Summary of the invention
Above-mentioned deficiency at prior art, the objective of the invention is existing compound Nanxing pain paste pharmaceutical composition is improved, a kind of new compound Nanxing pain paste compositions is provided, and the pharmaceutical pack capacity of said composition will increase exponentially, to strengthen the topical administration amount; And raising is strengthened medication effect to the penetration of skin layer; Also will reduce the zest of medicine significantly, obviously reduce clinical irritated rate, eliminate severe anaphylactic reaction; And solve the problem that viscosity problem and oleaginous base pollute skin and medicated clothing that sticks of plaster simultaneously.The present invention also will provide this compound Nanxing pain paste preparation of compositions method.
The technical scheme of finishing the foregoing invention task is: a kind of compound Nanxing pain paste and cataplasm combination is characterized in that the component and the weight of said composition crude drug are as follows:
Arisaema Cum Bile 260g~275g Radix Aconiti 190g~210g
Flos Caryophylli 160g~175g Cortex Cinnamomi 160g~175g
Radix Angelicae Dahuricae 285g~315g Herba Asari 120g~130g
Rhizoma Chuanxiong 190g~210g Radix Cynanchi Paniculati 120g~130g
Olibanum (system) 80g~86g Myrrha (processed) 80g~86g
Camphora 60g~66g Borneolum Syntheticum 60g~66g;
Simultaneously, the component of said composition adjuvant and weight are as follows:
NP-700 1.4kg~1.6kg dihydroxyaluminum aminoacetate 55g~65g
Tartaric acid 55g~65g EDTA 12g~18g
Glycerol 5.5kg~6.5kg polyvinylpolypyrrolidone 350g~374g
Purified water 10kg~12kg gelatin 90g~110g
Azone 90g~110g propylene glycol 190g~210g
Methyl salicylate 150g~170g
Make 1000 cataplasma ointment altogether.
The application recommends following optimized proportion, and the component and the weight of said composition crude drug are as follows:
Arisaema Cum Bile 267g Radix Aconiti 200g
Flos Caryophylli 167g Cortex Cinnamomi 167g
Radix Angelicae Dahuricae 300g Herba Asari 125g
Rhizoma Chuanxiong 200g Radix Cynanchi Paniculati 125g
Olibanum (system) 83g Myrrha (processed) 83g
Camphora 63g Borneolum Syntheticum 63g
Simultaneously, the component of said composition adjuvant and weight are as follows:
NP-700 1.5kg dihydroxyaluminum aminoacetate 60g
Tartaric acid 60g EDTA 15g
Glycerol 6kg polyvinylpolypyrrolidone 362g
Purified water 11kg gelatin 100g
Azone 100g propylene glycol 200g
Methyl salicylate 160g
Make 1000 cataplasma ointment altogether.
Further optimization of the present invention has following prioritization scheme,
In the said composition, described each crude drug component is meant:
Rhizoma Arisaematis, Radix Aconiti, the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum and Myrrha are respectively, the concentrated extract of the ethanol extract of Rhizoma Arisaematis, Radix Aconiti, the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum and Myrrha;
Camphora, Borneolum Syntheticum are the powder of Camphora, Borneolum Syntheticum.
More optimize and more particularly, described each crude drug component is meant:
Rhizoma Arisaematis, Radix Aconiti, the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum and Myrrha are measured 85% alcohol reflux 2 times with 6 times, each 1.5 hours, filter, merging filtrate, recovery ethanol to density is about 1.05~1.15 extractum (50 ℃); Camphora, Borneolum Syntheticum are: pulverize, cross the powder of 80 mesh sieves.
The preparation method of above compound Nanxing pain paste and cataplasm combination is: crude drug and adjuvant are got the raw materials ready according to aforementioned proportion respectively, it is characterized in that, step is as follows:
Rhizoma Arisaematis, Radix Aconiti, the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum, Myrrha ten flavor medical materials are measured 85% alcohol reflux 2 times, each 1.5 hours with 6 times;
Backflow filters, and merging filtrate reclaims ethanol, is about 1.05~1.15 (50 ℃) to extractum density, and is standby;
Camphora, Borneolum Syntheticum powder are broken to 80 orders, sieve, and be standby;
Dihydroxyaluminum aminoacetate is added in the glycerol, and fully mixing adds above-mentioned extractum, EDTA, polyvinylpolypyrrolidone, azone, propylene glycol, Camphora, Borneolum Syntheticum, methyl salicylate respectively, continues to stir;
Put into NP-700, stir;
Tartaric acid, gelatin are put into the glycerol phase after dissolving with purified water, continuously evacuation stir about 30min;
Take out mastic, coating, covering adherent layer, section, promptly.
The purified water consumption is 11kg in the above step.
