Summary of the invention
The object of the invention is to provide a kind of preparation method and content assaying method thereof of five tastes manna dipping powder.Technical solution of the present invention is as follows:
Get Juniperus oxycedrus 1-10 weight portion, Cacumen Myricariae Germanicae 1-10 weight portion, Rhododendron 1-10 weight portion, Artemisia sieversiana Willd. 1-10 weight portion and Herba Ephedrae 1-10 weight portion, will carry out pretreated medical material mix homogeneously, superfine powder is broken into 0.01-100 μ m, and packing gets final product.
Wherein can also add Cortex Eucommiae 1-3 weight portion or Radix Lamiophlomidis Rotatae 1-3 weight portion in the above-mentioned raw materials medicine, also can add 0.001-1 weight portion suspending agent or 0.001-0.1 parts by volume spice in the above-mentioned raw materials medicine;
Wherein suspending agent is silicon Bentonite, carbomer, aluminium silicate or sodium acrylate (NP-700), sodium carboxymethyl cellulose (CMC-Na) Michelia leap oil, Flos micheliae Albae oil, Flos Jasmini Sambac oil, Oleum menthae, Radix Angelicae Dahuricae oil, Oleum Rosae Rugosae, foenugreek oil, rhododendron oil, oleum lini or Semen Sesami wet goods.
Wherein micronizing is ground into 0.01-0.5 μ m, 0.4-1.2 μ m, 1-50 μ m, 40-80 μ m, 60-100 μ m.
The pairing unit of wherein wt part/parts by volume is g/ml.
Content assaying method in the method for quality control of this pharmaceutical composition is as follows:
Chromatographic condition and system suitability test are filler with octadecyl silane; With 2-5: 98-95: 0.1-0.5 methanol-water-triethylamine is a mobile phase, with the phosphoric acid adjust pH to 2-6; The detection wavelength is 210nm; Column temperature: 15-25 ℃; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the ephedrine hydrochloride reference substance, accurate claims surely, and the methanol solution that adds methanol solution or contain 2-10% ammonia is made the solution that every 1ml contains 0.1mg, promptly;
The preparation of need testing solution: get this pharmaceutical composition 1.0-1.5g that is ground into 0.01-150 μ m, the accurate title, decide, and puts in the 100ml tool plug conical flask, the accurate methanol solution 25ml that adds methanol solution or contain 2-10% ammonia, claim to decide weight, reflux, extract, or supersound extraction 30-60 minute, put coldly, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, promptly;
Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
This pharmaceutical composition contains ephedrine in ephedrine hydrochloride C10H15NOHCl, must not be less than 0.05%.
Traditional grinding and processing method makes drug particle size the thinnest between 180-150 μ m in Chinese medicine, the Tibetan medicine.And of the present invention the crude drug superfine powder is broken into 0.01-100 μ m, and no longer need lixiviate, heating to decoct, therefore the concrete advantage of method of the present invention is as follows:
1, simple, the easy row of production and processing of the present invention, having changed needs numerous and diverse program of decocting in the former using method, meet modern's allegro life; Simultaneously can realize suitability for industrialized production, product also can be transported for long-distance and enter the international market towards huge numbers of families' patient.
2, the five tastes manna dipping soup after the change looses, and the drug effect mode is simple, convenient, need not add the stabilizing agent of any chemosynthesis, so safe in utilization, convenient.In daily process of having a bath, both can use simultaneously, evident in efficacy.
3, the present invention adopts a day modern advanced, keeps the active substance in the medical material and the active ingredient of medical material effectively, has improved the dissolution of crude drug composition again greatly, improves bioavailability simultaneously, makes curative effect better;
4, prolong the medicine resting period greatly, easy to use and carry.Adopt accelerated test method, new five tastes manna dipping powder of the present invention was observed 24 months under the condition of 37-40 ℃ and relative humidity 75%, measured every, physicochemical character does not all change, every index illustrates that it has good stability all by experiment.
5, the present invention adopts the main effective ingredient ephedrine in the high-efficient liquid phase technique mensuration five tastes manna bath powder, and the result shows that method is easy, and repeatability and stability improve than thin slice scan determination, can be used as quality control index.
Following experimental example is used to further specify but is not limited to the present invention.
