CN106377583A - Gansu genuine medicinal material radix aconiti penduli processing technology and quality detection method of Gansu genuine medicinal material radix aconiti penduli - Google Patents
Gansu genuine medicinal material radix aconiti penduli processing technology and quality detection method of Gansu genuine medicinal material radix aconiti penduli Download PDFInfo
- Publication number
- CN106377583A CN106377583A CN201611012596.2A CN201611012596A CN106377583A CN 106377583 A CN106377583 A CN 106377583A CN 201611012596 A CN201611012596 A CN 201611012596A CN 106377583 A CN106377583 A CN 106377583A
- Authority
- CN
- China
- Prior art keywords
- processed product
- aconitum szechenyianum
- szechenyianum gay
- methanol
- aconitum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
- A61K36/714—Aconitum (monkshood)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7023—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms
- A61K9/703—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms characterised by shape or structure; Details concerning release liner or backing; Refillable patches; User-activated patches
- A61K9/7038—Transdermal patches of the drug-in-adhesive type, i.e. comprising drug in the skin-adhesive layer
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7023—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms
- A61K9/703—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms characterised by shape or structure; Details concerning release liner or backing; Refillable patches; User-activated patches
- A61K9/7038—Transdermal patches of the drug-in-adhesive type, i.e. comprising drug in the skin-adhesive layer
- A61K9/7076—Transdermal patches of the drug-in-adhesive type, i.e. comprising drug in the skin-adhesive layer the adhesive comprising ingredients of undetermined constitution or reaction products thereof, e.g. rosin or other plant resins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Dermatology (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the field of Chinese herbal medicine processing, and relates to a Gansu genuine medicinal material radix aconiti penduli processing technology and a quality detection method thereof. The invention provides a radix aconiti penduli processing product, which is prepared by the following steps that radix aconiti penduli medicinal materials are taken and are cut into slices; rice vinegar is used for stewing and moistening for 12 to 36h; drying is performed after the stewing and moistening; 8 to 12 times of the quantity of medium sand is added; stir-frying is performed for 5 to 8 minutes at 255 to 265 DEG C; cooling is performed to obtain the radix aconiti penduli processing product. The invention also provides a method for simultaneously measuring the content of three kinds of ingredients in the radix aconiti penduli processing product by a high-performance liquid chromatography method, so that the quality of the radix aconiti penduli processing product can be effectively controlled.
Description
Technical field
The invention belongs to Chinese medicine preparation field, it is related to processing procedure and its quality inspection of a kind of Gansu genunie medicinal materials Aconitum Szechenyianum Gay
Survey method.
Background technology
Aconitum Szechenyianum Gay is for Ranunculaceae aconitum plant Aconitum Szechenyianum GayAconitum pendulum Busch [A.szecheny - ianum Gay.]With Aconitum flavum Hand-Mazz A.flavum Hand.-Mazz.[A.anthora L.var.gilvum Maxim.]'s
Root tuber.Aconitum Szechenyianum Gay is Gansu Province's genunie medicinal materials, the property of medicine：It is bitter, pungent, temperature.It is very toxic.With promoting blood circulation, dispelling wind and eliminating dampness, detumescence stops
The effect of pain.Cure mainly traumatic injury.The swollen pain of fracture silt, lumbago due to wind-wetness evil, Carbuncle swells dislikes sore, nameless sores or boils, the not routed person of scrofula, poisonous snake
Bite, pernio.
At present, method is scalded using sand more than the processing mode of Aconitum Szechenyianum Gay, Aconitum Szechenyianum Gay main pharmacodynamics composition is alkaloids, wherein newly
Black peaceful alkali, 3-acetylaconitine, 12- table acetyl dehydrogenations aconitine are main alkaloid, and now there are some researches show Xin Wuning alkali toxicity
Larger, 3-acetylaconitine and 12- table acetyl dehydrogenation aconitine toxicity are smaller, and have stronger bioactivity, and research shows,
Heating, which is processed, can reduce the content of Xin Wuning alkali, but as heating is processed, its bioactive ingredients 3-acetylaconitine and 12- tables
Acetyl dehydrogenation aconitine is also in reduction, and this is a problem of industry.In addition, existing study Aconitum Szechenyianum Gay concocting process,
Processing parameter is described, but it is empirical too strong, and concocting process is difficult to hold, and manufacturing parameter is also indefinite, and in many reports
To passing on experiential operating the measurement and process control of temperature, without controllability more.
Based on this, the important unit Gansu university of TCM that the present inventor studies as Gansu Province's Aconitum Szechenyianum Gay medicinal material
Scientific research personnel, Aconitum Szechenyianum Gay has been carried out years of researches, inventor is by experimental study repeatedly, it is determined that Aconitum Szechenyianum Gay
Processing procedure, be its production and quality control accurate experimental basis is provided.
Also, had been surprisingly found that in Aconitum Szechenyianum Gay lasting research for many years, Aconitum Szechenyianum Gay has to asthma certain controls curative effect
Really, but scientific research personnel's research finds after only being processed to Aconitum Szechenyianum Gay using specific concocting method, have necessarily to asthma
Therapeutic effect, raw Aconitum Szechenyianum Gay, fry that other Aconitum Szechenyianum Gay processed products such as Aconitum Szechenyianum Gay are substantially invalid, inventor has carried out many to this problem
The research in year, has invented a kind of processed product pharmaceutical preparation of Aconitum Szechenyianum Gay, the medicine has extraordinary therapeutic effect to asthma,
After a series of experimental study for carrying out science, specification, technical scheme has been obtained.
The content of the invention
It is an object of the invention to provide a kind of Aconitum Szechenyianum Gay processed product.
It is a further object of the present invention to provide the preparation method of the Aconitum Szechenyianum Gay processed product.
The quality determining method that the Aconitum Szechenyianum Gay processed product is also provided of the present invention.
The preparation method that the Aconitum Szechenyianum Gay processed product rubber ointment is also provided of the present invention
The present invention's also provides application of the Aconitum Szechenyianum Gay processed product rubber ointment in treatment asthmatic medicament is prepared.
The purpose of the present invention is achieved in the following ways：
A kind of Aconitum Szechenyianum Gay processed product, the Aconitum Szechenyianum Gay processed product is to adopt to prepare with the following method：Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into,
12~36 h are stewed to moisten with rice vinegar, are dried after stewing to moisten, plus 8~12 times amount middle sand, at 255~265 DEG C, the min of frying 5~8,
Cooling, produces Aconitum Szechenyianum Gay processed product.
Described Aconitum Szechenyianum Gay processed product, is preferably, adopts and prepare with the following method：Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, with rice
Vinegar stews to moisten 24h, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C, and frying 6min, cooling produces Aconitum Szechenyianum Gay processed product.