According to traditional theory, improve the curative effect of medicine or reduce toxicity, side effect, mainly should consider to change the component and the part by weight of crude drug.Change technology, adjuvant (substrate) effect therein are very accessory; Only change technology, adjuvant and can't reach the curative effect of improving medicine or the purpose that reduces toxicity, side effect.And the present invention has overcome above-mentioned traditional prejudice, the solution of the present invention is on the basis that absorbs conventional pastes YAOJING China, the component and the part by weight that keep the crude drug of compound Nanxing pain paste rubber-emplastrum in the prior art, utilize modern patch technology and promote transdermal drag delivery, overcome the many shortcoming of traditional plaster in matrix formulations, preparation technology, clinical efficacy, use, have that drug loading is big, water conservation, preserve moisture, good permeability, transdermal characteristic are good, repeat noticing property good, can use repeatedly; Not having side effect such as irritated, non-stimulated, is a kind of ideal Chinese medicine for external application.Concrete advantage has:
1, contains dose bigger (40%): about 2 to 5 millimeters of the stromal thickness of cataplasma, and general ventilative rubber plaster catablasm base material and medicine layer 0.1 millimeter thickness only, compare the substrate that increased more than tens of times and the thickness of medicine layer, the appearance dose is increased exponentially, strengthen the topical administration amount, strengthened medication effect.(compound Nanxing pain paste prescription drug total amount is the 1843g/1000 sheet)
2, transdermal effect is stronger: contain water-soluble macromolecule in the crust cloth cream and help as substrate and ooze promoter, greatly strengthened transdermal effect, realized the strong osmotic to skin layer.
3, breathability is better: adopt the large-mesh non-woven fabrics, permeability is better.
4, can not produce stimulation: crust cloth cream base matter uniqueness does not produce stimulation to skin.And rubber plaster sticks for a long time owing to adopt rubber mass, and skin is produced to stimulate, and scratchy even erythema etc. occur.
5, can take off subsides repeatedly: it is strong and immutable that the common drawback of general external use plaster is that viscosity is crossed, and in a single day takes off during use, and easy adhesive tape chaeta not only also is difficult to keep and recovers viscosity and continue to use.And the substrate of cataplasma can be regulated viscosity as required, and finished product can be taken off subsides in use repeatedly, can the adhesive tape chaeta, and be to paste with comfortable exterior-applied formulation.
Below be a large amount of contrast test data of the technology of the present invention effect.
One, prescription
Arisaema Cum Bile 267g Radix Aconiti 200g
Flos Caryophylli 167g Cortex Cinnamomi 167g
Radix Angelicae Dahuricae 300g Herba Asari 125g
Rhizoma Chuanxiong 200g Radix Cynanchi Paniculati 125g
Olibanum (system) 83g Myrrha (processed) 83g
Camphora 63g Borneolum Syntheticum 63g
Substrate:
NP-700 1.5kg dihydroxyaluminum aminoacetate 60g
Tartaric acid 60g EDTA 15g
Glycerol 6kg polyvinylpolypyrrolidone 362g
Purified water 11kg gelatin 100g
Azone 100g propylene glycol 200g
Methyl salicylate 160g makes 1000 altogether
Two, medicinal material extract technology
For monarch drug Rhizoma Arisaematis, Radix Aconiti, contained alkaloid is the main effective ingredient in analgesic, at present resulting information spinner will be with the composition of aconitine for can survey index in the document, and the composition of aconitine has the analgesic effect really, therefore the inventor determines with the aconitine tentatively to be carried out Preliminary screening with sour water (with hydrochloric acid adjusting pH value to 1.5), aqueous alkali (with sodium hydroxide adjusting pH value to 9), low-concentration ethanol (30% ethanol), four kinds of extraction schemes of high concentration ethanol (85% ethanol) by surveying index components according to document.Concrete grammar is for getting the crude drug (Rhizoma Arisaematis 26.7g, Radix Aconiti 20g) of 1/10th recipe quantities, extract 8 times of amounts of solvent with above-mentioned four kinds respectively and carry out reflux, extract,, 1.5 hours extraction times, extraction time 3 times, difference merge extractive liquid,, high speed centrifugation (5000r/m), get supernatant concentration to 200ml, get 1ml high speed centrifugation (12000r/m) respectively, filter, enter high performance liquid chromatograph analysis with the 0.45um microporous filter membrane.
Chromatographic condition: Agilent 1200 high performance liquid chromatographs
Chromatographic column: Dalian Yi Lite Hypersil BDS C18,4.6 * 250mm, 5 μ m
Mobile phase: methanol: water: chloroform: diethylamine (80: 38: 2: 0.1)
Detect wavelength: 232nm
Column temperature: 35 ℃
Flow velocity: 0.8ml/min
Sample size: 20 μ l.
Test sample is handled: get 1ml sample concentration liquid respectively and be loaded on centrifuge tube, high speed centrifugation (12000r/min) 5min gets supernatant and filters the back direct injected with 0.45 μ m microporous filter membrane.
The reference substance preparation: precision takes by weighing aconitine 4.94mg, mesaconitine 5.04mg, hypaconitine 5.04mg, and is in same 100ml volumetric flask, standby with methanol constant volume.
Result: through high-efficient liquid phase analysis, though the impurity peaks that occurs is more, characteristic peak has been produced considerable influence, but can find that total aconitine is relative higher with yield in the high concentration alcohol extraction at acid extraction, compared with other two kinds of extracting method and have explicitly difference.Therefore the inventor extracts solvents to these two kinds and carries out orthogonal test more respectively, the parameter that makes extraction process more reasonable economization in large-scale production process from now on.