Experimental example 1: the clinical efficacy comparative test of micronizing and the diffusing decoction of tradition
Diagnostic criteria: (1) main clinical manifestation: positions such as joint, skin, muscles and bones pain, or swelling deadlock in morning, numbness is weighing or joint stuffiness, joint swelling deformation very then, tetanic not stretching or amyotrophy etc.; (2) characteristics of incidence: how relevant with climate change; (3) sex age characteristics: good sending out in person between twenty and fifty, the woman is more than the man; (3) sex age characteristics: good sending out in person between twenty and fifty, the woman is more than the man.The diagnosis of the corresponding disease of doctor trained in Western medicine: the diagnosis of (1) rheumatoid arthritis: must possess following 4 or 4 above standards: a. deadlock in morning at least 1 hour continued more than 6 weeks; B.3 the individual or arthroncus more than 3 continues more than 6 weeks at least; ) c. carpal joint, metacarpophalangeal joints (MCP) or proximal interphalangeal joint (PIP) be more than 6 weeks of swelling; D. symmetry arthroncus; E. subcutaneous rheumatoid nodules; F. the rheumatoid factor positive; G. finger-joint X line changes confirmation.(2) rheumatic arthritis diagnostic criteria: a. clinical manifestation: the streptococcal infection medical history is arranged, and acute stage, can have multiple and the migration joint aches more, mostly occurs in big joint, can accompany red and swollen pain or tuberosity or erythema, and chronic phase can only be felt joint aches.B. chemical examination: anti-" 0 " 1: 500 or more or antistreptokinase surpass 80 units, or antihyaluronidase is above 128 units, erythrocyte sedimentation rate speeds or c reactive protein (CRP)>10%; C.X line sheet: sclerotin is harmless.
Checking case standard: 1, include the case standard in: (1) meets Tibetan medicine diagnostic criteria person; (2) age is 18~65 years old patient.2, get rid of the case standard: late deformity, maimed person, the disability person of (1) rheumatoid arthritis; (2) age under-18s, over-65s, anemia of pregnant woman or women breast-feeding their children; (3) though be primary disease, take Western medicine or other drug person for a long time, must stop using, otherwise should get rid of; (4) merge serious primary disease such as cardiovascular, liver, harmonization of the stomach hemopoietic system, psychotic; (5) do not meet the standard of including in, not medication in accordance with regulations can't judge that curative effect or data are not congruent, affects the treatment or safety judgement person.
Administrated method: treatment group: get Juniperus oxycedrus 1 weight portion, Cacumen Myricariae Germanicae 1 weight portion, Rhododendron 1 weight portion, Artemisia sieversiana Willd. 1 weight portion and Herba Ephedrae 1 weight portion, will carry out pretreated medical material mix homogeneously, superfine powder is broken into 50 μ m, is packed as the 500g/ bag.Each one bag, inject bathtub, 40 ℃ of water temperatures stir evenly and soak whole body or disease sites, every day 2 times, each 15--30 minute, bed diaphoresis; Matched group: get Juniperus oxycedrus 1 weight portion, Cacumen Myricariae Germanicae 1 weight portion, Rhododendron 1 weight portion, Artemisia sieversiana Willd. 1 weight portion and Herba Ephedrae 1 weight portion, will carry out pretreated medical material mix homogeneously, superfine powder is broken into 200 μ m, is packed as the 500g/ bag.Each one bag, to fry in shallow oil soup and pour bathtub into, 40 ℃ of water temperatures are soaked whole body or disease sites, every day 2 times, each 15--30 minute, bed diaphoresis.
The course of treatment: two groups was a course of treatment with 7 days all, can repeat 1-2 the course of treatment in case of necessity.Observation index: 1, local symptoms such as joint, muscle, muscles and bones, as pain, swelling degree change; 2, the recovery situation of function of joint; The improvement situation of the General Symptoms of 3, following; 4, the tongue arteries and veins changes; 5, erythrocyte sedimentation rate, anti-" 0 ".Curative effect determinate standard: 1, recovery from illness: symptom all disappears, and it is normal that functional activity recovers, and it is normal that main doctor trained in Western medicine detects index; 2, produce effects: symptomatology is eliminated or cardinal symptom is eliminated, and function of joint is recovered substantially, can participate in operate as normal and work, and it is normal substantially to detect index; 3, effective: cardinal symptom is eliminated substantially, and main function of joint is recovered substantially or had made marked progress, and can't take care of oneself to transfer to can take care of oneself, or loses the job and work capacity transfers work to and ability to work is recovered to some extent, mainly detects index and makes moderate progress.