A kind of detection method of Aconitum Szechenyianum Gay processed product, the Aconitum Szechenyianum Gay processed product is adopted to be prepared with the following method：Take iron staff
Hammer into shape medicinal material, be cut into sheet, 12~36 h stewed to moisten with rice vinegar, dried after stewing to moisten, plus 8~12 times amount middle sand, at 255~265 DEG C
Under, the min of frying 5~8, cooling produces Aconitum Szechenyianum Gay processed product；
Gone using Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl in hplc simultaneous determination Aconitum Szechenyianum Gay processed product
The content of hydrogen rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 30~40 DEG C of column temperature, 0.5~1.5mLmin of flow velocity-1, the μ of sample size 5~20
L；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to alkali containing Xin Wuning, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines
Mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Accurately weighed Aconitum Szechenyianum Gay processed product powder, puts in conical flask with cover, and precision addition 65~
75% methanol, close plug, weighed quality is heated to reflux, let cool, then weighed quality, and the quality of less loss is supplied with 65~75% methanol, is shaken
It is even, filtered with miillpore filter, take subsequent filtrate, produced；
（4）Determine：Each 5~20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
The detection method of described Aconitum Szechenyianum Gay processed product, be preferably, the Aconitum Szechenyianum Gay processed product is to adopt to prepare with the following method
's：Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C, fry
6min processed, cooling, produces Aconitum Szechenyianum Gay processed product；
Preferably, using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Accurately weighed Aconitum Szechenyianum Gay processed product powder 0.8g of the present invention, puts in conical flask with cover, essence
70% methanol 25mL of close addition, close plug, weighed quality is heated to reflux 1h, let cool, then weighed quality, and less loss is supplied with 70% methanol
Quality, shake up, filtered with 0.22 μm of miillpore filter, take subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
A kind of Aconitum Szechenyianum Gay processed product rubber ointment, the Aconitum Szechenyianum Gay processed product rubber ointment is to adopt to prepare with the following method：
（1）Take Aconitum Szechenyianum Gay medicinal material, be cut into sheet, 12~36 h are stewed to moisten with rice vinegar, dried after stewing to moisten, plus 8~12 times amount middle sand,
At 255~265 DEG C, the min of frying 5~8, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add the conventional substrate of rubber ointment, mix, be made coating, carry out painting cream, segment,
Lid lining, is cut into small pieces, produces.
Described Aconitum Szechenyianum Gay processed product rubber ointment, is preferably that the Aconitum Szechenyianum Gay processed product rubber ointment is to use such as lower section
It is prepared by method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced.
A kind of application of Aconitum Szechenyianum Gay processed product rubber ointment in treatment asthmatic medicament is prepared, the Aconitum Szechenyianum Gay processed product rubber
Paste is to adopt to prepare with the following method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced；
Using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product rubber ointment
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Aconitum Szechenyianum Gay processed product rubber ointment 1 is taken, it is accurately weighed, it is cut into the broken of 1cm × 1cm
Piece, puts in 50mL round-bottomed flasks, and precision adds 70% methanol 25mL, and close plug, weighed quality is heated to reflux 1h, let cool, then weighed
Quality, the quality of less loss is supplied with 70% methanol, is shaken up, and is filtered with 0.22 μm of miillpore filter, is taken subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
A kind of application of Aconitum Szechenyianum Gay processed product rubber ointment in treatment of arthritis thing is prepared, the Aconitum Szechenyianum Gay processed product rubber
Paste is to adopt to prepare with the following method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced；
Using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product rubber ointment
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Aconitum Szechenyianum Gay processed product rubber ointment 1 is taken, it is accurately weighed, it is cut into the broken of 1cm × 1cm
Piece, puts in 50mL round-bottomed flasks, and precision adds 70% methanol 25mL, and close plug, weighed quality is heated to reflux 1h, let cool, then weighed
Quality, the quality of less loss is supplied with 70% methanol, is shaken up, and is filtered with 0.22 μm of miillpore filter, is taken subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
The technique effect of the present invention is verified by following experimental study：
Experiment one：The experimental study that Aconitum Szechenyianum Gay processed product rubber ointment is acted on arthritis treatment
1 materials and methods
1.1 experimental animal
SD rats 60, male, body weight（100±20）G, Gansu university of TCM Medical experimental center is provided, the animal quality certification
Number：SCXK（It is sweet）2011-0001-0001011；Experimental Establishment quality certification number：SYXK（It is sweet）2011-0001-0000314, is raised
In Gansu university of TCM Medical experimental center.
1.2 Experimental agents
1.2.1 Aconitum Szechenyianum Gay processed product rubber ointment A
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 260 DEG C, frying 6min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.2.2 Aconitum Szechenyianum Gay processed product rubber ointment B
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet, plus 10 times of middle sand measured are cut into, at 260 DEG C, frying 6min,
Cooling, produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.2.3 Aconitum Szechenyianum Gay processed product rubber ointment C
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 220 DEG C, frying 3min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.2.4 Aconitum Szechenyianum Gay processed product rubber ointment D
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with yellow rice wine, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 300 DEG C, frying 6min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.3 modeling
Take the appropriate Collagen Type VI of ox II（Beijing Bo Leide bio tech ltd）It is dissolved in 0.01mol/L acetic acid solution
（2mg collagens are dissolved in 1ml acetic acid solutions）, equivalent freund 's incomplete adjuvant is added after fully being dissolved overnight at 4 DEG C, in ice bath
Under fully emulsified emulsion is made.The intracutaneous multi-point injection collagen emulsion 1.5ml/100g of rat back under intraperitoneal anesthesia.
1.4 packets and treatment
Modeling is i.e. random after terminating to be divided into 5 groups by rat, every group 12, is respectively：Aconitum Szechenyianum Gay processed product rubber ointment A groups, iron
Rod hammer processed product rubber ointment B groups, Aconitum Szechenyianum Gay processed product rubber ointment C groups, Aconitum Szechenyianum Gay processed product rubber ointment D groups, blank base
Matter control group.
Application group takes Shenzhu, Zhiyang and MINGMEN acupoint with reference to " rat acupuncture point collection of illustrative plates ", and the medicine of 1cm × 1cm sizes is applied per cave
Thing rubber ointment paster；The paster of bare substrate control group is in addition to not drug containing, remaining composition and Experimental agents rubber ointment group
Identical, acupoint selection and paster area are with application group.Each group rat is treated since after modeling the 12nd day, rubber ointment paster, often
Day 1 time, application 12 hours, continuous to treat 12 days every time.
1.5 swelling degree of the paw are determined
Using homemade plethysmometer, its swelling degree of the paw is observed according to the Volume Changes of rat paw.Each group rat is distinguished
The 12nd day after modeling（Before treatment）, the 18th day after modeling（After treatment 6 days）And the 24th day after modeling（After treatment 12 days）Determine
The volume of left and right rear vola pedis, and calculate the average swelling of every rat hindleg plantar.
1.6 arthritis index are determined
Using WoodShi arthritis score standards, according to each rat redness and swelling of joints degree and involve joint number, score and compare.Specifically
It is described as follows.
0 point：Normally；1 point：Redness is related to 1 toe joint；2 points：Redness is related to 2 and above toe joint or whole vola pedis
Mild redness；3 points：Vola pedis redness is more serious；4 points：Vola pedis severe is red and swollen, lacks flexibility.Every vola pedis in each 4 vola pedis of rat
All remember 0~4 point, calculate total mark.Daily observed and recorded 1 time.
1.7 PGE2The measure of concentration
The 29th day eye socket takes blood about 1.5mL, centrifuging and taking serum to each rat after the 15th day and modeling before modeling, after modeling respectively.
Last time takes each rat after blood and takes off cervical vertebra execution, cuts left and right rear vola pedis at 1cm on ankle-joint, divests skin, weigh
After be cut into small pieces, by 1mL/300mg dosage add physiological saline, homogenate is made, 37 DEG C of warm bath are after 15 minutes, centrifuging and taking supernatant
Liquid.According to the explanation of kit, PGE in serum and vola pedis homogenate is determined respectively with radioimmunology2Concentration.
The measure each group rat of 1.8 index and spleen index wins spleen after execution, weighs, and calculates index and spleen index：Spleen refers to
Number=spleen weight（mg）/ body weight（g）.
All data informations of 1.9 statistical procedures are measurement data, with mean ± standard deviation（±s）Represent.Make
Handled with SPSS10.0 statistical softwares, comparing two-by-two between carrying out each group with t inspections, with P<0.05 is to have significant difference, P<
0.01 is to have significant differences.
