CN102218095B - Processing method of myrobalan-processed kusnezoff monkshood root as mongolian medicine - Google Patents
Processing method of myrobalan-processed kusnezoff monkshood root as mongolian medicine Download PDFInfo
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- radix aconiti
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- fructus chebulae
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Abstract
The invention relates to a method for processing kusnezoff monkshood root. In the invention, on the basis of the traditional method for processing the kusnezoff monkshood root, main factors, such as, the soaking time, the proportion of the influencing myrobalan system wild aconite to myrobalan, the frequency of changing solvents, the proportion of medicinal material to the solvents, and the like, influencing myrobalan-processed kusnezoff monkshood root are specifically and deeply researched, the optimal technological condition is determined, and the safety of products is researched; and the processing method specifically comprises the following steps of: putting the kusnezoff monkshood root and the myrobalan in the mass ratio of 1:0.8 in an irony container, adding water three times more than medicinal materials by weight, changing water one time every day, soaking for 7 days, taking out and drying in the air.
Description
Technical field
The present invention relates to the concocting method of Radix Aconiti Kusnezoffii, especially, the present invention relates to a kind of concocting method of Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata.
Background technology
Radix Aconiti Kusnezoffii is the dried root of ranunculaceae plant Aconitum kusnezoffii Reichb Aconitum-kusnezoffii Reichb., and mongolian medicine name pump Ah, is Mongolian medicine's common medicine.Traditional Mongolian medicine thinks that Radix Aconiti Kusnezoffii is hot, temperature, gently, very toxic, function is killed " gluing ", pain relieving, dry " Xieri Wusu Symptom " is used for pestilence, sticking, neat Kazakhstan, twinge, Adam's apple, acute filthy disease, gout, trip pain disease, joint " Xieri Wusu Symptom ", rheumatism, the heart " conspicuous complying with ", toothache etc., and the prescription that clinical use contains Radix Aconiti Kusnezoffii accounts for 20% of prescribed preparation, because it is the toxicity medical material, so be used as medicine mainly with processed product.The tradition of mongolian medicine Radix Aconiti Kusnezoffii (experience) concocting method has the several different methods such as Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata, urine of baby boys Radix Aconiti Kusnezoffii Preparata, yoghourt Radix Aconiti Kusnezoffii Preparata, and wherein Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata is the most commonly used.But owing to there is no so far unified mongolian medicine Radix Aconiti Kusnezoffii concocted specification and quality standard, the custom that the concocting method of each mongolian medicine manufacturing enterprise and Mongolian medicine medical institutions and degree depend on constituent parts and operator's personal experience, so that the quality of mongolian medicine Radix Aconiti Kusnezoffii Preparata and safety can't be effectively controlled and guarantee, become the bottleneck of restriction Mongolia Medicine development.
For this practical problem, the present invention take the Mongolia Medicine theory as guidance, carries out systematic study to this concocting method take the traditional concocting method of mongolian medicine-Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata as object of study.At first by literature research (comprise historical document, modern study document and use present situation), clearly the Historical Development of this medicinal material processing is used for reference existing research, determines research starting point of the present invention.
Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata, the Four-Volume Medical Code begins to be stated from: " method of preparation garidi-5 ball-Fructus chebulae immaturus 20g, Radix Aucklandiae 5g, Rhizoma Acori Graminei 3.5g, Radix Aconiti Kusnezoffii 2.5g are ground into respectively fine powder; insert in the ironware; add an amount of urine of baby boys and stirred 3 days; add the Moschus 1.5g that is soaked in water when black and glossy, make the watered pill "." Mongolian medicine's prescription choosing commonly used " (Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata): " Radix Aconiti Kusnezoffii was soaked 7 days at glycyrrhiza decoction or Fructus Chebulae Tang Nei, changed soup every day one time, took out to get final product "." China's book on Chinese herbal medicine " (mongolian medicine volume) (Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata): " Radix Aconiti Kusnezoffii is scraped off feeler, earth, put equivalent Fructus Chebulae Tang Nei and soaked 3～7, must stir 1 every day between soak period, takes out to dry to get final product "." the mongolian medicine process of preparing Chinese medicine is learned " (Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata): " get clean medical material, soaked 1～3 with Fructus Chebulae's soup, taste when slightly numb feeling in the tongue being arranged to mouth, take out, dry.(the clean Radix Aconiti Kusnezoffii of every 10kg adds 3kg Fructus Chebulae and boils into the about 30L of soup, and must stir 1～2 every day between soak period) ".
