CN104833736A - Fast qualitative and quantitative detection method for 25-ingredient coral pills through quantitative analysis of multi-components by single-marker - Google Patents
Fast qualitative and quantitative detection method for 25-ingredient coral pills through quantitative analysis of multi-components by single-marker Download PDFInfo
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Abstract
The invention relates to a fast qualitative and quantitative detection method for 25-ingredient coral pills through quantitative analysis of multi-components by single-marker, and is characterized in that a thin layer chromatography method is adopted, a same sample testing solution is adopted, on five thin-layer plates, 9 traditional Chinese medicines comprising safflower, stigma croci, flos pyrethri tatsienensis, liquorice, swertia bimaculata, fructus chebulae, radix aucklandiae, clove and rhizoma acori calami are identified, pretreatment of a sample and a control medicinal material only requires 30 minutes, the amount of a solvent is 30 ml, spots are clear, and the method is quick; the quantitative method for eugenol, costunolide and dehydrocostus lactone through quantitative analysis of multi-components by single-marker is established, double channels are adopted, eugenol is detected at the detection wavelength of 280 nm, costunolide and dehydrocostus lactone are detected at the detection wavelength of 210 nm, and thus not only can the problems that an eugenol baseline does not return to zero and is interfered be solved, but also the problems that costunolide and dehydrocostus lactone are low in content, have no absorption peak and cannot be detected are solved. The wave peak sizes are appropriate and are not interfered with each other, quantitative analysis of multi-components by single-marker is achieved, the efficiency is high, and the cost is low.
Description
Technical field
Fast qualitative and quantitative detecting method are commented in the survey that the present invention relates to a kind of 25-component coral ball more.Qualitative rapid techniques refers to employing thin-layer identification method, to the quick TLC distinguish of safflower, west safflower, DAJIANJU, Radix Glycyrrhizae, Swertia patens, myrobalan, the banksia rose, cloves and Rhizoma Acori Calami 9 taste Chinese medicine; Rapid Quantification refers to efficient liquid-phase chromatography method, the different determined wavelength of binary channels, surveys three evaluation quantitative determinations to one of eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b.
Background technology
In compound Chinese medicinal preparation, the particularly compound preparation of the above prescription of 25-component, because flavour of a drug are many, the dosage of each flavour of a drug is low, and numerous micro-effective constituent is disturbed mutually, and TLC distinguish rate is low, quantitative measurement index is few, for version Chinese Pharmacopoeia in 2010, the compound preparation more than 25-component recorded, rough calculating about has 14 kinds, its TLC distinguish records rate average out to about 12%, quantitative measurement index, calculates, on average less than a kind of composition by the composition measured.That is in the preparation of an above prescription of 25-component, it is with or without feeding intake to only have 3-4 taste medicinal material to identify, a kind of composition can control its content, and numerous medicinal materials and effective constituent are uncontrollable its feeds intake situation and content, quality standard identification level is too low, cannot provide strong support to quality supervision.
Even if in the kind that quality detecting index is relatively many, TLC distinguish is also is mostly kinds, and differentiate if any N item, will prepare N kind need testing solution, N block thin layer plate, launches N time, differentiates traditional differential mode of N taste medicinal material.For exclusive PCR, the many complexity of pre-treatment program of sample, loaded down with trivial details, need by a large amount of organic reagents purification process repeatedly, effort, time-consuming, take reagent, contaminated environment, health risk, sense cycle is long.Like this, a quality standard containing 6 ~ 7 TLC distinguish and binomial assay has detected, and generally will spend the time of one week, as retrial, the time is double, and detection speed seriously governs modernization of Chinese medicine speed of production.So find simple, fast detection method, raising detection efficiency, reduction testing cost, it is also the difficulty that traditional Chinese medicine quality controls to break through.The capsule with pseudo-ginseng and Chinese fanpalm seed quality standard recorded for Chinese Pharmacopoeia version one in 2010 dissects as follows.This kind is made up of 24 taste Chinese medicines, has recorded 7 TLC distinguish and two assays under its normal term.Concrete grammar is as follows:
This product content 4g is got in [discriminating] (1), adds methyl alcohol 20ml, floods 10 minutes, filter, get filtrate 10ml, evaporate to dryness, the residue 10ml that adds water makes dissolving, then adds hydrochloric acid 1ml, puts in water-bath and adds hot reflux 30 minutes, cool immediately, extract 2 times with ether jolting, each 10ml, merge ether solution, evaporate to dryness, residue adds methenyl choloride 1ml makes dissolving, as need testing solution.Separately get rheum officinale control medicinal material 0.1g, be made in the same way of control medicinal material solution.Draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be that the upper solution of the sherwood oil-ethyl formate-formic acid of 15: 5: 1 30 ~ 60 DEG C is for developping agent with volume ratio, launch, take out, dry, inspect under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious five identical orange-yellow fluorescence spots; Put after smoking in ammonia steam, spot becomes red.
(2) get this product content 4g, add strong ammonia solution 1ml and methenyl choloride 20ml, flood 1 hour, jolting constantly, filter, filtrate evaporate to dryness, residue adds ethanol 5ml makes dissolving, as need testing solution.Separately get tetrahydropalmatine reference substance, add ethanol and make the solution of every 1ml containing 1mg, product solution in contrast.Draw need testing solution 20 μ l, reference substance solution 1 μ l, put respectively on same silica gel g thin-layer plate, with volume ratio be the normal hexane-methenyl choloride-methyl alcohol of 10: 6: 1 for developping agent, launch, take out, dry, put in iodine vapor and smoke to spot development clear, wave the iodine that most thin layer plate adsorbs, inspect under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color.