2 results
The influence of the average swelling of 2.1 pairs of collagen induced-arthritis rat paws
This experimental result is shown in Table 1, as a result shows, Aconitum Szechenyianum Gay processed product rubber ointment A is applied to collagen induced-arthritis rat
The inflammatory reaction of pedal swelling there is obvious inhibitory action, and the action effect of Aconitum Szechenyianum Gay processed product rubber ointment A groups
It is substantially better than Aconitum Szechenyianum Gay processed product rubber ointment B groups, C groups and D groups.
The average swelling of the vola pedis of each group rat different times of table 1 compares（±s）
Compared with same period blank group, * P<0.05, * * P<0.01；
2.2 the influence to collagen induced-arthritis rat arthritis index
This experimental result is shown in Table 2, as a result shows, Aconitum Szechenyianum Gay processed product rubber ointment A is applied to collagen induced-arthritis rat
Arthritis index there is obvious inhibitory action, and the action effect of Aconitum Szechenyianum Gay processed product rubber ointment A groups is substantially better than
Aconitum Szechenyianum Gay processed product rubber ointment B groups, C groups and D groups.
The comparison of the arthritis index of each group rat different times of table 2（±s）
Compared with same period blank group, * P<0.05, * * P<0.01；
PGE in 2.3 pairs of collagen induced-arthritis rat paw inflammatory tissues2Influence
This experimental result is shown in Table 3, as a result shows, Aconitum Szechenyianum Gay processed product rubber ointment A applications can significantly reduce collagen Ⅱ induced pass
PGE after the scorching rat two of section in vola pedis inflammatory tissue2Concentration, points out the therapy may be by suppressing PGE2Generation and effectively
Local inflammation is controlled, and the action effect of Aconitum Szechenyianum Gay processed product rubber ointment A groups is substantially better than Aconitum Szechenyianum Gay processed product rubber paste
Agent B groups, C groups and D groups.
PGE in each group rat different times vola pedis inflammatory tissue of table 32The comparison of concentration（± s, pg/mL）
Compared with blank group, * P<0.05, * * P<0.01；
2.4 the influence to collagen induced-arthritis Rats Spleen index
This experimental result is shown in Table 4, as a result shows, the spleen of collagen induced-arthritis animal model has different degrees of enlargement.
In this experiment, the index and spleen index of Aconitum Szechenyianum Gay processed product rubber ointment A group rats is significantly less than bare substrate control group, shows
Aconitum Szechenyianum Gay processed product rubber ointment A can suppress the enlargement of spleen, and then point out the therapy to have immune response hyperfunction in vivo
Inhibitory action, and the action effect of Aconitum Szechenyianum Gay processed product rubber ointment A groups is substantially better than Aconitum Szechenyianum Gay processed product rubber ointment B
Group, C groups and D groups.
The comparison of index and spleen index after the treatment of each group rat of table 4（±s）
Compared * * P with blank group<0.01
3 conclusions
Experimental study shows that Aconitum Szechenyianum Gay processed product rubber ointment A has significant therapeutic effect, and Aconitum Szechenyianum Gay big gun to arthritis
Product rubber ointment A action effect is substantially better than Aconitum Szechenyianum Gay processed product rubber ointment B, C and D.Illustrate only to carry in the present invention
The Aconitum Szechenyianum Gay processed product rubber ointment prepared under the specific concocting method supplied just controls curative effect to arthritis with particularly pertinent
Really.
Experiment three：The experimental study of rubber ointment application treatment Experimental Asthma In Guinea-pigs of the present invention
1 materials and methods
Experimental animal：Healthy guinea pig 60（Male and female half and half）300 ± 30g of body weight.Gansu university of TCM Medical experimental center is carried
For animal quality certification number：SCXK（It is sweet）2011-0001-0001011；Experimental Establishment quality certification number：SYXK（It is sweet）2011-0001-
0000314, raise in Gansu university of TCM Medical experimental center.
1.1 Aconitum Szechenyianum Gay processed product rubber ointment A
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 260 DEG C, frying 6min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.2 Aconitum Szechenyianum Gay processed product rubber ointment B
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet, plus 10 times of middle sand measured are cut into, at 260 DEG C, frying 6min,
Cooling, produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.3 Aconitum Szechenyianum Gay processed product rubber ointment C
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 220 DEG C, frying 3min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.4 Aconitum Szechenyianum Gay processed product rubber ointment D
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with yellow rice wine, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 300 DEG C, frying 6min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.5 GUILONG KECHUANNING JIAONANG：Osmanthus dragon medicine company（Anhui）Co., Ltd produces, Chinese medicines quasi-word Z20053135.
1.6 reagents and instrument：
Egg protein dry powder formulations（Wuhan Boster Biological Technology Co., Ltd. produces）, IL-4 radioimmunological kits（The inscription life of Suzhou section
Thing Technology Co., Ltd. produces）, IgG radioimmunological kits（Shanghai Hu Yu bio tech ltd is produced）, CX5 Biochemical Analyzers
（U.S. Bake Man produces, and core dept. of radiology of the People's Hospital of Gansu Province provides）, CW-20D ultrasonic atomizatio instrument（German Bry company
Manufacture）, calculating instrument is immunized in the full-automatic C of FJ-2008（Generation connection is rich to grind the manufacture of (Beijing) Science and Technology Ltd.）.
2 EXPERIMENTAL METHODS Animals are grouped：
2.1 packet
With reference to " rat acupuncture point collection of illustrative plates ", from Shenzhu, Zhiyang and MINGMEN acupoint.By 84 cavys with the nursing of balanced feed adaptability
6 days, it is divided into according to " the digital random table method of dividision into groups "：Aconitum Szechenyianum Gay processed product rubber ointment A groups, Aconitum Szechenyianum Gay processed product rubber ointment B
Group, Aconitum Szechenyianum Gay processed product rubber ointment C groups, Aconitum Szechenyianum Gay processed product rubber ointment D groups, blank control group, model control group, Gui Long
Methoxyphenamini hydrochloridum control group, totally 7 groups, every group 12.
2.2 modeling
In addition to blank control group, to Aconitum Szechenyianum Gay processed product rubber ointment A groups, Aconitum Szechenyianum Gay processed product rubber ointment B groups, Aconitum Szechenyianum Gay big gun
Product rubber ointment C groups, Aconitum Szechenyianum Gay processed product rubber ointment D groups, model control group, Guilong Kechuanning Capsule control group, modeling.To every
The egg protein physiological saline 1mL of intraperitoneal injection 10% of cavy carries out sensitization, is repeated once within 6 days.Cavy is put into atomization after 12 days
In case, with the egg protein physiological saline 15min of constant pressure 400mmHg Neulized inhalations 1%.Observation occurs dehiscing to pant to cavy, tripe abdomen drum
Untill, i.e. modeling success.
2.3 administration：
Aconitum Szechenyianum Gay processed product rubber ointment A groups：Cavy, is carried out with 2% sodium thiosulfate at its Shenzhu, Zhiyang and MINGMEN acupoint position
Depilation, depilation area 2cm × 2cm or so.After 75% alcohol disinfecting, Aconitum Szechenyianum Gay processed product rubber ointment A is spread at depilation.
Rubber ointment of the present invention is taken down after 18h.1 time a day, stick altogether 6 days.
Aconitum Szechenyianum Gay processed product rubber ointment B groups：Cavy, with 2% sodium thiosulfate at its Shenzhu, Zhiyang and MINGMEN acupoint position
Lost hair or feathers, depilation area 2cm × 2cm or so.After 75% alcohol disinfecting, Aconitum Szechenyianum Gay processed product rubber ointment B is spread on into depilation
Place.Rubber ointment of the present invention is taken down after 18h.1 time a day, stick altogether 6 days.