1, Rhizoma Arisaematis, Radix Aconiti acid extraction orthogonal test
Get Rhizoma Arisaematis 26.7g, Radix Aconiti 20g (1/10th recipe quantities), parallelly take by weighing 9 parts and launch test by design in the following orthogonal table
Attached quadrature factor level table:
Factor level |
Sour water (PH) |
Extraction time (h) |
Extraction time |
The solvent multiple |
1 |
1.0 |
1 |
1 |
4 |
2 |
1.5 |
1.5 |
2 |
6 |
3 |
2. |
2.5 |
3 |
8 |
Chromatographic condition:
Chromatographic condition: Agilent 1200 high performance liquid chromatographs
Chromatographic column: Dalian Yi Lite Hypersil BDS C18,4.6 * 250mm, 5 μ m
Mobile phase: methanol: water: chloroform: diethylamine (65: 38: 2: 0.1)
Detect wavelength: 232nm
Column temperature: 35 ℃
Flow velocity: 0.8ml/min
Sample size: 20 μ l.
Test sample is handled: above-mentioned 9 kinds of acid water extracting liquids are got supernatant after centrifugal heat respectively, being concentrated into volume is 200ml, precision is measured 20ml, pours in the separatory funnel after being diluted with water to 40ml, carries out 3 extractions with chloroform 50ml, 50ml, 50ml, the combined chloroform layer, with the natrium carbonicum calcinatum dehydration, filter the reclaim under reduced pressure evaporate to dryness, be settled in the 5ml volumetric flask with dichloromethane, liquid phase is standby.
The reference substance preparation: precision takes by weighing mesaconitine 3.64mg, aconitine 3.88mg, hypaconitine 3.66mg reference substance, is settled in the 25ml volumetric flask with dichloromethane.Precision is measured 1.5ml and is dissolved in the 5ml dichloromethane, is made into concentration and is followed successively by 0.04368mg/ml, 0.04656mg/ml, 0.04392mg/ml.Through the quadrature statistical analysis, the result is as follows for experimental data (concentration is calculated gained for being settled to the 5ml volumetric flask in the table):
Interpretation of result:
By the intuitive analysis of R value and The results of analysis of variance as can be known: each factor is A>C>D>B to the size order that influences that extracts, the A factor is first factor, the C factor is second factor, the D factor is the 3rd factor, the B factor is the 4th factor, but there was no significant difference between the R value of each factor, and it is less to influence each other, table is arranged as seen, optimal level is A
1B
2C
3D
3Consider cost and other factors, the inventor has done further screening to extraction conditions.It is big more to extract solvent acidity, and extraction effect is good more, so the ph value is to select 1 to be advisable; Extraction time is best with 1.5h; Best on the extraction time with 3 times, but 2 extraction effects are more or less the same with it; Best with 8 times of dose-effects fruits on the extraction multiple, comprehensive above each side factor has finally been selected following excellent level: A
1B
2C
2D
3
2, Rhizoma Arisaematis, Radix Aconiti alcohol extraction orthogonal test
Get Rhizoma Arisaematis 26.7g, Radix Aconiti 20g (1/10th recipe quantities), parallelly take by weighing 9 parts by design test in the following orthogonal table, concrete condition is as follows:
Attached quadrature factor level table:
Factor level |
Concentration of alcohol (%) |
Extraction time (h) |
Extraction time |
The solvent multiple |
1 |
75 |
1 |
1 |
4 |
2 |
85 |
1.5 |
2 |
6 |
Chromatographic condition: the same.
Sample treatment: above-mentioned 9 kinds of extracting solution are carried out decompression recycling ethanol respectively, being concentrated into volume is 100ml, precision is poured in the separatory funnel after measuring 25ml, carry out 3 extractions with chloroform 30ml, 30ml, 30ml, the combined chloroform layer dewaters with natrium carbonicum calcinatum, filter, the reclaim under reduced pressure evaporate to dryness is settled in the 5ml volumetric flask with dichloromethane, and liquid phase is standby.In extraction process, find, because still there is part ethanol, consider that this solvent both had been dissolved in chloroform and has also had in the water of being partially soluble in, so remaining solution concentration is to 25ml, after getting 5ml and being diluted with water to 10ml, with chloroform 15,15,15ml extraction 3 times, collect chloroform solution, dewater with natrium carbonicum calcinatum, filter, gained chloroform solution reclaim under reduced pressure is to evaporate to dryness, and the cooling back is settled to the 5ml volumetric flask with dichloromethane, and liquid phase is standby.
The reference substance preparation: precision takes by weighing mesaconitine 3.64mg, aconitine 3.88mg, hypaconitine 3.66mg reference substance, is settled in the 25ml volumetric flask with dichloromethane.Get precision and measure 1.5ml and be dissolved in the 5ml dichloromethane, be made into concentration and be followed successively by 0.04368mg/ml, 0.04656mg/ml, 0.04392mg/ml.