(3) get this product content 3g, add methenyl choloride 25ml, add hot reflux 5 hours, filter, add activated charcoal 0.3g, jolting, place 30 minutes in filtrate, filter, filtrate is concentrated into dry, and residue adds methenyl choloride 1ml makes dissolving, as need testing solution.Separately get Cinobufagin reference substance, add methenyl choloride and make the solution of every 1ml containing 2mg, product solution in contrast.Draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate, is that the cyclohexane-methenyl choloride-acetone of 4: 3: 3 is for developping agent with volume ratio, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 DEG C to be heated to spot development clear.In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
(4) get this product content 6g, add 80% acetone soln 20ml, ultrasonic process 30 minutes, leave standstill, get supernatant, as need testing solution.Separately get safflower control medicinal material 0.5g, be made in the same way of control medicinal material solution.Draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio be the acetate-methanol-formic acid-water of 7: 0.4: 2: 3 for developping agent, launch, take out, dry.In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color.
(5) get this product content 6g, add methyl alcohol 50ml, add hot reflux 1 hour, let cool, filter, filtrate evaporate to dryness, the residue 30ml that adds water makes dissolving, extracts 2 times, each 30ml with water saturated normal butyl alcohol jolting, merge normal butyl alcohol liquid, add 1% sodium hydroxide solution and wash 2 times, each 20ml, get normal butyl alcohol liquid, with normal butyl alcohol saturated be washed to neutrality, get normal butyl alcohol liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Separately get Astragaloside IV reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Draw need testing solution 10 μ l, reference substance solution 5 μ l, putting respectively on same silica gel g thin-layer plate, is that lower floor's solution of the methenyl choloride-methanol-water of 13: 7: 2 is for developping agent with volume ratio, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 DEG C to be heated to spot development clear.In test sample chromatogram, on the position corresponding to reference substance chromatogram, aobvious same color spot; Inspect under putting ultraviolet lamp 365nm, the fluorescence spot of aobvious same color.
(6) get this product content 5g, add methenyl choloride 20ml, ultrasonic process 30 minutes, filter, filtrate evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Separately get cholic acid reference substance, add ethanol and make the solution of every 1ml containing 1mg, product solution in contrast.Draw need testing solution 10 μ l, reference substance solution 4 μ l, putting respectively on same silica gel g thin-layer plate, is that the isooctane-ethyl acetate-glacial acetic acid of 15: 7: 5 is for developping agent with volume ratio, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 DEG C to be heated to spot development clear.In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
(7) get this product content 10g, add methenyl choloride 30ml, add hot reflux 4 hours, let cool, filter, filtrate evaporate to dryness, residue adds methenyl choloride 1ml makes dissolving, as need testing solution.Separately get pangolin control medicinal material 2g, be made in the same way of control medicinal material solution.Draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio be the toluene-acetone of 20: 1 for developping agent, launch, take out, dry.Spray take volume ratio as the mixed solution of the aceticanhydride-sulfuric acid of 9: 1, and 80 DEG C to be heated to spot development clear.In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color.
Kuh-seng assay
Chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filling agent, be that the phosphate buffer-triethylamine of the acetonitrile-methanol-pH6.8 of 18: 18: 70: 0.1 is for mobile phase with volume ratio, determined wavelength is 220nm, number of theoretical plate, calculates be not less than 8000 by matrine peak.
It is appropriate, accurately weighed that matrine reference substance is got in reference substance solution preparation, adds methyl alcohol and make the solution of every 1ml containing 60 μ g, to obtain final product.
This product content under content uniformity item is got in need testing solution preparation, and porphyrize, gets about 0.5g, accurately weighed, put in tool plug conical flask, add liquor ammoniae fortis 2ml and make moistening, add methenyl choloride 25ml again, with power 250w, frequency 33kHz, ultrasonic process 20 minutes, filter, get filtrate, then wash residue, container and filter 4 times with methenyl choloride, each 5ml, filter, merging filtrate, puts evaporate to dryness in water-bath, residue methyl alcohol dissolves and is transferred in 10ml measuring bottle, add methyl alcohol to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
This product every contains kuh-seng with matrine C
16h
24n
2o counts, and must not be less than 0.16mg.
Dried venom of toads assay
Chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filling agent, with volume ratio be the acetonitrile-water of 43.5: 56.5 for mobile phase, determined wavelength is 292nm, number of theoretical plate, by Cinobufagin peak calculate should be not less than 4000.
It is appropriate, accurately weighed that Cinobufagin reference substance is got in reference substance solution preparation, adds methyl alcohol and make the solution of every 1ml containing 0.16mg, to obtain final product.
The content of this product 40 is got in need testing solution preparation, accurately weighed, mixing, and porphyrize, gets about 5g, accurately weighed, put in apparatus,Soxhlet's, add methenyl choloride appropriate, add hot reflux 5 hours, filter, filtrate adds activated charcoal 0.3g, jolting, places 5 minutes, filter, use methenyl choloride 10ml, gradation washing nozzle and filter residue, merging filtrate, evaporate to dryness, residue adds methyl alcohol makes dissolving in right amount, is transferred in 5ml measuring bottle, adds methyl alcohol to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
This product every contains the dried venom of toads with Cinobufagin C
26h
34o
6meter, must not be less than 25 μ g.
