CN105758950A - Quality control method based on dose-effect color card for anti-inflammatory and analgesia action of qi-stagnation and stomachache granules - Google Patents
Quality control method based on dose-effect color card for anti-inflammatory and analgesia action of qi-stagnation and stomachache granules Download PDFInfo
- Publication number
- CN105758950A CN105758950A CN201610106193.8A CN201610106193A CN105758950A CN 105758950 A CN105758950 A CN 105758950A CN 201610106193 A CN201610106193 A CN 201610106193A CN 105758950 A CN105758950 A CN 105758950A
- Authority
- CN
- China
- Prior art keywords
- inflammatory
- effect
- qizhi weitong
- weitong granules
- antalgic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a quality control method based on a dose-effect color card for anti-inflammatory and analgesia action of qi-stagnation and stomachache granules. According to the quality control method, correlation between the quality and the medicine effect of the qi-stagnation and stomachache granules is evaluated through construction of a fingerprint of the qi-stagnation and stomachache granules and evaluation of the anti-inflammatory and analgesia action of the qi-stagnation and stomachache granules; a dose-effect relationship equation is constructed; the dose-effect color card for the anti-inflammatory and analgesia action of the qi-stagnation and stomachache granules is successfully constructed; and the anti-inflammatory and analgesia action can be intuitively reflected in a form of the 'color card' by detecting the relative peak area of specific components in the qi-stagnation and stomachache granules. The quality control method can be used for evaluating the anti-ulcer action degree and the quality of the qi-stagnation and stomachache granules.
Description
Technical field
The present invention relates to Control of drug quality method, particularly relate to the qizhi weitong granules anti-inflammatory and antalgic method of quality control of a kind of " colour atla " software reflection quantity effect relationship directly perceived.
Background technology
Compound Chinese medicinal preparation quality control is always up puzzlement Chinese medicine preparation quality monitoring and moves towards difficult point and the hot issue of international market, is also important foundation and the key of the modernization of Chinese medicine.China's compound Chinese medicinal preparation quality standard experienced by grow out of nothing, from simple to the process of gradual perfection, now still among constantly exploring.In the version Pharmacopoeias of the People's Republic of China in 2005, thin layer chromatography is widely used in the discriminating of compound preparation；High performance liquid chromatography becomes the main flow of assay.In quantizating index, also by testing index composition to measuring active component transition, by measuring single component to measuring Multiple components transition.Wherein, the appearance of fingerprint pattern technology, bring hope for solving compound preparation complicated component problem.But, the effective active of disengaging composition and safe activity consider the fingerprint atlas detection method of index components assay, although the method for analysis is advanced, but due to the drug effect information of its be beyond expression Chinese crude drug and Chinese medicine compound, thus most of research worker only using it as the simple quality discrimination means of one.
Qizhi weitong granules is made up of Radix Bupleuri, Rhizoma Corydalis (processing), Fructus Aurantii, Rhizoma Cyperi (processing), the Radix Paeoniae Alba, Radix Glycyrrhizae Preparata 6 taste Chinese medicine, has liver-smoothing, qi-regulating and the effect of stomach and alleviating pain.It is clinically used for stagnation of QI due to depression of the liver, feeling of stuffiness in chest distension, gastralgia.The clinical experiment of Yuan's Polygonum etc. shows that qizhi weitong granules has good antiinflammatory action, Zhang Yabing etc. to treat reflux gastritis with qizhi weitong granules, also achieves good therapeutic effect.At present, the quality control of qizhi weitong granules is had been carried out more research, applicant's early stage has been for the method for quality control of qizhi weitong granules and has carried out patent protection (qizhi weitong granules all the period of time multi-wavelength fusion fingerprint pattern quality control method, the patent No. 201210223306.4), but it is only carried out specification by simple method of quality control its chemical composition, and whether these chemical compositions are relevant to its drug effect and unknowable, therefore this patent is on its basis, for qizhi weitong granules anti-inflammatory and antalgic drug effect, establish a kind of qizhi weitong granules anti-inflammatory and antalgic method of quality control based on " dose-effect colour atla ", adopt " colour atla " this more intuitive way, the quality of qizhi weitong granules is evaluated from the angle of drug effect.
Summary of the invention
For the problems referred to above, qizhi weitong granules anti-inflammatory and antalgic method of quality control based on " dose-effect colour atla ", the drug effect of the evaluation of result qizhi weitong granules performance anti-inflammatory and analgesic effect of high performance liquid chromatography detection can be applied by the method, open up new thinking for Chinese medicine compound quality control.
For realizing the above-mentioned purpose of the present invention, the present invention adopts the following technical scheme that.
The present invention provides a kind of qizhi weitong granules anti-inflammatory and antalgic method of quality control based on " dose-effect colour atla ", concretely comprises the following steps:
Step one, qizhi weitong granules finger printing foundation:
The preparation of need testing solution: in qizhi weitong granules, 6 taste medical material Radix Bupleuri, Rhizoma Corydalis, Fructus Aurantii, Rhizoma Cyperi, Radix Glycyrrhizae are by 0～10g, consumption is carried out 20 Latin hypercubes by 0～15g and randomly draws by the Radix Paeoniae Alba, obtain the medical material compatibility group of 20 different proportions, each compatibility group medical material is mixed, put in round-bottomed flask, 20 times of weight water reflux, extract, 2h, filter, volatilize, it is settled to 250mL, taking 2mL need testing solution, adding acetaminophen reference substance 200 μ L(concentration is 0.1240mg/mL), as interior mark liquid.All the other need testing solutions are put in water-bath and are volatilized, and after calculating paste-forming rate, give over to the experiment of anti-inflammatory and antalgic drug effect and use.
Chromatographic condition: chromatographic column AgilentTC-C18(4.6mm × 250mm, 5 μm)；Mobile phase 0.2 ‰ formic acid water (A)-acetonitrile (B)；Flow velocity 1.0mL min-1；Column temperature is 25 DEG C；Detector wavelength 230nm, 254nm, 283nm, 365nm；Sample size 15 μ L；
Eluent gradient elution parameters is:
Time 0min, mobile phase A 95%, Mobile phase B 5%；
Time 10min, mobile phase A 90%, Mobile phase B 10%；
Time 61min, mobile phase A 63%, Mobile phase B 27%；
Time 70min, mobile phase A 60%, Mobile phase B 40%；
Time 90min, mobile phase A 50%, Mobile phase B 50%.
Test sample all the period of time four wavelength merges collection of illustrative plates to be set up: takes 2mL need testing solution, adds 200 μ L acetaminophen as interior mark liquid (concentration is 0.1240mg/mL).With acetaminophen for internal standard substance, adopt high performance liquid chromatography, utilize diode array (DAD) detector, set up the finger printing of 20 compatibility groups, adopt data processing software to 210nm, 230nm, tetra-wavelength graph modal data of 254nm, 283nm, 365nm process, obtain and reflect that all the period of time four wavelength of four wavelength information merges collection of illustrative plates and one group of collection of illustrative plates file simultaneously, and determine 38 total peaks.
Step 2, qizhi weitong granules anti-inflammatory and analgesic effect evaluating drug effect: mouse monokaryon macrophage RAW264.7 give plasma containing drug, with cell culture supernatant TNF-α, IL-6 and NO content for Testing index, calculate drug effect comprehensive grading.