From successive dynasties mongolian medicine document record as seen, Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata is for the process of preparing Chinese medicine of preparation Mongolia patent drug " garidi-5 ball " Radix Aconiti Kusnezoffii the earliest, develops into gradually afterwards the process of preparing Chinese medicine of Radix Aconiti Kusnezoffii, and various document record concocting method is not quite similar.Although substantially all be to finish through the method for soaking about Kusnezoff monkshood root processing method in the prior art, but they do not make research to soaking the factors such as raw material usage ratio, immersion quantity of solvent, soak time, soak cycle to the combined influence of concocting absolutely, do not have these factors that the combined influence of concocting product quality is made research yet, therefore do not put down in writing accordingly for these influence factors' optimal processing parameter yet.
Summary of the invention
The present invention adopts chemical composition analysis method, pharmacodynamic experiment, myocardial toxicity experiment to Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata tradition concocting method, by orthogonal test research, many indexs are carried out overall merit, determine best processing procedure condition, clear and definite concrete technology parameter is formulated concocted specification; Adopt the chemical composition analysis method, set up the quality standard of concocting medical material; Feed intake with the Radix Aconiti Kusnezoffii Preparata of concocting by the processing procedure of determining and be up to the standards by the quality standard draft, prepare representative Mongolia patent drug preparation " garidi-5 "; Radix Aconiti Kusnezoffii Preparata list medical material and patent medicine garidi-5 are done long term toxicity test, and research is to degree of impairment and single medical material and the prescribed preparation difference each other of animal tissue's organ; With Mongolia patent drug preparation " garidi-5 " clinical administration, the toxic component in experimenter's blood is analyzed; Clinical poisoning indication is observed, do the clinical assay that can reflect visceral organ injury, overall merit is carried out in the safety of medication.
The present invention is take traditional concocting method of Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata as the basis, select to affect the principal element of Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata, be soak time (A), Radix Aconiti Kusnezoffii and Fructus Chebulae's ratio (B), change the solvent frequency (C) and medical material and solvent ratios (D) is carried out orthogonal, can reflect the total alkaloid content of processed product safety and effectiveness, diester-type alkaloids content, the result of analgesic experiment and antiinflammatory experiment and myocardial toxicity experiment is as selection indicators, adopt many index comprehensives of orthogonal design point system to optimize Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata optimal processing parameter, and by the quadrature demonstration test, prove conclusively feasibility and the stability of best processing procedure.
Method
1 Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata processing procedure Orthogonal Experiment and Design
1.1 traditional concocting method of Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata
Get equivalent Fructus Chebulae and Radix Aconiti Kusnezoffii, put in the same ironware, add water to till the submergence medical material, soaked 5-7 days under the room temperature condition, changed water one time in two days, stir 1-2 every day.
1.2 orthogonal design
Concoct consulting of condition according to tradition, this research selects to affect the principal element of Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata, be soak time (A), Radix Aconiti Kusnezoffii and Fructus Chebulae's ratio (B), change the solvent frequency (C) and medical material and solvent ratios (D) and carry out orthogonal design and investigate, respectively get 3 levels, with L
9(3
4) each experiment of orthogonal table arrangement, make 9 kinds of processed products.The factor level design is such as table 1.
Table 1 factor level table
2, Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata processed product is investigated the mensuration of index
2.1 the mensuration of total alkaloid content
According to " content assaying method under Radix Aconiti Preparata item of Chinese Pharmacopoeia 2005 version is measured 9 kinds of processed products, the results are shown in Table 3.
2.2 the mensuration of diester-type alkaloids content
2.2.1 chromatographic condition and system suitability
Take octadecylsilane chemically bonded silica as filler; Take acetonitrile-oxolane (25: 15) as mobile phase A, take 0.1mol/L ammonium acetate (1000ml contains glacial acetic acid 0.5ml) as Mobile phase B, carry out gradient elution by table 2 program; The detection wavelength is 235nm; Flow velocity 0.8ml/min; Number of theoretical plate calculates by the hypaconitine peak should be not less than 5000.
