CN105983021A - External-use medicinal composition as well as preparation method and applications thereof - Google Patents
External-use medicinal composition as well as preparation method and applications thereof Download PDFInfo
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Abstract
The invention relates to an external-use medicinal composition for treating solar dermatitis as well as a preparation method and applications of the external-use medicinal composition, belonging to the field of medicinal preparations. The medicinal composition is definite in curative effects, uniform in content, good in stability and suitable for industrial production. The external-use medicinal composition is characterized by being prepared from the following medicinal materials in parts by weight: 4-8 parts of radix sophorae flavescentis, 4-8 parts of wild chrysanthemum flowers, 4-8 parts of dandelion, 4-8 parts of herba violae, 4-8 parts of honeysuckle flowers, 2-6 parts of cortex phellodendri, 2-6 parts of rhizoma polygoni cuspidate, 2-6 parts of cortex dictamni, 2-6 parts of fructus cnidii, and 0.5-1 part of borneol. The preparation method comprises the following steps: taking the six medicinal materials including radix sophorae flavescentis, adding with 12-fold amount of water for decocting and extracting for three times, wherein each time lasts for 1h, mixing the obtained three extracting solutions, concentrating the mixed extracting solution till the concentration that each milliliter of the concentrated solution is equivalent to 0.5g of crude drug, and placing the concentrated solution for later use; taking the three medicinal materials including cortex phellodendri, adding with 8-fold amount of 60% of alcohol, carrying out reflux extraction for three times, mixing the extracting solutions, recycling alcohol, concentrating the mixed extracting solution till the concentration that each milliliter of the concentrated solution is equivalent to 0.5g of crude drug, and placing the concentrated solution for later use; and adding with borneol and pharmaceutically acceptable auxiliary materials, and preparing various external-use preparations.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, be specifically related to a kind of treat the externally-applied medicinal composition of solar dermatitis, Preparation Method And The Use.
Background technology
Medicine currently for solar dermatitis is less, and curative effect is the most definite, and clinical report is less；Traditional treatment many employings hormone medicine, antihistaminic and antibiotic are treated, and hormone medicine there will be the side effect such as local skin atrophy, telangiectasis, pigment alteration, local infection, even causes steroid-dependent dermatitis；The side effect such as its drowsiness, weak, xerostomia of antihistaminic.
Chinese medical discrimination: solar dermatitis is similar with " allergic dermatitis " in traditional Chinese medical science ancient literature.The Ming Dynasty " surgery living person fixed this allergic dermatitis " records: " this disease is born on front, red and swollen and float.Just feel preferably dissipates ... if becoming big poison the latest, eye is remained silent swollen, and cheekbone height chin or cheek is convex, very can fear also." primary disease is weak to due to natural endowment, accumulates in heat in blood, stimulating food of taking food, taste are lacked of proper care, damp and hot interior life, and multiple because of Exposure to Sunlight, light poison exposes photograph, and pyretic toxicity attacks skin, sends out suddenly.Analyzing primary disease pathogenesis, it is believed that primary disease natural endowment is weak to, accumulate in heat in blood for endogenous cause of ill, feed stimulating food, to experience light poison be exopathogenic factor, and wherein a large amount of feeds or contact certain plants are inducement, stands solar exposure, to experience light poison be topmost pathogenic factors.Light poison is caused a disease, few described in traditional Chinese medical science classical literature.Qing Dynasty's Chen Shiduo " Qingnang Mijue " is write " solar dermatitis, be expose summer expose strong day and winner also, must first ache and then break, be that exterior-heat is hindered, non-interior-heat is damaged also." feature caused a disease of pictute light poison, and point out that primary disease " exterior-heat " is main cause, endogenous cause of ill is not occupied an leading position, but does not suggests that the name of " light poison "." light poison " this concept is to be proposed according to the feature of Photodermatoses with post-modernism doctor in liberation, but the discussion to its system is the rarest." light poison " is one of poison heresy, caused by daylight.Sun belonging to YANG, moon belonging to YIN, the initial standard distinguishing negative and positive in Basic Theories of Chinese Medicine is i.e. as sun at sun exposure, is cloudy at backlight, therefore the character of daylight should belong to sun heat undoubtedly.In light poison pathogenic course, patient stands solar exposure, scorchingly hot skin of attacking, therefore sees that skin is red and swollen, scorching hot, and the damp and hot outgoing in evil poison priming body is in skin, therefore sees that swelling vesicle, erosion are oozed out.Outer photosensitive poison is the primary outer reason of primary disease morbidity, and the morbidity of primary disease is also relevant with the inherent physical factors of patient, is i.e. " natural endowment is weak to " described in the traditional Chinese medical science, for these pathogenetic important intrinsic factor.Therefore, body constitution in the pathogenesis of primary disease, Exposure to Sunlight the two factor are indispensable.
Solar dermatitis is commonly called as " sunburn ", and the traditional Chinese medical science is considered owing to skin " interspaces of skin and muscles being loose ", " poison of suffering from heatstroke outward " cause, and its Therapeutic Principle is mainly with clearing away heat and cooling blood, and removing toxic substances rash, reducing swelling and alleviating pain are main.Flos Chrysanthemi Indici toil in side, is slightly cold, heat-clearing and toxic substances removing, and the property of bitter cold is better than Flos Chrysanthemi and Herba Tagetis Patulae, solely arrogates to oneself the merit of heat clearing away, and Compendium of Material Medica claims it " to control carbuncle furunculosis "；Herba Taraxaci is cold in nature, sweet in the mouth, micro-hardship, wherein bitter with drop-down, and sweet with removing toxic substances, the heat clearing away of cold energy is held concurrently and dissipated stagnant gas, and for heat-clearing and toxic substances removing, the good merchantable brand of dispersing swelling and dissipating binds, both share, are monarch drug altogether.Cortex Phellodendri bitter in the mouth cold in nature, can heat clearing and damp drying, with can eliminating fire and detoxication, " medicineization justice ": " Cortex Phellodendri, bitter in the mouth to the marrow, is can be capit ad calcem with pathogenic fire reducing "；Auxiliary monarch drug heat-clearing and toxic substances removing, closes the poison that drives away summer heat, so being ministerial drug altogether.Solar dermatitis gargalesthesia after illumination increases the weight of, therefore needs killing parasites for relieving itching.In side, Radix Sophorae Flavescentis bitter in the mouth is cold in nature, has heat clearing and damp drying, the merit of killing parasites for relieving itching." book on Chinese herbal medicine justice ": " Radix Sophorae Flavescentis, big bitter Great Cold, drop-down of bringing down a fever, cleanse wet fire "；Fructus Cnidii killing parasites for relieving itching, dispeiling pathogenic wind and removing dampness, " book on Chinese herbal medicine is newly organized ": " Fructus Cnidii, function is quite strange, and inside and outside tool can treat, and external treatment is excellent ", with Radix Sophorae Flavescentis compatibility, mutual reinforcement between is use, and antipruritic dynamics strengthens；Rhizoma Polygoni Cuspidati has removing heat from blood to expel the heat-evil, the merit of astringing to arrest bleeding；Borneolum Syntheticum work hard taste tremble with fear, there is heat-clearing and toxic substances removing, the merit of promoting tissue regeneration and ulcer healing, owing to property is the most cold and cool, being good at clearing away heat to alleviate pain, amplification on Canon of Materia Medica: " Borneolum Syntheticum; big tonneau closes heat of the diaphragm plug, its delicate fragrance is that hundred medicines are first, the medicine of right non-informal dress; then gesture of walking alone is weak, and assistant makes, and gains merit ", above three tastes share, removing dampness is antipruritic, clearing away heat to alleviate pain, helps principal drug assistance, so being adjuvant drug altogether.Wherein Borneolum Syntheticum can reach diseased region with priming, so holding concurrently as making medicine.All medicines share, and play heat-clearing and toxic substances removing altogether, the merit that removing heat from blood drives away summer heat.Dispelling in order to pathogenic heat, yang-energy must be answered, and space between skin and muscles obtains close, and all cards can be healed.This prescription carries out scientific composition according to theory of Chinese medical science, meets the traditional Chinese medical science Therapeutic Principle to solar dermatitis.