The preparation method of described plasma containing drug: by cleaning grade SD rat 100 (200 ~ 220g), be randomly divided into 20 groups, often group 5.Compatibility group gavage gives the suspension solution (gavage amount=each group crude drug amount × paste-forming rate × 70kg × 0.018/ rat body weight) that Latin hypercube designs 20 groups of different ratio medical materials, every day twice, every minor tick 12h, continuous 3d.Before rat extracting blood, water is can't help in 12h fasting, and after last gavage 1h, aseptic rat eye of winning takes blood in right amount, puts in 2mL centrifuge tube, stands 30min, in high speed centrifuge, and 3000r min-1Centrifugal 15min, aseptic separation serum, merge with organizing serum, mixing.Through 56 DEG C, after 30min inactivation treatment, 0.22 μm of filtering with microporous membrane is degerming, puts-20 DEG C and saves backup.
Described MTT Composition analyzed method: experimental cell is divided into: negative control group (not modeling, be not administered), model group (modeling, be not administered), qizhi weitong granules group, 20 compatibility groups (modeling, to Contained Serum) and blank well (zeroing hole), often group sets 5 multiple holes.Processing procedure is as follows: 1 bottle of the cell that trophophase of taking the logarithm, growth conditions are good, and adjusting cell concentration is 1 × 105Individual mL-1, it is inoculated in 96 well culture plates, every hole 100 μ L, continues to cultivate about 24h, treat that cell attachment is complete.Matched group and model group add 10% blank serum 100 μ L and cultivate, and 20 compatibility groups are separately added into respective 10% Contained Serum 100 μ L.After administration 2h, model group and compatibility group add LPS (final concentration of 1mg L-1) build inflammatory model, in 37 DEG C, 5%CO after each group cell is processed separately2Continuing in incubator to cultivate 24h, collect supernatant ,-20 DEG C of Refrigerator stores are standby.Cell adopts mtt assay, and microplate reader 492nm place is scanned, and measures light absorption value (OD).
Step 3, qizhi weitong granules quality and drug effect relativity evaluation.
Utilize Grey Incidence Analysis, quantization peak area and anti-inflammatory and antalgic drug effect degree of being associated analysis to 38 total peaks of 20 medical material compatibility groups, obtain 17 chromatographic peaks, get rid of the chromatographic peak peak area chromatographic peak lower than 100, filter out 11 total peaks relevant to drug effect.
Step 4, qizhi weitong granules anti-inflammatory and analgesic effect " dose-effect colour atla " structure.
The relative retention time at 11 total peaks relevant to drug effect that calculating sifting goes out, relative peak area, determine that peak belongs to, calculate the relative peak area of each chromatographic peak relevant to drug effect and the product of the degree of association, and add and, being scaled in percentage with maximum for 10, setting up dose-effect relationship equation is: YDrug effect is marked=(0.9951X1+0.9872X2+0.9823X3+0.9756X4+0.9722X5+0.9612X6+0.9530X7+0.9432X8+0.9387X9+0.9375X10+0.9168X11)/10 × 100.Wherein YDrug effect is markedDrug effect for qizhi weitong granules anti-inflammatory and antalgic is marked, X1-11The respectively relative peak area of 11 chromatographic peaks.Application VisualBasic(VB) above procedure carries out programming by programming language, works out " dose-effect colour atla " software.Software colour atla can show 6 color regions divided equally (red, orange, yellow, green, blue, purple), totally 100 points, indicates the final drug effect scoring of qizhi weitong granules with pointer.
The high performance liquid chromatography testing result of 11 anti-inflammatory and antalgic effective ingredient of qizhi weitong granules is inputted " dose-effect colour atla " software, select through medicine to be checked, internal standard substance retention time selects with peak area setting, reading data, standard efficacy data, read efficacy data, anti-inflammatory and antalgic drug effect detection process, can intuitively be reflected the drug effect of qizhi weitong granules anti-inflammatory and antalgic by pointer.
Compared with prior art, the positive effect of the present invention is in that.
(1) the invention discloses a kind of method being associated by qizhi weitong granules active constituent content with its anti-inflammatory and analgesic effect.Although Chinese medicine fingerprint can indicate the multiple chemical composition in Chinese medicine, it is possible to control Chinese medicine quality on the whole, but its chemical composition embodied be whether active ingredient and with the degree of correlation of drug effect indefinite.Therefore, finger printing is utilized merely to there is also certain limitation to the quality evaluating Chinese medicine quality.The present invention is by quality and drug effect correlation research, illustrate the mutual relation of qizhi weitong granules Fingerprints and its anti-inflammatory and analgesic effect drug effect, so that the quality of the qizhi weitong granules more targeted control qizhi weitong granules of anti-inflammatory and antalgic drug effect finger printing built.
(2) present invention is first by the comprehensive grading of qizhi weitong granules anti-inflammatory and antalgic drug effect, shows with the form of " dose-effect colour atla ".Applicant is for the dependency of finger printing and drug action, set up corresponding dose-effect relationship equation, develop corresponding software on this basis, by the drug effect of qizhi weitong granules anti-inflammatory and antalgic by " dose-effect colour atla " this intuitively form show, it is achieved " visualization " of Curing Effect of Chinese medicine prediction.
(3) 10 batches of qizhi weitong granules anti-inflammatory and analgesic effects have been evaluated by first Application of the present invention " dose-effect colour atla " software, namely by inputting finger printing data, directly calculate the drug action of this medicine, the method is easy and simple to handle, the prediction to drug action and calculating can be realized, and then evaluate the quality of qizhi weitong granules, the method is applied to enterprise practical produce, its drug effect can be intuitively reflected by measuring active constituent content, drug quality is ensured for enterprise, stablize clinical efficacy, establish public's praise significant, there is good actual application value.
Accompanying drawing explanation
Fig. 1 is six single medicinal material compatibility group all the period of time multi-wavelength information fusion chromatograms in qizhi weitong granules, and wherein S is acetaminophen.
Fig. 2 is colour atla software operation interface.
Fig. 3 is that No. 1 qizhi weitong granules fingerprint merges collection of illustrative plates.
Fig. 4 is that No. 2 qizhi weitong granules fingerprints merge collection of illustrative plates.
Fig. 5 is that No. 3 qizhi weitong granules fingerprints merge collection of illustrative plates.
Fig. 6 is that No. 4 qizhi weitong granules fingerprints merge collection of illustrative plates.
Fig. 7 is that No. 5 qizhi weitong granules fingerprints merge collection of illustrative plates.
Fig. 8 is that No. 6 qizhi weitong granules fingerprints merge collection of illustrative plates.
Fig. 9 is that No. 7 qizhi weitong granules fingerprints merge collection of illustrative plates.
Figure 10 is that No. 8 qizhi weitong granules fingerprints merge collection of illustrative plates.
Figure 11 is that No. 9 qizhi weitong granules fingerprints merge collection of illustrative plates.
Figure 12 is that No. 10 qizhi weitong granules fingerprints merge collection of illustrative plates.
Detailed description of the invention:
1. experiment material.