Behind positive statistical analysis, the result is as follows for experimental data (concentration is calculated gained for being settled to the 5ml volumetric flask in the table):
Interpretation of result:
By the intuitive analysis of R value and The results of analysis of variance as can be known: each factor is B>D>A>C to the size order that influences that extracts, and the B factor has significant differences, and D also has significant difference.By table as seen, optimal level is A
3B
1C
2D
2Consider cost and other factors, we have done further screening to extraction conditions.Best behind the analysis-by-synthesis between three levels of extraction solvent with 95% alcohol extraction effect, but being more or less the same with it, 85% alcohol extraction select the latter from still the consideration of safety aspect is all more suitable economically; On extraction time, best with 1h; Best on the extraction time with 2 times, but diversity is little; Best on the extraction multiple with 6 times of amounts, so final optimum level is A
2B
1C
2D
2
3, two kinds of medical material orthogonal experiments are analyzed and are discussed
In the comparison of high concentration alcohol extraction and two kinds of technologies of acid extraction, from the extraction yield of total aconitine, acid extraction is than the summary height of high concentration alcohol extraction.But the inventor finds the extracting solution of high concentration alcohol extraction gained and relatively clarifies, starch and mucilaginous amount are many in the sour water gained extracting solution, it is centrifugal to cause extracting solution to be difficult to, and places and still have precipitation to separate out after spending the night, and may produce considerable influence for from now on production.
4, eight flavor medical material orthogonal experiments:
The medical material amount of getting (prescription 1/10th): Radix Angelicae Dahuricae 30g, Rhizoma Chuanxiong 20g, Cortex Cinnamomi 16.7g, Flos Caryophylli 16.7g, Herba Asari 12.5g, Radix Cynanchi Paniculati 12.5g, Olibanum 8.3g, Myrrha 8.3g gross weight: 125g
Attached quadrature factor level table:
Factor level |
Concentration of alcohol (%) |
Extraction time (h/ time) |
Extraction time |
The solvent multiple |
1 |
65 |
1 |
1 |
6 |
2 |
75 |
1.5 |
2 |
8 |
3 |
85 |
2.5 |
3 |
10 |
Chromatographic condition: Agilent 1200 high performance liquid chromatographs
Chromatographic column: phenomenex. C18,4.6 * 150mm, 5 μ m
Mobile phase: methanol: water: phosphoric acid (50: 50: 0.2)
Detect wavelength: 276nm
Column temperature: 30 ℃
Flow velocity: 1.0ml/min
Sample size: 5 μ l
The test liquid preparation: with above-mentioned 9 kinds of extracting solution difference decompression recycling ethanol, being concentrated into volume is 100ml, and precision is measured 1ml, be settled to 5ml with 50% ethanol, get wherein sample segment and be positioned over high speed centrifugation (12000r/min) 5min in the centrifuge tube, running finishes to get supernatant, and liquid phase is standby.
The reference substance preparation: get paeonol, eugenol reference substance, being mixed with concentration respectively with methanol is 0.019mg/ml, 0.221mg/ml.
(1) through the quadrature statistical analysis, the result is as follows for paeonol experimental data (concentration is calculated gained for being settled to the 5ml volumetric flask in the table):
By the intuitive analysis of R value and The results of analysis of variance as can be known: each factor is A>D>C>B to the size order that influences that extracts, and between three levels of A factor significant difference is arranged, and other factor significances are little, from the final excellent horizontal A of data analysis
3B
3C
3D
1Adopt 85% ethanol extraction effect best as seen from the table; Adopt on extraction time 2.5 hours best, be more or less the same with it but extracted in 1.5 hours, should select for use 1.5 hours.Same extraction time select for use 2 times best; Extract the solvent load aspect, consider extraction effect and cost problem, adopt six times of amounts more suitable, the finally selected excellent level of comprehensive above each side factor is A
3B
2C
2D
1
(2) through the quadrature statistical analysis, the result is as follows for eugenol experimental data (concentration is calculated gained for being settled to the 5ml volumetric flask in the table):
By the intuitive analysis of R value and The results of analysis of variance as can be known: each factor is A>D>C>B to the size order that influences that extracts, and between three levels of A factor significant difference is arranged, and diversity is less between other factors, data intuitive analysis from table, and optimal level is A
3B
3C
3D
1Extracting solvent adopts 85% ethanol effect best; Adopt on extraction time 2.5 hours best, be more or less the same with it but extracted in 1.5 hours, should select for use 1.5 hours; Same extraction time should be selected for use 2 times; Extract the solvent load aspect, consider the active constituent content extraction rate and in conjunction with cost, adopt 6 times of amounts more suitable, the finally selected excellent level of comprehensive above each side factor is A
3B
2C
2D
1
Quadrature is summed up:
Following extraction scheme has finally been selected in the extraction of eight flavor medical materials: the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum, Myrrha are measured 85% alcohol reflux 2 times with 6 times, each 1.5 hours, collect and extract medicinal liquid, decompression recycling ethanol to concentrated solution density is (50 ℃) about 1.05-1.15.