Summary of the invention
It is an object of the invention to provide a kind of externally-applied medicinal composition, Preparation Method And The Use, the externally-applied medicinal composition of the present invention, the purposes in preparation treatment solar dermatitis disease medicament.
The externally-applied medicinal composition content of the treatment solar dermatitis of the present invention uniformly, have good stability, good effect.
It is a further object to provide the preparation method of a kind of externally-applied medicinal composition being suitable for industrial, easy treatment solar dermatitis.
Concrete technical scheme is as follows:
The present invention provides a kind of externally-applied medicinal composition, it is characterised in that described pharmaceutical composition is mainly made up of the crude drug of following weight portion: Radix Sophorae Flavescentis 4-8 part, Flos Chrysanthemi Indici 4-8 part, Herba Taraxaci 4-8 part, Herba Violae 4-8 part, Flos Lonicerae 4-8 part, Cortex Phellodendri 2-6 part, Rhizoma Polygoni Cuspidati 2-6 part, Cortex Dictamni 2-6 part, Fructus Cnidii 2-6 part, Borneolum Syntheticum 0.5-1 part.
Preferably, the externally-applied medicinal composition of the present invention, it is characterised in that be made up of the crude drug of following weight portion: Radix Sophorae Flavescentis 5-7 part, Flos Chrysanthemi Indici 5-7 part, Herba Taraxaci 5-7 part, Herba Violae 5-7 part, Flos Lonicerae 5-7 part, Cortex Phellodendri 3-5 part, Rhizoma Polygoni Cuspidati 3-5 part, Cortex Dictamni 3-5 part, Fructus Cnidii 3-5 part, Borneolum Syntheticum 0.6-0.9 part.
Preferred, the externally-applied medicinal composition of the present invention, it is characterised in that be made up of the crude drug of following weight portion: Radix Sophorae Flavescentis 6 parts, Flos Chrysanthemi Indici 6 parts, Herba Taraxaci 6 parts, Herba Violae 6 parts, Flos Lonicerae 6 parts, Cortex Phellodendri 4.5 parts, Rhizoma Polygoni Cuspidati 4.5 parts, Cortex Dictamni 4.5 parts, Fructus Cnidii 4.5 parts, Borneolum Syntheticum 0.75 part.
The present invention provides a kind of externally-applied medicinal composition, the purposes in preparation treatment solar dermatitis disease medicament.
The present invention treats the externally-applied medicinal composition of solar dermatitis, it includes above-mentioned externally-applied medicinal composition and pharmaceutically acceptable adjuvant, for exterior-applied formulation, including solution, lotion, oil preparation, tincture, liniment, ointment, Emulsion, paste, liniment, spray, membrane, spirit, mucilage, paste, gel.
The present invention provides a kind of method of externally-applied medicinal composition preparing treatment solar dermatitis, it is characterised in that comprise the steps:
1) weigh described weight proportion takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；
2) separately take the Cortex Phellodendri of described weight proportion, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby；
3) by 1) and 2) merging of gained solution, separately take Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add 5% propylene glycol, 1% azone, 3% carbomer, stir evenly, with triethylamine tune pH value to 6.5,1000ml the most processed, obtain gel.
The preparation method of the externally-applied medicinal composition of above-mentioned treatment solar dermatitis, described step 2) after, by 1) and 2) merging of gained solution, separately take Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, stir evenly, 1000ml the most processed, obtain solution.
The preparation method of the externally-applied medicinal composition of above-mentioned treatment solar dermatitis, described step 2) after, separately prepare emulsion type ointment base: oil-phase component (stearic acid 7.5g, Sorbitan Oleate 2g, glyceryl monostearate 7.5g, liquid paraffin 12g and vaseline 7g) and water-phase component (polyoxyethylene sorbitan monoleate: 6g, glycerol 12.5, sorbic acid 1g and water 75g) are separately heated to 80, oil phase is added in aqueous phase, stirring while adding, to being condensed into emulsion-type substrate.By 1) and 2) after gained solution merges, be slowly added in the emulsion-type substrate prepared, stirring while adding, to be completely uniformly to, obtain ointment.
The preparation method of the externally-applied medicinal composition of above-mentioned treatment solar dermatitis, described step 2) after, by 1) and 2) merging of gained solution, separately take Borneolum Syntheticum to add with after appropriate anhydrous alcohol solution, add polysorbate-8020ml, glycerol 60ml, azone 15ml, adjustment volume, to 1000ml, obtains liniment.
The preparation method of the externally-applied medicinal composition of above-mentioned treatment solar dermatitis, described step 2) after, separately take sodium carboxymethyl cellulose 30g, polyvinyl alcohol 22g and add water respectively and appropriate soaks swelling and make to dissolve, stir evenly, filter, obtain filtrate standby；By filtrate, 1) and 2) gained solution merges, separately takes Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add glycerol 30g, adjustment volume, to 1000ml, obtains liniment.
Accompanying drawing explanation
Fig. 1 is Cortex Phellodendri TLC chromatogram, wherein: 1-berberine hydrochloride；2-Cortex Phellodendri；3,4,5-compound recipe ginseng gold gel；6-negative control.
Fig. 2 is Fructus Cnidii TLC chromatogram, wherein: 1-Fructus Cnidii；2-Fructus Cnidii；3,4,5-compound recipe ginseng gold gel；6-negative control.
Fig. 3 is Radix Sophorae Flavescentis TLC chromatogram, wherein: 1-oxymatrine；2-matrine；3,4,5-compound recipe ginseng gold gel；6-Radix Sophorae Flavescentis；7-negative control.
Fig. 4 is chlorogenic acid reference substance HPLC chromatogram.
Fig. 5 is test sample HPLC chromatogram.
Fig. 6 is negative control HPLC chromatogram.
Detailed description of the invention
Below by way of the description of detailed description of the invention, the invention will be further described, but this is not limitation of the present invention, those skilled in the art are according to the basic thought of the present invention, various modifications may be made or improves, but without departing from the basic thought of the present invention, the most within the scope of the present invention.
Embodiment 1 gel
Prescription forms:
Preparation method: above ten tastes, takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.Above-mentioned two parts of solution are merged, separately takes Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add 5% propylene glycol, 1% azone, 3% carbomer, stir evenly, with triethylamine tune pH value to 6.5,1000ml the most processed, to obtain final product.
Embodiment 2 solution
Prescription forms:
Preparation method: above ten tastes, takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.Above-mentioned two parts of solution are merged, separately takes Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, stir evenly, 1000ml the most processed, to obtain final product.
Embodiment 3 ointment
Prescription forms:
Preparation method: above ten tastes, takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.The preparation of emulsion type ointment base, oil-phase component (stearic acid 7.5g, Sorbitan Oleate 2g, glyceryl monostearate 7.5g, liquid paraffin 12g and vaseline 7g) and water-phase component (polyoxyethylene sorbitan monoleate: 6g, glycerol 12.5, sorbic acid 1g and water 75g) are separately heated to 80, Shan is added in aqueous phase, stirring while adding, to being condensed into emulsion-type substrate.After two kinds of extracting solution are merged, be slowly added in the emulsion-type substrate prepared, stirring while adding, to be the most uniformly to, to obtain final product.
Embodiment 4 liniment
Prescription forms:
Above ten tastes, take Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, add 12 times amount water boiling and extraction 3 times, each 1 hour, merge three extracting solution, and are concentrated into every milliliter and are equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.Being merged by above-mentioned two parts of solution, separately take Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add polysorbate-80 20ml, glycerol 60ml, azone 15ml, adjustment volume, to 1000ml, to obtain final product.