1.1 medical materials and reagent: Radix Bupleuri, the Radix Paeoniae Alba, Radix Glycyrrhizae, Rhizoma Corydalis, Fructus Aurantii, Rhizoma Cyperi medical material (Liaoning Benxi Sanyao Co., Ltd.)；Acetaminophen reference substance (medicine bioassay institute of China, lot number: 018-8905)；Acetonitrile (TEDIA company of the chromatographically pure U.S.)；Formic acid (chromatographically pure Tianjin Kermel Chemical Reagent Co., Ltd.)；Qizhi weitong granules (Liaoning Huarun Benxi Third Pharmaceutical Co., Ltd., lot number: No. 1: 20110710；No. 2: 20110903；No. 3: 20110413；No. 4: No. 201106035: 20110525；No. 6: 20111224；No. 7: 20111218；No. 8: No. 201112239: 20110910；No. 10: 20110713)；Rat prostate element E2Test kit (Lang Dun bio tech ltd, Shanghai, lot number: 201501013)；Interleukin 8 (IL-8) test kit (Lang Dun bio tech ltd, Shanghai, lot number: 201501017GH)；Carrageenin (Yuan Ye bio tech ltd, Shanghai, lot number: J08J6R2)；Murine tumor necrosis factor (TNF-α), mouse nitrous oxide (NO), mouse interleukin-6 (IL-6) test kit (all purchased from Yuan Ye bio tech ltd, Shanghai)；Water is ultra-pure water.
1.2 instrument and equipments: Agilent-1290 high performance liquid chromatograph (Agilent Technologies of the U.S.)；ACCULABALC-11C.4 type electronic balance (Sai Duolisi group of Germany)；DZTW type regulating temp. electrothermal cover (Beijing is bright Medical Instruments factory forever)；SHZ-D III type circulating water type vacuum pump (Yuhua Instrument Co., Ltd., Gongyi City)；Milli-Q ultrapure water treating device (Millipore company of the U.S.).
1.3 cell strains: mouse monokaryon macrophage strain RAW264.7 is provided by Shanghai Inst. of Life Science, CAS cellular resources center.
1.4 laboratory animals: healthy cleaning grade SD rat, body weight (200 ~ 220g), female, Dalian Medical Univ's Experimental Animal Center provide, animal credit number SCXK(the Liao Dynasty) 2011-0002.Animal feeding is in air conditioning chamber, and room temperature 20 ± 2 DEG C, relative humidity 50%-60%, pellet is fed, and freely drinks water.
2. experimental technique and result
2.1 qizhi weitong granules finger printing researchs
2.1.1 the preparation of need testing solution: in application Isight software qizhi weitong granules, 6 taste medical material Radix Bupleuri, Rhizoma Corydalis, Fructus Aurantii, Rhizoma Cyperi, Radix Glycyrrhizae are by 0～10g, consumption is carried out 20 Latin hypercubes by 0～15g and randomly draws by the Radix Paeoniae Alba, obtaining the medical material compatibility group of 20 different proportions, result is in Table 1.Each compatibility group medical material is mixed, puts in round-bottomed flask, 20 times of weight water reflux, extract, 2h, filter, volatilize, be settled to 250mL, take 2mL need testing solution, adding acetaminophen reference substance 200 μ L(concentration is 0.1240mg/mL), as interior mark liquid.All the other need testing solutions are put in water-bath and are volatilized, and after calculating paste-forming rate, give over to the experiment of anti-inflammatory and antalgic drug effect and use.
Table 1 Latin hypercube stochastic sampling result (g).
| Factor | Radix Bupleuri | Rhizoma Cyperi | The Radix Paeoniae Alba | Fructus Aurantii | Rhizoma Corydalis | Radix Glycyrrhizae |
| 1 | 3.7 | 8.5 | 11.0 | 5.8 | 2.9 | 7.2 |
| 2 | 6.7 | 9.2 | 13.0 | 1.5 | 3.6 | 5.2 |
| 3 | 5.2 | 2.5 | 11.3 | 2.7 | 1.1 | 4.8 |
| 4 | 1.5 | 4.0 | 8.5 | 6.7 | 7.2 | 2.4 |
| 5 | 5.7 | 5.7 | 4.0 | 4.2 | 4.8 | 7.6 |
| 6 | 6.0 | 6.3 | 4.8 | 7.2 | 1.7 | 6.2 |
| 7 | 7.5 | 8.2 | 7.3 | 0.5 | 0.5 | 9.7 |
| 8 | 3.4 | 1.4 | 1.6 | 9.7 | 6.6 | 2.6 4 --> |
| 9 | 4.7 | 7.7 | 13.6 | 1.6 | 9.2 | 3.7 |
| 10 | 1.3 | 7.1 | 10.0 | 3.7 | 6.2 | 5.5 |
| 11 | 8.5 | 0.8 | 9.2 | 9.4 | 0.5 | 1.0 |
| 12 | 8.5 | 0.5 | 12.3 | 6.0 | 2.3 | 3.0 |
| 13 | 2.2 | 9.5 | 14.4 | 2.3 | 7.7 | 9.2 |
| 14 | 0.1 | 3.2 | 3.0 | 0.1 | 4.1 | 6.8 |
| 15 | 4.1 | 3.7 | 7.6 | 3.1 | 8.7 | 0.2 |
| 16 | 9.2 | 2.2 | 5.5 | 8.7 | 8.2 | 8.5 |
| 17 | 0.8 | 6.7 | 6.3 | 5.0 | 5.5 | 1.5 |
| 18 | 2.7 | 1.6 | 0.2 | 8.2 | 9.5 | 8.1 |
| 19 | 9.6 | 5.1 | 0.8 | 7.6 | 5.2 | 4.1 |
| 20 | 7.1 | 4.6 | 3.4 | 4.5 | 3.0 | 1.2 |
2.1.2 test sample all the period of time four wavelength merges collection of illustrative plates foundation:
(1) chromatographic condition: chromatographic column: AgilentTC-C18(4.6mm × 250mm, 5 μm) chromatographic column；Mobile phase: 0.2 ‰ formic acid water (A)-acetonitrile (B)；Flow velocity: 1.0mL min-1；Column temperature: 25 DEG C；Detector wavelength: 230nm, 254nm, 283nm, 365nm；Sample size: 15 μ L.Eluent gradient eluting table is in Table 2.
Table 2 eluent gradient eluting table.
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 95 | 5 |
| 10 | 90 | 10 |
| 61 | 63 | 27 |
| 70 | 60 | 40 |
| 90 | 50 | 50 |
(2) test sample all the period of time four wavelength merges collection of illustrative plates foundation:
With acetaminophen for interior mark, by DAD detector, 20 compatibility group chromatograms are carried out ultraviolet all-wave length (200-400nm) and scan.Determine that fusion wavelength is 230nm, 254nm, 283nm, 365nm according to scanning result.230nm is derived respectively from Agilent1100 chromatographic work station, 254nm, 283nm, the dif formatted data file of 365nm, use Matlab software programming, dif formatted data is carried out all the period of time multi-wavelength information fusion, obtains a chromatogram simultaneously reflecting four wavelength information and one group of collection of illustrative plates file, and determine 38 total peaks.For compatibility 7, all the period of time multi-wavelength information fusion chromatogram is shown in Fig. 1.