So the alcohol extraction of two flavor medical materials considers that the selected excellent level of cost problem is A
2B
1C
1D
1, but according to data analysis, the inventor also can consider to carry out according to eight flavor medicinal material extract methods.Whether above-mentioned ten kinds of medical materials are considered to mix is together carried out alcohol extraction: ten flavor medical materials are measured 85% alcohol reflux 2 times with 6 times, and each 1.5 hours, to collect and extract medicinal liquid, decompression recycling ethanol to concentrated solution density is (50 ℃) about 1.05-1.15.
Comprehensive above quadrature result, Rhizoma Arisaematis, Radix Aconiti, the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum, Myrrha ten flavor medical materials are measured 85% alcohol reflux 2 times with 6 times, each 1.5 hours, filter, merging filtrate, reclaim ethanol to extractum density and be (50 ℃) about 1.05-1.15, standby; Can take into account the proposition of effective ingredient of ten flavor principal agents, again can energy savings, resource and enhance productivity.
We have selected following several extraction assembled schemes to make Transdermal absorption and the pharmacological testing sample reasonability with checking technology:
1, full extractum (ten flavor medical material alcohol-extracted extracts):
Medical material is handled: above Rhizoma Arisaematis, Radix Aconiti, the Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati, Olibanum, Myrrha ten flavor medical materials are measured 85% alcohol reflux 2 times with 6 times, each 1.5 hours, filter, merging filtrate, reclaim ethanol to extractum density and be (50 ℃) about 1.05-1.15, standby; Camphora, Borneolum Syntheticum powder are broken to 80 orders, sieve, and be standby.
Forming processes: dihydroxyaluminum aminoacetate is added in the glycerol, and fully mixing adds above-mentioned extractum, EDTA, polyvinylpolypyrrolidone, propylene glycol, Camphora, Borneolum Syntheticum, methyl salicylate respectively, continues to stir, and puts into NP-700 at last, stirs.Tartaric acid, gelatin are put into the glycerol phase after dissolving with purified water, behind the evacuation stir about 30min, take out, are coated with, cover adherent layer, section, promptly continuously.
In forming process, add the cutaneous permeable agent azone by this technology, prepare a duplicate samples and test.
2, powder, four Chinese medicine material extractum, volatile oil clathrate compound
Medical material is handled: Rhizoma Arisaematis, Radix Aconiti pulverize at low temperature, cross 120 mesh sieves, and standby.The Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Cortex Cinnamomi, Herba Asari, Flos Caryophylli, Radix Cynanchi Paniculati carry out CO
2The supercritical fluid extraction volatile ingredient, and volatile ingredient carried out cyclodextrin inclusion compound, 50~60 ℃ of enclose temperature, the time is 4~5h, the gained clathrate is standby.The Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Olibanum, Myrrha 80% ethanol extraction reclaims ethanol, and extractum is condensed into density and is 1.10~1.2 (25 ℃), and is standby.
Forming processes: dihydroxyaluminum aminoacetate is added in the glycerol, and fully mixing adds above-mentioned medicated powder and extractum, EDTA, polyvinylpolypyrrolidone, azone, propylene glycol, Camphora, Borneolum Syntheticum, methyl salicylate respectively, continues to stir, and puts into NP-700 at last, stirs.Tartaric acid, gelatin are put into the glycerol phase after dissolving with purified water, behind the evacuation stir about 30min, take out, are coated with, cover adherent layer, section, promptly continuously.
3, medicated powder and eight kinds of medicinal material extract extractum
Medical material is handled: Rhizoma Arisaematis, Radix Aconiti are pulverized, and cross 120 mesh sieves, and be standby.The Radix Angelicae Dahuricae, Rhizoma Chuanxiong, Herba Asari, Radix Cynanchi Paniculati, Flos Caryophylli, Cortex Cinnamomi, Olibanum, Myrrha eight flavor medical materials are with 85% alcohol reflux 2 times, each 1.5h, and recovery ethanol, extractum is condensed into density and is 1.10~1.20 (25 ℃), and is standby.
Forming processes: dihydroxyaluminum aminoacetate is added in the glycerol, and fully mixing adds above-mentioned medicated powder and extractum, EDTA, polyvinylpolypyrrolidone, propylene glycol, Camphora, Borneolum Syntheticum, methyl salicylate respectively, continues to stir, and puts into NP-700 at last, stirs.Tartaric acid, gelatin are put into the glycerol phase after dissolving with purified water, behind the evacuation stir about 30min, take out, are coated with, cover adherent layer, section, promptly continuously.
In forming process, add the cutaneous permeable agent azone by this technology, prepare a duplicate samples and test.
