CN104666700A - Traditional Chinese medicine composition for treating fracture and quality detection method of traditional Chinese medicine composition - Google Patents
Traditional Chinese medicine composition for treating fracture and quality detection method of traditional Chinese medicine composition Download PDFInfo
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Abstract
Aiming at the defects in the background technology, the provides a traditional Chinese medicine composition which is exact in curative effect and is capable of quickly treating fracture and a quality detection method of the traditional Chinese medicine composition. According to the technical scheme provided by the invention, the traditional Chinese medicine composition for promoting growth of bone fracture is characterized by being prepared from the following traditional Chinese medicines in parts by weight: 28-32 parts of caulis spatholobi, 13-17 parts of flos carthami, 28-32 parts of salviae miltiorrhizae, 18-22 parts of ligusticum wallichii, 28-32 parts of hematoxylon, 13-17 parts of native copper, 1-5 parts of borneol, 13-17 parts of radix aconite, 8-12 parts of fried nux vomica, 4-8 parts of dragon blood, 28-32 parts of calcined gypsum, 28-32 parts of honeysuckle stems, 13-17 parts of ground beetles and 18-22 parts of angelica sinensis.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and the quality determining method thereof that are used for the treatment of fracture, belong to Chinese prescription field.
Background technology
Fracture refers to a kind of disease because the reason such as wound or pathology causes bone parts or ruptures completely.It is use plaster fixing again by after fracture manual reduction that the rear conventional treatments of fracture occurs, and severe patient need carry out operative treatment, is fixed by fracture, be equipped with liniment with steel nail or steel plate, relies on the regeneration capacity of skeleton self to make its normal healing afterwards.This Therapeutic Method utilizing operation and medicine, weak point is that patient easily occurs that growth of spur is slow, the problems such as long-term disunion, treatment cycle is longer, medical expense is expensive of fracturing, and causes soft tissue complication thus, brings inconvenience to patient.
Treatment by Chinese herbs fracture in China's application for a long time, has fast fracture-setting, promotes the feature of growth of spur, compared with treating, has obvious advantage with fracture routine operation.But the Chinese medicine preparation of at present treatment fracture is less, exploitation dynamics is inadequate, and there is uncertain therapeutic efficacy and cut, and preparation method is indefinite, and cost is higher, the problem such as limited in Clinical practice.Therefore, be badly in need of a kind of therapeutic effect at present definite, rapid healing is fractured, and cheap Chinese medicine.
Summary of the invention
The present invention is intended to the deficiency for existing in background technology, and the Chinese medicine composition of a kind of determined curative effect provided, rapid healing fracture and quality determining method thereof.
The present invention is achieved through the following technical solutions:
Promote to it is characterized in that the Chinese medicine composition that bone injury affected part grows, be made up of the Chinese medicine of following weight ratio: Caulis Spatholobi 28-32 part, Flos Carthami 13-17 part, Radix Salviae Miltiorrhizae 28-32 part, Rhizoma Chuanxiong 18-22 part, Lignum Sappan 28-32 part, Pyritum 13-17 part, Borneolum Syntheticum 1-5 part, Radix Aconiti 13-17 part, Semen Strychni (parched) 8-12 part, Sanguis Draxonis 4-8 part, Gypsum Fibrosum Preparatum 28-32 part, Caulis Lonicerae 28-32 part, soil suppresses worm 13-17 part, Radix Angelicae Sinensis 18-22 part.
The Chinese medicine composition of aforesaid promotion bone injury affected part growth, it is characterized in that, the weight ratio of each Chinese medicine is specifically: Caulis Spatholobi 30 parts, 15 parts, Flos Carthami, Radix Salviae Miltiorrhizae 30 parts, Rhizoma Chuanxiong 20 parts, Lignum Sappan 30 parts, Pyritum 15 parts, Borneolum Syntheticum 3 parts, Radix Aconiti 15 parts, Semen Strychni (parched) 10 parts, Sanguis Draxonis 6 parts, Gypsum Fibrosum Preparatum 30 parts, Caulis Lonicerae 30 parts, soil suppresses worm 15 parts, Radix Angelicae Sinensis 20 parts, after it is pulverized and mixed, add the Olibanum 1 part being equivalent to the same weight ratio again, Myrrha 1 part, with aforesaid Caulis Spatholobi after being pulverized, Flos Carthami, Radix Salviae Miltiorrhizae, Rhizoma Chuanxiong, Lignum Sappan, Pyritum, Borneolum Syntheticum, Radix Aconiti, Semen Strychni (parched), Sanguis Draxonis, Gypsum Fibrosum Preparatum, Caulis Lonicerae, soil suppresses worm, the ground product mix homogeneously of Radix Angelicae Sinensis, add batching, be made into externally used paste.