Treatment group and matched group data situation: sex: in 82 examples, the male: 23 examples are organized in treatment, matched group 13 examples; The women: 28 examples are organized in treatment, matched group 18 examples.Age distribution sees Table 1; The course of disease distributed and sees Table 2 when each organized patient admission; Following clinical data shows that treatment group, matched group are isostatic at aspects such as sex, age, the courses of disease substantially, have comparability.Pattern of syndrome: select case treatment group and matched group to be cold disease.
Table 1: each organizes the patient age distribution situation
Table 2: each is organized the course of disease and distributes
Two groups of therapeutic effect highly significants, recovery from illness, produce effects, total effective rate are through X 2 test, and P<0.05 proves that two groups of curative effects have significant difference.
Table 3: each organizes curative effect relatively
The present invention bathes on the diffusing basis of soup at former five tastes manna to develop through process modification, when keeping original drug form curative effect, has kept the characteristic of Tibetan medicine's bathing treatment, and the national rarity of this motherland of Tibetan medicine's dipping has been obtained developing fully.Prove that by clinical verification five tastes manna dipping soup diffusing (promptly being broken into 50 μ m through superfine powder) has significant therapeutical effect, treatment group cure rate 33.3%, recovery from illness obvious effective rate 62.7%, total effective rate 94.1%, matched group cure rate 9.7%, recovery from illness obvious effective rate 32.3%, total effective rate 87.1%.Treatment group curative effect is higher than matched group, and group there is significant difference P<0.05, two.
Experimental example 2: particle size is to the influence test of release
The preparation mean diameter is 10 μ m, 50 μ m, three kinds of microgranules of 200 μ m.
Quicken release experiment: the microgranule 100mg of each particle diameter of precision weighing, place 6 100ml culture bottles, accurately add the dissolution fluid 100ml of 0.1M, tighten with the lid of band aluminum film, at once after jolting mixes, use the culture bottle traverse, be fixed on the fixed station, be dipped in advance thermoregulation in 37 ℃ water-bath, 100 vibrations of per minute, culture bottle is taken out in vibration beginning 1,2,3,4,5,6 hour, extracts each 10ml of turbid solution in each container.
Chromatographic condition and system suitability test are filler with octadecyl silane; With 2-5: 98-95: 0.3 methanol-water-triethylamine is a mobile phase, with the phosphoric acid adjust pH to 3-4; The detection wavelength is 210nm; Column temperature: 15-25 ℃; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the ephedrine hydrochloride reference substance, accurate claims surely, and the methanol solution that adds methanol solution or contain 8% ammonia is made the solution that every 1ml contains 0.1mg, promptly;
The preparation of need testing solution: extract that each 10ml of turbid solution puts in the 100ml tool plug conical flask in each container, the accurate methanol solution 25ml that adds methanol solution or contain 8% ammonia, claim to decide weight, reflux, extract, or supersound extraction 30-60 minute, put coldly, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, promptly;
Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
This pharmaceutical composition contains ephedrine in ephedrine hydrochloride C10H15NOHCl
Table 4
Table 4 explanation granularity is more little, and the effective ingredient release concentration is big more.The degree that general mechanical activation comminution reaches at most is 200 μ m, and of the present invention being improved to is crushed to 0.01-100 μ m, and the release of effective ingredient increases.
Experimental example 3: linear relationship is investigated test
Accurate ephedrine hydrochloride reference substance solution (0.022mg/ml) 1,4,8,12,16, the 20 μ l that draw, sample introduction is measured the peak area integrated value respectively.Sample size (μ g) with ephedrine is an abscissa, is vertical coordinate with the peak area integrated value, gets regression equation, sees Fig. 1.
Table 5 ephedrine hydrochloride linear relationship is investigated
Sampling volume (μ l) |
Sample size μ g |
Peak area |
1 |
0.022 |
118 |
4 |
0.088 |
490 |
8 |
0.176 |
980 |
12 |
0.264 |
1439 |
16 |
0.352 |
1924 |
20 |
0.440 |
2389 |
Obtain the result by regression equation and show, in the scope of 0.022~0.440 μ g, have favorable linearity.
Experimental example 4: different solvent extraction effect comparative test
Get same batch sample (lot number, 20010301) and adopt ether, methanol, 8% ammonia methanol solution as extracting solvent respectively, gained solution is measured content respectively in accordance with the law, the results are shown in Table 6.