7 TLC distinguish in above-mentioned example, calculate roughly, 7 TLC distinguish only time for sample pretreatment, 20 hours will be spent, sample 38g, organic solvent 370ml, add developping agent 70ml and duration of run 7 hours, the TLC distinguish of 1 batch sample, will spend time and the 440ml organic solvent of about 4 days.Add the 3 days quantitative measurement time of kuh-seng and the dried venom of toads, sample preparation solvent 165ml, the organic phase 330ml in mobile phase, TLC distinguish and the quantitative measurement of a batch sample complete, and take time 7 days, organic reagent 770ml.As retrial, obtain testing result, just need half month, detection speed cannot match with the large production of mechanization.The method of quality control of the compound Chinese medicinal preparation as anatomy example can be found everywhere, so improve detection efficiency, reduction testing cost, minimizing environmental pollution, become the instant objective of the struggle of testing staff, we are exactly under this background condition, the discriminating recorded for version in 2010 pharmacopeia less, again without Tibetan's medicine 25-component coral ball of quantitative measurement index, the raising having carried out quality standard is studied with simplification, has invented this survey and has commented fast qualitative and quantitative detecting method more.
Its prescription composition and preparation method repeat no more under seeing the 25-component coral ball item that Chinese Pharmacopoeia version one in 2010 is recorded.
Summary of the invention
Adopt thin layer chromatography, same need testing solution, on five blocks of thin layer plates, authenticated safflower, west safflower, DAJIANJU (Fig. 1-1,1-2); Radix Glycyrrhizae and Swertia patens (Fig. 2-1,2-2); Myrobalan (Fig. 3-1,3-2); The banksia rose (Fig. 4); Cloves and Rhizoma Acori Calami (Fig. 5-1,5-2) 9 taste Chinese medicine, the pre-treatment of sample and control medicinal material only needs 30 minutes, solvent 30ml, clear spot, and method is quick; One of the eugenol set up, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b is surveyed and is evaluated metering method more, with binary channels, eugenol detects at 210nm at 280nm, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b, both eugenol baseline not back to zero, interference problem had been solved, solve again costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b without inhaling crest, a unmeasured difficult problem.Make each crest suitable size, do not interfere with each other, one surveys three comments, and efficiency is high, and cost is low.
The TLC distinguish of this invention 9 taste medicinal material, and the quantitative measurement of three kinds of effective constituents, need test sample 3.6g be used altogether, 40 minutes pre-treatment time, add the time of thin-layer developing and quantitative measurement, solvent, total cost solvent 330ml, 2 days time, compares with above-mentioned capsule with pseudo-ginseng and Chinese fanpalm seed, when the TLC distinguish of 2 taste medicinal materials more than capsule with pseudo-ginseng and Chinese fanpalm seed and a kind of component quantifying measure, TLC distinguish and the quantitative measurement of one batch sample complete, and also can save time 5 days, organic reagent 440ml.So the present invention is increasing substantially on quality of the pharmaceutical preparations controllability social benefit basis, and then also improve detection efficiency more than 5 times, save detection time 80%, reduce testing cost 80%, reduce environmental pollution 90%, form the modernization standard of a set of easy, quick, efficient, low consumption, low stain, multi-objective control drug quality.
By methodological study, eugenol sample size is at 0.1879 ~ 1.879 μ g, and be good linear relationship with peak area, regression equation is: Y=989573.4X+435, γ=0.99999 (table 1, Fig. 6); Costunolide sample size is at 0.03272 ~ 0.3272 μ g, and be good linear relationship with peak area, regression equation is: Y=2736197.6X-1870, γ=0.99999 (table 2, Fig. 7); Decahydro-3,6,9-tris(methylene)azuleno[4,5-b sample size is at 0.04778 ~ 0.4778 μ g, and be good linear relationship with peak area, regression equation is: Y=3193542.3X-2071, γ=0.99999 (table 3, Fig. 8); Adopt application of sample recovery experiment, result shows: the average recovery rate that eugenol measures for 6 times is 99.98%, RSD is 0.88% (table 4); The average recovery rate of costunolide is 99.11%, RSD is 1.85% (table 5); The average recovery rate of Decahydro-3,6,9-tris(methylene)azuleno[4,5-b is 100.21%, RSD is 1.20% (table 6); Precision (table 7), stability (table 8), repeatability (table 9), specificity (Fig. 9-1,9-2,9-3, Figure 10-1,10-2,10-3) and post durability (table 10, Figure 11-1,11-2, Figure 12-1,12-2, Figure 13-1,13-2) data all meet methodology requirement.Be applicable to one of eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b in 25-component coral ball and survey three evaluation quantitative determination requirements.