Aconitum Szechenyianum Gay processed product rubber ointment C groups：Cavy, with 2% sodium thiosulfate at its Shenzhu, Zhiyang and MINGMEN acupoint position
Lost hair or feathers, depilation area 2cm × 2cm or so.After 75% alcohol disinfecting, Aconitum Szechenyianum Gay processed product rubber ointment C is spread on into depilation
Place.Rubber ointment of the present invention is taken down after 18h.1 time a day, stick altogether 6 days.
Aconitum Szechenyianum Gay processed product rubber ointment D groups：Cavy, with 2% sodium thiosulfate at its Shenzhu, Zhiyang and MINGMEN acupoint position
Lost hair or feathers, depilation area 2cm × 2cm or so.After 75% alcohol disinfecting, Aconitum Szechenyianum Gay processed product rubber ointment D is spread on into depilation
Place.Rubber ointment of the present invention is taken down after 18h.1 time a day, stick altogether 6 days.
Guilong Kechuanning Capsule control group：Cavy, capsule is removed with GUILONG KECHUANNING JIAONANG, built-in medicinal powder is with 1.0 g/kg/d use
Amount, 1 is pressed with distilled water：8 ratio is tuned into water pasty state, carries out gavage treatment.
Blank group, model group animal do not make any processing in addition to daily crawl is stimulated.
Collection of specimens and detection method EOS detections：To every group of cavy, cavy tail point blood is drawn using capillary acupoint selection pipe
20 μ L, add in 0.38mL Yihong-formaldehyde dilution, gently shake up, and stand 5min, are detected using conventional counting method.
IgG, IL-4 are detected：To every group of cavy, with 3.5% sodium phenobarbital 0.1g/kg intraperitoneal injection of anesthesia, eyeball vein
Take a blood sample 6mL, dispenses 3 test tubes.Each 2mL it is to be solidified after separation serum, -20 DEG C preservation, according to kit illustration method and will
Operation is asked, IgG, IL-4 are detected using the method for exempting from point low temperature refrigerator is put.
Statistical procedures：Handled using SPSS11.0 system softwares, all data mean ± standard deviation（±s）Table
Show, compare between each group and use variance analysis.
3 experimental results
Experimental result is shown in Table 5.As can be seen that the EOS of model group animal is counted after modeling and IL-4 content significantly rises（P<
0.05, P<0.01）, IgG is decreased obviously（P<0.05）, illustrate modeling success.
After treatment, Aconitum Szechenyianum Gay processed product rubber ointment A groups are compared with model group, and EOS is significantly reduced（P<0.01）,
IgG significantly increases（P<0.01）, there is significant differences；IL-4 is decreased obviously（P<0.05）.Aconitum Szechenyianum Gay processed product rubber
Glue paste B groups, C groups, D groups are compared with model group, and EOS is not significantly reduced, and IgG does not increase significantly, and IL-4 is also without obvious
Decline.
EOS and the results contrast of serum total Ig G, IL-4 in each group guinea pig blood of table 5（±s）
Note：Compared with blank group, * P<0.05, * * P<0.01；
The comparison of model group, #P<0.05, ##P<0.01.
4 conclusions
This experimental result confirms that Aconitum Szechenyianum Gay processed product rubber ointment A can substantially reduce EOS in asthmatic guinea pigs blood and count, and can carry
The content of high serum IgG, reduces IL-4 contents, absolutely proves that Aconitum Szechenyianum Gay processed product rubber ointment A can improve the immune work(of body
Can, there is preferable resolving sputum and relieving asthma to act on, its curative effect is substantially better than Aconitum Szechenyianum Gay processed product rubber ointment B groups, C groups and D groups.Illustrate only
There is the Aconitum Szechenyianum Gay processed product rubber ointment prepared under the specific concocting method that the present invention is provided that just there is asthma especially to dash forward
The therapeutic effect gone out.
Experiment four：Safety research of the Aconitum Szechenyianum Gay processed product rubber ointment through percutaneous drug delivery
In order to ensure the clinical security through percutaneous drug delivery of Aconitum Szechenyianum Gay processed product rubber ointment, inventor carries out related experiment evaluation
The security of Aconitum Szechenyianum Gay processed product rubber ointment.
1 material
1.1 instrument
TCS-30 electronic platform scales（Xiamen Yi Heng electronic-weighings Co., Ltd）；TP-2101 electronic balances（The virtuous permanent electronic-weighing in Xiamen
Co., Ltd）.
1.2 medicines and reagent
Aconitum Szechenyianum Gay processed product rubber ointment：
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of amounts
Middle sand, at 260 DEG C, frying 6min, cooling, produce Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced.
1.3 animal
Regular grade new zealand rabbit, ♀ ♂ half and half, 2.0~2.5kg of weight, Gansu university of TCM Medical experimental center is provided,
Animal quality certification number：SCXK（It is sweet）2011-0001-0001011；Experimental Establishment quality certification number：SYXK（It is sweet）2011-0001-
0000314, raise in Gansu university of TCM Medical experimental center.
Regular grade FMMU Albino guinea pigs, ♀ ♂ half and half, average weight is 270g, Gansu university of TCM when experiment starts
Medical experimental center is provided, animal quality certification number：SCXK（It is sweet）2011-0001-0001011；Experimental Establishment quality certification number：SYXK
（It is sweet）2011-0001-0000314, is raised in Gansu university of TCM Medical experimental center.
2 methods
2.1 acute toxicity testing
Rabbit 20 is taken, ♀ ♂ half and half are randomly divided into intact skin group and damaged skin group, every group 10.24h before administration, electricity consumption
Clipper is in rabbit back depilation, and lose hair or feathers area about 9cm × 9cm, checks whether skin of unhairing is injured and rejects injured family after unhairing
Rabbit.Damaged skin group 75% ethanol disinfection of rabbit back hair removal section, then with disposable syringe syringe needle in depilation zoning " well "
Word otch, not damage hypodermis and have oozing of blood as standard.After each group rabbit patch correspondence patch 24h, remained with warm water cleaning
Medicine, starts the indexs such as observer's rabbit skin, eyes, mucous membrane, breathing, diet, stool and urine, autonomic activities, behavior change at once；
When there is toxicity reaction, then the order of severity and Disappearance Scenarios of toxic reaction are observed.Continuous Observation 12 days, observation 2 times daily, and
In the 0th, 1,6,12 days measurement weights.If any dead or dying rabbit, or when there is macroscopic lesion, then carry out in time
Dissection and histopathologic examination.
2.2 single primary cutaneous stimulation tests
Rabbit 20 is taken, ♀ ♂ half and half are randomly divided into intact skin group and damaged skin group, every group 10.Each group rabbit uses
Consubstantiality itself left and right sides method of comparison, vaseline is given on right side（0.5g/ is only）, correspondence medicine is given in left side（2cm × 2cm/ is only）, it is single
Medicine is removed after secondary administration, fixed 6h, warm water cleaning left drug is used.Rabbit is recorded before experiment starts and at the end of the observation period
Weight and the erythema and oedema situation for removing coating position after medicine the 1st, 24,48,72h.