Three, Transdermal absorption experiment
Experimental apparatus: TP-5 intelligence Transdermal absorption instrument
Test method: with fixing the giving coyote hole and accept between the pond of diffusion cell of big Corium Mus, the cataplasma sample is affixed on the horny layer surface of isolated skin, makes it to contact closely with skin.Corium one side contacts with acceptable solution, the emptying bubble.Bath temperature is (37 ± 0.5) ℃, and rotating speed is 300r/min, accepts that cell body is long-pending to be 15.5ml, is sampled as 1ml at every turn.It is as follows that acceptable solution is joined by institute:
Acceptable solution one: contain 40% alcoholic acid normal saline;
Two: 0.9% normal saline of acceptable solution;
Three: 0.9% normal saline of acceptable solution: 10% PEG400=1: 1 (containing 40% ethanol);
Four: 0.9% normal saline of acceptable solution: 10% PEG400=1: 1
Acceptable solution |
Sampling amount g |
Peak area |
See through rate of transform % |
Acceptable solution one |
0.3307 |
161063 |
44.8 |
Acceptable solution two |
0.2573 |
62928.5 |
24.9 |
Acceptable solution three |
0.3074 |
341635.81 |
58.41 |
Acceptable solution four |
0.2769 |
145604 |
24.27 |
The result: above-mentioned four kinds of acceptable solutions through prerun relatively, found that 40% ethanol and 0.9% normal saline: two kinds of acceptable solution acceptable effects of 10% PEG400=1: 1 (containing 40% ethanol) are better, take the thickness similarity of imitation human body fluid into consideration, select 0.9% normal saline for use: 10% PEG400=contain 40% ethanol this acceptable solution test at 1: 1.
The inventor adopts the cataplasma of five kinds of different supplementary material proportionings to carry out the transmitance test altogether, respectively sampling in 1,3,5,8,10,12,24 hour, as investigating index, measure its medicine transmitance with high-efficient liquid phase technique with paeonol and methyl salicylate, it is as follows to measure chromatographic condition:
Mobile phase: methanol: water: phosphoric acid (50: 50: 0.2), the detection wavelength is 276nm, and column temperature is 30 ℃, and flow velocity is 1.0ml/min.
The Transdermal absorption test data gathers
(0.9% normal saline: 10% PEG400=1: 1 contains 40% ethanol) |
Peak area (dpf) |
Content (dpf) μ g/g |
See through rate of transform % |
Peak area (sysjz) |
Content (sysjz) mg/g |
See through rate of transform % |
Finished product 1h (dz070330) |
79506.50 |
43.52 |
12.43 |
248389.50 |
1.81 |
5.23 |
Finished product 3h (dz070329) |
140503.41 |
76.91 |
21.97 |
504080.47 |
3.67 |
10.62 |
Finished product 5h (dz070327) |
219591.77 |
120.20 |
34.34 |
793871.31 |
5.79 |
16.72 |
Finished product 8h (dz070327) |
256536.45 |
140.43 |
40.12 |
1023126.81 |
7.46 |
21.55 |
Finished product 10h (dz070327) |
288675.88 |
158.02 |
45.15 |
1125219.00 |
8.20 |
23.70 |
Finished product 12h (dz070326) |
322797.84 |
176.70 |
50.49 |
1292270.38 |
9.42 |
27.22 |
Finished product 24h (dz070326) |
464916.84 |
254.49 |
72.71 |
1738799.38 |
12.67 |
36.63 |
Full extractum 1h (dz070330) |
17957.51 |
9.68 |
2.76 |
32462.10 |
0.23 |
0.67 |
Full extractum 3h (dz070329) |
23768.10 |
13.05 |
3.73 |
68799.60 |
0.50 |
1.44 |
Full extractum 5h (dz070329) |
37533.71 |
20.61 |
5.89 |
51558.20 |
0.37 |
1.08 |
Full extractum 8h (dz070327) |
45485.84 |
24.84 |
7.10 |
84136.55 |
0.59 |
1.72 |
Full extractum 10h (dz070327) |
50498.72 |
27.57 |
7.88 |
60077.40 |
0.42 |
1.23 |
Full extractum 12h (dz070326) |
60280.30 |
32.87 |
9.39 |
46853.90 |
0.34 |
0.97 |
Full extractum 24h (dz070326) |
60204.24 |
32.82 |
9.38 |
54912.64 |
0.39 |
1.14 |
Full extractum N1h (dz070329) |
31275.40 |
18.59 |
5.31 |
171075.70 |
1.34 |
3.89 |
Full extractum N3h (dz070329) |
63456.55 |
37.73 |
10.78 |
209295.33 |
1.64 |
4.75 |
Full extractum N5h (dz070329) |
71536.66 |
42.53 |
12.15 |
238117.09 |
1.87 |
5.41 |
Full extractum N8h (dz070327) |
92965.46 |
54.97 |
15.71 |
324541.78 |
2.48 |
7.17 |
Full extractum N10h (dz070327) |
99854.10 |
59.04 |
16.87 |
319829.63 |
2.44 |
7.06 |
Full extractum N12h (dz070326) |
111086.98 |
65.59 |
18.74 |
336836.91 |
2.62 |
7.56 |
Full extractum N24h (dz070326) |
146964.27 |
86.77 |
24.79 |
368570.06 |
2.86 |
8.28 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 1h |
17134.91 |
10.80 |
3.09 |
77032.20 |
0.65 |
1.87 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 3h |
30245.60 |
19.42 |
5.55 |
100848.50 |
0.79 |
2.29 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 5h |
43968.71 |