The present invention solves the scheme that its technical matters adopts:
(1) quick thin-layer identification method
1. get 25-component coral ball 3g, porphyrize, add methyl alcohol 10ml, ultrasonic process 20 minutes, filter, filtrate evaporate to dryness, residue adds methyl alcohol 1ml, makes dissolving, as need testing solution; Separately get safflower control medicinal material 0.2g, west safflower control medicinal material 0.04g, DAJIANJU control medicinal material 0.2g, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw need testing solution and each 5 ~ 8 μ l of above-mentioned control medicinal material solution, put respectively in same silica G F
254on thin layer plate, with volume ratio be the acetic ether-methanoic acid-water-methanol of 10: 2: 2: 0.4 for developping agent, launch, take out, hot blast drying, inspects under putting daylight; In test sample chromatogram, on the position corresponding to west safflower control medicinal material chromatogram, aobvious identical yellow principal spot; Inspect under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding with DAJIANJU control medicinal material chromatogram to safflower, the fluorescence principal spot of aobvious same color respectively, verify through blank, blank sample (not containing the sample of safflower or west safflower or DAJIANJU) is noiseless, differentiates exclusive;
2. extracting Radix Glycyrrhizae and each 0.1g of Swertia patens control medicinal material, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, and supernatant is medicinal material solution in contrast; Draw control medicinal material solution and differentiate 1. each 5 ~ 8 μ l of need testing solution under item, putting respectively in same silica G F
254on thin layer plate, with volume ratio be the petroleum ether-ethyl acetate-formic acid of 60-90 DEG C of 10: 4: 0.5 for developping agent, launch, take out, hot blast drying, inspect under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding with Swertia patens control medicinal material chromatogram to Radix Glycyrrhizae, aobvious same color fluorescence principal spot respectively, through blank checking, blank sample (not containing the sample of Radix Glycyrrhizae or Swertia patens) is noiseless, differentiates exclusive;
3. get myrobalan's control medicinal material 0.2g, add methyl alcohol 2ml, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw control medicinal material solution 3-5 μ l, differentiate 1. item under need testing solution 8 ~ 10 μ l, put respectively in same silica G F
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate-formic acid of 5.5: 4.5: 0.5 for developping agent, launch, take out, hot blast drying, inspects under putting ultraviolet lamp 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious same color principal spot; Spray 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, inspect under daylight, in test sample chromatogram, on the position corresponding to myrobalan's control medicinal material chromatogram, aobvious same color principal spot, verifies through blank, blank sample (not containing the sample of myrobalan) is noiseless, differentiates exclusive;
4. get banksia rose control medicinal material 0.2g, add methyl alcohol 2ml, ultrasonic process 20 minutes, in contrast medicinal material solution; Draw control medicinal material solution and differentiate 1. each 5 ~ 8 μ l of need testing solution, respectively same silica G F under item
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate-formic acid of 8: 2: 0.1 for developping agent, launch, take out, hot blast drying, spraying with volume ratio is the 5% vanillin-sulfuric acid solution of 1: 6 and the mixed solution of ethanolic solution, and 105 DEG C to be heated to spot development clear, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious same color principal spot, verifies through blank, blank sample (not containing the sample of the banksia rose) is noiseless, differentiates exclusive;
5. get Rhizoma Acori Calami control medicinal material 0.2g, cloves control medicinal material 0.1g, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; 1. need testing solution and each 5 ~ 8 μ l of above-mentioned control medicinal material solution under item are differentiated in absorption, put respectively in same silica G F
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate of 10: 2 for developping agent, launch, take out, dry, inspect under putting ultraviolet lamp 254nm, in test sample chromatogram, on the position corresponding with cloves control medicinal material chromatogram to Rhizoma Acori Calami, aobvious same color principal spot respectively; Spray 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, inspect under putting ultraviolet lamp 365nm immediately, in test sample chromatogram, on the position corresponding to cloves control medicinal material chromatogram, aobvious same color fluorescence principal spot, verifies through blank, blank sample (not containing the sample of cloves or Rhizoma Acori Calami) is noiseless, differentiates exclusive;
Eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b one is surveyed three and is commented rapid assay methods
Chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filling agent; Be that acetonitrile-methanol-0.1% phosphoric acid of 37: 18: 45 is for mobile phase with volume ratio; Determined wavelength: eugenol is 280nm, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b is 210nm; Column temperature is 30 DEG C; Number of theoretical plate calculates by Decahydro-3,6,9-tris(methylene)azuleno[4,5-b peak and is not less than 2000;
It is appropriate that eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b reference substance are got in the preparation of reference substance solution, accurately weighed, add 70% methyl alcohol and make every 1ml containing eugenol 90 μ g, costunolide 16 μ g, each 24 μ g mixed solutions of Decahydro-3,6,9-tris(methylene)azuleno[4,5-b, to obtain final product;
The preparation of need testing solution gets 25-component coral ball in right amount, and porphyrize, gets 0.6g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with the ultrasonic process of power 250W, frequency 25kHz 30 minutes, let cool, weighed weight again, supplies the weight of less loss, shakes up with methyl alcohol, filter, to obtain final product;
Determination method is accurate respectively draws reference substance solution 5 μ l, need testing solution 8 μ l, injects liquid phase color development spectrometer, measures, to obtain final product.
Principle of the present invention is as follows:
1. different with polarity according to the chemical constitution of each effective constituent of Chinese medicine, along with the movement of developping agent, the ability that the Adsorption and desorption on thin layer plate is attached and different, makes the spot of each effective constituent be separated.Again by each compositional polarity size, choose the effective constituent that polarity is approximate, under different conditions of inspecting, present spot colors different separately, overlapped, but do not interfere with each other, same thin layer plate detects several medicinal materials simultaneously, same need testing solution, for multinomial discriminating application.
2. pass through the component of adjustment mobile phase, ratio and determined wavelength, make different in kind, effective constituent that content differs greatly can in same mobile phase, under different determined wavelength, can present crest is separated good, noiseless, peak area is suitable for, and its PeakArea and content are good linear relation, and be able to quantitative measurement.