Scores of erythema：Without 0 point of erythema meter, slight erythema（In pale red）1 point of meter, moderate erythema（In cerise）2 points of meter,
Severe erythema（In kermesinus）3 points of meter, Severe erythema（Aubergine erythema is formed to slight eschar）4 points of meter；Oedema scores：It is anhydrous
Swollen 0 point of meter, Mild edema（It is visible reluctantly）1 point of meter, intermediate edema（Skin substantially swells）2 points of meter, severe edema（Cutaneous protuberance
1mm, profile understands）3 points of meter, Severe edema（Cutaneous protuberance ＞ 1mm simultaneously have expansion）4 points of meter.Observe each group rabbit erythema and water
The average value of swollen integration, by skin irritatin intensity evaluation standard, carries out stimulus intensity evaluation, total score is 0~8 point, wherein 0~
0.49 point be it is nonirritant, 0.5~2.99 point be slight stimulation, 3.0~5.99 points be moderate excitant, 6.0~8.0 points be
Strong and stimulating.
2.3 sensitizations of skin are tested
FMMU cavys, ♀ ♂ half and half are randomly divided into intact skin group and damaged skin group, every group 10.Carried out in the 0th, 6,12d
Induction exposure, medicine-feeding part is that Guinea Pig Left side rib belly removes hair-fields skin；Contact is excited in 24d rows, medicine-feeding part is cavy
Right side rib belly removes hair-fields skin, and dosage and Induction exposure are consistent.Observe the 0th, 24,48, local skin is administered after 72h
Erythema, oedema situation, while observing animal whether there is the systemic anaphylaxis such as asthma, astasia or shock.Erythema, oedema evaluation
With method under " 2.2 " item, allergic reaction incidence=group in occur the number of animals of erythema, oedema or systemic anaphylaxis/
This group of animal number × 100%.Allergic reaction incidence is evaluated as evaluating when no sensitization, 10%~30% when being 0~10%
Be evaluated as when being evaluated as moderate sensitization, 60%~80% during for slight sensitization, 30%~60% height sensitization, 80%~
Extreme sensitization is evaluated as when 100%.
2.4 statistical method
Weight data are handled using SPSS18.0 statistical softwares.
3 results
3.1 acute toxicity testing results
During observation, each group rabbit diet drinking-water is normal, is showed no perpendicular hair, secretion and excreta, eyelid exception.Indivedual rabbit
It can be seen that slight red trace, suspected of patch itself stickiness with caused by artificial tear, 3d or so can disappear.Damaged skin each group rabbit is formed a scab
Outer to be showed no exception, the state of mind is good, and rabbit outward appearance and behavioral activity are normal, and damaged skin can be voluntarily cured at 6 days or so
Close, it is dead without rabbit at the end of experiment.0th, 1,6,12 days each group rabbit body mass ratios are compared with no significant difference.
3.2 single primary cutaneous stimulation test results
Aconitum Szechenyianum Gay processed product rubber ointment damaged skin group rabbit shows slight stimulation after medicine 1,24h is removed, and removes
Excitant disappears after medicine 48h.Aconitum Szechenyianum Gay processed product rubber ointment intact skin group rabbit is removing medicine 1,24,48,72h
Afterwards, skin irritatin average strength is 0 point, is evaluated as nonirritant.Refer to table 6.
The each group rabbit single primary cutaneous stimulation test result of table 6
3.3 sensitization of skin experimental results
Aconitum Szechenyianum Gay processed product rubber ointment intact skin group and Aconitum Szechenyianum Gay processed product rubber ointment damaged skin group cavy excite portion
Position skin does not occur erythema, oedema, and the systemic anaphylaxis such as no asthma, astasia or shock, allergic reaction incidence is equal
For 0, it is evaluated as having no cutaneous anaphylaxis.As a result show, Aconitum Szechenyianum Gay processed product rubber ointment is made to cavy without sensitization of skin
With.
4 conclusions
Aconitum Szechenyianum Gay processed product rubber ointment is non-toxic to skin, nonirritant, without sensitization, shows that it uses safe and nontoxic pair
Effect.In summary, Aconitum Szechenyianum Gay processed product rubber ointment is preferable through percutaneous drug delivery security.
Experiment five：HPLC determines the content of 3 kinds of compositions in Aconitum Szechenyianum Gay processed product of the present invention simultaneously
The present invention switches method on the basis of prior art research, using dual wavelength and determined simultaneously in Aconitum Szechenyianum Gay processed product of the present invention
Xin Wuning alkali, 3-acetylaconitine, the content of 12- table acetyl dehydrogenation aconitines, set up the stronger detection method of specificity, be
The quality of control Aconitum Szechenyianum Gay processed product of the present invention provides guarantee comprehensively.
1 instrument and reagent
The series of high efficiency liquid chromatographs of Agilent 1200, including G1322A types on-line degassing machine, G1311A types quaternary pump,
G1316A types column oven, G1314A variable-wavelenght detectors（VWD）, BS124S type electronic analytical balances（Sartorius）.
Xin Wuning alkali（Guangzhou Juan Mu bio tech ltd, lot number 151023,151102, through HPLC purity tests, is used
Normalization method is calculated, purity >=99%）, 3-acetylaconitine reference substance（Wuhan Rostand Chemical Industry Science Co., Ltd, lot number
151203rd, 151228, through HPLC purity tests, purity >=99%）, 12- table acetyl dehydrogenation aconitine reference substances（Nanjing jam this
Pharmaceutical Technology Co., Ltd, lot number 150926,151021, through HPLC purity tests, purity >=99%）, methanol（Chromatographic grade,
TEDIA companies）, water is distilled water, and the analysis of other reagents position is pure.
Aconitum Szechenyianum Gay processed product：Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10
The middle sand measured again, at 260 DEG C, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product.
2 methods and result
2.1 chromatographic condition
Chromatographic condition：Syncronis C chromatographic columns, ladder is carried out by mobility of methanol -0.05mol/L biphosphates sodium water solution
Degree elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at 85%；16
~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to 80%；21
~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to 60%；Inspection
Survey wavelength 235nm（0~20min, detection Xin Wuning alkali, 3-acetylaconitine）And 365nm（21~30min, detects 12- table second
Acyl dehydrogenation aconitine）, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20.
The preparation of 2.2 solution
2.2.1 the preparation of mixed reference substance solution
Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines reference substance appropriate, it is accurately weighed, dissolved with methanol
And it is diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg, 12- table acetyl dehydrogenation aconitine 0.220mg
Mixed reference substance solution, produce.
2.2.2 the preparation of need testing solution
Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines reference substance appropriate, it is accurately weighed, dissolved with methanol
And it is diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg, 12- table acetyl dehydrogenation aconitine 0.220mg
Mixed reference substance solution, produce.
2.3 methodological study
2.3.1 system suitability test
Precision draws each 20 μ L injections liquid chromatograph of mixed reference substance solution, need testing solution, enters by 2.1 lower chromatographic conditions
Sample is determined, and sees accompanying drawing 1 and accompanying drawing 2.Wherein Xin Wuning alkali, 3-acetylaconitine, the retention time of 12- table acetyl dehydrogenation aconitines
Respectively 8.15,22.85,46.64min, separating degree is good, is not disturbed by other components.
2.3.2 linear relationship is investigated
Precision draws 0.2,0.4,0.6,0.8,1.0,1.2mL mixed reference substance solution, is placed in 10mL measuring bottles, dilute with distilled water
Constant volume is released, the reference substance solution of 6 different quality concentration is made.Above-mentioned reference substance solution is taken, by 2.1 lower chromatographic condition sample introductions
Analysis, determines peak area, with integrating peak areas value（Y）For ordinate, reference substance mass concentration（X）It is bent that standard is drawn for abscissa
Line, calculates regression equation.It the results are shown in Table 7.
The linear relationship of table 7
2.3.3 precision is investigated
Precision draws same need testing solution by 2.1 lower chromatographic condition continuous sample introductions 6 times, records peak area, as a result Xin Wuning
Alkali, 3-acetylaconitine, the peak area RSD of 12- table acetyl dehydrogenation aconitines are respectively 0.26%, 0.29%, 0.32%, show instrument
The precision of device is good.