28.08 |
8.02 |
115133.20 |
0.88 |
2.54 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 8h |
56323.87 |
35.98 |
10.28 |
120087.46 |
0.92 |
2.65 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 10h |
63450.60 |
40.53 |
11.58 |
113228.30 |
0.87 |
2.50 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 12h |
86865.54 |
55.48 |
15.85 |
148019.30 |
1.13 |
3.27 |
Powder and four Chinese medicine material extractum, volatile oil clathrate compound 24h |
91588.50 |
58.41 |
16.69 |
118163.40 |
0.92 |
2.65 |
Powder and eight flavor medical material extractum 1h |
38380.41 |
22.39 |
6.40 |
114705.70 |
0.88 |
2.56 |
Powder and eight flavor medical material extractum 3h |
66842.35 |
38.99 |
11.14 |
159965.67 |
1.23 |
3.57 |
Powder and eight flavor medical material extractum 5h |
120821.11 |
70.47 |
20.14 |
215473.59 |
1.66 |
4.80 |
Powder and eight flavor medical material extractum 8h |
180510.63 |
104.72 |
29.92 |
292596.13 |
2.19 |
6.34 |
Powder and eight flavor medical material extractum 10h |
192796.77 |
111.85 |
31.96 |
300588.81 |
2.25 |
6.51 |
Powder and eight flavor medical material extractum 12h |
239846.03 |
138.93 |
39.70 |
335984.75 |
2.56 |
7.40 |
Powder and eight flavor medical material extractum 24h |
341635.81 |
198.20 |
56.63 |
404291.41 |
3.03 |
8.76 |
Powder and eight flavor medical material extractum N1h |
46483.50 |
27.51 |
7.86 |
87753.40 |
0.69 |
1.98 |
Powder and eight flavor medical material extractum N3h |
53131.18 |
31.27 |
8.93 |
199489.19 |
1.52 |
4.38 |
Powder and eight flavor medical material extractum N5h |
81120.43 |
48.00 |
13.72 |
213100.50 |
1.67 |
4.82 |
Powder and eight flavor medical material extractum N8h |
92821.78 |
54.63 |
15.61 |
255207.64 |
1.94 |
5.61 |
Powder and eight flavor medical material extractum N10h |
92701.38 |
54.56 |
15.59 |
249096.61 |
1.89 |
5.48 |
Powder and eight flavor medical material extractum N12h |
106043.06 |
62.32 |
17.81 |
281361.63 |
2.18 |
6.29 |
Powder and eight flavor medical material extractum N24h |
147114.36 |
86.46 |
24.70 |
296011.78 |
2.29 |
6.62 |
Four, the compound Nanxing pain paste and cataplasm test of pesticide effectiveness---Different Preparation anti-inflammatory analgesic effect relatively
1. experiment purpose
Observe of the influence of compound Nanxing pain paste of the present invention (cataplasma) Different Preparation to swelling of rat toes and tenderness.
2. experiment material
2.1 medicine:
2.1.1 be subjected to reagent:
(1) compound Nanxing pain paste: lot number: 051217, provide by Jiangsu Nanxing Pharmaceutical Co., Ltd; (being equivalent to contain 1.84 gram crude drug/sheets).
(2) compound Nanxing pain paste (cataplasma): numbering: 070501, provide by Jiangsu Nanxing Pharmaceutical Co., Ltd; (be equivalent to contain 1.84g crude drug/sheet; Medicated powder and eight kinds of medicinal material extract extractum prescriptions).
(3) compound Nanxing pain paste (cataplasma): numbering: 070502, provide by Jiangsu Nanxing Pharmaceutical Co., Ltd; (be equivalent to contain 1.84g crude drug/sheet; Medicated powder and eight kinds of medicinal material extract extractum prescriptions add 1% azone in the substrate simultaneously).
(4) compound Nanxing pain paste (cataplasma): numbering: 070503, provide by Jiangsu Nanxing Pharmaceutical Co., Ltd; (be equivalent to contain 1.84g crude drug/sheet; Full extractum prescription).
(5) compound Nanxing pain paste (cataplasma): numbering: 070504, provide by Jiangsu Nanxing Pharmaceutical Co., Ltd; (be equivalent to contain 1.84g crude drug/sheet; Full extractum prescription adds 1% azone in the substrate simultaneously).
(6) indometacin enteric-coated tablet (indometacin): the 25mg/ sheet, lot number: 0609018, produce by Jiangsu Yabang Aipusen Pharmaceutical Co., Ltd..
2.1.2 reagent: trypsin: the 25g/ bottle, lot number: 1837A33 is produced by Amresco company
2.2 instrument: YLS-3E electronics tenderness instrument is produced by medical science equipment station, Shandong
YLS-7A toes volumetric measurement instrument is produced by medical science equipment station, Shandong
2.3 laboratory animal: the male 140-170g of SD rat
Provide by Nanjing University of Traditional Chinese Medicine's Experimental Animal Center
3. experiment is divided into groups and dosage:
Experiment is divided into 6 of blank groups, and all the other each groups are 12.
Model group: externally applied plaster substrate
Positive controls: indometacin enteric-coated tablet 6.75mg/kg is made into 0.675mg/ml solution with distilled water.