The quality determining method of the Chinese medicine composition of aforesaid promotion bone injury affected part growth, is characterized in that:
It comprises respectively to the thin-layer chromatographic analysis determination step of Flos Carthami, Radix Salviae Miltiorrhizae, Lignum Sappan, Caulis Lonicerae;
1) be contrast Flos Carthami sample for reference substance to the thin-layer chromatographic analysis of Flos Carthami, be that the acetic ether-methanoic acid-water-methanol mixture of 7:2:3:0.4 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel H lamellae launches:
Get Flos Carthami 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Flos Carthami sample 0.5g, add 80% aqueous acetone solution 5ml, sealing of jumping a queue, jolting 15min, leave standstill, get supernatant medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel H lamellae, take volume ratio as the acetic ether-methanoic acid-water-methanol mixture of 7:2:3:0.4 be developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Flos Carthami sample in thin layer chromatography;
2) be contrast Radix Salviae Miltiorrhizae sample for reference substance to the thin-layer chromatographic analysis of Radix Salviae Miltiorrhizae, be that the petroleum ether-ethyl acetate of 60-90 DEG C of 4:1 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel g thin-layer plate launches:
Get Radix Salviae Miltiorrhizae 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Radix Salviae Miltiorrhizae sample 1g, add diethyl ether 5ml, jolting, places 1 hour, filters, and after filtrate volatilization completely, residue adds ethyl acetate 1ml and dissolves, and gained solution is medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, volume ratio is the petroleum ether-ethyl acetate of 60-90 DEG C of 4:1 is developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Radix Salviae Miltiorrhizae sample in thin layer chromatography;
3) being contrast Lignum Sappan sample for reference substance to the thin-layer chromatographic analysis of Lignum Sappan, is that the chloroform-acetone-formic acid of 8:4:1 is developing solvent with volume ratio, with silica gel G F
254the thin layer chromatography that lamellae launches carries out thin layer chromatography check analysis:
Get Lignum Sappan 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate.After evaporate to dryness, add methanol 2ml and make dissolving, as need testing solution;
Separately get contrast Lignum Sappan sample 1g, add methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
According to thin layer chromatography test, draw need testing solution and each 3 μ l of control medicinal material solution, put respectively in same silica gel G F
254on lamellae, take volume ratio as the chloroform-acetone-formic acid of 8:4:1 be developing solvent, launch, taking-up is dried, put in exsiccator and place 12 hours, inspect under being placed on 254nm ultraviolet light, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Lignum Sappan sample in thin layer chromatography;
4) be contrast Caulis Lonicerae sample for reference substance to the thin-layer chromatographic analysis of Caulis Lonicerae, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 65:35:10 is placed below 10 DEG C be developing solvent, so that silica gel g thin-layer plate to launch, the tlc identification method being developer with 10% sulphuric acid ethanol:
Get Caulis Lonicerae 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate.After evaporate to dryness, add methanol 2ml and make dissolving, as need testing solution;
Separately get Caulis Lonicerae control medicinal material 1g, add 50% methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 10 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 65:35:10 is placed below 10 DEG C be developing solvent, launch, taking-up is dried, and sprays with 10% ethanol solution of sulfuric acid, be heated to colour developing at 105 DEG C clear, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Caulis Lonicerae sample in thin layer chromatography;
And
5) detection of HPLC finger printing is carried out to Caulis Spatholobi, detect the effective peak area of Caulis Spatholobi used at 11 chromatographic fingerprint peaks place;
6) mensuration pipe is carried out to the ferulaic acid content in Rhizoma Chuanxiong, contrast with the content contrasted with sample ferulic acid;
7) X-ray diffraction finger printing carried out to Pyritum, analyze the quantity at total peak, contrast the raw product of Pyritum, forge the X-ray diffraction finger printing of product sample, judge that Pyritum is raw product, forges product or be mixed in proportion thing;
8) near infrared spectrum detection is carried out to Borneolum Syntheticum, itself and the Borneolum Syntheticum sample contrasted all are made into 100mg/ml, contrast with regard to itself and five the strongest peaks of sample in 4000-9000 wave number, do semi-quantitative analysis qualitatively simultaneously;
9) for Radix Aconiti, Semen Strychni (parched), Sanguis Draxonis, Gypsum Fibrosum Preparatum, soil suppresses worm, Radix Angelicae Sinensis can carry out visual comparison's detection with standard sample;
In embodiments of the present invention, can take the above-mentioned each taste Chinese medicinal powder pulverized, mixing, then carries out ethanol extraction and water extraction, through to Rabbits with Fracture clinical effect trial, selects the optimum extraction mode of Chinese medicine composition.And then the extract of Chinese medicine composition is prepared into required dosage form.
Weight portion of the present invention can be the known unit of weights of field of medicaments such as μ g, mg, g or kg.
Chinese medicine composition dosage form of the present invention is external application paste, and its preparation method is as follows:
Get the raw Oleum Sesami of Chinese medicine gross weight 5 times, stir, boil 2-4 hour.
In medicinal liquid, add Plumbum preparatium, then boil 10-60min.
Add Chinese medicine composition, stir, dispel fire-toxin, receive cream and get final product.