Embodiment 5 liniment
Prescription forms:
Preparation method: above ten tastes, takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.Separately take sodium carboxymethyl cellulose 30g, polyvinyl alcohol 22g to add water respectively and appropriate soaks swelling and make to dissolve, stir evenly, filter, filtrate is standby, is merged by above-mentioned three parts of solution, separately takes Borneolum Syntheticum and adds with after appropriate anhydrous alcohol solution, adding glycerol 30g, adjustment volume, to 1000ml, to obtain final product.
Embodiment 6 gel
Prescription forms:
Preparation method: above ten tastes, takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.Above-mentioned two parts of solution are merged, separately takes Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add 5% propylene glycol, 1% azone, 3% carbomer, stir evenly, with triethylamine tune pH value to 6.5,1000ml the most processed, to obtain final product.
Embodiment 7 solution
Prescription forms:
Preparation method: above ten tastes, takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；Separately take Cortex Phellodendri, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby.Above-mentioned two parts of solution are merged, separately takes Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, stir evenly, 1000ml the most processed, to obtain final product.
Embodiment 8 orthogonal test preferred compound recipe ginseng gold gel extraction process by water
1 instrument and reagent
High performance liquid chromatograph (power & light company of the U.S.), joins P4000 pump, UV1000 detector, N 2000 data workstation；Kromasi C18(4.6 × 250mm, 5 μm)；RE52AA type Rotary Evaporators (Shanghai Yarong Biochemical Instrument Plant)；HS3120 ultrasonic cleaner (Tianjin Heng Ao technology & development Co.)；Chlorogenic acid reference substance (for assay, 0753-200111) (Nat'l Pharmaceutical & Biological Products Control Institute)；The medical materials such as Flos Lonicerae are purchased from medical material company of Shaanxi Province, the Xi'an medicine inspecting institute bright professor of pharmacy of expert Xie Zhi identify；Methanol (chromatographically pure, TEDIA company of the U.S.), remaining reagent is analytical pure.
2 methods and result
The selection of 2.1 factor levels
Flos Lonicerae is mainly composed of chlorogenic acid, for water soluble ingredient, therefore selecting water is to extract solvent, according to preliminary result, selecting reflow's cycle (A), return time (B), 3 factors of amount of water (C) is investigation factor, with monarch drug Chlorogenic Acid of Flos Lonicerae content and yield of extract as index, use L9(34) orthogonal table carries out experimental design, factor level table is shown in Table 1.
Table 1 factor level table
2.2 test method
Weighing the medicinal material coarse powder such as Flos Lonicerae in prescription ratio and put in reflux, with water for extracting solvent, extract by Orthogonal Experiment and Design condition, medicinal liquid filters, and it is standby that centrifuging and taking supernatant concentration is settled to 50ml.
2.3 yield of extract measure
The above-mentioned medicinal liquid 10ml of accurate absorption, puts and is dried to the evaporating dish of constant weight, after water bath method, in 105 DEG C of dry 3h, in dislocation exsiccator, cools down 30min, rapid accurately weighed weight, calculates yield of extract.
2.4 assay
2.4.1 chromatographic condition
Chromatographic column Kromasi C18(4.6 × 150mm, 5 μm)；With methanol one 0.4% phosphoric acid solution (13:87) for flowing phase；Detection wavelength is 327nm.Number of theoretical plate is calculated by chlorogenic acid peak should be not less than 3000.
2.4.2 the preparation of reference substance solution
Take chlorogenic acid reference substance appropriate, accurately weighed, add 50% methanol and make every 1ml solution containing 80 μ g, to obtain final product.
2.4.3 the preparation of test sample
Accurate absorption adds first 50% alcohol to scale in orthogonal test gained solution each 2ml to 25ml measuring bottle, shake up, to obtain final product.
2.4.4 linear relationship is investigated
Precision draws reference substance solution respectively: 1,3,5,10,15,20 μ l, with sample size C (μ g) as abscissa, peak area A is vertical coordinate, calculates regression equation and is: Y=3151711.36X-755.70, r=0.9999；Show chlorogenic acid in 0.08～1.60 μ g range, peak area and sample size are good linear relationship.
2.4.5 assay
Accurate chlorogenic acid reference substance solution and each 10 μ l of need testing solution of drawing, note people's chromatograph of liquid, measure, to obtain final product.
2.5 orthogonal experiments and variance analysis
Carry out orthogonal test by 2.2 experimental techniques and obtain 9 parts of need testing solutions, record the experimental data of each need testing solution.Orthogonal experiments is shown in Table 2, and variance analysis is shown in Table 3.
From table 2, table 3, the optimum extraction process with chlorogenic acid as index is as A3B1C3；Optimum extraction process with yield of extract as index is as A3B2C3, and A there were significant differences, optimised process be 12 times amount water reflux 3 times, each 1h.The factors such as comprehensive 2 evaluation indexes also consider in actual production cost-effective, the saving time, final selection optimised process is A3B1C3I.e. 12 times amount water reflux 3 times, each 1h.
Table 2 orthogonal experiments table
Table 3 analysis of variance table
2.6 optimised process checkings
3 parts of medical materials are weighed, the optimised process A obtained according to screening in prescription ratio is parallel3B1C3I.e. 12 times amount water reflux 3 times, each 1h.Carrying out checking test, and measure yield of extract and chlorogenic acid content respectively, fruit is shown in Table 4.Experiment shows that optimum process condition also indicates that this technique has preferable stability the most simultaneously.
Table 4 optimised process confirmatory experiment
Embodiment 9 orthogonal test preferred compound recipe ginseng gold gel alcohol extraction process
1 instrument and reagent
High performance liquid chromatograph (power & light company of the U.S.), joins P4000 pump, UV1000 detector, N 2000 data workstation；Kromasi C18(4.6 × 250mm, 5 μm)；HS3120 ultrasonic cleaner (Tianjin Heng Ao technology & development Co.)；Berberine hydrochloride reference substance (for assay, 0713-200107)；Osthole reference substance (for assay, 110822-200406)；Methanol (chromatographically pure, TEDIA company of the U.S.), remaining reagent is analytical pure.
2 methods and result
The selection of 2.1 factor levels
Cortex Phellodendri is mainly composed of berberine hydrochloride, Fructus Cnidii is mainly composed of osthole, it is ester soluble components, therefore selecting different concentration ethanol is to extract solvent, according to preliminary result, selecting reflow's cycle (A), return time (B), 3 factors of amount of water (C) is investigation factor, in side ministerial drug Cortex Phellodendri, Fructus Cnidii active ingredient hydrochloric acid berberine, osthole and yield of extract as index, use L9(34) orthogonal table carries out experimental design, factor level table is shown in Table 1.
Table 1 factor level table
2.2 test method
Weighing the medicinal material coarse powder such as Cortex Phellodendri, Fructus Cnidii in prescription ratio and put in reflux, with different concentration ethanol for extracting solvent, extract by Orthogonal Experiment and Design condition, medicinal liquid filters, and it is standby that centrifuging and taking supernatant concentration is settled to 50ml.
2.3 yield of extract measure
The above-mentioned medicinal liquid 10ml of accurate absorption, puts and is dried to the evaporating dish of constant weight, after water bath method, in 105 DEG C of dry 3h, in dislocation exsiccator, cools down 30min, rapid accurately weighed weight, calculates yield of extract.2.4 assay
2.4.1 Cortex Phellodendri assay
2.4.1.1 Cortex Phellodendri fixation spectral condition
Chromatographic column Kromasi C18(4.6 × 150mm, 5 μm), with acetonitrile one 0.1% phosphoric acid solution (56:44) (every 100ml adds dodecyl sodium sulfate 0.1g) for flowing phase, detection wavelength is 265nm, and number of theoretical plate is calculated by berberine hydrochloride should be not less than 3000.
2.4.1.2 the preparation of reference substance solution
Take berberine hydrochloride reference substance appropriate, accurately weighed, add methanol and make every 1ml solution containing 48 μ g, to obtain final product.