(3) synergy analysis: in qizhi weitong granules based on chemical composition contained by six single medical materials, by its crude drug source of each synergy in compatibility group, and belongs to its chemical composition further, makes effective ingredient definitely, in Table 3.
Table 3 compatibility group synergy table.
| Peak number | Retention time (min) | Medical material belongs to | Perk purity |
| 1 | 7.60 | Rhizoma Corydalis | |
| 2 | 8.13 | The Radix Paeoniae Alba | Gallic acid |
| 3 | 10.03 | Rhizoma Corydalis | |
| 4 | 13.72 | 5 --> | |
| 5 | 14.95 | Rhizoma Cyperi | |
| 6 | 18.21 | ||
| 7 | 20.40 | The Radix Paeoniae Alba | Oxypaeoniflorin |
| 8 | 21.05 | Rhizoma Cyperi | |
| 9 | 22.52 | The Radix Paeoniae Alba | Paeoniflorin sulfonate |
| 10 | 28.13 | The Radix Paeoniae Alba | Lactone glucoside of Radix Paeoniae |
| 11 | 29.28 | Fructus Aurantii | |
| 12 | 31.23 | The Radix Paeoniae Alba | Peoniflorin |
| 13 | 36.63 | Fructus Aurantii | Eriocitrin |
| 14 | 38.19 | The Radix Paeoniae Alba | Lactone glucoside of Radix Paeoniae analog |
| 15 | 39.07 | Radix Glycyrrhizae | Celery glycosyl liquirtin |
| 16 | 40.39 | Radix Glycyrrhizae | Liquirtin |
| 17 | 41.52 | The Radix Paeoniae Alba | |
| 18 | 42.42 | Fructus Aurantii | New eriodictin |
| 19 | 46.59 | Fructus Aurantii | Folium Symplocoris Caudatae naringin |
| 20 | 47.20 | Fructus Aurantii | |
| 21 | 49.23 | Fructus Aurantii | Naringin |
| 22 | 50.79 | Fructus Aurantii | Hesperidin |
| 23 | 53.37 | Fructus Aurantii | Neohesperidin |
| 24 | 56.10 | Fructus Aurantii | |
| 25 | 57.03 | Radix Glycyrrhizae | |
| 26 | 58.46 | Fructus Aurantii | |
| 27 | 59.21 | Radix Glycyrrhizae | Isoliquiritin |
| 28 | 62.19 | Radix Glycyrrhizae | Glycyrrhizin |
| 29 | 67.14 | ||
| 30 | 68.24 | Fructus Aurantii | Poncirin |
| 31 | 71.59 | The Radix Paeoniae Alba | Benzoylpaeoniflorin |
| 32 | 72.74 | The Radix Paeoniae Alba | |
| 33 | 73.74 | Radix Bupleuri | |
| 34 | 74.63 | Radix Glycyrrhizae | |
| 35 | 76.85 | Radix Glycyrrhizae | Isoliquiritigenin |
| 36 | 77.49 | Rhizoma Cyperi | |
| 37 | 79.32 | Radix Glycyrrhizae | Monoammonium glycyrrhizinate |
| 38 | 81.18 | Radix Bupleuri | Saikoside A |
Application Matlab software programming, 20 compatibility group chromatograms are carried out all the period of time multi-wavelength information fusion, obtain the data of each chromatographic peak peak area in 20 compatibility groups, the peak area of chromatographic peak is changed into chemical composition content (every day is to the content of certain chemical composition in the medicine of rat oral gavage), carries out grey correlation analysis with pharmacy in vitro data.
2.2 qizhi weitong granules anti-inflammatory and analgesic effect evaluating drug effects.
2.2.1 the preparation of plasma containing drug.
By cleaning grade rat 100 (200 ~ 220g), it is randomly divided into 20 groups, often group 5.Compatibility group gavage gives to design the suspension solution (gavage amount=each group crude drug amount × paste-forming rate × 70kg × 0.018/ rat body weight) of 20 groups of different ratio medical materials, every day twice, every minor tick 12h, continuous 3d by table 1 Latin hypercube.Before rat extracting blood, water is can't help in 12h fasting, and after last gavage 1h, aseptic rat eye of winning takes blood in right amount, puts in 2mL centrifuge tube, stands 30min, in high speed centrifuge, and 3000r min-1Centrifugal 15min, aseptic separation serum, merge with organizing serum.Through 56 DEG C, after 30min inactivation treatment, 0.22 μm of filtering with microporous membrane is degerming, puts-20 DEG C and saves backup.
2.2.2MTT method detection drug effect.
Being divided into by experimental cell: negative control group (not modeling is not administered), model group (modeling is not administered), 20 compatibility groups (modeling, to Contained Serum) and blank group (zeroing hole), often group sets 5 multiple holes.Processing procedure is as follows: 1 bottle of the cell that trophophase of taking the logarithm, growth conditions are good, and adjusting cell concentration is 1 × 105Individual mL-1, it is inoculated in 96 well culture plates, every hole 100 μ L, continues to cultivate about 24h, treat that cell attachment is complete.Matched group and model group add 10% blank serum 100 μ L and cultivate, and 20 compatibility groups are separately added into respective 10% Contained Serum 100 μ L.After administration 2h, model group and compatibility group add LPS (final concentration of 1mg L-1) build inflammatory model, in 37 DEG C, 5%CO after each group cell is processed separately2Continuing in incubator to cultivate 24h, collect supernatant ,-20 DEG C of Refrigerator stores are standby.Cell adopts mtt assay, and microplate reader 492nm place is scanned, and measures light absorption value (OD).
2.2.3 TNF-α, IL-6 and NO content detection in cell conditioned medium liquid.
By cell conditioned medium liquid, by TNF-α, the operation of IL-6 and NO test kit description, measure OD value by microplate reader, calculate the content of each group of TNF-α, IL-6 and NO.
TNF-α suppression ratio=(1-TNF-alpha contentTest group/ TNF-α contentMatched group) × 100%.
IL-6 suppression ratio=(1-IL-6 contentTest group/ IL-6 contentMatched group) × 100%.
NO suppression ratio=(1-NO contentTest group/ NO contentMatched group) × 100%.
Making overall merit with TNF-α suppression ratio, IL-6 suppression ratio, NO suppression ratio for index, total score is 100 points, and TNF-α suppression ratio, IL-6 suppression ratio respectively account for 35 points, and NO suppression ratio accounts for 30 points.With the maximum of each index for best result, (if TNF-α suppression ratio maximum is 72.58, it is chosen as 35 points, all the other scoring Y respectively organized by that analogyi,TNF-α=35×Xi,TNF-α/ 72.58), result is in Table 4.