The function of each taste Chinese medicine is as follows:
Caulis Spatholobi can wind-damp dispelling, relaxing muscles and tendons network, blood circulation promoting and enriching; Flos Carthami: invigorate blood circulation, blood in heat of dehematizing, dispelling the stagnated QI, remove congestion; Radix Salviae Miltiorrhizae: blood circulation promoting and blood stasis dispelling, calming heart and tranquilizing mind, evacuation of pus pain relieving; Rhizoma Chuanxiong: blood circulation promoting and blood stasis dispelling, activating QI to alleviate the depression, wind-expelling pain-stopping;
Lignum Sappan: scattered silt is long-pending and continue wound; Pyritum: eliminating stasis to stop pain, reunion of bone; Borneolum Syntheticum: sensible scattered silt, reducing swelling and alleviating pain; Sanguis Draxonis: dissipating blood stasis analgesic therapy, hemostasia and promoting granulation;
Radix Aconiti: expelling wind and removing dampness, antalgic; Semen Strychni (parched): activating spleen and strengthening stomach, external qualcomm meridian, mass dissipating and swelling eliminating;
Gypsum Fibrosum Preparatum: promoting tissue regeneration and ulcer healing;
Caulis Lonicerae: heat-clearing and toxic substances removing, dredge the meridian passage; Soil suppresses worm: dissipating blood stasis is long-pending and continue wound; Radix Angelicae Sinensis: enrich blood invigorate blood circulation, granulation promoting pain relieving;
Olibanum: there is regulating qi and activating blood, analgesic therapy, chases after effect of poison;
Myrrha: there is the effect such as promoting blood circulation and stopping pain, detumescence and promoting granulation.
The invention has the beneficial effects as follows: this Chinese medicine composition can treat various fracture, promote growth of spur and healing, quick reducing swelling and alleviating pain, promoting blood circulation by removing blood stasis, therapeutic effect is definite, and effective percentage is high, instant effect.Dosage form of the present invention is externally used paste, easy to use, and direct external application is in affected part, and therapeutic effect is good, can rapid recovery patient suffering, belongs to external treatment, thus avoids the toxic and side effects of endo-medicine.The usage of plaster and consumption: fracturing the swelling phase, ointment is directly spread upon affected part, external splint is fixed, change a medicine, to porotic stage, plaster used fiery molten, is affixed on affected part for 24 hours, within 7 days, changes once.Within 4 weeks, be a course for the treatment of, use 1-3 the course for the treatment of.
Detailed description of the invention
Following examples will contribute to those of ordinary skill in the art and understand the present invention further, but not limit the present invention in any form.
Embodiment 1
1. treat the extraction of the Chinese medicine composition effective ingredient of fracture for one kind:
Experimental apparatus and reagent: Chinese medicine grinder, Rotary Evaporators, vacuum drying oven; Ethanol, distilled water.
Experimental technique:
By the Caulis Spatholobi in Chinese prescription 30 parts, 15 parts, Flos Carthami, Radix Salviae Miltiorrhizae 30 parts, Rhizoma Chuanxiong 20 parts, Lignum Sappan 30 parts, Pyritum 15 parts, Borneolum Syntheticum 3 parts, Radix Aconiti 15 parts, Semen Strychni (parched) 10 parts, Sanguis Draxonis 6 parts, Gypsum Fibrosum Preparatum 30 parts, Caulis Lonicerae 30 parts, soil suppresses worm 15 parts, Radix Angelicae Sinensis 20 parts.Pulverize, be mixed in proportion, carry out alcohol extraction or carry out water extraction.
Its concrete steps are:
Alcohol extraction is followed the example of: by the Chinese medicinal powder 90% ethanol cooling for reflux that is pulverized and mixed 3 times, the amount at every turn added is 1100ml, each two hours, and each backflow end carries out sucking filtration, merging filtrate.Obtained filtrate is obtained concentrated extracting solution by Rotary Evaporators at 55 DEG C.After the Chinese medicine extraction liquid obtaining concentrating, add Olibanum and each portion of Myrrha more successively, put into vacuum drying oven inner drying after stirring and evenly mixing, to obtain Chinese medicinal powder.
Through the Chinese medicine of water extraction and alcohol extraction, dry fine powder can be obtained after vacuum drying oven inner drying, use for the preparation of next step plaster.
Prepared by alcohol extraction group Chinese medicine composition plaster
Step 1: the raw Oleum Sesami getting whole component weight 5 times, stirs, boils 2-4 hour, boil temperature at 280-320 DEG C;
Step 2: the Plumbum preparatium of then dropping into Chinese medicine gross weight 2 times, mixes well, uniform stirring 20-25 minute.Boil 10-60 minute;
Step 3: continue to boil the raw Oleum Sesami being mixed with Plumbum preparatium, until after ointment drips into pearl, be cooled to 80-85 DEG C while stirring;
Step 4: add each Chinese medicine composition fine powder through ethanol extraction, continues stirring and makes temperature be down to 45-50 DEG C, spread upon on plaster paper for subsequent use, be cooled to room temperature, namely obtain Chinese medicine composition Dressed plaster after ointment stirs.