2.3.4 study on the stability
Same need testing solution is drawn by 2.1 lower chromatographic condition precisions to analyze in 0,3,6,9,12,15h sample introduction, records peak face
Product, as a result Xin Wuning alkali, 3-acetylaconitine, the peak area RSD of 12- table acetyl dehydrogenation aconitines are 1.65%, 1.38%,
1.39%, show that need testing solution is stable in 15h.
2.3.5 repeatability is investigated
6 parts of the Aconitum Szechenyianum Gay processed product powder of the present invention of same lot number is taken, by need testing solution is made under 2.2.2 respectively, by 2.1
Chromatographic condition sample introduction is analyzed under, as a result Xin Wuning alkali, 3-acetylaconitine, the peak area RSD of 12- table acetyl dehydrogenation aconitines
Respectively 1.41%, 1.32%, 1.45%, show repeated good.
2.3.6 recovery test
Precision weighs 6 parts of the Aconitum Szechenyianum Gay processed product powder of the present invention of known content, and every part of 0.5g is accurate respectively to add mixing control
Product storing solution 1.0mL, is analyzed by 2.1 lower chromatographic condition sample introductions, records peak area, as a result Xin Wuning alkali, 3-acetylaconitine,
The average recovery rate of 12- table acetyl dehydrogenation aconitines is respectively 98.23%, 101.25%, 102.34%, and rate of recovery RSD is respectively
1.65%, 1.48%, 1.52%, experimental result is shown in Table 8.
Each composition sample-adding recovery test in the Aconitum Szechenyianum Gay processed product of table 8
2.4 sample sizes are determined
3 batches of samples are taken respectively, and by 2.2.2 lower section legal system available test sample solutions, Xin Wuning alkali, 3- acetyl are calculated using external standard method
The content of aconitine, 12- table acetyl dehydrogenation aconitines.It the results are shown in Table 9.
The Aconitum Szechenyianum Gay processed product sample size of the present invention of table 9 is determined（n=3） mg·g-1
3 discuss and conclusion
3.1 the selection of Detection wavelength
Determine, respectively 3 are compareed while Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines in order to realize
Product solution carries out UV scanning in 200~700nm, and wherein Xin Wuning alkali, 3-acetylaconitine has maximum suction in 233,240nm
Receive, but 12- table acetyl dehydrogenation aconitines maximum absorption wavelength in 365nm, it is difficult to select single wavelength to enter this 3 compounds
Row ultraviolet detection.Therefore according to appearance time, change detection wavelength is measured, the absorption maximum of each composition is taken into account, inspection is improved
Survey sensitivity.
The selection of 3.2 mobile phases
Methanol-water, methanol -0.05mol/L biphosphates sodium water solution, methanol-water, 4 kinds of the acetic acid of methanol -0.1% water are investigated in experiment
Mobile phase, as a result with the elution best results of methanol -0.05mol/L sodium dihydrogen phosphate aqueous solution systems, and has attempted the flowing
A variety of linear gradient elutions of phase, obtained chromatographic peak peak shape and separating degree is all preferable, baseline stability.
This experiment set up method can determine simultaneously Xin Wuning alkali in Aconitum Szechenyianum Gay processed product of the present invention, 3-acetylaconitine,
12- table acetyl dehydrogenation Content of Aconitine, this method is easy, reliable, reproducible, can thoroughly evaluating preparation quality.Pass through inspection
The Aconitum Szechenyianum Gay processed product of the present invention of different batches production is surveyed, it is found that the uniformity of 3 kinds of active constituent contents is preferable, this more illustrates to survey
They fixed content either still ensures that drug quality all has for control production technology and is of great significance.
Experiment six：Xin Wuning alkali in each group Aconitum Szechenyianum Gay processed product, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines
Assay
1 testing sample
（1）Aconitum Szechenyianum Gay processed product A：Aconitum Szechenyianum Gay medicinal material 100g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10
The middle sand measured again, at 260 DEG C, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product A, is ground into fine powder, standby.
（2）Aconitum Szechenyianum Gay processed product B：Aconitum Szechenyianum Gay medicinal material 100g is taken, sheet, plus 10 times of middle sand measured are cut into, at 260 DEG C,
Frying 6min, cooling, produces Aconitum Szechenyianum Gay processed product B, is ground into fine powder, standby.
（3）Aconitum Szechenyianum Gay processed product C：Aconitum Szechenyianum Gay medicinal material 100g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten,
Plus the middle sand of 10 times of amounts, at 220 DEG C, frying 3min, cooling produces Aconitum Szechenyianum Gay processed product C, is ground into fine powder, standby.
（4）Aconitum Szechenyianum Gay processed product D：Aconitum Szechenyianum Gay medicinal material 100g is taken, sheet is cut into, 24h is stewed to moisten with yellow rice wine, is dried after stewing to moisten,
Plus the middle sand of 10 times of amounts, at 300 DEG C, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product D, is ground into fine powder, standby.
2 assay methods
The high performance liquid chromatographies provided using present invention experiment four, Xin Wuning alkali in Simultaneous Determination each group Aconitum Szechenyianum Gay processed product,
The content of 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines.
3 measurement results
Measurement result, is shown in Table 10.
The sample size measurement result mgg of table 10-1
4 discuss
As can be seen from Table 10, the content of the higher Xin Wuning alkali of toxicity in Aconitum Szechenyianum Gay processed product A is significantly lower than Aconitum Szechenyianum Gay
Processed product B, C and D, and drug activity composition 3-acetylaconitine and the content of 12- table acetyl dehydrogenation aconitines that it contains are bright
Aobvious to be higher than other Aconitum Szechenyianum Gay processed products, this may be also that effects of this Aconitum Szechenyianum Gay processed product A in terms for the treatment of of arthritis, asthma is excellent
In other Aconitum Szechenyianum Gay processed products, and its toxicity is the reason for be greatly lowered again.
Brief description of the drawings：
Fig. 1：Mix reference substance high-efficient liquid phase chromatogram
Fig. 2：Aconitum Szechenyianum Gay processed product test sample high-efficient liquid phase chromatogram
Wherein：No. 1 peak is that Xin Wuning alkaline colors spectral peak, No. 2 peaks are that 3-acetylaconitine chromatographic peak, No. 3 peaks are 12- table acetyl dehydrogenations
Aconitine chromatographic peak
Embodiment：
Embodiment 1：Aconitum Szechenyianum Gay processed product
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：Aconitum Szechenyianum Gay medicinal material 1000 is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus in 10 times of amounts
Sand, at 260 DEG C, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product.
Using Xin Wuning alkali, 3-acetylaconitine, 12- table second in hplc simultaneous determination Aconitum Szechenyianum Gay processed product
The content of acyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
Gone using Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl in hplc simultaneous determination Aconitum Szechenyianum Gay processed product
The content of hydrogen rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Accurately weighed Aconitum Szechenyianum Gay processed product powder 0.8g of the present invention, puts in conical flask with cover, essence
70% methanol 25mL of close addition, close plug, weighed quality is heated to reflux 1h, let cool, then weighed quality, and less loss is supplied with 70% methanol
Quality, shake up, filtered with 0.22 μm of miillpore filter, take subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
（5）Measurement result：Xin Wuning alkali, 3-acetylaconitine, the 12- table acetyl dehydrogenation rhizome of Chinese monkshoods in every gram of Aconitum Szechenyianum Gay processed product
The content of alkali composition is respectively 1.12mg, 2.33mg, 0.31mg.