420 compatibility group Pharmacodynamics in vitro measurement results of table.
| Group | TNF-α suppression ratio/% | Score value/point | IL-6 suppression ratio/% | Score value/point | NO suppression ratio/% | Score value/point | Total score |
| 1 | 61.75 | 29.77 | 42.71 | 16.22 | 25.67 | 30.00 | 75.99 |
| 2 | 72.58 | 35.00 | 76.87 | 29.19 | 10.02 | 11.71 | 75.90 |
| 3 | 62.61 | 30.19 | 91.21 | 34.64 | 16.73 | 19.55 | 84.38 |
| 4 | 61.58 | 29.70 | 62.64 | 23.79 | 10.20 | 11.92 | 65.41 |
| 5 | 56.45 | 27.22 | 32.90 | 12.49 | -2.07 | -2.41 | 37.30 |
| 6 | 61.41 | 29.62 | 47.56 | 18.06 | 0.81 | 0.94 | 48.62 |
| 7 | 59.84 | 28.86 | 64.09 | 24.34 | 14.49 | 16.93 | 70.13 |
| 8 | 64.23 | 30.97 | 74.88 | 28.43 | 1.70 | 1.99 | 61.39 |
| 9 | -5.25 | -2.53 | 60.04 | 22.80 | 0.09 | 0.10 | 20.37 |
| 10 | 48.10 | 23.19 | 73.26 | 27.82 | 0.27 | 0.31 | 51.32 |
| 11 | 51.78 | 24.97 | 57.61 | 21.88 | 2.50 | 2.93 | 49.78 |
| 12 | 8.27 | 3.99 | 62.00 | 23.55 | 0.81 | 0.94 | 28.48 |
| 13 | 33.13 | 15.97 | 58.36 | 22.16 | 7.25 | 8.47 | 46.60 |
| 14 | 13.44 | 6.48 | 92.16 | 35.00 | 12.16 | 14.22 | 55.70 |
| 15 | -13.48 | -6.50 | 66.48 | 25.25 | 2.06 | 2.40 | 21.15 7 --> |
| 16 | 33.79 | 16.29 | 64.08 | 24.33 | 8.23 | 9.62 | 50.24 |
| 17 | 16.58 | 8.00 | 83.42 | 31.68 | 4.11 | 4.81 | 44.49 |
| 18 | 1.57 | 0.76 | 71.09 | 27.00 | 5.46 | 6.38 | 34.14 |
| 19 | 21.40 | 10.32 | 63.50 | 24.11 | 14.79 | 17.28 | 51.71 |
| 20 | 10.42 | 5.03 | 37.50 | 14.24 | 12.40 | 14.49 | 33.76 |
2.3 qizhi weitong granules quality and drug effect relativity evaluation.
Application gray system theory modeling software (GTMS3.0), to the quantization peak area at 38 total peaks of 20 different proportion medical material groups and qizhi weitong granules anti-inflammatory and antalgic drug effect degree of being associated analysis, chooses the degree of association total peak more than 0.9, and result is in Table 5.
The each total peak of table 5 and anti-inflammatory and antalgic drug effect dependency.
| Ranking | Peak number | Ownership medical material | Coefficient of association | Perk purity |
| 1 | 11 | Fructus Aurantii | 0.9969 | |
| 2 | 16 | Radix Glycyrrhizae | 0.9951 | Liquirtin |
| 3 | 19 | Fructus Aurantii | 0.9872 | Folium Symplocoris Caudatae naringin |
| 4 | 38 | Radix Bupleuri | 0.9823 | Saikoside A |
| 5 | 35 | Radix Glycyrrhizae | 0.9756 | Isoliquiritigenin |
| 6 | 25 | Radix Glycyrrhizae | 0.9722 | |
| 7 | 2 | The Radix Paeoniae Alba | 0.9612 | Gallic acid |
| 8 | 34 | Radix Glycyrrhizae | 0.9612 | |
| 9 | 15 | Radix Glycyrrhizae | 0.955 | Celery glycosyl liquirtin |
| 10 | 4 | 0.9546 | ||
| 11 | 6 | 0.953 | ||
| 12 | 26 | Fructus Aurantii | 0.9432 | |
| 13 | 37 | Radix Glycyrrhizae | 0.9387 | Monoammonium glycyrrhizinate |
| 14 | 12 | The Radix Paeoniae Alba | 0.9375 | Peoniflorin |
| 15 | 14 | The Radix Paeoniae Alba | 0.9292 | Lactone glucoside of Radix Paeoniae analog |
| 16 | 13 | Fructus Aurantii | 0.9212 | Eriocitrin |
| 17 | 10 | The Radix Paeoniae Alba | 0.9168 | Lactone glucoside of Radix Paeoniae |
Obtain in qizhi weitong granules through grey correlation analysis and associate comparatively close chromatographic peak with anti-inflammatory and antalgic drug effect, consider that indivedual chromatographic peak peak area is relatively low, the contribution of relevant drug effect is less, screen for result above, get rid of the chromatographic peak peak area chromatographic peak lower than 100, finally give the total peak relevant to anti-inflammatory and antalgic drug effect as follows.
The each total peak of table 6 and anti-inflammatory and antalgic drug effect dependency.
| Peak number | Ownership medical material | Coefficient of association | Perk purity |
| 16 | Radix Glycyrrhizae | 0.9951 | Liquirtin |
| 19 | Fructus Aurantii | 0.9872 | Folium Symplocoris Caudatae naringin |
| 38 | Radix Bupleuri | 0.9823 | Saikoside A |
| 35 | Radix Glycyrrhizae | 0.9756 | Isoliquiritigenin |
| 25 | Radix Glycyrrhizae | 0.9722 | |
| 2 | The Radix Paeoniae Alba | 0.9612 | Gallic acid |
| 6 | 0.953 | 8 --> | |
| 26 | Fructus Aurantii | 0.9432 | |
| 37 | Radix Glycyrrhizae | 0.9387 | Monoammonium glycyrrhizinate |
| 12 | The Radix Paeoniae Alba | 0.9375 | Peoniflorin |
| 10 | The Radix Paeoniae Alba | 0.9168 | Lactone glucoside of Radix Paeoniae |
The programming of 2.4 dose-effect colour atla softwares
Application VisualBasic(VB) programming language card software of checking colors carries out programming, with acetaminophen for interior mark, calculate each chromatographic peak relative retention time and relative peak area, determine that peak belongs to relative retention time, calculate the relative peak area of each chromatographic peak relevant to drug effect and the product of the degree of association, and add and, be scaled in percentage with maximum for 10, setting up dose-effect relationship equation is: YDrug effect is marked=(0.9951X1+0.9872X2+0.9823X3+0.9756X4+0.9722X5+0.9612X6+0.9530X7+0.9432X8+0.9387X9+0.9375X10+0.9168X11)/10 × 100.Wherein YDrug effect is markedDrug effect for qizhi weitong granules anti-inflammatory and antalgic is marked, X1-11The respectively relative peak area of 11 chromatographic peaks.Finally give the drug effect scoring that anti-inflammation analgesia medicine is imitated by qizhi weitong granules.