The different solvents of table 6 extract the result
The result shows, extracts ephedrine hydrochloride with methanol and the methanol that contains 8% ammonia as solvent and does than ether that to extract solvent suitable.
Experimental example 5: the selection of extracting method, time test in the assay:
To same batch sample (20010301), be that solvent adopts apparatus,Soxhlet's reflux extraction and ultrasonic processing method respectively with methanol, 8% ammonia methanol solution, and extraction time investigated that measurement result sees Table 7.
The selection of table 7 extracting method
The result shows, there is not significant difference with reflux, extract,, methanol and the methanol solution supersound extraction ephedrine hydrochloride content that contains 8% ammonia, 30,45,60 minutes no significant differences of supersound extraction, but, extracting with the methanol solution of 8% ammonia, the sample impurity peaks is less, simultaneously, in order to extract fully, adopt sample with the methanol of 8% ammonia as extracting solution, supersound extraction 45 minutes.
Experimental example 6: precision test
Accurate ephedrine hydrochloride standard solution (0.022mg/ml) the 10 μ l that draw repeat sample introduction 5 times, and the peak area value of ephedrine hydrochloride is 1201.1, and RSD is 1.37%.
Table 8 Precision test result
Sequence number |
Peak area |
On average |
RSD(%) |
1 2 3 4 5 6 |
1222.3 1178.1 1213.4 1189.1 1208.3 1195.3 |
1201.1 |
1.37 |
Precision test result shows that the RSD% of 5 sample introductions is 1.37%, shows that precision is good.
Experimental example 7: stability test
Accurate sample (lot number 20010321) the solution 10 μ l that draw, respectively at 0,2, sample introduction was measured in 4,6,8 hours, the chromatographic peak area value of ephedrine hydrochloride in the record sample, peak area value is stable in 8 hours.The results are shown in Table 9.
Table 9 stability test result
Time (h) |
Peak area |
On average |
RSD(%) |
0.0 2.0 4.0 6.0 8.0 |
1584.1 1605.0 1568.2 1558.3 1552.8 |
1573.7 |
1.35 |
Experimental example 8: repeatability test
Get 5 parts of same batch samples (lot number 20010321), press chromatographic condition mensuration under the assay item, repeatability is good as a result, sees Table 10.
Table 10 reproducible test results
Sequence number |
Example weight |
Peak area |
Sample size (%) |
On average (%) |
RSD(%) |
1 2 3 4 5 |
1.2045 1.2098 1.2032 1.2006 1.2013 |
1348.5 1359.2 1375.2 1386.6 1384.4 |
0.0584 0.0583 0.0567 0.0574 0.0594 |
0.0580 |
1.78 |
Experimental example 9: application of sample reclaims experiment
Precision takes by weighing sample (lot number 20010321, Content of Ephedrine With the are 0.06%) 0.6g of known content, and accurate the title decides, accurate reference substance solution (0.45mg/ml) 1ml that adds, methanol 24ml claims to decide weight, supersound process 60 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, get filtrate, measure by above-mentioned sample size assay method, experimental result sees Table 11.
Table 11 application of sample reclaims experimental result
Sequence number |
Sample heavy (g) |
Sample epheday intermedia alkali number (mg) |
Add Herba Ephedrae alkali number (mg) |
Record peak area |
Measurement result (mg) |
The response rate (%) |
Average recovery rate (%) |
RSD (%) |
1 2 3 4 5 |
0.6012 0.6100 0.6058 0.6047 0.6146 |
0.3601 0.3660 0.3635 0.3628 0.3688 |
0.45 0.45 0.45 0.45 0.45 |
4415.3 4418.6 4473.7 4391.2 4407.2 |
0.8122 0.8127 0.8229 0.8077 0.8106 |
100.3 99.6 101.2 99.4 99.0 |
99.88 |
0.7 1 |
The average recovery rate of 5 average recovery tests is 99.88%, and RSD is 0.71%, shows that the response rate is good.
Experimental example 10: sample determination experiment
Accurate reference substance solution and each 10 μ l of sample solution of drawing, sample introduction is measured respectively, measures peak area, with ephedrine hydrochloride content in the external standard method calculation sample, sees Table 12.