Table 2 eluent gradient elution program
2.2.2 the preparation of reference substance solution
It is an amount of that precision takes by weighing aconitine reference substance, hypaconitine reference substance and mesaconitine, adds isopropyl alcohol-chloroform (1: 1) and make the mixed solution that every 1ml contains 0.1mg, shakes up, and get final product.
2.2.3 the preparation of need testing solution and mensuration
Get different each 2g of processed product of Radix Aconiti Kusnezoffii, accurately weighed, put respectively in the tool plug conical flask, add strong ammonia solution 1ml, place a moment, add isopropyl alcohol-ethyl acetate (1: 1) 50ml, weighed weight, supersound process 30 minutes lets cool, more weighed weight, supply the weight that subtracts mistake with isopropyl alcohol-ethyl acetate (1: 1), shake up, filter, precision is measured subsequent filtrate 25ml, put in the rotary evaporator, decompression and solvent recovery below 40 ℃ is to doing, and residue dissolves with isopropyl alcohol-chloroform (1: 1) and is settled in the 5ml measuring bottle, filters with microporous filter membrane (0.45 μ m).Precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and the injection liquid chromatography is measured, and the results are shown in Table 3.
2.3 the mensuration of writhing number of times
Get mice, female, be divided at random 9 groups, 15 every group, oral administration 6 days.Each Mus lumbar injection 0.6% glacial acetic acid 0.2ml, the number of times of each Mus writhing response in the record 20min.The results are shown in Table 3.
2.4 Ovum Gallus domesticus album causes the mensuration of rat toes swelling
Get 100 of healthy rats, male, be divided at random 10 groups, i.e. blank group (5 ‰ CMC) and administration group.Oral administration 5 days.At the 5th day, every rat is rule with marking pen along prominence, right back ankle joint before the administration, survey its normal right back sufficient volume with the toes volumetric type meter, then administration, after the administration 1 hour, in Rat Right metapedes plantar subcutaneous injection Fresh Egg clear liquid 0.1ml, treat to measure in 1 hour right back sufficient volume, with the difference of right back sufficient volume before and after the administration as swelling, and the difference between administration group and blank group relatively respectively.The results are shown in Table 3.
2.5 the mensuration of rat heart muscle TnT
Get 120 of rats, be divided at random 11 groups of administrations.Oral administration 12 days, 1h after the administration in the 12nd day, large rathole frame venous blood sampling, the centrifugal 10min of 3000rpm gets the content that serum Elisa method is measured serum cardiac troponin T.The results are shown in Table 3.
3, aggregative indicator evaluation result
The present invention selects L
9(3
4) the orthogonal table experiment arrangement, with the result of the total alkaloid content in each processed product, diester-type alkaloids content, writhing number of times, paw swelling and cardiac troponin content as investigating index, utilize many index comprehensives method of weighting scores deal with data, determine best processing procedure.Scoring method is: total alkaloid content the highest be decided to be 100 minutes (" Gao You " index), what diester-type alkaloids content was minimum is decided to be 100 minutes (" low excellent " index), what writhing number of times (analgesic experiment) was minimum is decided to be 100 minutes (" low excellent " index), (the antiinflammatory experiment) of paw swelling minimum is decided to be 100 minutes (" low excellent " index), cardiac troponin content (cardiac toxicity experiment) is minimum is decided to be 100 minutes (" low excellent " index), the total alkaloid content of all the other each time experiments, diester-type alkaloids content, the writhing number of times, the result of paw swelling and cardiac troponin content is converted into corresponding mark successively.The results of analysis of variance according to each index shows that the D factor of total alkaloids and the A factor of diester-type alkaloids have significant difference.Through considering, weight coefficient is specially: total alkaloids 35%, diester-type alkaloids 35%, writhing number of times 10%, paw swelling 10% and cardiac troponin content 10%, then five weighted scoring results added are asked its comprehensive grading value Y
Always, such as No. 2 total points be: Y
Always=2.73/6.34 * 100 * 35%+ (1-0.260/0.505) * 100 * 35%+ (1-32.15/39.43) * 100 * 10%+ (1-0.7275/1.0786) * 100 * 10%+ (1-11.38/19.89) * 100 * 10%=41.44 carries out quadrature analysis and variance analysis with integrate score.Its orthogonal experiment arrangement and each index determining result and intuitive analysis the results are shown in Table 3, and each index and comprehensive grading the results of analysis of variance see Table 4, table 5, table 6, table 7, table 8, table 9.