Embodiment 2：Aconitum Szechenyianum Gay processed product rubber ointment
Prescription：Aconitum Szechenyianum Gay medicinal material 1000g
Preparation method：
（1）Aconitum Szechenyianum Gay medicinal material 1000g is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured,
At 260 DEG C, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber 10kg, rosin
10kg, lanolin 3kg, vaseline 1.0kg, atoleine 0.5kg, zinc oxide 10kg, gasoline 30kg, mix, coating are made, enters
Row applies cream, and segment, lid lining is cut into 5cm × 7cm 1000 pieces of fritter, produced；
Using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product rubber ointment
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Aconitum Szechenyianum Gay processed product rubber ointment 1 is taken, it is accurately weighed, it is cut into the broken of 1cm × 1cm
Piece, puts in 50mL round-bottomed flasks, and precision adds 70% methanol 25mL, and close plug, weighed quality is heated to reflux 1h, let cool, then weighed
Quality, the quality of less loss is supplied with 70% methanol, is shaken up, and is filtered with 0.22 μm of miillpore filter, is taken subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined；
（5）Measurement result：Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenations in every Aconitum Szechenyianum Gay processed product rubber ointment
The content of rhizome of Chinese monkshood alkali composition is respectively 1.05mg, 2.20mg, 0.24mg.
Claims (9)
1. a kind of Aconitum Szechenyianum Gay processed product, it is characterised in that the Aconitum Szechenyianum Gay processed product is to adopt to prepare with the following method：
Take Aconitum Szechenyianum Gay medicinal material, be cut into sheet, 12~36 h are stewed to moisten with rice vinegar, dried after stewing to moisten, plus 8~12 times amount middle sand,
At 255~265 DEG C, the min of frying 5~8, cooling produces Aconitum Szechenyianum Gay processed product.
2. Aconitum Szechenyianum Gay processed product according to claim 1, it is characterised in that the Aconitum Szechenyianum Gay processed product is to adopt with the following method
Prepare：Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C,
Frying 6min, cooling, produces Aconitum Szechenyianum Gay processed product.
3. a kind of detection method of Aconitum Szechenyianum Gay processed product, the Aconitum Szechenyianum Gay processed product is adopted to be prepared with the following method：Take Aconitum Szechenyianum Gay
Medicinal material, is cut into sheet, and 12~36 h are stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 8~12 times amount middle sand, at 255~265 DEG C,
The min of frying 5~8, cooling, produces Aconitum Szechenyianum Gay processed product；Characterized in that, using hplc simultaneous determination iron staff
Hammer Xin Wuning alkali, 3-acetylaconitine, the content of 12- table acetyl dehydrogenation rhizome of Chinese monkshood alkali compositions in processed product into shape, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 30~40 DEG C of column temperature, 0.5~1.5mLmin of flow velocity-1, the μ of sample size 5~20
L；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to alkali containing Xin Wuning, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitines
Mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Accurately weighed Aconitum Szechenyianum Gay processed product powder, puts in conical flask with cover, and precision addition 65~
75% methanol, close plug, weighed quality is heated to reflux, let cool, then weighed quality, and the quality of less loss is supplied with 65~75% methanol, is shaken
It is even, filtered with miillpore filter, take subsequent filtrate, produced；
（4）Determine：Each 5~20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
4. the detection method of Aconitum Szechenyianum Gay processed product as claimed in claim 3, the Aconitum Szechenyianum Gay processed product is to adopt to make with the following method
Standby：Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C,
Frying 6min, cooling, produces Aconitum Szechenyianum Gay processed product；Characterized in that, using hplc simultaneous determination Aconitum Szechenyianum Gay big gun
Xin Wuning alkali, 3-acetylaconitine, the content of 12- table acetyl dehydrogenation rhizome of Chinese monkshood alkali compositions in product, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Accurately weighed Aconitum Szechenyianum Gay processed product powder 0.8g, is put in conical flask with cover, and precision is added
70% methanol 25mL, close plug, weighed quality is heated to reflux 1h, let cool, then weighed quality, and the quality of less loss is supplied with 70% methanol,
Shake up, filtered with 0.22 μm of miillpore filter, take subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
5. a kind of Aconitum Szechenyianum Gay processed product rubber ointment, it is characterised in that the Aconitum Szechenyianum Gay processed product rubber ointment is to use such as lower section
It is prepared by method：
（1）Take Aconitum Szechenyianum Gay medicinal material, be cut into sheet, 12~36 h are stewed to moisten with rice vinegar, dried after stewing to moisten, plus 8~12 times amount middle sand,
At 255~265 DEG C, the min of frying 5~8, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add the conventional substrate of rubber ointment, mix, be made coating, carry out painting cream, segment,
Lid lining, is cut into small pieces, produces.
6. Aconitum Szechenyianum Gay processed product rubber ointment as claimed in claim 5, it is characterised in that the Aconitum Szechenyianum Gay processed product rubber ointment
It is to adopt to prepare with the following method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced.
7. a kind of application of Aconitum Szechenyianum Gay processed product rubber ointment in treatment asthmatic medicament is prepared, it is characterised in that the Aconitum Szechenyianum Gay
Processed product rubber ointment is to adopt to prepare with the following method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced；
Using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product rubber ointment
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Aconitum Szechenyianum Gay processed product rubber ointment 1 is taken, it is accurately weighed, it is cut into the broken of 1cm × 1cm
Piece, puts in 50mL round-bottomed flasks, and precision adds 70% methanol 25mL, and close plug, weighed quality is heated to reflux 1h, let cool, then weighed
Quality, the quality of less loss is supplied with 70% methanol, is shaken up, and is filtered with 0.22 μm of miillpore filter, is taken subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
8. a kind of application of Aconitum Szechenyianum Gay processed product rubber ointment in treatment of arthritis thing is prepared, it is characterised in that the Aconitum Szechenyianum Gay
Processed product rubber ointment is to adopt to prepare with the following method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced；
Using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product rubber ointment
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Aconitum Szechenyianum Gay processed product rubber ointment 1 is taken, it is accurately weighed, it is cut into the broken of 1cm × 1cm
Piece, puts in 50mL round-bottomed flasks, and precision adds 70% methanol 25mL, and close plug, weighed quality is heated to reflux 1h, let cool, then weighed
Quality, the quality of less loss is supplied with 70% methanol, is shaken up, and is filtered with 0.22 μm of miillpore filter, is taken subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
9. a kind of application of Aconitum Szechenyianum Gay processed product rubber ointment in treatment rhinitis thing is prepared, it is characterised in that the Aconitum Szechenyianum Gay big gun
Product rubber ointment is to adopt to prepare with the following method：
（1）Aconitum Szechenyianum Gay medicinal material is taken, sheet is cut into, 24h is stewed to moisten with rice vinegar, is dried after stewing to moisten, plus 10 times of middle sand measured, at 260 DEG C
Under, frying 6min, cooling produces Aconitum Szechenyianum Gay processed product；
（2）Take step（1）Aconitum Szechenyianum Gay processed product, be ground into fine powder, add water stirring be made relative density be 1.10 liquid extract；