2. alcohol extraction group Chinese medicine composition effective constituent determination
Step 1: to the thin-layer chromatographic analysis of Flos Carthami specifically:
To contrast Flos Carthami sample for reference substance, be that the acetic ether-methanoic acid-water-methanol mixture of 7:2:3:0.4 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel H lamellae launches:
Get Flos Carthami 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Flos Carthami sample 0.5g, add 80% aqueous acetone solution 5ml, sealing of jumping a queue, jolting 15min, leave standstill, get supernatant medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel H lamellae, take volume ratio as the acetic ether-methanoic acid-water-methanol mixture of 7:2:3:0.4 be developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Flos Carthami sample in thin layer chromatography; Dry chromatographic sheet at 80-100 DEG C, can according to test sample with contrast the different of speckle displacement that Flos Carthami sample shows, to judge test sample flos carthami character;
Step 2: to the thin-layer chromatographic analysis of Radix Salviae Miltiorrhizae specifically:
To contrast Radix Salviae Miltiorrhizae sample for reference substance, be that the petroleum ether-ethyl acetate of 60-90 DEG C of 4:1 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel g thin-layer plate launches:
Get Radix Salviae Miltiorrhizae 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Radix Salviae Miltiorrhizae sample 1g, add diethyl ether 5ml, jolting, places 1 hour, filters, and after filtrate volatilization completely, residue adds ethyl acetate 1ml and dissolves, and gained solution is medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, volume ratio is the petroleum ether-ethyl acetate of 60-90 DEG C of 4:1 is developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Radix Salviae Miltiorrhizae sample in thin layer chromatography; Dry chromatographic sheet at 100-110 DEG C, inspect with the uviol lamp of about 365nm wavelength, can according to test sample with contrast the different of speckle displacement that Radix Salviae Miltiorrhizae sample shows, to judge test sample red rooted salvia character;
Step 3: to the thin-layer chromatographic analysis of Lignum Sappan specifically:
To contrast Lignum Sappan sample for reference substance, be that the chloroform-acetone-formic acid of 8:4:1 is developing solvent with volume ratio, with silica gel G F
254the thin layer chromatography that lamellae launches carries out thin layer chromatography check analysis:
Get Lignum Sappan 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate.After evaporate to dryness, add methanol 2ml and make dissolving, as need testing solution;
Separately get contrast Lignum Sappan sample 1g, add methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
According to thin layer chromatography test, draw need testing solution and each 3 μ l of control medicinal material solution, put respectively in same silica gel G F
254on lamellae, take volume ratio as the chloroform-acetone-formic acid of 8:4:1 be developing solvent, launch, taking-up is dried, put in exsiccator and place 12 hours, inspect under being placed on 254nm ultraviolet light, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Lignum Sappan sample in thin layer chromatography;
Step 4: to the thin-layer chromatographic analysis of Lignum Sappan specifically:
To contrast Caulis Lonicerae sample for reference substance, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 65:35:10 is placed below 10 DEG C be developing solvent, so that silica gel g thin-layer plate to launch, the tlc identification method being developer with 10% sulphuric acid ethanol:
Get Caulis Lonicerae 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate.After evaporate to dryness, add methanol 2ml and make dissolving, as need testing solution;
Separately get Caulis Lonicerae control medicinal material 1g, add 50% methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 10 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 65:35:10 is placed below 10 DEG C be developing solvent, launch, taking-up is dried, and sprays with 10% ethanol solution of sulfuric acid, be heated to colour developing at 105 DEG C clear, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Caulis Lonicerae sample in thin layer chromatography;
Step 5:
The detection of HPLC finger printing is carried out to Caulis Spatholobi, detects the effective peak area of Caulis Spatholobi used at 23 chromatographic fingerprint peaks place; Through carrying out sounding out the people in a given scope one by one in order to break a criminal case of chromatographic fingerprint peak to the Caulis Spatholobi of a large amount of batches, wherein the content of seven kinds is larger, corresponding chromatographic peak is larger, be easier to carry out correspondence calculate, these seven kinds is protocatechuic acid, epicatechin, catechin, daidzein, calycosin, genistein, isoliquiritigenin, can only carry out calculating for the chromatographic fingerprint peak of these seven kinds of materials and with the contrast of standard sample; Preferably, can only carry out calculating for the chromatographic fingerprint peak of three kinds of materials and with the contrast of standard sample, protocatechuic acid, epicatechin, catechin that namely chromatographic peak is maximum;
Step 6:
Mensuration pipe is carried out to the ferulaic acid content in Rhizoma Chuanxiong, contrasts with the content contrasted with sample ferulic acid; Here, think that effective ingredient that Rhizoma Chuanxiong mainly utilizes is exactly the ferulic acid of the inside, HPLC method can be utilized to detect ferulaic acid content, with C18 chromatographic column, methanol-2% acetum (25:75) is mobile phase, and flow velocity is 1ml/min, determined wavelength 251nm, column temperature 18-25 DEG C, by measuring, the ferulic acid characteristic peak area between contrast test sample and control sample and intensity;
Step 7:
X-ray diffraction finger printing carried out to Pyritum, analyzes the quantity at total peak, contrast the raw product of Pyritum, forge the X-ray diffraction finger printing of product sample, judge that Pyritum is raw product, forges product or be mixed in proportion thing; This mainly considers that the raw product of Pyritum on market forge product confusion, and often doping causes, and raw product, forges product or be mixed in proportion thing difference, and the active constituent content difference in Unit Weight is comparatively large, affects compositions practical application effect;
Step 8:
Near infrared spectrum detection is carried out to Borneolum Syntheticum, itself and the Borneolum Syntheticum sample contrasted all is made into 100mg/ml, contrasts with regard to itself and five the strongest peaks of sample in 4000-9000 wave number, do semi-quantitative analysis qualitatively simultaneously;
Step 9:
For Radix Aconiti, Semen Strychni (parched), Sanguis Draxonis, Gypsum Fibrosum Preparatum, soil suppresses worm, Radix Angelicae Sinensis can carry out visual comparison's detection with standard sample; This several Chinese medicine does not generally adulterate problem, and only only have whether certified products, whether process is suitable, can only judge with general general knowledge;
Step 10: described inspection should meet pertinent regulations under " Chinese Pharmacopoeia " version in 2010 first annex emplastrum item;
Step 11: the mensuration of described extractum: get this medicament content 5g, accurately weighed, use 100% ethanol as solvent, measure according to the cold-maceration under ethanol-soluble extractives algoscopy item, must not 75% be less than.