Table 3 orthogonal test and experimental result and intuitive analysis table
Annotate: represent optimum condition * number
Table 4 total alkaloids analysis of variance table
(F
0.05＝19.0)
Table 5 diester-type alkaloids analysis of variance table
(F
0.05＝19.0)
Table 6 writhing number of times analysis of variance table
(F
0.05＝19.0)
Table 7 paw swelling analysis of variance table
(F
0.05＝19.0)
Table 8 cardiac troponin analysis of variance table
(F
0.05＝19.0)
Table 9 comprehensive grading is analysis of variance table as a result
(F
0.05＝19.0)
4, interpretation of result and checking
Intuitive analysis result by table 3 sees that the best process of preparing Chinese medicine condition during singly take total alkaloid content as index is A
1B
3C
1D
1, affecting order is D＞C＞A＞B; The best process of preparing Chinese medicine condition during singly take diester-type alkaloids content as index is A
3B
2C
2D
2(D item extreme difference minimum is made as the error row), affecting order is A＞C＞B＞D; The best process of preparing Chinese medicine condition during singly take the writhing number of times as index is A
2B
1C
2D
3, affecting order is C＞B＞A＞D; The best process of preparing Chinese medicine condition during singly take paw swelling as index is A
1B
1C
2D
1, affecting order is A＞C＞B＞D; The best process of preparing Chinese medicine condition during singly take cardiac troponin content as index is A
1B
2C
3D
3, affecting order is B＞D＞A＞C; If take the comprehensive grading result as standard, best processing procedure condition is A
3B
2C
2D
1, affecting order is A＞D＞C＞B.Can find out that from six group analysis results it is basically identical that single the best during take diester-type alkaloids content as standard is concocted the best processing procedure condition of condition with take the comprehensive grading result as standard the time, and these two groups of results more can reflect the quality of concocting effect.
Variance analysis (table 4～9): the results of analysis of variance during singly take total alkaloid content as standard with singly take diester-type alkaloids content as standard shows, factor D (medical material and solvent ratios) and A (soak time) are significance impact (P＜0.05) to experimental result, variance analysis during take other indexs as standard shows, each investigation factor is on experimental result there are no significant impact (P＞0.05).Show through weighted comprehensive scoring intuitive analysis and variance analysis: each investigation factor to the quality influence size order of Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata is: A＞D＞C＞B, wherein the B factor affecting is minimum, so consider that from the angle of energy savings selecting Fructus Chebulae's addition is 1: 0.8 (B
1).By range analysis as can be known, the best experimental program of orthogonal design is: A
3B
1C
2D
1, namely soak time is 7 days, and Radix Aconiti Kusnezoffii and Fructus Chebulae's ratio are 1: 0.8, change solvent one time in one day, and medical material and solvent ratios are 1: 3.
Demonstration test: for further verifying feasibility and the stability of gained technique, get parallel three parts of a collection of Radix Aconiti Kusnezoffii, concoct by the best process of preparing Chinese medicine scheme.That is: soaked 7 days, Radix Aconiti Kusnezoffii and Fructus Chebulae's ratio are 1: 0.8, changed solvent in one day one time, medical material and solvent ratios are that 1: 3 condition is concocted, the measurement result of investigating index is seen Table 10, the minima of each time experiment in the equal near orthogonal design of diester-type alkaloids content, writhing number of times and paw swelling index, the result is satisfied, illustrates and optimizes successfully.