Application VB program development tools design software operation interface, and the scoring way of output.This project VB engineering comprises 2 forms altogether, 2 modules and 1 user control, wherein forms comprise welcome interface and primary operational forms, shown in Fig. 2, Main form has 5 click events and 7 change events, they are controlled by corresponding programming language, owing to instrument and equipment is different, under same chromatographic condition, the retention time of spectral peak is likely to difference, therefore the retention time and the peak area that are manually entered interior mark peak are adopted during this programming, to avoid the deviation owing to instrument difference causes.2 modules have input the public function used in program, process, constant, self-defined structure, global variable etc..As seen from Figure 2, colour atla is equally divided into six color regions (red, orange, yellow, green, blue, purple), totally 100 points, indicates the final drug effect scoring of qizhi weitong granules with pointer.Selected by detected sample, import detected sample and merge spectrum data, input internal standard substance retention time and peak area, read detected sample data, data to be tested capacity showing, detected sample has peak number amount, selection standard efficacy data, the spectrum data at the total peak, 11 relevant to anti-inflammatory and antalgic namely filtered out, standard drug effect capacity is shown as 11, in standard efficacy data, relative retention time is as index, detected sample efficacy data is read by dose-effect relationship equation, finally detection anti-inflammatory and antalgic drug effect, the drug effect of qizhi weitong granules anti-inflammatory and antalgic can be intuitively reflected by pointer directional point.
2.510 batches of qizhi weitong granules finger printing.
2.5.1 the preparation of need testing solution.
nullTake the qizhi weitong granules of 10 batches，Each 2.5g，Mixing，Finely ground，Precision weighs 2.5g，Put in tool plug conical flask，Accurate addition distilled water 100mL，Close plug，Weighed weight，Supersound process (power 250W，Frequency 33kHz) 60 minutes，Take out，Let cool，Weighed weight again，The weight of less loss is supplied with distilled water，Shake up，Filter，Accurate absorption subsequent filtrate 25mL，It is added in AB-8 resin column (0.3-1.25mm，20g，Internal diameter 1.5cm) on，Wash with distilled water 150mL，Use 200mL95% ethanol elution again，Collect ethanol elution，It is evaporated，Residue adds 20% acetonitrile to be made dissolving and is transferred in 10mL volumetric flask，Add 20% acetonitrile and be settled to scale，Shake up，Filter，Take subsequent filtrate and get final product，Add acetaminophen as interior mark，Final concentration 0.02067mg mL-1。
2.5.2 chromatographic condition.
Chromatographic condition under same Part I " 2.1.2 " item.Using Matlab software programming, the finger printing under 4 wavelength is carried out multi-wavelength fusion, fusion results is shown in Fig. 3-12.
2.5.310 the scoring of qizhi weitong granules drug effect is criticized.
Result after merging is derived, operates by colour atla software operating process, obtain each group of anti-inflammatory and antalgic drug effect scoring, as shown in table 7.
Table 7 respectively organizes drug effect scoring and comprehensive grading.
| Numbering | No. 1 | No. 2 | No. 3 | No. 4 | No. 5 | No. 6 | No. 7 | No. 8 | No. 9 | No. 10 |
| Scoring | 70.01 | 65.69 | 108.74 | 67.55 | 51.97 | 61.53 | 56.84 | 61.11 | 53.53 | 67.24 |
Drug effect checking in 2.610 batches of qizhi weitong granules anti-inflammatory and antalgic bodies.
2.6.1 animal packet and administration.
Healthy SD rat is randomly divided into 12 groups by body weight, often group 10, respectively blank group, model group, 10 batches of qizhi weitong granules groups.Except blank group, model group, all the other rat gastric infusions every day 1 time, 0.15g d-1.Blank group, model group gavage equal-volume distilled water.Respectively organize after rat was administered in the 3rd day after 40min, model group injecting normal saline 15mL kg-1, all the other respectively organize lumbar injection 0.7% glacial acetic acid 15mL kg-1, there is writhing sum after record injection in 20min, result is in Table 8.
Table 8 glacial acetic acid causes rat writhing response.
| Group/index | Writhing number of times (± S) | Suppression ratio (%) |
| Blank | — | — |
| Model | 16.10±2.19 | — |
| No. 1 | 4.50±2.38** | 75 |
| No. 2 | 12.30±3.33 | 25 |
| No. 3 | 4.60±2.04** | 75 |
| No. 4 | 9.50±2.66* | 43.75 |
| No. 5 | 11.40±2.56 | 31.25 |
| No. 6 | 5.70±3.52** | 68.75 |
| No. 7 | 13.60±4.23 | 18.75 |
| No. 8 | 6.10±1.11** | 62.5 |
| No. 9 | 7.90±3.89** | 56.25 |
| No. 10 | 8.20±3.78* | 50 |
Note: compare with model group, * P < 0.05, * * P < 0.01
Rat successive administration 7 days, before last is administered, adopts self-control foot volume measurement device to measure the left back sufficient volume of rat record, 30min after administration, except blank group, cause inflammation, blank group injection equal-volume normal saline in rat left hind foot plantar subcutaneous injection carrageenin, after 2h, rat is plucked eyeball and takes blood, separates serum standby, and de-cervical vertebra puts to death rat, with same method, measure the left back sufficient volume of rat record.Left sufficient sample and rat stomach is stayed to be organized in-80 DEG C of preservations.Result is in Table 9.
Table 9 respectively organize inhibitory rate of intumesce (± S).
| Group | Dosage (g d-1) | Paw swelling | Inhibitory rate of intumesce (%) |
| Blank | — | 0.250±0.014** | — |
| Model | — | 0.848±0.076 | — |
| No. 1 | 0.15 | 0.650±0.082 | 23.22 |
| No. 2 | 0.15 | 0.576±0.019* | 32.05 |
| No. 3 | 0.15 | 0.541±0.011* | 36.18 |
| No. 4 | 0.15 | 0.536±0.010** | 36.77 |
| No. 5 | 0.15 | 0.725±0.008 | 14.38 |
| No. 6 | 0.15 | 0.727±0.021 | 14.15 |
| No. 7 | 0.15 | 0.573±0.095* | 32.39 |
| No. 8 | 0.15 | 0.820±0.036 | 3.21 |
| No. 9 | 0.15 | 0.608±0.007 | 28.26 10 --> |
| No. 10 | 0.15 | 0.515±0.078** | 39.19 |
Note: compare with model group, * P < 0.05, * * P < 0.01
2.6.2 rat blood serum PGE2, IL-8 content detection.
After each group rat extracting blood, standing the centrifugal 15min of 30min, 3000r, separate serum, detect by test kit operating instruction, result is in Table 10.
Table 10 is to PGE2And the impact of IL-8 element (± S).
| Group | Sample size | PGE2(ng L-1) | IL-8(ng L-1) |
| Blank group | 10 | 337.38±4.70** | 236.83±2.12** |
| Model group | 10 | 403.86±2.11 | 305.08±6.26 |
| No. 1 | 10 | 359.55±2.43* | 280.73±4.50 |
| No. 2 | 10 | 351.93±3.11** | 263.88±3.48* |
| No. 3 | 10 | 352.77±3.16** | 258.59±2.85** |
| No. 4 | 10 | 373.93±6.09 | 277.06±6.67 |
| No. 5 | 10 | 347.22±3.96** | 271.19±7.79* |
| No. 6 | 10 | 359.08±2.34* | 298.91±8.73 |
| No. 7 | 10 | 376.70±7.65 | 267.76±3.62* |
| No. 8 | 10 | 384.35±3.12 | 259.92±3.90** |
| No. 9 | 10 | 379.56±3.47 | 268.97±2.14* |
| No. 10 | 10 | 381.79±4.28 | 282.68±6.95 |
Note: compare with model group, * P < 0.05, * * P < 0.01
2.6.310 drug effect overall merit in qizhi weitong granules anti-inflammatory and antalgic body is criticized.