Innovative point of the present invention and beneficial effect as follows:
(1) breaching a herbal species has N item to differentiate, N kind need testing solution will be prepared, N block thin layer plate, launch N time, differentiate traditional identification method of N taste medicinal material, establish and adopt same need testing solution and the ultrasonic each control medicinal material supernatant of methyl alcohol, on 5 blocks of thin layer plates, complete the quick thin-layer identification method of 9 taste Chinese medicines.Both had and broken through conventional novelty, there is again raising detection efficiency, saved testing cost, reduced the practicality of environmental pollution.
(2) on same thin layer plate, different inspect condition under, detect safflower, west safflower and DAJIANJU simultaneously; The discrimination method of Radix Glycyrrhizae and Swertia patens and Rhizoma Acori Calami and cloves, yet there are no identical report, tool novelty and practicality.
(3) prescription of the present invention is made up of 25-component Chinese medicine, and the myrobalan of most high-load, converts in every 1g pill, just containing 0.087g medicinal material, the muscone of minimum content, converts in every 1g pill, only containing 0.0017g medicinal material, prescription is large, complicated component, and each flavour of a drug disturb mutually, but content is too low again separately, define TLC distinguish and revise and enlarge difficulty greatly, record rate low, be difficult to the present situation controlling drug quality.As the Ershiwuwei zhenzhu wan that Chinese Pharmacopoeia version in 2010 is recorded, Chaihushugan Pill, the kind of 14 above flavour of a drug compositions of 25 taste such as children-welfare tablet, its TLC distinguish revises and enlarges rate on average about 12-13%.The present invention only uses a kind of need testing solution, five blocks of thin layer plates are differentiated 9 taste medicinal materials, the discriminating rate of revising and enlarging is brought up to about about 36%, and method is easy, quick, 3g sample, 30ml methyl alcohol Extraction solvent, 60ml developping agent, spend 2 hours, can complete the TLC distinguish of 9 taste medicinal materials, its agility, novelty, practicality are that discrimination method conventional at present does not possess.
(4) the present invention also for more than 25 tastes large prescription, multiple medicines taste composition its quantitative measurement index of preparation on average less than the present situation of a kind of composition, 3 kinds of compositions are brought up in the quantitative measurement of 25-component coral ball, and be in the mode of the most frequently used isocratic elution, same mobile phase, the Simultaneously test under different determined wavelength.Determined wavelength: eugenol 280nm, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b 210nm, both eugenol crest value had at 210 nm been solved too large, baseline is back to zero, interference problem not, solve again costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b under 280nm without absorbing, and under the 225nm of bibliographical information, crest value is too little, the drawback that the reappearance of data is not good enough.Under different wavelength, detect different compositions, each crest suitable size, does not interfere with each other, and one surveys three comments, and easy, quick, accurate, cost is low, efficiency is high, is conducive to market surveillance.
(5) the present invention is compared with the quality determining method of the Chinese medicine preparation of the above prescription of current 25 taste, not only the quality controllability of preparation increases substantially, also improve detection efficiency more than 5 times, save detection time 80%, reduce testing cost 80%, reduce environmental pollution 90%, define the modernization standard of a set of easy, quick, efficient, low consumption, low stain, multi-objective control drug quality.
Accompanying drawing explanation
Fig. 1-1,1-2 are the TLC distinguish figure of safflower, west safflower and DAJIANJU.
Fig. 2 is the TLC distinguish figure of Radix Glycyrrhizae and Swertia patens.
Fig. 3-1,3-2 are the TLC distinguish figure of myrobalan.
Fig. 4 is banksia rose TLC distinguish figure.
Fig. 5-1,5-2 are cloves and Rhizoma Acori Calami TLC distinguish figure.
Fig. 6 eugenol reference substance linear relationship chart
Fig. 7 costunolide reference substance linear relationship chart
Fig. 8 Decahydro-3,6,9-tris(methylene)azuleno[4,5-b reference substance linear relationship chart
The HPLC chromatogram of Fig. 9-1 eugenol reference substance
Fig. 9-2 25-component coral ball sample HPLC chromatogram
Fig. 9-3 lacks the blank sample HPLC chromatogram of cloves
The HPLC chromatogram of Figure 10-1 costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b reference substance
Figure 10-2 25-component coral ball sample HPLC chromatogram
Figure 10-3 lacks the blank sample HPLC chromatogram of the banksia rose
Eugenol HPLC chromatogram in Figure 11-1 Shimadzu-SP-ODS post (4.6 × 150mm) coral ball
Costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b HPLC chromatogram in Figure 11-2 Shimadzu-SP-ODS post (4.6 × 150mm) coral ball
Eugenol HPLC chromatogram in Figure 12-1 enlightening horse dimonsil chromatographic column (4.6 × 150mm) coral ball
Costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b HPLC chromatogram in Figure 12-2 enlightening horse dimonsil chromatographic column (4.6 × 150mm) coral ball
Eugenol HPLC chromatogram in Figure 13-1 Shimadzu-VP-ODS post (4.6 × 150mm) coral ball
Costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b HPLC chromatogram in Figure 13-2 Shimadzu-VP-ODS post (4.6 × 150mm) coral ball