2.4.1.3 the preparation of test sample
Accurate absorption adds methanol to scale in orthogonal test gained solution each 2ml to 25ml measuring bottle, shake up, to obtain final product.
2.4.1.4 linear relationship is investigated
Precision absorption reference substance solution respectively: 2.5,5,7.5,10,12.50 μ l, with sample size C (μ g) as abscissa, peak area A is vertical coordinate, calculating regression equation is: Y=491922X+125468, r=0.9999, show berberine hydrochloride in 0.12 μ g～0.6 μ g range, peak area and sample size are good linear relationship.
2.4.1.5 assay
Accurate berberine hydrochloride reference substance solution and each 10 μ l of need testing solution of drawing, injection chromatograph of liquid, measure, to obtain final product.
2.4.2 Fructus Cnidii assay
2.4.2.1 Fructus Cnidii chromatographic condition
Chromatographic column Kromasi C18(4.6 × 150mm, 5 μm), with acetonitrile one water (65:35) for flowing phase, detection wavelength is 322nm, and number of theoretical plate is calculated by osthole should be not less than 3000.
2.4.2.2 the preparation of reference substance solution
Take osthole reference substance appropriate, accurately weighed, add ethanol and make every 1ml solution containing 56 μ g, to obtain final product.
2.4.1.3 the preparation of test sample
Accurate absorption adds methanol to scale in orthogonal test gained solution each 2ml to 25ml measuring bottle, shake up, to obtain final product.
2.4.1.4 linear relationship is investigated
Precision absorption reference substance solution respectively: 1,2,3,4,5 μ l, with sample size C (μ g) as abscissa, peak area A is vertical coordinate, calculating regression equation is: Y=232254X+11805, r=0.9997, show osthole in 0.056 μ g～0.28 μ g range, peak area and sample size are good linear relationship.
2.4.1.5 assay
Accurate osthole reference substance solution and each 10 μ l of need testing solution of taking, note people's chromatograph of liquid, measure, to obtain final product.
2.5, orthogonal experiments and variance analysis
Carry out orthogonal test by 2.2 experimental techniques and obtain 9 parts of need testing solutions, record the experimental data of each need testing solution.Orthogonal experiments is shown in Table 2, and variance analysis is shown in Table 3.
From table 2, table 3, the optimum extraction process with berberine hydrochloride as index is as A3B1C3D3And A, C there were significant differences；Optimum extraction process with osthole as index is as A3B2C2(C3)D2And A there were significant differences；Optimum extraction process with yield of extract as index is as A3B1C3D3, and A, C factors such as there were significant differences, and comprehensive 3 evaluation indexes also consider in actual production cost-effective, the saving time, final selection optimised process is A3B2C2D2I.e. 8 times amount 60% alcohol refluxs 3 times, each 1h.
Table 2 orthogonal experiments table
Table 3 analysis of variance table
2.6 optimised process checkings
3 parts of medical materials are weighed, the optimised process A obtained according to screening in prescription ratio is parallel3B2C2D2I.e. 8 times amount 60% alcohol refluxs 3 times, each 1h carries out checking test, and measures content of berberine hydrochloride, Content of Osthole and yield of extract respectively and the results are shown in Table 4.Experiment shows that optimum process condition also indicates that this technique has preferable stability the most simultaneously.
Table 4 optimised process confirmatory experiment
The ginseng gold Process for preparing hydrogels research of embodiment 10 orthogonal test preferred compound recipe
1, formulation factors is investigated
Evaluation index is with glossiness, stretchability, the uniformity and centrifugum comprehensive grading as inspection target, and standards of grading are full marks 100 points, glossiness, stretchability, the uniformity and centrifugum respectively account for 25 points, and concrete standards of grading are shown in Table 1.
Table 1 gel prescription screening and the scoring of preparation technology index
2, orthogonal test takes the extractum of recipe quantity, carries out L with glossiness, stretchability, the uniformity and centrifugum comprehensive grading for index9(34) orthogonal test, factor level table is shown in Table 2, the results are shown in Table 3,4.
Table 2 gel prescription preparation technology orthogonal test factor level
Table 3 gel orthogonal test arranges and data analytical table
Table 4 analysis of variance table
*F0.05(2 , 2)=19.00
Analysis result directly perceived is it can be seen that influence factor A ＞ C ＞ B, and A2B1C2Being preferred, the results of analysis of variance shows that A, C factor has a significant impact, and B factor affects without significance, it is thus determined that gel prescription is carbomer 3%, propylene glycol 8%, pH value is 6.5.
The TCL identification research of embodiment 11 compound recipe ginseng gold gel
1 instrument and reagent
1.1 instrument ZF-2 types three are with ultraviolet devices (Town in Shanghai booth Electronic Instruments Plant)；101-OA electric drying oven with forced convection (Tianjin Stettlen Instrument Ltd.)；HS3120 ultrasonic cleaner (Tianjin Heng Ao technology & development Co.).
1.2 reagent berberine hydrochloride reference substances (for assay, 0713-200107)；Osthole reference substance (for assay, 110822-200406)；Matrine reference substance (for assay, 805-200005)；Oxymatrine reference substance (for assay, 0780-200004)；Cortex Phellodendri control medicinal material (for differentiating, 121510-201105) is purchased from Nat'l Pharmaceutical & Biological Products Control Institute；Reagent is analytical pure.
2 methods and result
2.1 thin layer chromatography qualitative identification
2.1.1 the discriminating of Cortex Phellodendri takes this product 5g, adds methanol 25ml, supersound process 30 minutes, filters, and filtrate is evaporated, and residue adds 2ml methanol makes dissolving, as need testing solution.Separately take Cortex Phellodendri control medicinal material 0.5g, be made in the same way of control medicinal material solution.Take berberine hydrochloride reference substance, add methanol and make every 1ml solution containing 0.5mg, as reference substance solution.Separately press the prescription tissue preparation blank sample without Cortex Phellodendri, and press need testing solution preparation method negative control solution.Test according to thin layer chromatography (2010 editions annex VIB of Chinese Pharmacopoeia), draw each 5 μ L of above-mentioned four kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-isopropanol-methanol-water-triethylamine (3: 3.5: 1: 1.5: 0.5: 1) as developing solvent, put with in the strong ammonia solution presaturation expansion cylinder of 20 minutes, launch, take out, dry, put and inspect under ultra-violet lamp (365nm).In test sample chromatograph, with on control medicinal material chromatograph relevant position, show identical yellow fluorescence speckle, with on reference substance chromatograph relevant position, show an identical yellow fluorescence speckle.Negative noiseless (see Figure of description 1).
2.1.2 the discriminating of Fructus Cnidii takes this product 2g, adds ethyl acetate 5ml, supersound process 30 minutes, filters, as need testing solution.Separately take Fructus Cnidii control medicinal material 1.0g, be made in the same way of control medicinal material solution.Take osthole reference substance, add ethanol and make every 1ml solution containing 1mg, as reference substance solution.Separately press prescription tissue preparation and do not contain the blank sample of son on cnidium monnieri, and press need testing solution preparation method negative control solution.Test according to thin layer chromatography (2010 editions-portion of Chinese Pharmacopoeia annex VIB), draw each 5 μ L of above-mentioned four kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (9: 1) as developing solvent, launch, take out, dry, put and inspect under ultra-violet lamp (365nm).In test sample chromatograph, with in control medicinal material chromatograph and reference substance chromatograph relevant position, show the fluorescence speckle of same color.Negative noiseless (see Figure of description 2).