This quality determining method adopts thin layer chromatography to carry out TLC distinguish to the Flos Carthami in preparation, Radix Salviae Miltiorrhizae, Lignum Sappan and Caulis Lonicerae, and result all significantly can identify the existence of above 4 kinds of medicines.Adopt ethanol-soluble extractives algoscopy, take straight alcohol as solvent, the ethanol-soluble extractives in preparation is measured, and according to the measured data of 10 batch sample determinations of extractives, work out the content limit of preparation ethanol-soluble extractives.More than to differentiate and the quality standard research result such as mensuration of ethanol-soluble extractives shows, this preparing process fully remains effective ingredient and the main component of each medicine of prescription, through the quality inspection to many batches of pilot scale preparation samples, the quality of the pharmaceutical preparations all meets appended " quality standard " and " Chinese Pharmacopoeia " to the pertinent regulations of medicine sanitary standard, the quality of the pharmaceutical preparations is stablized, and this shows that the preparing process of this preparation is scientific and reasonable.
Embodiment 2
Experiment conditions etc. are basic identical with embodiment 1, wherein:
Experimental apparatus and reagent: Chinese medicine grinder, Rotary Evaporators, vacuum drying oven; Ethanol, distilled water.
Experimental technique:
By the Caulis Spatholobi in Chinese prescription 28 parts, 14 parts, Flos Carthami, Radix Salviae Miltiorrhizae 28 parts, Rhizoma Chuanxiong 20 parts, Lignum Sappan 28 parts, Pyritum 14 parts, Borneolum Syntheticum 5 parts, Radix Aconiti 14 parts, Semen Strychni (parched) 8 parts, Sanguis Draxonis 8 parts, Gypsum Fibrosum Preparatum 28 parts, Caulis Lonicerae 28 parts, soil suppresses worm 14 parts, Radix Angelicae Sinensis 18 parts.Pulverize, be mixed in proportion, carry out alcohol extraction or carry out water extraction.
Its concrete steps are:
Water extraction method: take the above-mentioned Chinese medicine having pulverized mixing, with distilled water cooling for reflux 5 times, the amount at every turn added is 800ml, each two hours, and each backflow end carries out sucking filtration, merging filtrate, by Rotary Evaporators at 68 DEG C of concentrated extracting solutions.After the Chinese medicine extraction liquid obtaining concentrating, then add Olibanum successively and do not have each 1 part of process and above same step alcohol extraction, stir and put into vacuum drying oven inner drying, to obtain Powdered Chinese medicine extract.
Through the Chinese medicine of water extraction, dry fine powder can be obtained after vacuum drying oven inner drying, use for the preparation of next step plaster.
Prepared by water extraction group Chinese medicine composition plaster
Step 1: the raw Oleum Sesami getting whole component weight 5 times, stirs, boils 5 hours, boil temperature at 270 DEG C;
Step 2: the Plumbum preparatium of then dropping into Chinese medicine gross weight 2 times, mixes well, uniform stirring 30 minutes.Boil 80 minutes;
Step 3: continue to boil the raw Oleum Sesami being mixed with Plumbum preparatium, until after ointment drips into pearl, be cooled to 65 DEG C while stirring;
Step 4: add each Chinese medicine composition fine powder through water extraction, continues stirring and makes temperature be down to 45-50 DEG C, spread upon on plaster paper for subsequent use, be cooled to room temperature, namely obtain Chinese medicine composition Dressed plaster after ointment stirs.