Fig. 1-1,1-2 are same thin layer plate, and Fig. 1-1 is the yellow spotting inspecting west safflower in the sunlight, and Fig. 1-2 is the fluorescence spot inspecting safflower and DAJIANJU under uviol lamp 365nm.In figure, 1 safflower; 2 safflowers are blank; 3 west safflowers; 4 west safflowers are blank; 5,6,7 coral ball samples; 8 DAJIANJUs; 9 DAJIANJUs are blank
In Fig. 2,1 Swertia patens is blank; 2 Radix Glycyrrhizaes; 3 Radix Glycyrrhizaes are blank; 4,5,6 coral ball samples; 7 Swertia patens
Fig. 3-1,3-2 are same thin layer plate, and Fig. 3-1 is the myrobalan's taupe characteristic spots inspected under ultraviolet lamp 254nm, and Fig. 3-2 is another characteristic spots inspected under daylight after the colour developing of spraying 10% ethanol solution of sulfuric acid.In figure, 1. blank 2.3.4 coral ball sample 5. myrobalan
In Fig. 4,1.6 banksia rose; 2 is blank; 3.4.5 coral ball sample
Fig. 5-1,5-2 are same thin layer plate, and Fig. 5-1 is the cloves and Rhizoma Acori Calami color characteristic spot of inspecting under ultraviolet lamp 254nm, and Fig. 5-2 is after the colour developing of spraying 10% ethanol solution of sulfuric acid, the cloves fluorescent characteristics spot inspected under ultraviolet lamp 365nm.In figure, blank 6. Rhizoma Acori Calami 7. Rhizoma Acori Calamis of 1.2.3 sample 4. cloves 5. cloves are blank
In Fig. 6, Fig. 7, Fig. 8, horizontal ordinate is sample size μ g, and ordinate is peak area
In Fig. 9-1, Fig. 9-2, Fig. 9-3, Figure 10-1, Figure 10-2, Figure 10-3, Figure 11-1, Figure 11-2, Figure 12-1, Figure 12-2, Figure 13-1, Figure 13-2, be 1. eugenol, 2. costunolide 3. Decahydro-3,6,9-tris(methylene)azuleno[4,5-b
The specific embodiment of the invention is as follows:
(1) quick thin-layer identification method
1. get 25-component coral ball 3g, porphyrize, add methyl alcohol 10ml, ultrasonic process 20 minutes, filter, filtrate evaporate to dryness, residue adds methyl alcohol 1ml, makes dissolving, as need testing solution; Separately get safflower control medicinal material 0.2g, west safflower control medicinal material 0.04g, DAJIANJU control medicinal material 0.2g, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw need testing solution and each 5 ~ 8 μ l of above-mentioned control medicinal material solution, put respectively in same silica G F
254on thin layer plate, with volume ratio be the acetic ether-methanoic acid-water-methanol of 10: 2: 2: 0.4 for developping agent, launch, take out, hot blast drying, inspects under putting daylight; In test sample chromatogram, on the position corresponding to west safflower control medicinal material chromatogram, aobvious identical yellow principal spot; Inspect under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding with DAJIANJU control medicinal material chromatogram to safflower, the fluorescence principal spot of aobvious same color respectively;
2. extracting Radix Glycyrrhizae and each 0.1g of Swertia patens control medicinal material, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, and supernatant is medicinal material solution in contrast; Draw control medicinal material solution and differentiate 1. each 5 ~ 8 μ l of need testing solution under item, putting respectively in same silica G F
254on thin layer plate, be that the petroleum ether-ethyl acetate-formic acid of 60-90 DEG C of 10: 4: 0.5 is for developping agent with volume ratio, launch, take out, hot blast drying, inspects under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding with Swertia patens control medicinal material chromatogram to Radix Glycyrrhizae, aobvious same color fluorescence principal spot respectively;
3. get myrobalan's control medicinal material 0.2g, add methyl alcohol 2ml, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw control medicinal material solution 3-5 μ l, differentiate 1. item under each 8 ~ 10 μ l of need testing solution, put respectively in same silica G F
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate-formic acid of 5.5: 4.5: 0.5 for developping agent, launch, take out, hot blast drying, inspects under putting ultraviolet lamp 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious same color principal spot; Spray 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, inspects under daylight, in test sample chromatogram, on the position corresponding to myrobalan's control medicinal material chromatogram, and aobvious same color principal spot;
4. get banksia rose control medicinal material 0.2g, add methyl alcohol 2ml, ultrasonic process 20 minutes, in contrast medicinal material solution; Draw control medicinal material solution and differentiate 1. each 5 ~ 8 μ l of need testing solution, respectively same silica G F under item
254on thin layer plate, be that the cyclohexane-ethyl acetate-formic acid of 8: 2: 0.1 is for developping agent with volume ratio, launch, take out, hot blast drying, spraying with volume ratio is the 5% vanillin-sulfuric acid solution of 1: 6 and the mixed solution of ethanolic solution, 105 DEG C to be heated to spot development clear, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious same color principal spot;
5. get Rhizoma Acori Calami control medicinal material 0.2g, cloves control medicinal material 0.1g, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; 1. need testing solution and each 5 ~ 8 μ l of above-mentioned control medicinal material solution under item are differentiated in absorption, put respectively in same silica G F
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate of 10: 2 for developping agent, launch, take out, dry, inspect under putting ultraviolet lamp 254nm, in test sample chromatogram, on the position corresponding with cloves control medicinal material chromatogram to Rhizoma Acori Calami, aobvious same color principal spot respectively; Spray 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp 365nm immediately, in test sample chromatogram, on the position corresponding to cloves control medicinal material chromatogram, and aobvious same color fluorescence principal spot.
Eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b one is surveyed three and is commented rapid assay methods
Chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filling agent; Be that acetonitrile-methanol-0.1% phosphoric acid of 37: 18: 45 is for mobile phase with volume ratio; Determined wavelength: eugenol is 280nm, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b is 210nm; Column temperature is 30 DEG C; Number of theoretical plate calculates by Decahydro-3,6,9-tris(methylene)azuleno[4,5-b peak and is not less than 2000;
It is appropriate that eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b reference substance are got in the preparation of reference substance solution, accurately weighed, add 70% methyl alcohol and make every 1ml containing eugenol 90 μ g, costunolide 16 μ g, each 24 μ g mixed solutions of Decahydro-3,6,9-tris(methylene)azuleno[4,5-b, to obtain final product;
The preparation of need testing solution gets 25-component coral ball in right amount, and porphyrize, gets 0.6g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with the ultrasonic process of power 250W, frequency 25kHz 30 minutes, let cool, weighed weight again, supplies the weight of less loss, shakes up with methyl alcohol, filter, to obtain final product;
Determination method is accurate respectively draws reference substance solution 5 μ l, need testing solution 8 μ l, injects liquid phase color development spectrometer, measures, to obtain final product.Determine the content of eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b in many batches of 25-component coral balls, result (table 11).
Table 1 eugenol sample size and peak area
Table 2 costunolide sample size and peak area
Table 3 Decahydro-3,6,9-tris(methylene)azuleno[4,5-b sample size and peak area
Eugenol recovery test result in table 4 sample
Costunolide recovery test result in table 5 sample
Decahydro-3,6,9-tris(methylene)azuleno[4,5-b recovery test result in table 6 sample
Note: for saving reference substance, the cumulative volume of sample is not 25ml, is 15ml, in the consistent situation of concentration, sampling amount reduces.
Table 7 Precision Experiment result
Table 8 stability test result
Table 9 replica test result (mg/g)
Table 10 different chromatographic column serviceability test result (mg/g)
Three component content measurement result (mg/g) (n=2) in table 11 25-component coral ball
Claims (2)
1. fast qualitative and quantitative detecting method are commented in the survey that the present invention relates to a kind of 25-component coral ball more, it is characterized in that:
Fast qualitative discrimination method is commented in (1) one survey more
1. get 25-component coral ball 3g, porphyrize, add methyl alcohol 10ml, ultrasonic process 20 minutes, filter, filtrate evaporate to dryness, residue adds methyl alcohol 1ml, makes dissolving, as need testing solution; Separately get safflower control medicinal material 0.2g, west safflower control medicinal material 0.04g, DAJIANJU control medicinal material 0.2g, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw above-mentioned need testing solution and each 5 ~ 8 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, with volume ratio be the acetic ether-methanoic acid-water-methanol of 10: 2: 2: 0.4 for developping agent, launch, take out, hot blast drying, inspects under putting daylight; In test sample chromatogram, on the position corresponding to west safflower control medicinal material chromatogram, aobvious identical yellow principal spot; Inspect under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding with DAJIANJU control medicinal material chromatogram to safflower, aobvious identical fluorescence principal spot respectively;
2. extracting Radix Glycyrrhizae and each 0.1g of Swertia patens control medicinal material, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, and supernatant is medicinal material solution in contrast; Draw control medicinal material solution and differentiate 1. each 5 ~ 8 μ l of need testing solution under item, putting respectively in same silica G F
254on thin layer plate, be that the petroleum ether-ethyl acetate-formic acid of 60-90 DEG C of 10: 4: 0.5 is for developping agent with volume ratio, launch, take out, hot blast drying, inspects under putting ultraviolet lamp 365nm, in test sample chromatogram, on the position corresponding with Swertia patens control medicinal material chromatogram to Radix Glycyrrhizae, aobvious same color fluorescence principal spot respectively;
3. get myrobalan's control medicinal material 0.2g, add methyl alcohol 2ml, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw control medicinal material solution 3-5 μ l, differentiate 1. item under each 8 ~ 10 μ l of need testing solution, put respectively in same silica G F
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate-formic acid of 5.5: 4.5: 0.5 for developping agent, launch, take out, hot blast drying, inspects under putting ultraviolet lamp 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious same color principal spot; Spray 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, inspects under daylight, in test sample chromatogram, on the position corresponding to myrobalan's control medicinal material chromatogram, and aobvious same color principal spot;
4. get banksia rose control medicinal material 0.2g, add methyl alcohol 2ml, ultrasonic process 20 minutes, in contrast medicinal material solution; Draw control medicinal material solution and differentiate 1. each 5 ~ 8 μ l of need testing solution under item, putting respectively in same silica G F
254on thin layer plate, be that the cyclohexane-ethyl acetate-formic acid of 8: 2: 0.1 is for developping agent with volume ratio, launch, take out, hot blast drying, spraying with volume ratio is the 5% vanillin-sulfuric acid solution of 1: 6 and the mixed solution of ethanolic solution, 105 DEG C to be heated to spot development clear, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, aobvious same color principal spot;
5. get Rhizoma Acori Calami control medicinal material 0.2g, cloves control medicinal material 0.1g, add methyl alcohol 2ml respectively, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; 1. need testing solution and each 5 ~ 8 μ l of above-mentioned control medicinal material solution under item are differentiated in absorption, put respectively in same silica G F
254on thin layer plate, with volume ratio be the cyclohexane-ethyl acetate of 10: 2 for developping agent, launch, take out, dry, inspect under putting ultraviolet lamp 254nm, in test sample chromatogram, on the position corresponding with cloves control medicinal material chromatogram to Rhizoma Acori Calami, aobvious same color principal spot respectively; Spray with 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp 365nm immediately, in test sample chromatogram, on the position corresponding to cloves control medicinal material chromatogram, and aobvious same color fluorescence principal spot.