2.1.3 the discriminating of Radix Sophorae Flavescentis takes this product 2g, adds strong ammonia solution 0.3ml, chloroform 25ml, stands overnight, and filters, and filtrate is evaporated, and residue adds 2ml chloroform makes dissolving, as need testing solution.Separately take Radix Sophorae Flavescentis control medicinal material 0.5g, be made in the same way of control medicinal material solution.Take matrine reference substance, oxymatrine reference substance, respectively add ethanol and make every 1ml solution containing 0.2mg, as respective reference substance solution.Separately press the prescription tissue preparation blank sample without Radix Sophorae Flavescentis, and press need testing solution preparation method negative control solution.Test according to thin layer chromatography (2010 editions annex VIB of Chinese Pharmacopoeia), draw each 5 μ L of above-mentioned five kinds of solution, put respectively on silica gel g thin-layer plate prepared by same 2% sodium hydroxide solution, with 10 DEG C of upper solution arranged below of toluene-ethyl acetate-methanol-water (2: 4: 2: 1) as developing solvent, launch, take out, dry, spray bismuth potassium iodide test solution, in test sample chromatograph, with in control medicinal material chromatograph and reference substance chromatograph relevant position, show the fluorescence speckle feminine gender noiseless (see Figure of description 3) of same color.
Embodiment 12 HPLC method measures the content of compound recipe ginseng gold gel Content of Chlorogenic Acid
1 instrument and reagent
High performance liquid chromatograph (U.S.'s thermoelectricity), joins P4000 pump, UV1000 detector, N 2000 data workstation；Kromasi C18(4.6 × 250mm, 5 μm)；HS3120 ultrasonic cleaner (Tianjin Heng Ao technology & development Co.)；Chlorogenic acid reference substance (for assay, 110753-200212) (Nat'l Pharmaceutical & Biological Products Control Institute)；Acetonitrile (chromatographically pure, TEDIA company of the U.S.), remaining reagent is analytical pure.
2 methods and result
2.1 chromatographic condition chromatographic column Kromasi C18(4.6 × 150mm, 5 μm)；Flow velocity: 1.0mL/min；Flowing is methanol-0.4% phosphoric acid solution (13: 87) mutually；Detection wavelength is 327nm；Column temperature: room temperature；Sample size: 10 μ L.
The preparation of 2.2 solution
2.2.1 the preparation of need testing solution
Precision weighs in compound recipe ginseng gold gel 2g to 25ml measuring bottle and adds 50% methanol to scale, shakes up, supersound process (150W, 20kHZ) 20 minutes, lets cool, filters, to obtain final product.
2.2.2 the preparation of reference substance solution
Take chlorogenic acid reference substance appropriate, accurately weighed, add 50% methanol and make every 1ml solution containing 80 μ g, to obtain final product.
2.2.3 the preparation of negative control solution
By the recipe quantity preparation negative sample without Chinese medicine honeysuckle, prepare by need testing solution method, to obtain final product.
2.3 specificity test precisions respectively draw above-mentioned reference substance solution, need testing solution and each 10 μ L of negative controls solution, inject chromatograph of liquid, measure, record chromatogram, result is shown in Figure of description 4-6, in negative control chromatograph, on position corresponding with chlorogenic acid reference substance, noiseless peak.
2.4 linear relationships are investigated and chlorogenic acid reference substance are configured to the 1ml 50% methanol solution containing 0.080mg, accurate sample introduction 1,3,5,10,15,20 μ l successively, with sample size C (μ g) as abscissa, peak area A is vertical coordinate, calculating regression equation is: Y=3151711.36X-755.70, r=0.9999；Show chlorogenic acid in 0.08～1.60 μ g range, peak area and sample size are good linear relationship.
2.5 precision tests precision respectively draws reference substance solution and each 10 μ l of need testing solution, repeats sample introduction 6 times respectively, and the RSD of result chlorogenic acid integrating peak areas value is respectively 1.13%；1.08%.
2.6 stability test precision draws chlorogenic acid reference substance solution 10 μ l, need testing solution 10 μ l, respectively at 0, within 2,4,6,8,10,12,24 hours, measuring, result chlorogenic acid peak area value RSD is respectively 0.93%, 1.10%.Show that chlorogenic acid was stable in 24 hours.
2.7 replica tests take same sample lots (lot number 20111115) 5 parts, prepare method operation by need testing solution respectively, measure by chromatographic condition, result chlorogenic acid average content be 2.043mg/ml, RSD be 1.21%.
2.8 average recovery tests take 5 parts of the sample (lot number 20111115) of known content, each 0.2g, accurately weighed, each addition chlorogenic acid reference substance solution (0.080mg/mL) is appropriate, prepare method operation according to need testing solution, measure by chromatographic condition, calculate the response rate, result average recovery rate is 98.99%, and RSD is 0.71%.It is shown in Table 1.
Table 1 compound recipe ginseng gold gel Content of Chlorogenic Acid average recovery
2.9 assays prepare reference substance solution and the compound recipe ginseng gold gel need testing solution of chlorogenic acid according to the method described above, take reference substance solution respectively and need testing solution 10 μ L measures, with external standard method calculated by peak area in accordance with the law.Determine the content of 3 batch samples (20120909,20120915,20120919), the results are shown in Table 2.
Table 2 compound recipe ginseng gold gel Content of Chlorogenic Acid measurement result (n=3)
Embodiment 13 compound recipe SJ gel Pharmacodynamic test of active extract
Experiment purpose: cause the impact of Mice Auricle inflammation, the impact on mice granulomatous inflammation by investigating compound recipe SJ gel xylol, on the impact of mice acetic acid abdominal cavity induced pain effect, impact on hot plate method in mice induced pain effect, specifying the pharmacology pharmacodynamic effect of compound recipe SJ gel, the further investigation further for this product provides experimental basis.
Experiment content
1 experimental chemical and major experimental facility
1.1 compound recipe SJ gels
Title: compound recipe SJ gel.Function cures mainly: solar dermatitis.Specification: 10ml/ props up (0.5g crude drug/ml).Character: sepia gel.Lot number: 20120828.Usage: external preparation for skin medicine.Draft clinical usage and consumption: smear in right amount according to skin solar dermatitis area.Banking system: stored refrigerated.Offer unit: No.323 Hospital, PLA.
1.2 comparison medicines
1.2.1 indomethacin ointment, lot number: 1100910.Specification: 1%.Shengyang City Xingqi Pharmaceutical Co.
1.2.2 injection of morphia liquid, lot number: 20101101.Specification: 25mg/ props up.Qinghai Dadi Pharmacy Co., Ltd.1.2.3 diphenhydramine hydrochloride injection, lot number: 20110621.Specification: 1ml:20mg, Wuhan pharmacy Group Plc.
The feeding and management of 1.3 laboratory animals
1.3.1 laboratory condition
Xi'an Jiaotong University Medical College's Experimental Animal Center, laboratory temperature: 18～26 DEG C, relative humidity: 40～70%, illumination condition: 12h/12h light and shade replaces, ventilation situation: all-fresh air.
1.3.2 Drinking Water freely drunk by drinking-water.
1.3.3 feedstuff Mus full-valence pellet feed, nutritional labeling meets GB14924.3-2001, production unit: Xi'an Jiaotong University Medical College's Experimental Animal Center.
2 compound recipe SJ gel xylol cause the impact of Mice Auricle inflammation
2.1 experiment material
2.1.1 compound recipe SJ gel compound recipe SJ gel (referring to 1.1.1).
2.1.2 reference substance indomethacin ointment (referring to 1.2.1).
2.1.3 major experimental instrument and reagent FA1104 type trace electronic analytical balance, Shanghai Precision Scientific Apparatus Co., Ltd.
Dimethylbenzene, lot number: 20110401, Shanghai organic chemical industry's reagent institute.
2.1.4 laboratory animal ICR mice, one-level, 60,18～22g, female, male half and half.Credit number: SCXK (Shan): No. 2007-001.Supplying unit: Xi'an Jiaotong University Medical College's Experimental Animal Center.
2.2 experimental technique
2.2.1 packet and dosage arrange animal and are respectively adopted completely random method by sex, are divided into 5 groups, and male and female half and half see table 1.