Compound Nanxing pain paste group: 0.1656g crude drug/kg
Compound Nanxing pain paste (cataplasma) medicated powder group: 0.1803g crude drug/kg
Compound Nanxing pain paste (cataplasma) medicated powder group (N): 0.1803g crude drug/kg
Compound Nanxing pain paste (cataplasma) extractum group: 0.1803g crude drug/kg
Compound Nanxing pain paste (cataplasma) extractum group (N): 0.1803g crude drug/kg
4. method:
With the rat sub-cage rearing, 6 in every cage, adaptability was fed three days.Cause normal pain threshold of the right back foot of scorching before measurement and toes volume, and after being divided into eight groups by pain threshold and toes volume at random by above-mentioned grouping, removing blank group does not inject and causes inflammation, all the other each groups are brought out local inflammation at the right back sufficient plantar subcutaneous injection 0.25% trypsin 0.1mL of every Mus and are taken place, be subjected to cause the stickup administration of scorching position with the adhesive plaster winding immediately after the injection of reagent group, positive group gastric infusion, respectively at causing scorching back 15min, 30min, 1h, the 1.5h the (the 3rd, 4 batches of rats are in 15min to inflammation, 30min, 1h, 1.5h, 2h, 3h, 4h, 6h) utilize YLS-3E type electronics tenderness instrument rat inflammatory foot to measure pain threshold, the weight of being given with shouting of causing of pain behind the rat inflammatory foot pressurized or when struggling is as pain threshold, and 1h behind the Yu Zhiyan, 2h, 3h, 4h, 6h measures toes volume calculations swelling rate and inhibitory rate of intumesce with YLS-7A toes volumetric measurement instrument.Computational methods are as follows:
5. experimental result:
5.1 the compound Nanxing pain paste different dosage form causes the influence of rat toes tenderness to pancreatin: the pain threshold of indometacin group, compound recipe compound Nanxing pain paste group and the remarkable rising 15min of extractum (N) group, 30min, 1h, 1.5h, 2h, 3h, 4h, significantly the raise pain threshold of 30min, 1h, 3h, 4h of medicated powder group, the pain threshold of the remarkable rising 30min of medicated powder (N) group, 1h, 1.5h, 2h, then significantly the raise pain threshold of 30min, 1.5h, 3h of extractum group relatively has significant difference (table 1) with model group.Indometacin group, compound recipe compound Nanxing pain paste group and extractum (N) group significantly increase the tenderness difference of 15min, 30min, 1h, 1.5h, 2h, 3h, the medicated powder group significantly increases the tenderness difference of 15min, 30min, 1h, 3h, medicated powder (N) group significantly increases the tenderness difference of 30min, 1h, 1.5h, 2h, then significantly the raise tenderness difference of 30min, 3h of extractum group relatively has significant difference (table 2) with model group.
Table 1. pair pancreatin causes the influence (X ± SD) of rat toes tenderness
With the model control group ratio,
*P<0.05;
*P<0.01 (down together)
Table 2 pair pancreatin causes the influence (X ± SD) of rat toes tenderness difference
5.2 the compound Nanxing pain paste different dosage form causes the influence of rat toes swelling to pancreatin: compare with the blank group, model group toes volume significantly increases.Compare with model group, the indometacin group significantly reduces at 3h toes volume, and compound recipe compound Nanxing pain paste group, extractum (N) group, medicated powder (N) group significantly reduce at 2h, 3h toes volume, and medicated powder (N) group also significantly reduces (table 3) at 4h, 6h toes volume.Compare with model group, the indometacin group significantly reduces in 2h, 3h, 4h, 6h toes difference, compound recipe compound Nanxing pain paste group and extractum (N) group significantly reduces in 2h, 3h, 4h toes difference, medicated powder (N) group significantly reduces in 1h, 3h, 4h, 6h toe difference, and respectively organizes toes swelling rate and also significantly reduce (table 4, table 5) than model group in the corresponding time period.
Table 3 pair rat pancreatin causes the volumetrical influence of toes (X ± SD)
Table 4 pair rat pancreatin causes the influence (X ± SD) of toes volume difference
Table 5 pair rat pancreatin causes the influence (X ± SD) of toes swelling rate
With the model control group ratio,
*P<0.05;
*P<0.01
6. conclusion:
Compound Nanxing pain paste, compound Nanxing pain paste (cataplasma) Different Preparation be rat toes swelling pain threshold value due in various degree the raising trypsin all, alleviates toes swelling rate due to the trypsin, has certain anti-inflammatory analgesic effect.Wherein for analgesic effect and, compound Nanxing pain paste group, compound Nanxing pain paste (cataplasma) extractum group (N), compound Nanxing pain paste (cataplasma) medicated powder group (N) effect are quite; Compound Nanxing pain paste (cataplasma) medicated powder group and compound Nanxing pain paste (cataplasma) extractum group are slightly poor.Be more or less the same for antiinflammatory action compound Nanxing pain paste group, compound Nanxing pain paste (cataplasma) extractum group (N), compound Nanxing pain paste (cataplasma) medicated powder group (N) effect; Compound Nanxing pain paste (cataplasma) extractum group and compound Nanxing pain paste (cataplasma) medicated powder group are not then seen the obvious anti-inflammatory and anti effect.Increase the effect of cutaneous permeable agent (N) anti-inflammatory analgesic and all be better than not adding group.