(2). eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b one is surveyed three and is commented rapid assay methods
A. chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filling agent; Be that acetonitrile-methanol-0.1% phosphoric acid of 37: 18: 45 is for mobile phase with volume ratio; Determined wavelength: eugenol is 280nm, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b is 210nm; Column temperature is 30 DEG C; Number of theoretical plate calculates by Decahydro-3,6,9-tris(methylene)azuleno[4,5-b peak and is not less than 2000;
B. reference substance solution preparation gets eugenol, costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b reference substance in right amount, accurately weighed, adds 70% methyl alcohol and makes every 1ml containing eugenol 90 μ g, costunolide 16 μ g, each 24 μ g mixed solutions of Decahydro-3,6,9-tris(methylene)azuleno[4,5-b, to obtain final product;
C. need testing solution preparation gets 25-component coral ball in right amount, and porphyrize, gets 0.6g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with the ultrasonic process of power 250W, frequency 25kHz 30 minutes, let cool, weighed weight again, supplies the weight of less loss, shakes up with methyl alcohol, filter, to obtain final product;
D. determination method is accurate respectively draws reference substance solution 5 μ l, need testing solution 8 μ l, injection liquid chromatography, measures, to obtain final product.
2. a kind of 25-component coral ball one according to claim 1 is surveyed and is commented fast qualitative and quantitative detecting method more, it is characterized in that described qualitative rapid techniques refers to thin-layer identification method; Rapid Quantification refers to efficient liquid-phase chromatography method, and adopts binary channels, and different wave length detects or instantly changing wavelength detecting.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106706766A (en) * | 2015-08-12 | 2017-05-24 | 上海百岁行药业有限公司 | Method for detecting newly increased detection components in traditional Chinese medicinal Tang Herb for treating AIDS |
| CN106950289A (en) * | 2016-01-07 | 2017-07-14 | 陕西步长制药有限公司 | A kind of coronary disease treatment capsule one is surveyed comments detection method of content more |
| CN109085284A (en) * | 2018-08-28 | 2018-12-25 | 王智森 | A kind of quick thin-layer identification method of multiple medicine taste of Ershiwuwei zhenzhu wan |
| CN110031588A (en) * | 2019-03-26 | 2019-07-19 | 浙江金大康动物保健品有限公司 | An a kind of quick thin-layer identification method of plate multiple medicine taste of livestock and poultry antiviral granule |
| CN111830187A (en) * | 2020-06-02 | 2020-10-27 | 鉴甄检测技术（上海）有限公司 | Rapid thin-layer identification method for multiple medicines in small radix bupleuri granule finished product |
-
2015
- 2015-04-20 CN CN201510200425.1A patent/CN104833736B/en active Active
Non-Patent Citations (4)
| Title |
|---|
| 刘刚等: "变换波长HPLC测定七味广枣丸中丁香酚、木香烃内酯和去氢木香内酯的含量", 《中国药学杂志》 * |
| 孙全明等: "HPLC测定木香分气丸中5种成分的含量", 《中成药》 * |
| 德吉措姆等: "HPLC 法测定藏药二十五味珊瑚丸中乌头碱限量", 《中国药品标准》 * |
| 杜连平等: "《藏药二十五味肺病胶囊质量标准研究》", 《食品与药品》 * |
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|---|---|---|---|---|
| CN106706766A (en) * | 2015-08-12 | 2017-05-24 | 上海百岁行药业有限公司 | Method for detecting newly increased detection components in traditional Chinese medicinal Tang Herb for treating AIDS |
| CN106706766B (en) * | 2015-08-12 | 2019-04-16 | 上海百岁行药业有限公司 | A kind of measuring method of the newly-increased detection ingredient of Chinese medicine Tang blade that treating AIDS |
| CN106950289A (en) * | 2016-01-07 | 2017-07-14 | 陕西步长制药有限公司 | A kind of coronary disease treatment capsule one is surveyed comments detection method of content more |
| CN106950289B (en) * | 2016-01-07 | 2019-10-29 | 陕西步长制药有限公司 | A kind of coronary disease treatment capsule one is surveyed comments detection method of content more |
| CN109085284A (en) * | 2018-08-28 | 2018-12-25 | 王智森 | A kind of quick thin-layer identification method of multiple medicine taste of Ershiwuwei zhenzhu wan |
| CN110031588A (en) * | 2019-03-26 | 2019-07-19 | 浙江金大康动物保健品有限公司 | An a kind of quick thin-layer identification method of plate multiple medicine taste of livestock and poultry antiviral granule |
| CN111830187A (en) * | 2020-06-02 | 2020-10-27 | 鉴甄检测技术（上海）有限公司 | Rapid thin-layer identification method for multiple medicines in small radix bupleuri granule finished product |
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Effective date of registration: 20190102 Address after: No. 18 Puzhang Road, Xikaze City, Tibet Autonomous Region Patentee after: Tibet Tibetan pharmaceutical Limited by Share Ltd Address before: Room 1112, Science and Technology Center, 136 Huanghe Avenue, Shijiazhuang Hi-tech Development Zone, Hebei Province, 050035 Patentee before: Wang Zhisen |