With internal inhibitory rate, foot swelling suppression ratio, PGE2Relative amount, IL-8 relative amount are index, and drug effect in 10 batches of qizhi weitong granules anti-inflammatory and antalgic bodies is carried out overall merit, according to scoring formula: comprehensive grading=[administration group inhibitory rate/inhibitory ratemas+ administration group foot swelling suppression ratio/foot swelling suppression ratiomas+ (1-administration group PGE2Content/PGE2Contentmas)+(1-administration group IL-8 content/IL-8 contentmas)], during each indicator-specific statistics, person's relative inhibition (or content) gives actual ratio to have significant difference, and person's score is 0 not to have significant difference, and result is in Table 11.
Drug effect comprehensive grading result in table 11 qizhi weitong granules anti-inflammatory and antalgic body.
| Group/index | Inhibitory rate | Foot swelling suppression ratio | PGE2Relative amount | IL-8 relative amount | Comprehensive grading |
| No. 1 | 1 | 0 | 0.06 | 0 | 1.06 |
| No. 2 | 0 | 0.82 | 0.08 | 0.12 | 1.02 |
| No. 3 | 1 | 0.92 | 0.08 | 0.13 | 2.14 |
| No. 4 | 0.58 | 0.94 | 0 | 0 | 1.52 |
| No. 5 | 0 | 0 | 0.1 | 0.09 | 0.19 |
| No. 6 | 0.92 | 0 | 0.07 | 0 | 0.98 |
| No. 7 | 0 | 0.83 | 0 | 0.1 | 0.93 |
| No. 8 | 0.83 | 0 | 0 | 0.13 | 0.96 |
| No. 9 | 0.75 | 0 | 0 | 0.1 | 0.85 |
| No. 10 | 0.67 | 1 | 0 | 0 | 1.67 |
Application " dose-effect colour atla " software can obtain the scoring of 10 batches of qizhi weitong granules anti-inflammatory and antalgic drug effects, compare with effect experiment comprehensive grading result, basically identical through software gained appraisal result and drug effect appraisal result, illustrate that this software can react the drug effect of qizhi weitong granules to a certain extent, it is possible in the middle of qizhi weitong granules quality control.
Claims (8)
1. based on the qizhi weitong granules anti-inflammatory and antalgic method of quality control of dose-effect colour atla, it is characterised in that the method carries out as follows:
(1) foundation of qizhi weitong granules finger printing: the consumption of Six-element Chinese medicine in qizhi weitong granules is carried out stochastic sampling, there are 20 groups of mixing medical materials, it is extracted, adopt high performance liquid chromatography detection；
(2) qizhi weitong granules anti-inflammatory and analgesic effect evaluating drug effect: mouse monokaryon macrophage RAW264.7 give plasma containing drug, with cell culture supernatant TNF-α, IL-6 and NO content for Testing index, calculates drug effect comprehensive grading；
(3) qizhi weitong granules quality and drug effect relativity evaluation: quantization peak area and qizhi weitong granules anti-inflammatory and antalgic drug effect degree of the being associated analysis to the total peak of 20 different proportion medical material groups, obtains the correlation coefficient of different compounds and its anti-inflammatory and analgesic effect in qizhi weitong granules；
(4) structure of qizhi weitong granules anti-inflammatory and analgesic effect " dose-effect colour atla ": screen and play, to qizhi weitong granules, the compound that anti-inflammatory and analgesic effect is relevant, application programming language card software of checking colors carries out programming, builds qizhi weitong granules anti-inflammatory and analgesic effect dose-effect colour atla.
2. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterized in that, the preparation method of need testing solution is as follows: in qizhi weitong granules, 6 taste medical material Radix Bupleuri, Rhizoma Corydalis, Fructus Aurantii, Rhizoma Cyperi, Radix Glycyrrhizae are by 0～10g, consumption is carried out 20 Latin hypercubes by 0～15g and randomly draws by the Radix Paeoniae Alba, obtain the medical material compatibility group of 20 different proportions, each compatibility group medical material is mixed, put in round-bottomed flask, 20 times amount water reflux, extract, 2h, filter, volatilize, be settled to 250mL, to obtain final product.
3. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterised in that chromatographic column is AgilentTC-C18(4.6mm × 250mm, 5 μm) chromatographic column；Mobile phase: 0.2 ‰ formic acid water (A)-acetonitrile (B)；Flow velocity: 1.0mL min-1；Column temperature: 25 DEG C；Detector wavelength: 230nm, 254nm, 283nm, 365nm；Sample size: 15 μ L；
Eluent gradient elution parameters is:
Time 0min, mobile phase A 95%, Mobile phase B 5%；
Time 10min, mobile phase A 90%, Mobile phase B 10%；
Time 61min, mobile phase A 63%, Mobile phase B 27%；
Time 70min, mobile phase A 60%, Mobile phase B 40%；
Time 90min, mobile phase A 50%, Mobile phase B 50%.
4. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterized in that, to 230nm, 254nm, 283nm, 365nm detects the collection of illustrative plates of wavelength and carries out multi-wavelength fusion, obtains a chromatogram simultaneously reflecting four wavelength information and one group of collection of illustrative plates file.
5. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterised in that the preparation method of its plasma containing drug is, different rat gavages respectively give suspension solution every day twice of 20 groups of different ratio medical materials, continuous 3d, takes blood, separated plasma.
6. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterized in that, application gray system theory modeling software (GTMS3.0) is to the quantization peak area at 38 total peaks of 20 different proportion medical material groups and qizhi weitong granules anti-inflammatory and antalgic drug effect degree of being associated analysis, and through screening, finally give 11 effective ingredient relevant with anti-inflammatory and antalgic.
7. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterized in that, with acetaminophen for interior mark, calculate each chromatographic peak relative retention time and relative peak area, determine that peak belongs to relative retention time, the relative peak area of 11 each chromatographic peaks relevant to drug effect that calculating sifting goes out and the product of the degree of association, and add and, being scaled in percentage with maximum for 10, setting up dose-effect relationship equation is YDrug effect is marked=(0.9951X1+0.9872X2+0.9823X3+0.9756X4+0.9722X5+0.9612X6+0.9530X7+0.9432X8+0.9387X9+0.9375X10+0.9168X11)/10 × 100, wherein YDrug effect is markedDrug effect for qizhi weitong granules anti-inflammatory and antalgic is marked, X1-11The respectively relative peak area of 11 chromatographic peaks, above procedure is carried out programming by application programming language, establishment " dose-effect colour atla " software, software colour atla can show 6 color regions divided equally (red, orange, yellow, green, blue, purple), totally 100 points, indicate the final drug effect scoring of qizhi weitong granules with pointer.