Table 1 compound recipe SJ gel xylol causes the experiment packet of Mice Auricle inflammatory effect and dose design
2.2.2 compound recipe SJ gel is applied in the basic, normal, high each dosage group mice bilateral auricle portion of compound recipe SJ gel, every secondary amounts about 0.25ml by operating procedure equably, and model control group smears equal-volume normal saline, and positive controls smears indomethacin ointment.Every day 2 times, successive administration 7 days, 30min after last administration, inflammation is caused at each mouse right ear uniform application analytical pure dimethylbenzene 50 μ l, after causing inflammation, 1h takes off cervical vertebra execution mice, cuts left and right two ears, punches along left and right auricle same position homalographic with the card punch of diameter 8mm, sweep away both sides auricle to weigh respectively, record.Two auricle weight difference are swelling, and result carries out statistical analysis, and calculates inhibitory rate of intumesce.
2.2.3 data processing method SPSS13.0 statistical analysis software carries out t inspection or variance analysis.
2.3 experimental result
2.3.1 the impact of compound recipe SJ gel xylol cause Mice Auricle inflammation is shown in Table 2.
The impact of table 2 compound recipe SJ gel xylol induced mice auricle edema ()
Note: compare with model control group, * P ＜ 0.05, * * P ＜ 0.01
From upper table result, indomethacin ointment group xylol causes mice auricle swelling obvious inhibitory action, compares with model control group, and the difference of auricle swelling degree has significance (P ＜ 0.01).Compound recipe SJ gel delivery group xylol causes mice auricle swelling obvious inhibitory action, comparing with model control group, compound recipe SJ gel high dose group, the difference of middle dosage group auricle swelling degree have significance (respectively P ＜ 0.01 and P ＜ 0.05).
2.3.2 experiment brief summary mice administered locally to compound recipe SJ gel through 7 days, and middle and high dosage group all can significantly alleviate mice caused by dimethylbenzene xylene auricle inflammation, shows that compound recipe SJ gel has certain antiinflammatory action.
The impact on mice granulomatous inflammation of the 3 compound recipe SJ gels
3.1 experiment material
3.1.1 compound recipe SJ gel compound recipe SJ gel (referring to 1.1.1).
3.1.2 reference substance indomethacin ointment (referring to 1.2.1).
3.1.3 major experimental instrument and reagent
FA1104 type trace electronic analytical balance, Shanghai Precision Scientific Apparatus Co., Ltd.Filter paper (diameter about 8mm, thickness about 1.5mm, weight 1.9mg), dry sterilization 30 minutes.Pentobarbital sodium, lot number: F20091012, Chemical Reagent Co., Ltd., Sinopharm Group.
3.1.4 laboratory animal
ICR mice, one-level, 50,18～22g, female.Credit number: SCXK (Shan): No. 2007-001.Supplying unit: Xi'an Jiaotong University Medical College's Experimental Animal Center.
3.2 experimental technique
3.2.1 packet and dosage arrange animal and are respectively adopted completely random method by sex, are divided into 5 groups, and male and female half and half see table 3.
Table 3 compound recipe SJ gel xylol causes the experiment packet of Mice Auricle inflammatory effect and dose design
3.2.2 operating procedure
3.2.2.1 modeling
Under 0.3% pentobarbital sodium anesthesia, the scraps of paper are implanted respectively 50 left and right shoulders of animal or subcutaneous each 1 of upper wrist (totally 2), sews up.
3.2.2.2 medication
It is randomly divided into 5 groups by body weight after all mice modeling 24h.Compound recipe SJ gel group gives the compound recipe SJ gel partial smearing of various dose, and positive controls gives isopyknic indomethacin ointment partial smearing, and the every bu of total dosage gives for 3 times, successive administration 7 days.After last is administered, 30min puts to death animal, is extracted in the lump with granulation tissue by the scraps of paper, claims dry weight in 60 DEG C of dry 24h.Dry weight deducts scraps of paper weight, is granulation net weight.
3.2.3 data processing method uses SPSS13.0 statistical analysis software to carry out t inspection or variance analysis.
3.3 experimental result
3.3.1 compound recipe SJ gel is shown in Table 4 to the impact of mice granulomatous inflammation.
Table 4 compound recipe SJ gel on the impact of mice granulomatous inflammation ()
Note: compare with model control group, * P ＜ 0.05, * * P ＜ 0.01
From upper table result, mice granulomatous inflammation is had obvious inhibitory action, granulation net weight to compare with model control group by positive controls, and difference has significance (P ＜ 0.01).Compound recipe SJ gel delivery group has obvious inhibitory action to mice granuloma, granulation net weight compares with model control group, and the difference of compound recipe SJ gel high, medium and low dosage group has significance (respectively P ＜ 0.01, P ＜ 0.05 and P ＜ 0.05).
3.3.2 experiment brief summary is under this experiment condition, and mice gave compound recipe SJ gel through 7 days, and high, medium and low dosage group can significantly inhibit the granulomatous formation of mice, showed that compound recipe SJ gel has certain antiinflammatory action.
4 compound recipe SJ gels cause the impact that mouse writhing reacts to acetic acid
4.1 experiment material
4.1.1 compound recipe SJ gel compound recipe SJ gel (referring to 1.1.1).4.1.2 reference substance injection of morphia liquid (referring to 1.2.2).
4.1.3 laboratory animal
Animal: ICR mice, grade: one-level, quantity: 60, weight: 18-22g, sex: female.Supplying unit: Xi'an Jiaotong University Medical College's Experimental Animal Center.Credit number: SCXK (Shan): No. 2007-001.
4.2 experimental technique
4.2.1 packet and dosage arrange animal and are respectively adopted completely random method by sex, are divided into 5 groups, and often group 12, gives for each group
Pharmaceutical quantities sees table 5.
Table 5 compound recipe SJ gel causes mouse writhing reaction test packet and dose design to acetic acid
4.2.2 operating procedure:
Except morphine group is in addition to experimental day 0.02g/kg subcutaneous injection, remaining respectively group the most according to dosage topical every day or normal saline 1 time, continuous 7d, last be administered after 45min, every mouse peritoneal injects 0.7% acetic acid 0.01ml/g body weight, starts to record mouse writhing number of times in 10min after 5min.Result carries out statistical analysis.
4.2.3 data processing method SPSS11.0 statistical software carries out one factor analysis of variance.
4.3 experimental result
4.3.1 the impact that acetic acid is caused mouse writhing to react by compound recipe SJ gel is shown in Table 6.
Table 6 compound recipe SJ gel on acetic acid cause mouse writhing reaction impact ()
Note: compare with blank group, * P ＜ 0.05, * * * P ＜ 0.001.
From upper table result, positive controls causes mouse writhing number of times to acetic acid obvious minimizing effect, and writhing number of times compares with model control group, and difference has significance (P ＜ 0.01).Compound recipe SJ gel delivery group causes mouse writhing number of times to Mouse Acetic Acid obvious minimizing effect, writhing number of times compares with model control group, and the difference of compound recipe SJ gel dosage group high, middle has significance (respectively P ＜ 0.01 and P ＜ 0.05).
4.3.2 experiment brief summary is under this experiment condition, and mice gives compound recipe SJ gel dosage group high, middle has significant inhibitory action to acetic acid cause mouse writhing reaction times, shows that compound recipe SJ gel has certain analgesic activity.
5 compound recipe SJ gel impact preclinical on burning pain irritant reaction (hot plate method)
5.1 experiment material
5.1.1 compound recipe SJ gel compound recipe SJ gel (referring to 1.1.1).5.1.2 reference substance injection of morphia liquid (referring to 1.2.2).
5.1.3 laboratory animal
Animal: ICR mice, grade: one-level, quantity: 60, weight: 18-22g, sex: female.Supplying unit: Xi'an Jiaotong University Medical College's Experimental Animal Center.Credit number: SCXK (Shan): No. 2007-001.