The confirmatory experiment result of the best process of preparing Chinese medicine condition of table 10
5, safety evaluatio
Carry out pilot scale according to the process of preparing Chinese medicine scheme after optimizing and amplify, prepare the Radix Aconiti Kusnezoffii processed product, and feed intake with it, prepare representative Mongolia patent drug preparation " garidi-5 flavor ball ".Medicinal material processing product and patent medicine " garidi-5 flavor ball " are done long term toxicity test; " garidi-5 flavor ball " is used for clinical trial, the safety of its clinical administration prescribed dose scope is investigated.
Long term toxicity test is the result show, Radix Aconiti Kusnezoffii Preparata (high dose 2.4g/kg is that 8 times of quantity, low dosage 0.3g/kg are clinical dosage), garidi-5 ball (1.05g/kg is that 11.7 times of quantity, 0.09g/kg are clinical dosage) be in 3 middle of the month of rat administration, on its food-intake, body weight, hematology, blood biochemical learn, organ coefficient all has impact in various degree.Learn from blood biochemical, two medicines cause that the main target organ of pathological changes may be heart, kidney and liver, and the damage of Radix Aconiti Kusnezoffii Preparata lungs and liver is heavy than garidi-5.Radix Aconiti Kusnezoffii Preparata can not recover in convalescent period the damage of rat liver.The prompting clinical administration should strictly be controlled dosage.
Test is investigated in the safety that contains Radix Aconiti Kusnezoffii Preparata preparation " garidi-5 flavor ball " clinical administration prescribed dose scope, screening obtains 32 routine health volunteers altogether, single-dose enters group 24 examples, high (1.0g), in (0.6g), every group of 8 examples of low (0.4) dosage group; Successive administration (dosage is 0.6g, every day 1 time, successive administration 7 days) group 8 routine experimenters.Duration of test is observed different time points heart rate, the rhythm of the heart, breathing, blood pressure, body temperature, nervous system, respiratory system, cardiovascular system, digestive system, change of skin and perspiration situation, pupil size, consciousness and the mental status etc., and any untoward reaction does not appear in whole duration of test 32 routine health volunteers.Single-dose enters to organize in each experimenter of the each 4ml of rear blood sample collection totally 13 times ((0 o'clock) and take medicine rear 15min and 30min before the administration, 45min, 1h, the 75min of taking medicine, 90min, 2.0,3.0,4.0,6.0 8.0,12.0h gathers respectively experimenter's ulnar vein blood), detect blood drug level; Successive administration group took medicine in the 7th day each experimenter of the each 4ml of rear blood sample collection totally 13 times ((0 o'clock) and take medicine rear 15min and 30min before the administration in the 7th day, 45min, 1h, 75min, 90min, 2.0,3.0,4.0,6.0,8.0 12.0h gathers respectively experimenter's ulnar vein blood), detect blood drug level.The result shows that garidi-5 flavor ball may be influential to leukocyte differential count, but without clinical meaning.Garidi-5 flavor ball has good safety in the prescribed dose scope, and any adverse events does not occur.To 416 parts of blood examination results, 9 parts, 8 parts of blood samples of successive administration group detect the aconite alkaloids in 11 parts, middle dosage group in high dose group, all do not detect in all the other 388 parts of blood samples, and recall rate is 6.73%.
Safety testing is the result show, concocts Radix Aconiti Kusnezoffii according to the best processing procedure that this subject study is determined, and dosed administration in accordance with regulations clinically, can guarantee that mongolian medicine concocts the safety of Radix Aconiti Kusnezoffii medication.
The specific embodiment
Fructus Chebulae's Radix Aconiti Kusnezoffii Preparata processing procedure:
Get Radix Aconiti Kusnezoffii and Fructus Chebulae, with putting in the container made of iron, added the triplication water soaking 7 days, change water every day once, and fully stir twice, take out, dry.The clean Radix Aconiti Kusnezoffii of every 1kg is used Fructus Chebulae 0.8kg.
Claims (1)
1. a Fructus Chebulae concocts the method for Radix Aconiti Kusnezoffii, it is characterized in that: get weight ratio and be 1: 0.8 Radix Aconiti Kusnezoffii and Fructus Chebulae, place in the container made of iron, add the water soaking of medical material treble amount, change water every day once, soaked 7 days, take out, dry.
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