（3）Take step（2）Liquid extract, add rubber ointment conventional substrate, the conventional substrate be rubber, rosin, lanolin,
Vaseline, atoleine, zinc oxide, gasoline, are mixed, and coating is made, and carry out painting cream, and segment, lid lining is cut into small pieces, produced；
Using Xin Wuning alkali, 3-acetylaconitine, 12- in hplc simultaneous determination Aconitum Szechenyianum Gay processed product rubber ointment
The content of table acetyl dehydrogenation rhizome of Chinese monkshood alkali composition, determination step is：
（1）Chromatographic condition：Syncronis C chromatographic columns, are entered using methanol -0.05mol/L biphosphate sodium water solutions as mobility
Row gradient elution, eluting order：0~15min, methanol is maintained at 15%, 0.05mol/L biphosphate sodium water solutions and is maintained at
85%；16~20min, methanol from 15% linear rise be 20%, 0.05mol/L biphosphates sodium water solution from 85% linear decline to
80%；21~30min, methanol from 20% linear rise be 40%, 0.05mol/L biphosphates sodium water solution from 80% linear decline to
60%；During 0~20min, Detection wavelength 235nm, detection Xin Wuning alkali, 3-acetylaconitine；During 21~30min, Detection wavelength
365nm, detects 12- table acetyl dehydrogenation aconitines, 36 DEG C of column temperature, flow velocity 1.0mLmin-1, the μ L of sample size 20；
（2）The preparation of mixed reference substance solution：Take Xin Wuning alkali, 3-acetylaconitine, 12- table acetyl dehydrogenation aconitine reference substances
In right amount, it is accurately weighed, dissolved with methanol and be diluted to every 1mL alkali containing Xin Wuning 0.750mg, 3-acetylaconitine 0.800mg,
12- table acetyl dehydrogenation aconitines 0.220mg mixed reference substance solution, is produced；
（3）The preparation of need testing solution：Aconitum Szechenyianum Gay processed product rubber ointment 1 is taken, it is accurately weighed, it is cut into the broken of 1cm × 1cm
Piece, puts in 50mL round-bottomed flasks, and precision adds 70% methanol 25mL, and close plug, weighed quality is heated to reflux 1h, let cool, then weighed
Quality, the quality of less loss is supplied with 70% methanol, is shaken up, and is filtered with 0.22 μm of miillpore filter, is taken subsequent filtrate, produce；
（4）Determine：Each 20 μ L of mixed reference substance solution, need testing solution are taken, high performance liquid chromatograph is injected, determined.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611012596.2A CN106377583B (en) | 2016-11-17 | 2016-11-17 | A kind of processing procedure and its quality determining method of Gansu genunie medicinal materials Aconitum Szechenyianum Gay |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611012596.2A CN106377583B (en) | 2016-11-17 | 2016-11-17 | A kind of processing procedure and its quality determining method of Gansu genunie medicinal materials Aconitum Szechenyianum Gay |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106377583A true CN106377583A (en) | 2017-02-08 |
| CN106377583B CN106377583B (en) | 2017-10-03 |
Family
ID=57958812
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611012596.2A Active CN106377583B (en) | 2016-11-17 | 2016-11-17 | A kind of processing procedure and its quality determining method of Gansu genunie medicinal materials Aconitum Szechenyianum Gay |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106377583B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109856261A (en) * | 2018-12-29 | 2019-06-07 | 上海市儿童医院 | A method of effective component in " point application side " is analyzed using HPLC simultaneously |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1369278A (en) * | 2001-02-16 | 2002-09-18 | 中国科学院长春应用化学研究所 | Preparing process of hypertoxic Chinese-medicinal materials and as aconite root and Chinese monkshood rhizome |
| CN101591293A (en) * | 2009-07-02 | 2009-12-02 | 中国科学院长春应用化学研究所 | Diester-type and ester-type aconite alkaloids are converted into the method for monoester pyrolysis-type alkaloids |
| CN102078394A (en) * | 2010-12-24 | 2011-06-01 | 达吾列提别克·努尔沙汗 | Aconite extract-containing medicinal composition, preparation method and application thereof |
-
2016
- 2016-11-17 CN CN201611012596.2A patent/CN106377583B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1369278A (en) * | 2001-02-16 | 2002-09-18 | 中国科学院长春应用化学研究所 | Preparing process of hypertoxic Chinese-medicinal materials and as aconite root and Chinese monkshood rhizome |
| CN101591293A (en) * | 2009-07-02 | 2009-12-02 | 中国科学院长春应用化学研究所 | Diester-type and ester-type aconite alkaloids are converted into the method for monoester pyrolysis-type alkaloids |
| CN102078394A (en) * | 2010-12-24 | 2011-06-01 | 达吾列提别克·努尔沙汗 | Aconite extract-containing medicinal composition, preparation method and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| 张帆，等: "民族药铁棒锤不同炮制品中各元素的光谱分析", 《药物分析杂志》 * |
| 曾陈娟等: "羌药铁棒锤炮制品的抗炎镇痛作用及初步安全性评价", 《世界科学技术(中医药现代化)》 * |
| 杨威: "醋制中药初探", 《广西中医药》 * |
| 王毓杰，等: "民族药铁棒锤炮制减毒原理初步研究", 《中国中药杂志》 * |
| 谭睿，等: "藏药铁棒锤与同属中药基源及炮制的比较", 《时珍国医国药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109856261A (en) * | 2018-12-29 | 2019-06-07 | 上海市儿童医院 | A method of effective component in " point application side " is analyzed using HPLC simultaneously |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106377583B (en) | 2017-10-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104784270B (en) | A kind of swelling and pain relieving external preparation and preparation method thereof | |
| CN101773551B (en) | External-use medicine composition of traditional Mongolian medicine for curing soft tissue injury | |
| CN102380015B (en) | Medicinal composition for treating rheumatoid arthritis and preparation method and application thereof | |
| CN109568447A (en) | Improve the Chinese medicine composition and its preparation method and application of sleep quality | |
| CN102397462A (en) | Oral liquid used for resisting aging, preparation method thereof, and quality controlling method thereof | |
| CN102204994B (en) | Chinese medicinal preparation for treating osteoarthritis and preparation method thereof | |
| CN106377583B (en) | A kind of processing procedure and its quality determining method of Gansu genunie medicinal materials Aconitum Szechenyianum Gay | |
| WO2023197731A1 (en) | Traditional chinese medicine composition, preparation method therefor and use thereof | |
| CN105288418B (en) | A kind of micro emulsion gel paste and preparation method thereof for treating baby diarrhea | |
| CN109528917B (en) | Korean medicinal composition for treating rheumatoid arthritis and preparation method of cream thereof | |
| CN102861159B (en) | Medicine composition for treating dysmenorrheal as well as preparation method and application thereof | |
| CN104352633B (en) | A kind of preparation method of pharmaceutical composition that treating osteoarthropathy | |
| CN101584809A (en) | Medicine for treating cold grasserie and method of preparing the medicine | |
| CN108578487A (en) | The pharmaceutical composition and preparation and application and preparation for the treatment of psoriasis vulgaris | |
| CN109663100B (en) | Traditional Chinese medicine composition for treating optic neuritis and preparation method and application thereof | |
| CN105288501A (en) | Traditional Chinese medicine composition containing folium artemisiae argyi and treating obesity | |
| CN1927309B (en) | Traditional Chinese medicine composition detection method | |
| CN105687432A (en) | Application of traditional Chinese medicine composition in preparation of medicines for treating eczema | |
| CN101612246A (en) | A kind of pharmaceutical composition and preparation method and method of quality control for the treatment of nephritis | |
| CN104435314A (en) | Medicament composition for treating osteoarthrosis and application of medicament composition | |
| CN110624067A (en) | Traditional Chinese medicine composition with weight-losing effect and preparation method of ointment thereof | |
| CN111643606B (en) | Gel plaster for treating insomnia and preparation method thereof | |
| CN110812401A (en) | Traditional Chinese medicine composition for soft tissue injury and detection method thereof | |
| CN102590432B (en) | Detection method for medicine for curing cold nature yellow water disease | |
| CN110742888B (en) | Application of bulleyaconitine A |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |