion
Radix Rehmanniae Preparata 150-170 weight portion Colla Plastri Testudinis 30-40 weight portion
Fructus Rosae Laevigatae 120-140 weight portion Radix Astragali Preparata 100-110 weight portion
Rhizoma Dioscoreae 100-110 weight portion Fructus Rubi 210-230 weight portion.
2, the method for quality control of Chinese medicine composition as claimed in claim 1 is characterized in that the assay in this method is:
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With 755: 230: 10: 5 water-acetonitrile-oxolane-glacial acetic acid was a mobile phase; The detection wavelength is 270nm; Theory is pulled number should be not less than 3500 by the calculating of icariin peak; The preparation of reference substance solution: take by weighing the icariin reference substance, add methanol and make the solution that 1ml contains 20 μ g, promptly; The preparation of need testing solution: get 10 in this pharmaceutical composition tablet, remove coating, it is fixed to claim, get 2 amount, it is fixed to claim, puts in the tool plug conical flask, adding concentration is 50% ethanol 50ml, and close plug claims to decide weight, supersound process 1 hour is put coldly, claims to decide weight again, with concentration is that 50% ethanol is supplied the weight that subtracts mistake, shakes up, and filters, draw subsequent filtrate 25ml, put in the evaporating dish evaporate to dryness, residue adds water 5ml makes dissolving, is added in 13～80 orders of anticipating, 5g, internal diameter 1.5cm, on the polyamide column of dry column-packing, water 50ml eluting, discard eluent, continue, collect eluent with ethanol 80ml eluting, evaporate to dryness, residue adds dissolve with methanol and is transferred in the 5ml measuring bottle, adds methanol to scale, shakes up, filter, promptly; Algoscopy: draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly; This drug combination preparation per unit preparation contains Herba Epimedii by icariin C
33H
40O
15Meter must not be less than 0.05mg.
3, the method for quality control of Chinese medicine composition as claimed in claim 1 is characterized in that the crude drug of this Chinese medicine composition consists of:
Semen Cuscutae (parched) 128 weight portions steam Herba Epimedii 106 weight portions
Steam Radix Dipsaci 106 weight portions and steam Herba Cynomorii 128 weight portions
Steam Rhizoma Cibotii 160 weight portion Semen Ziziphi Spinosae (parched)s 106 weight portions
Radix Polygoni Multiflori Preparata 160 weight portion Radix Glycyrrhizae Preparatas 53 weight portions
Steam Pericarpium Citri Reticulatae 53 weight portions and fry Colla cornus cervi 23 weight portions
Radix Rehmanniae Preparata 160 weight portions are fried Colla Plastri Testudinis 34 weight portions
Steam Fructus Rosae Laevigatae 128 weight portion Radix Astragali Preparatas 106 weight portions
Rhizoma dioscoreae (parched) 106 weight portions steam Fructus Rubi 213 weight portions.
4,, it is characterized in that this method also comprises one or more in the following discriminating as the method for quality control of the arbitrary described Chinese medicine composition of claim 1-3:
A. get 10 in this pharmaceutical composition tablet, remove coating, porphyrize, add methanol 40-60ml, supersound process 10-20 minute, filter, the filtrate evaporate to dryness, residue adds water 20-40ml makes dissolving, uses ethyl acetate 20-40ml at every turn, and jolting is extracted 2-4 time, merge ethyl acetate liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Radix Polygoni Multiflori control medicinal material 0.5g, adds ethanol 40-60ml reflux 1-1.5 hour, filters, and filtrate is concentrated into 3ml, in contrast medical material solution; Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, control medicinal material and reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of binding agent, to make into ribbon with the carboxymethylcellulose sodium solution, with 10-20: 2-4: 1-2 toluene-ethyl acetate-formic acid is placed and was treated that stratified upper solution was developing solvent in 20-40 minute, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with control medicinal material and the corresponding position of contrast mouthful chromatograph on, show the fluorescence streak of same color; Behind the ammonia cure, become redness under the streak daylight;
B. get the aqueous solution behind the A item ethyl acetate extraction, use water saturated n-butyl alcohol 20-40ml at every turn, extract 2-4 time, merge n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, add the 0.5-1g neutral alumina and mix thoroughly, drying is added in 100～120 orders, 2g is on the neutral alumina post of internal diameter 10mm, with 30-50% methanol 20-40ml eluting, collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds water 30-40ml in addition, and reflux 1-1.5 hour, filter, the n-butyl alcohol that filtrate water is saturated shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, ethyl acetate-formic acid-glacial acetic acid-water with 15-20: 1-2: 1-2: 2-4 is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 100 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, on the corresponding position of control medicinal material chromatograph, daylight shows identical yellow principal spot down; Under the ultra-violet lamp, show identical yellow fluorescence principal spot;
C. get 10 in this pharmaceutical composition tablet, remove coating, porphyrize, add methanol 30-50ml, reflux 1-.5 hour, filter, filtrate evaporate to dryness, residue add water 30-40ml makes dissolving, uses ethyl acetate 20-40ml at every turn, extract 2-4 time, discard ethyl acetate liquid, water layer extracts 2-4 time with water saturated n-butyl alcohol 20-40ml at every turn, merge n-butanol extracting liquid, use 1% sodium hydroxide solution 20-40ml at every turn, wash 2-4 time, discard alkali wash water, each water 20-30ml washs 2-4 time, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, adding the 0.5-1g neutral alumina mixes thoroughly, drying is added in 100～120 orders, 2g, on the neutral alumina post of internal diameter 10mm, with 30-50% methanol 40-60ml eluting, collect eluent, evaporate to dryness, residue adds methanol 0.5ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 10 μ l, reference substance solution 5 μ l, put respectively on the same silica gel g thin-layer plate of thick 0.5mm, with 10-15: 20-25: 10-15: 5-10 chloroform-ethyl acetate-methanol-water, at lower floor's solution of placing 10-20 hour below 10 ℃ is developing solvent, launches; Take out, dry, spray is with the 10-15% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105-110 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, daylight shows down identical sepia speckle, ultra-violet lamp shows identical orange-yellow fluorescence speckle down.
5, the method for quality control of Chinese medicine composition as claimed in claim 4 is characterized in that this method also comprises one or more in the following discriminating:
A. get 10 in this pharmaceutical composition tablet, remove coating, porphyrize, add methanol 50ml, supersound process 15 minutes filters, the filtrate evaporate to dryness, residue adds water 30ml makes dissolving, adds the ethyl acetate jolting and extracts 2 times, each 30ml, merge ethyl acetate liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Radix Polygoni Multiflori control medicinal material 0.5g, adds ethanol 50ml reflux 1 hour, filters, and filtrate is concentrated into 3ml, in contrast medical material solution; Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, control medicinal material and reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of binding agent, to make into ribbon with the carboxymethylcellulose sodium solution, with 15: 2: 1 toluene-ethyl acetate-formic acid, place and treated that stratified upper solution was developing solvent in 30 minutes, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with control medicinal material and the corresponding position of contrast mouthful chromatograph on, show the fluorescence streak of same color; Behind the ammonia cure, become redness under the streak daylight;
B. get the aqueous solution behind the A item ethyl acetate extraction, with water saturated n-butanol extraction 2 times, each 30ml, merge n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, add an amount of neutral oxidation instruction aluminum and mix thoroughly, drying is added in 100～120 orders, 2g is on the internal diameter 10mm neutral alumina post, with 40% methanol 30ml eluting, collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice contrast medicine letter 0.3g adds water 30ml in addition, and reflux 1 hour filters, and the n-butyl alcohol that filtrate water is saturated shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 15: 1: 1: 2 ethyl acetates-formic acid-glacial acetic acid-water was developing solvent, launched, and took out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 100 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, on the corresponding position of control medicinal material chromatograph, daylight shows identical yellow principal spot down; Under the ultra-violet lamp, show identical yellow fluorescence principal spot;
C. get 10 in this pharmaceutical composition tablet, remove coating, porphyrize, add methanol 40ml, reflux 1 hour filters, filtrate evaporate to dryness, residue add water 30ml makes dissolving, uses ethyl acetate extraction 2 times, each 30ml discards ethyl acetate liquid, and water layer is with water saturated n-butanol extraction 2 times, each 30ml merges n-butanol extracting liquid, with 1% sodium hydroxide solution washing 2 times, each 30ml discards alkali wash water, washes with water 2 times, each 25ml discards water liquid, and n-butyl alcohol liquid steams, residue adds methanol 1ml makes dissolving, adds an amount of neutral alumina and mixes drying thoroughly, be added in 100～120 orders, 2g is on the internal diameter 10mm neutral alumina post, with 40% methanol 50ml eluting, collect eluent, evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 10 μ l, reference substance solution 5 μ l put respectively on the same silica gel g thin-layer plate of thick 0.5mm, with 10: 20: 11: lower floor's solution that 5 chloroforms-ethyl acetate-methanol-water was placed 12 hours below 10 ℃ was developing solvent, launched; Take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts respectively under daylight and the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, composing on the corresponding position with the reference substance bag, daylight shows identical sepia speckle down, and ultra-violet lamp shows identical orange-yellow fluorescence speckle down.
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| CN107412510B (en) * | 2017-04-28 | 2018-04-03 | 广东心宝药业科技有限公司 | A kind of Chinese medicine composition with tonifying both YIN and YANG effect and preparation method thereof |
| CN114306491B (en) * | 2021-12-22 | 2023-02-28 | 江苏七○七天然制药有限公司 | Fleece-flower root hair growth capsule and preparation, detection and component identification methods thereof |
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