Determination of Ephedrine Hydrochloride in table 12 sample
Lot number |
Content of Ephedrine With (%) |
On average (%) |
20010301 20010318 20010321 20010328 20010405 20010406 20010412 20010419 20010425 20010429 |
0.071 0.072 0.061 0.050 0.050 0.050 0.053 0.056 0.061 0.064 0.081 0.083 0.074 0.076 0.069 0.067 0.058 0.060 0.063 0.065 |
0.072 0.056 0.050 0.054 0.062 0.082 0.075 0.068 0.059 0.064 |
According to above 10 batch sample assay results, five tastes manna bath essence contains ephedrine and shows that for product mensuration this product contains Herba Ephedrae and counts 0.050%~0.082% with ephedrine hydrochloride, and on this basis, this product contains Herba Ephedrae with ephedrine hydrochloride (C
10H
15NOHCl) meter must not be less than 0.05%.
The specific embodiment:
Embodiment 1:
Get Juniperus oxycedrus 2kg, Cacumen Myricariae Germanicae 8kg, Rhododendron 3kg, Artemisia sieversiana Willd. 7kg, Herba Ephedrae 6kg adds Cortex Eucommiae 1kg, pretreated Tibetan medicine material mix homogeneously, superfine powder is broken into 1 μ m, and packing gets final product.
Embodiment 2:
Juniperus oxycedrus 3kg, Cacumen Myricariae Germanicae 5kg, Rhododendron 7kg, Artemisia sieversiana Willd. 4kg, Herba Ephedrae 9kg adds Radix Lamiophlomidis Rotatae 2kg, adds cosolvent carbomer 30g, perfuming foenugreek oil 5ml, pretreated Tibetan medicine material mix homogeneously, superfine powder is broken into 20 μ m, and packing gets final product.
Embodiment 3:
Juniperus oxycedrus 3kg, Cacumen Myricariae Germanicae 3kg, Rhododendron 3kg, Artemisia sieversiana Willd. 3kg, Herba Ephedrae 3kg adds Cortex Eucommiae 3kg, adds suspensoid aluminium silicate 180g, pretreated Tibetan medicine material mix homogeneously, superfine powder is broken into 50 μ m, and packing gets final product.
Embodiment 4:
Juniperus oxycedrus 6kg, Cacumen Myricariae Germanicae 2kg, Rhododendron 1kg, Artemisia sieversiana Willd. 2kg, Herba Ephedrae 6kg adds cosolvent carbomer 300g, perfuming Peppermint Oil 50 ml, pretreated Tibetan medicine material mix homogeneously, superfine powder is broken into 100 μ m, and packing gets final product.
Embodiment 5:
Get Juniperus oxycedrus 2kg, Cacumen Myricariae Germanicae 8kg, Rhododendron 3kg, Artemisia sieversiana Willd. 7kg, Herba Ephedrae 6kg adds solely-flavor 1kg, and perfuming Oleum sesami 100ml adds cosolvent silicon Bentonite 1kg, pretreated Tibetan medicine material mix homogeneously, superfine powder is broken into 0.1 μ m, and packing gets final product.
Embodiment 6:
Get Juniperus oxycedrus 2kg, Cacumen Myricariae Germanicae 8kg, Rhododendron 3kg, Artemisia sieversiana Willd. 7kg, Herba Ephedrae 6kg adds Cortex Eucommiae 1kg, adds the pretreated Tibetan medicine material of suspensoid aluminium silicate 3g mix homogeneously, and superfine powder is broken into 0.05 μ m, and packing gets final product.
Embodiment 7:The assay that five tastes manna is bathed smart (promptly being broken into the microgranule of 50 μ m through superfine powder)
Chromatographic condition and system suitability test: with octadecyl silane is filler; With 3: 97: 0.3 methanol-water-triethylamines was mobile phase, with phosphoric acid adjust pH to 3.5; The detection wavelength is 210nm; Column temperature: 20 ℃; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the ephedrine hydrochloride reference substance, and accurate the title decides, and adds the methanol solution that contains 8% ammonia and makes the solution that every 1ml contains 0.1mg, promptly;
The preparation of need testing solution: get this pharmaceutical composition 1.2g that is ground into 50 μ m, the accurate title, decide, and puts in the 100ml tool plug conical flask, the accurate methanol solution 25ml that contains 8% ammonia that adds, claim to decide weight, supersound process 45 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, promptly;
Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
This pharmaceutical composition contains ephedrine with ephedrine hydrochloride C
10H
15The NOHCl meter must not be less than 0.05%.