5.2 experimental technique
5.2.1 packet is arranged with dosage
Laboratory animal uses the intelligence hot-plate instrument screening threshold of pain less than 10 seconds or more than 30 seconds persons and Multiple hop person rejecting on hot plate, selects qualified mice 60, is randomly divided into 5 groups by body weight, often group 12, and each group dosage sees table 7.
Table 7 compound recipe SJ gel is to the packet of burning pain irritant reaction latency test and dose design
5.2.2 operating procedure:.
Except morphine group is in addition to experimental day 0.02g/kg subcutaneous injection, remaining respectively group the most according to dosage gastric infusion every day or water 1 time, 7d continuously, 45min, 90min and 120min after last administration, mice is put in intelligence hot-plate instrument, record mice is from putting into instrument to licking time of metapedes respectively, as the pain incubation period of mice.Data withRepresenting, t test between group, to judge the analgesic activity of medicine.Result carries out statistical analysis.
5.2.3 data processing method SPSS11.0 statistical software carries out one factor analysis of variance.
5.3 experimental result
5.3.1 the impact preclinical on burning pain irritant reaction of compound recipe SJ gel is shown in Table 8.
Table 8 compound recipe SJ gel on the impact of burning pain stimulation pain response latency (N=12)
Note: compare with blank group, * P ＜ 0.05, * * P ＜ 0.01, * * * P ＜ 0.001.
From upper table result, positive controls has obvious prolongation effect to the mice burning pain stimulation pain response latency, the burning pain stimulation pain response latency compares with blank group when 45min, 90min and 120min, and difference has significance (respectively P ＜ 0.01, P ＜ 0.01 and P ＜ 0.05).Compound recipe SJ gel delivery group has obvious prolongation effect to the mice burning pain stimulation pain response latency, the burning pain stimulation pain response latency compares with model control group, and the compound recipe SJ gel dosage group high, the middle difference when 45min and high dose group are at 90min has significance (respectively P ＜ 0.01, P ＜ 0.05 and P ＜ 0.05).
5.3.2 experiment brief summary is under this experiment condition, and mice gives compound recipe SJ gel dosage group high, middle has significant prolongation effect to the burning pain stimulation pain response latency, shows that compound recipe SJ gel has certain analgesic activity.
The ginseng gold gel acute toxicity test of embodiment 14 compound recipe
Test objective: observe animal intact skin and damaged skin contacts toxic reaction produced by tested material in a short time.
1. experimental condition
1.1 animal strains: KM mice, are provided by Xi'an Jiaotong University Medical College's Experimental Animal Center, the quality certification number: NO: 0014335, credit number: SCXK (Shan) 2007-001.Raise 1 week before experiment, select the normal mice of behavioral activity, diet, mental status to test.
1.2 test medicine: compound recipe ginseng gold gel is provided by hospital of PLA 323, lot number 20131101.External, is applied to affected part, and one for several times.
1.3 excipient: carbomer.
2. test method
2.1 animal subject Skin sensitization test
2.1.1 intact skin prepares: mice 12 hours before administration, by spinal column both sides, back unhairing about 2 × 2cm2, within after unhairing 24 hours, check skin of unhairing, the mice of compromised skin rejected.
2.1.2 damaged skin prepares: as stated above by test mice unhairing, after sterilization skin, with rubber paper friction hacking skin, slight oozing of blood occurs for degree with skin.
2.2 medications and observing time: rat is divided six groups at random by body weight, often group 10, male and female half and half, first group is intact skin matched group, being lost hair or feathers district in back part of animal by carbomer 1g even application, kraft paper covers also to be fixed with non-stimulated adhesive tape, and following coating fixed form is identical；Second group of intact skin group heavy dose group, is coated with compound recipe ginseng gold gel 1g；3rd group of intact skin small dose group, is coated with compound recipe ginseng gold gel 0.5g；4th group is damaged skin matched group, is coated with carbomer 1g；5th group of damaged skin heavy dose group, is coated with compound recipe ginseng gold gel 1g；6th group of damaged skin small dose group, is coated with compound recipe ginseng gold gel 0.5g.After being administered 24 hours, removing residual compound recipe ginseng gold gel with warm water, observe mouse systemic situation every day, crawler behavior changes, and breathes, circulation, and digestion and the toxic reaction of central nervous system and death condition record it day by day, continuous 14 days.And before weighing administration and be administered Mouse Weight after 14 days, become celestial after 14 days, the main organs heart, liver, spleen, lung, kidney, brain, pancreas, Stomach duodenum, rectum, bladder, testis, seminal vesicle, prostate, thymus, adrenal gland and body cavity are done gross examination of skeletal muscle, and perusal has obvious changer to make pathology microscopy.
3. result of the test
3.1 medication groups and control group mice are movable normal in 14 days, and irritant reaction is sensitive to external world, gloss, breathes steadily, and drinking-water of ingesting is normal, and two is the most without exception.Mouth, nose, eye secretions without exception.Without dead.
3.2 Mouse Weights increase, and no significant difference between group is shown in Table 1.
The organs such as 3.3 medication groups and the control rats heart, liver, spleen, lung, kidney analyse no abnormality seen.
4. conclusion:
Compound recipe ginseng gold gel does not finds that skin acute poisoning is reacted.
Table 1 anxious poison Mouse Weight (g) () n=10
| Group | Dosage | Body weight before coating | 7 days body weight after medicine | 14 days body weight after medicine |
| Intact skin matched group | 1g | 22.34±0.92 | 26.10±1.24 | 30.04±1.04 |
| Intact skin is heavy dose of | 1g | 22.51±0.84 | 26.37±1.19 | 30.19±1.14 |
| Intact skin is low dose of | 0.5g | 22.55±0.82 | 26.31±1.11 | 30.14±1.00 |
| Damaged skin matched group | 1g | 21.78±0.95 | 25.70±1.19 | 29.87±1.26 |
| Damaged skin is heavy dose of | 1g | 21.85±1.07 | 25.92±1.13 | 29.84±1.09 |
| Damaged skin is low dose of | 0.5g | 21.68±1.20 | 25.87±1.21 | 29.71±1.20 |
Note: P ＞ 0.05 compared with matched group
Embodiment 15 compound recipe ginseng gold gel hypersensitive test
Test objective: compound recipe ginseng gold gel repeats to contact guinea pig skin, observes with or without anaphylaxis.
1. test material
1.1 experimental animals: albino guinea-pig, the offer of big experimental animal center, body weight 200 ± 20g, male and female dual-purpose are handed in Xi'an.The quality certification number: NO: 0014340, credit number: SCXK (Shan) 2007-001.
1.2 tested materials: compound recipe ginseng gold gel, are provided by hospital of PLA 323, lot number 2013.12.1.External, is applied to affected part, and one for several times.
1.3 reagent: 2.4-dinitrochlorobenzene acetone be configured to 1% sensitization concentration and 0.1% excite concentration.
1.4 excipient: carbomer.
2. test method
2.1 tests prepare
In first 24 hours to tested material, guinea pig back diamond wool is taken off, remove the every side of hair-fields scope about 3cm × 3cm.Within after unhairing 24 hours, checking on skin of unhairing with or without injured because of unhairing, injured skin is unfit to do this test.
By body weight sex, Cavia porcellus being randomly divided into three groups, often 10 (male and female half and half) of group, first group to compound recipe ginseng gold gel；Second group of blank group is to carbomer；3rd group of positive controls DNFB.
2.2 process of the test
2.2.1 sensitization contact: compound recipe ginseng gold gel 0.2g is coated on the left of Cavia porcellus district of losing hair or feathers (cover kraft paper, then with immobilization with adhesive tape) lasting 6 hours, then cleans.Blank group carbomer 0.2g, positive controls is with 1%2, and 4-dinitrochlorobenzene 0.2ml, method of sensitization is ibid.7th day, sensitization 2 times again in the 14th day.