2. water extraction group Chinese medicine composition effective constituent determination
Step 1: to the thin-layer chromatographic analysis of Flos Carthami specifically:
To contrast Flos Carthami sample for reference substance, be that the acetic ether-methanoic acid-water-methanol mixture of 7:3:5:0.8 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel H lamellae launches:
Get Flos Carthami 5g, put in apparatus,Soxhlet's, add ethanol 150ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 5ml and makes dissolving, as need testing solution;
Separately get contrast Flos Carthami sample 0.5g, add 70% aqueous acetone solution 10ml, sealing of jumping a queue, jolting 10min, leave standstill, get supernatant medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel H lamellae, take volume ratio as the acetic ether-methanoic acid-water-methanol mixture of 7:3:5:0.8 be developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Flos Carthami sample in thin layer chromatography; Dry chromatographic sheet at 80-100 DEG C, can according to test sample with contrast the different of speckle displacement that Flos Carthami sample shows, to judge test sample flos carthami character;
Step 2: to the thin-layer chromatographic analysis of Radix Salviae Miltiorrhizae specifically:
To contrast Radix Salviae Miltiorrhizae sample for reference substance, be that the petroleum ether-ethyl acetate of 67-73 DEG C of 3:1 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel g thin-layer plate launches:
Get Radix Salviae Miltiorrhizae 5g, put in apparatus,Soxhlet's, add ethanol 150ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 5ml and makes dissolving, as need testing solution;
Separately get contrast Radix Salviae Miltiorrhizae sample 1g, add diethyl ether 8ml, jolting, places 1 hour, filters, and after filtrate volatilization completely, residue adds ethyl acetate 3ml and dissolves, and gained solution is medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, volume ratio is the petroleum ether-ethyl acetate of 67-73 DEG C of 3:1 is developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Radix Salviae Miltiorrhizae sample in thin layer chromatography; Dry chromatographic sheets at 106 DEG C, inspect with the uviol lamp of about 360nm wavelength, can according to test sample with contrast the different of speckle displacement that Radix Salviae Miltiorrhizae sample shows, to judge test sample red rooted salvia character;
Step 3: to the thin-layer chromatographic analysis of Lignum Sappan specifically:
To contrast Lignum Sappan sample for reference substance, be that the chloroform-acetone-formic acid of 6:3:1 is developing solvent with volume ratio, with silica gel G F
254the thin layer chromatography that lamellae launches carries out thin layer chromatography check analysis:
Get Lignum Sappan 5g, put in apparatus,Soxhlet's, add ethanol 150ml, reflux 2 hours, extract 3 times altogether, merging filtrate.After evaporate to dryness, add methanol 5ml and make dissolving, as need testing solution;
Separately get contrast Lignum Sappan sample 1g, add methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
According to thin layer chromatography test, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively in same silica gel G F
254on lamellae, take volume ratio as the chloroform-acetone-formic acid of 6:3:1 be developing solvent, launch, taking-up is dried, put in exsiccator and place 24 hours, inspect under being placed on 254nm ultraviolet light, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Lignum Sappan sample in thin layer chromatography;
Step 4: to the thin-layer chromatographic analysis of Lignum Sappan specifically:
To contrast Caulis Lonicerae sample for reference substance, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 55:30:15 is placed below 25 DEG C be developing solvent, so that silica gel g thin-layer plate to launch, the tlc identification method being developer with 18% sulphuric acid ethanol:
Get Caulis Lonicerae 5g, put in apparatus,Soxhlet's, add ethanol 150ml, reflux 2 hours, extract 3 times altogether, merging filtrate.After evaporate to dryness, add methanol 5ml and make dissolving, as need testing solution;
Separately get Caulis Lonicerae control medicinal material 1g, add 50% methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 10 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 55:30:15 is placed below 25 DEG C be developing solvent, launch, taking-up is dried, and sprays with 18% ethanol solution of sulfuric acid, be heated to colour developing at 108 DEG C clear, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Caulis Lonicerae sample in thin layer chromatography;
Step 5:
The detection of HPLC finger printing is carried out to Caulis Spatholobi, detects the effective peak area of Caulis Spatholobi used at 23 chromatographic fingerprint peaks place; Through carrying out sounding out the people in a given scope one by one in order to break a criminal case of chromatographic fingerprint peak to the Caulis Spatholobi of a large amount of batches, wherein the content of seven kinds is larger, corresponding chromatographic peak is larger, be easier to carry out correspondence calculate, these seven kinds is protocatechuic acid, epicatechin, catechin, daidzein, calycosin, genistein, isoliquiritigenin, can only carry out calculating for the chromatographic fingerprint peak of these seven kinds of materials and with the contrast of standard sample; Preferably, can only carry out calculating for the chromatographic fingerprint peak of four kinds of materials and with the contrast of standard sample, protocatechuic acid, epicatechin, catechin, daidzein that namely chromatographic peak is maximum;
Step 6:
Mensuration pipe is carried out to the ferulaic acid content in Rhizoma Chuanxiong, contrasts with the content contrasted with sample ferulic acid; Here, think that effective ingredient that Rhizoma Chuanxiong mainly utilizes is exactly the ferulic acid of the inside, HPLC method can be utilized to detect ferulaic acid content, with C18 chromatographic column, methanol-3% acetum (20:80) is mobile phase, and flow velocity is 1.2ml/min, determined wavelength 256nm, column temperature 25-30 DEG C, by measuring, the ferulic acid characteristic peak area between contrast test sample and control sample and intensity;
Step 7:
X-ray diffraction finger printing carried out to Pyritum, analyzes the quantity at total peak, contrast the raw product of Pyritum, forge the X-ray diffraction finger printing of product sample, judge that Pyritum is raw product, forges product or be mixed in proportion thing; This mainly considers that the raw product of Pyritum on market forge product confusion, and often doping causes, and raw product, forges product or be mixed in proportion thing difference, and the active constituent content difference in Unit Weight is comparatively large, affects compositions practical application effect;
Step 8:
Near infrared spectrum detection is carried out to Borneolum Syntheticum, itself and the Borneolum Syntheticum sample contrasted all is made into 120mg/ml, contrasts with regard to itself and six the strongest peaks of sample in 4000-9000 wave number, do semi-quantitative analysis qualitatively simultaneously;
Step 9:
For Radix Aconiti, Semen Strychni (parched), Sanguis Draxonis, Gypsum Fibrosum Preparatum, soil suppresses worm, Radix Angelicae Sinensis can carry out visual comparison's detection with standard sample; This several Chinese medicine does not generally adulterate problem, and only only have whether certified products, whether process is suitable, can only judge with general general knowledge;
Step 10: described inspection should meet pertinent regulations under " Chinese Pharmacopoeia " version in 2010 first annex emplastrum item;
Step 11: the mensuration of described extractum: get this medicament content 10g, accurately weighed, use 100% ethanol as solvent, measure according to the cold-maceration under ethanol-soluble extractives algoscopy item, must not 78% be less than.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.Constituent involved in the present invention and preparation method all can adjust accordingly according to embodiment, but can not exceed this invention protection domain.
Claims (3)
1. promote to it is characterized in that the Chinese medicine composition that bone injury affected part grows, be made up of the Chinese medicine of following weight ratio: Caulis Spatholobi 28-32 part, Flos Carthami 13-17 part, Radix Salviae Miltiorrhizae 28-32 part, Rhizoma Chuanxiong 18-22 part, Lignum Sappan 28-32 part, Pyritum 13-17 part, Borneolum Syntheticum 1-5 part, Radix Aconiti 13-17 part, Semen Strychni (parched) 8-12 part, Sanguis Draxonis 4-8 part, Gypsum Fibrosum Preparatum 28-32 part, Caulis Lonicerae 28-32 part, soil suppresses worm 13-17 part, Radix Angelicae Sinensis 18-22 part.
2. the Chinese medicine composition of promotion bone injury affected part according to claim 1 growth, it is characterized in that, the weight ratio of each Chinese medicine is specifically: Caulis Spatholobi 30 parts, 15 parts, Flos Carthami, Radix Salviae Miltiorrhizae 30 parts, Rhizoma Chuanxiong 20 parts, Lignum Sappan 30 parts, Pyritum 15 parts, Borneolum Syntheticum 3 parts, Radix Aconiti 15 parts, Semen Strychni (parched) 10 parts, Sanguis Draxonis 6 parts, Gypsum Fibrosum Preparatum 30 parts, Caulis Lonicerae 30 parts, soil suppresses worm 15 parts, Radix Angelicae Sinensis 20 parts, after it is pulverized and mixed, add the Olibanum 1 part being equivalent to the same weight ratio again, Myrrha 1 part, with aforesaid Caulis Spatholobi after being pulverized, Flos Carthami, Radix Salviae Miltiorrhizae, Rhizoma Chuanxiong, Lignum Sappan, Pyritum, Borneolum Syntheticum, Radix Aconiti, Semen Strychni (parched), Sanguis Draxonis, Gypsum Fibrosum Preparatum, Caulis Lonicerae, soil suppresses worm, the ground product mix homogeneously of Radix Angelicae Sinensis, add batching, be made into externally used paste.