2.2.2 contact is excited: the 28th day, by compound recipe ginseng gold gel 0.2g, carbomer 0.2g and 0.1%2.4-dinitrochlorobenzene 0.2ml, be coated in depilation district on the right side of Cavia porcellus, continue 6 hours, then clean.At once observing skin allergy situation, and skin allergy situation when 24h, 48h, 72h, skin allergy standards of grading are by shown in " new Chinese medicine development " page 110,111.
3. result of the test sensitization situation is shown in Table 1:
Table 3 is visible, and positive group performance extremely sensitization, compound recipe ginseng gold gel group and blank group are all without sensitization.
4. conclusion (of pressure testing): compound recipe joins gold gel to guinea pig skin without sensitization.
Table 1 anaphylaxis result of the test n=10
The clinical observation of embodiment 16 compound recipe ginseng gold gel for treating solar dermatitis
1 clinical data
1.1 physical data 189 examples are male, age 18～47 years old, 21.1 years old mean age, and sunburn to consultation time is 8～24h, average 16h.Injury red swelling of the skin, has an obvious burn feeling, face, cervical region and catch up with the exposure portion such as limb and erythema, pimple, mound kitchen rash occur, even oozes out, rotten to the corn.Average skin lesion area 9.8%TBSA wherein treatment group uses compound recipe ginseng gold gel for treating person 95 example,；Matched group uses Compound Calamine Lotion therapist 94 example.
The 1.2 simple debridements of Therapeutic Method Wound treatment, after normal saline cleans wound surface, wherein pollute obvious wound surface to sterilize with 1% benzalkonium bromide solution, compound recipe ginseng gold gel is directly uniformly sprayed on wound surface after dipping in dry wound surface by sterile gauze, every 4～6h are administered once, it is administered front cotton swab by wound surface exudate removal, to ensure that wound surface is clean, moistening, does not impregnates.To expose treatment, should not wrap up.It should be noted that not damaged principle in therapeutic process, do not make wound pain, hemorrhage, dry.Ibid, the course for the treatment of is 10d to the Therapeutic Method of matched group Compound Calamine Lotion.
1.3 statistical procedures: compare employing t inspection between two groups of groups.
2 result two groups analgesic time, healing times are shown in Table 1.
1 liang of table group solar dermatitis therapeutic effect compares
Note: compare with matched group**P ＜ 0.01, * P ＜ 0.05
3 discuss
Result shows herein, the compound recipe ginseng gold gel group treatment analgesic time (44 ± 15min) of solar dermatitis, healing time (6.1 ± 1.9min), cure rate (72.63%) and total effective rate (92.63%) the most a little higher than Compound Calamine Lotion (201 ± 19min, 8.1 ± 3.1min, 55.32%, 74.47%), there were significant differences for statistical procedures, there were significant differences for two groups of curative effects, treatment group is better than matched group, and treatment group action temperature and, without obvious adverse reaction.We apply compound recipe ginseng gold gel for treating solar dermatitis, achieve significant curative effect, have that relieving pain, anti-infectious function be strong, wound healing soon, more after the advantages such as cicatrix is light, its Therapeutic Method is simple, and curative effect is reliable.
Claims (6)
1. an externally-applied medicinal composition, is mainly made up of the crude drug of following weight portion: Radix Sophorae Flavescentis 4-8 part, Flos Chrysanthemi Indici 4-8 part, Herba Taraxaci 4-8 part, Herba Violae 4-8 part, Flos Lonicerae 4-8 part, Cortex Phellodendri 2-6 part, Rhizoma Polygoni Cuspidati 2-6 part, Cortex Dictamni 2-6 part, Fructus Cnidii 2-6 part, Borneolum Syntheticum 0.5-1 part.
2. externally-applied medicinal composition as claimed in claim 1, it is characterised in that be made up of the crude drug of following weight portion: Radix Sophorae Flavescentis 5-7 part, Flos Chrysanthemi Indici 5-7 part, Herba Taraxaci 5-7 part, Herba Violae 5-7 part, Flos Lonicerae 5-7 part, Cortex Phellodendri 3-5 part, Rhizoma Polygoni Cuspidati 3-5 part, Cortex Dictamni 3-5 part, Fructus Cnidii 3-5 part, Borneolum Syntheticum 0.6-0.9 part.
3. externally-applied medicinal composition as claimed in claim 1, it is characterised in that be made up of the crude drug of following weight portion: Radix Sophorae Flavescentis 6 parts, Flos Chrysanthemi Indici 6 parts, Herba Taraxaci 6 parts, Herba Violae 6 parts, Flos Lonicerae 6 parts, Cortex Phellodendri 4.5 parts, Rhizoma Polygoni Cuspidati 4.5 parts, Cortex Dictamni 4.5 parts, Fructus Cnidii 4.5 parts, Borneolum Syntheticum 0.75 part.
4. the externally-applied medicinal composition as described in any one of claim 1-3, the purposes in preparation treatment solar dermatitis disease medicament.
5. the externally-applied medicinal composition treating solar dermatitis, it includes the pharmaceutical composition described in any one of claim 1-3 and pharmaceutically acceptable adjuvant, for exterior-applied formulation, including solution, lotion, oil preparation, tincture, liniment, ointment, Emulsion, paste, liniment, spray, membrane, spirit, mucilage, paste, gel.
6. as described in claim 1 and 5, treat the externally-applied medicinal composition of solar dermatitis, it is characterised in that preparation method comprises the steps:
1) weigh described weight proportion takes Radix Sophorae Flavescentis, Flos Chrysanthemi Indici, Herba Taraxaci, Herba Violae, Flos Lonicerae, Cortex Dictamni Six-element, adds 12 times amount water boiling and extraction 3 times, each 1 hour, merges three extracting solution, and is concentrated into every milliliter and is equivalent to 0.5g crude drug, standby；
2) separately take the Cortex Phellodendri of described weight proportion, Rhizoma Polygoni Cuspidati, Fructus Cnidii three taste, add 8 times amount 60% alcohol reflux 3 times, united extraction liquid, reclaim ethanol and be concentrated into every milliliter and be equivalent to 0.5g crude drug, standby；
3) by 1) and 2) merging of gained solution, separately take Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add 5% propylene glycol, 1% azone, 3% carbomer, stir evenly, with triethylamine tune pH value to 6.5,1000ml the most processed, obtain gel；
4) in above-mentioned steps 2) after, by 1) and 2) merging of gained solution, separately take Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, stir evenly, 1000ml the most processed, obtain solution；
5) in above-mentioned steps 2) after, separately prepare emulsion type ointment base: oil-phase component (stearic acid 7.5g, Sorbitan Oleate 2g, glyceryl monostearate 7.5g, liquid paraffin 12g and vaseline 7g) and water-phase component (polyoxyethylene sorbitan monoleate: 6g, glycerol 12.5, sorbic acid 1g and water 75g) are separately heated to 80, oil phase is added in aqueous phase, stirring while adding, to being condensed into emulsion-type substrate, by 1) and 2) gained solution merge after, it is slowly added in the emulsion-type substrate prepared, stirring while adding, to be completely uniformly to, obtain ointment；
6) in above-mentioned steps 2) after, by 1) and 2) merging of gained solution, separately taking Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add polysorbate-8020ml, glycerol 60ml, azone 15ml, adjustment volume, to 1000ml, obtains liniment；
7) in above-mentioned steps 2) after, separately take sodium carboxymethyl cellulose 30g, polyvinyl alcohol 22g and add water respectively and appropriate soaks swelling and make to dissolve, stir evenly, filter, obtain filtrate standby；By filtrate, 1) and 2) gained solution merges, separately takes Borneolum Syntheticum and add with after appropriate anhydrous alcohol solution, add glycerol 30g, adjustment volume, to 1000ml, obtains liniment.
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| CN106692830A (en) * | 2017-03-03 | 2017-05-24 | 贵州都匀市剑江药业有限公司 | Traditional Chinese medicine composition for treating skin stubborn dermatitis and preparation method thereof |
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