3., for a quality determining method for the Chinese medicine composition of the promotion bone injury affected part growth described in claim 1 or 2, it is characterized in that:
It comprises respectively to the thin-layer chromatographic analysis determination step of Flos Carthami, Radix Salviae Miltiorrhizae, Lignum Sappan, Caulis Lonicerae;
1) be contrast Flos Carthami sample for reference substance to the thin-layer chromatographic analysis of Flos Carthami, be that the acetic ether-methanoic acid-water-methanol mixture of 7:2:3:0.4 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel H lamellae launches:
Get Flos Carthami 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Flos Carthami sample 0.5g, add 80% aqueous acetone solution 5ml, sealing of jumping a queue, jolting 15min, leave standstill, get supernatant medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel H lamellae, take volume ratio as the acetic ether-methanoic acid-water-methanol mixture of 7:2:3:0.4 be developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Flos Carthami sample in thin layer chromatography;
2) be contrast Radix Salviae Miltiorrhizae sample for reference substance to the thin-layer chromatographic analysis of Radix Salviae Miltiorrhizae, be that the petroleum ether-ethyl acetate of 60-90 DEG C of 4:1 is developing solvent with volume ratio, carry out thin layer chromatography check analysis with the thin layer chromatography that silica gel g thin-layer plate launches:
Get Radix Salviae Miltiorrhizae 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Radix Salviae Miltiorrhizae sample 1g, add diethyl ether 5ml, jolting, places 1 hour, filters, and after filtrate volatilization completely, residue adds ethyl acetate 1ml and dissolves, and gained solution is medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, volume ratio is the petroleum ether-ethyl acetate of 60-90 DEG C of 4:1 is developing solvent, launch, taking-up is dried, and test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Radix Salviae Miltiorrhizae sample in thin layer chromatography;
3) being contrast Lignum Sappan sample for reference substance to the thin-layer chromatographic analysis of Lignum Sappan, is that the chloroform-acetone-formic acid of 8:4:1 is developing solvent with volume ratio, with silica gel G F
254the thin layer chromatography that lamellae launches carries out thin layer chromatography check analysis:
Get Lignum Sappan 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get contrast Lignum Sappan sample 1g, add methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
According to thin layer chromatography test, draw need testing solution and each 3 μ l of control medicinal material solution, put respectively in same silica gel G F
254on lamellae, take volume ratio as the chloroform-acetone-formic acid of 8:4:1 be developing solvent, launch, taking-up is dried, put in exsiccator and place 12 hours, inspect under being placed on 254nm ultraviolet light, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Lignum Sappan sample in thin layer chromatography;
4) be contrast Caulis Lonicerae sample for reference substance to the thin-layer chromatographic analysis of Caulis Lonicerae, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 65:35:10 is placed below 10 DEG C be developing solvent, so that silica gel g thin-layer plate to launch, the tlc identification method being developer with 10% sulphuric acid ethanol:
Get Caulis Lonicerae 5g, put in apparatus,Soxhlet's, add ethanol 100ml, reflux 2 hours, extract 3 times altogether, merging filtrate, after evaporate to dryness, adds methanol 2ml and makes dissolving, as need testing solution;
Separately get Caulis Lonicerae control medicinal material 1g, add 50% methanol 10ml, supersound process 30min, filter, get filtrate medical material solution in contrast;
Test according to thin layer chromatography, draw need testing solution and each 10 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, take volume ratio as lower floor's solution that the chloroform-methanol-water mixtures of 65:35:10 is placed below 10 DEG C be developing solvent, launch, taking-up is dried, and sprays with 10% ethanol solution of sulfuric acid, be heated to colour developing at 105 DEG C clear, test sample demonstrates the different Development of Thin-Layer Chromatography situation for contrasting from contrast Caulis Lonicerae sample in thin layer chromatography;
And
5) detection of HPLC finger printing is carried out to Caulis Spatholobi, detect the effective peak area of Caulis Spatholobi used at 11 chromatographic fingerprint peaks place;
6) mensuration pipe is carried out to the ferulaic acid content in Rhizoma Chuanxiong, contrast with the content contrasted with sample ferulic acid;
7) X-ray diffraction finger printing carried out to Pyritum, analyze the quantity at total peak, contrast the raw product of Pyritum, forge the X-ray diffraction finger printing of product sample, judge that Pyritum is raw product, forges product or be mixed in proportion thing;
8) near infrared spectrum detection is carried out to Borneolum Syntheticum, itself and the Borneolum Syntheticum sample contrasted all are made into 100mg/ml, contrast with regard to itself and five the strongest peaks of sample in 4000-9000 wave number, do semi-quantitative analysis qualitatively simultaneously;
9) worm, Radix Angelicae Sinensis are suppressed for Radix Aconiti, Semen Strychni (parched), Sanguis Draxonis, Gypsum Fibrosum Preparatum, soil, itself and standard sample are carried out visual comparison's detection.
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| CN108169386A (en) * | 2018-03-15 | 2018-06-15 | 吉林修正药业新药开发有限公司 | A kind of construction method of the HPLC characteristic spectrums of neck waist recovering capsule |
| CN108310221A (en) * | 2018-05-03 | 2018-07-24 | 张双环 | A kind of plaster for treating fracture |
| CN108680673A (en) * | 2018-05-18 | 2018-10-19 | 南京中医药大学 | The method for building up and its finger-print of angelica sinensis finger-print |
| CN109374764A (en) * | 2018-10-19 | 2019-02-22 | 广西中医药大学 | Eight taste dragons of one kind bore particle HPLC finger-print and Contents of Main Components measuring method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108169386A (en) * | 2018-03-15 | 2018-06-15 | 吉林修正药业新药开发有限公司 | A kind of construction method of the HPLC characteristic spectrums of neck waist recovering capsule |
| CN108169386B (en) * | 2018-03-15 | 2020-09-18 | 吉林修正药业新药开发有限公司 | Method for constructing HPLC (high Performance liquid chromatography) characteristic spectrum of Jingyaokang capsule |
| CN108310221A (en) * | 2018-05-03 | 2018-07-24 | 张双环 | A kind of plaster for treating fracture |
| CN108680673A (en) * | 2018-05-18 | 2018-10-19 | 南京中医药大学 | The method for building up and its finger-print of angelica sinensis finger-print |
| CN109374764A (en) * | 2018-10-19 | 2019-02-22 | 广西中医药大学 | Eight taste dragons of one kind bore particle HPLC finger-print and Contents of Main Components measuring method |
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