8. the qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla as claimed in claim 1, it is characterized in that, the high performance liquid chromatography testing result of 11 anti-inflammatory and antalgic effective ingredient of qizhi weitong granules is inputted " dose-effect colour atla " software, select through medicine to be checked, internal standard substance retention time selects with peak area setting, reading data, standard efficacy data, read efficacy data, anti-inflammatory and antalgic drug effect detection process, can intuitively be reflected the drug effect of qizhi weitong granules anti-inflammatory and antalgic by pointer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610106193.8A CN105758950B (en) | 2016-02-26 | 2016-02-26 | Qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610106193.8A CN105758950B (en) | 2016-02-26 | 2016-02-26 | Qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105758950A true CN105758950A (en) | 2016-07-13 |
| CN105758950B CN105758950B (en) | 2018-02-06 |
Family
ID=56331327
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610106193.8A Active CN105758950B (en) | 2016-02-26 | 2016-02-26 | Qizhi weitong granules anti-inflammatory and antalgic method of quality control based on dose-effect colour atla |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105758950B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108195989A (en) * | 2018-02-01 | 2018-06-22 | 山西省医药与生命科学研究院 | A kind of yellow rose chemical composition evaluation method based on antithrombotic spectrum effect relationship |
| CN110806457A (en) * | 2019-11-29 | 2020-02-18 | 南京南医堂健康产业发展有限公司 | Detection method of fingerprint of Sijun manna drink |
| CN111801633A (en) * | 2018-06-14 | 2020-10-20 | 舍弗勒技术股份两合公司 | Production control method based on failure risk |
| CN112763597A (en) * | 2020-12-22 | 2021-05-07 | 葵花药业集团（贵州）宏奇有限公司 | Multi-index content determination method for astragalus root, dendrobium stem and hawthorn granules |
| CN113049711A (en) * | 2021-03-23 | 2021-06-29 | 宁夏医科大学 | Method for simultaneously determining multiple effective components in eight-treasure intelligence-benefiting mixture |
| CN113325115A (en) * | 2021-07-08 | 2021-08-31 | 辽宁华润本溪三药有限公司 | Method for establishing characteristic spectrum of qi stagnation stomachache granules and application thereof |
| CN113433244A (en) * | 2021-07-08 | 2021-09-24 | 辽宁华润本溪三药有限公司 | Method for determining tetrahydropalmatine content in qi stagnation stomachache granules and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101288699A (en) * | 2008-06-24 | 2008-10-22 | 中国医学科学院药用植物研究所 | Method for controlling quality of corydalis tuber and preparation thereof and drug effect thereof by using finger print |
| CN102759585A (en) * | 2012-07-02 | 2012-10-31 | 辽宁华润本溪三药有限公司 | Method for controlling full-time interval multi-wavelength fused fingerprint quality of qi stagnation and stomachache granules |
| CN102879486A (en) * | 2012-09-05 | 2013-01-16 | 山东大学 | Method for screening traditional Chinese medicine effect related ingredients and model building method |
| CN102980952A (en) * | 2012-11-19 | 2013-03-20 | 广州市谭山蜂业有限公司 | Spectrum-effect relationship-based propolis quality control method |
| CN104881579A (en) * | 2015-05-25 | 2015-09-02 | 长春康彼达科技有限公司 | Method for screening traditional Chinese medicine composition for treating qi stagnation type epigastric pain |
| CN104897837A (en) * | 2015-06-08 | 2015-09-09 | 辽宁华润本溪三药有限公司 | Detection method of pharmaceutical preparation for curing stomach pain caused by qi stagnation |
-
2016
- 2016-02-26 CN CN201610106193.8A patent/CN105758950B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101288699A (en) * | 2008-06-24 | 2008-10-22 | 中国医学科学院药用植物研究所 | Method for controlling quality of corydalis tuber and preparation thereof and drug effect thereof by using finger print |
| CN102759585A (en) * | 2012-07-02 | 2012-10-31 | 辽宁华润本溪三药有限公司 | Method for controlling full-time interval multi-wavelength fused fingerprint quality of qi stagnation and stomachache granules |
| CN102879486A (en) * | 2012-09-05 | 2013-01-16 | 山东大学 | Method for screening traditional Chinese medicine effect related ingredients and model building method |
| CN102980952A (en) * | 2012-11-19 | 2013-03-20 | 广州市谭山蜂业有限公司 | Spectrum-effect relationship-based propolis quality control method |
| CN104881579A (en) * | 2015-05-25 | 2015-09-02 | 长春康彼达科技有限公司 | Method for screening traditional Chinese medicine composition for treating qi stagnation type epigastric pain |
| CN104897837A (en) * | 2015-06-08 | 2015-09-09 | 辽宁华润本溪三药有限公司 | Detection method of pharmaceutical preparation for curing stomach pain caused by qi stagnation |
Non-Patent Citations (2)
| Title |
|---|
| 许雯雯 等: "神经网络结合灰色关联度法对气滞胃痛颗粒复方药材抗炎活性谱效关系研究", 《中国中药杂志》 * |
| 赵越 等: "基于谱效关系的灰色关联度分辨系数确定", 《太原师范学院学报（自然科学版）》 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108195989A (en) * | 2018-02-01 | 2018-06-22 | 山西省医药与生命科学研究院 | A kind of yellow rose chemical composition evaluation method based on antithrombotic spectrum effect relationship |
| CN108195989B (en) * | 2018-02-01 | 2021-04-20 | 山西省医药与生命科学研究院 | Method for evaluating chemical components of rosa davurica pall based on antithrombotic spectral efficiency relationship |
| CN111801633A (en) * | 2018-06-14 | 2020-10-20 | 舍弗勒技术股份两合公司 | Production control method based on failure risk |
| CN111801633B (en) * | 2018-06-14 | 2024-03-05 | 舍弗勒技术股份两合公司 | Production control method based on failure risk |
| CN110806457A (en) * | 2019-11-29 | 2020-02-18 | 南京南医堂健康产业发展有限公司 | Detection method of fingerprint of Sijun manna drink |
| CN110806457B (en) * | 2019-11-29 | 2021-10-08 | 南京南医堂健康产业发展有限公司 | Detection method of fingerprint of Sijun manna drink |
| CN112763597A (en) * | 2020-12-22 | 2021-05-07 | 葵花药业集团（贵州）宏奇有限公司 | Multi-index content determination method for astragalus root, dendrobium stem and hawthorn granules |
| CN112763597B (en) * | 2020-12-22 | 2023-02-24 | 葵花药业集团（贵州）宏奇有限公司 | Multi-index content determination method for astragalus root, dendrobium stem and hawthorn granules |
| CN113049711A (en) * | 2021-03-23 | 2021-06-29 | 宁夏医科大学 | Method for simultaneously determining multiple effective components in eight-treasure intelligence-benefiting mixture |
| CN113049711B (en) * | 2021-03-23 | 2022-09-23 | 宁夏医科大学 | Method for simultaneously determining multiple effective components in eight-treasure nootropic mixture |
| CN113325115A (en) * | 2021-07-08 | 2021-08-31 | 辽宁华润本溪三药有限公司 | Method for establishing characteristic spectrum of qi stagnation stomachache granules and application thereof |
| CN113433244A (en) * | 2021-07-08 | 2021-09-24 | 辽宁华润本溪三药有限公司 | Method for determining tetrahydropalmatine content in qi stagnation stomachache granules and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105758950B (en) | 2018-02-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105758950A (en) | Quality control method based on dose-effect color card for anti-inflammatory and analgesia action of qi-stagnation and stomachache granules | |
| CN106501434B (en) | A kind of HPLC finger print measuring methods of Double Harmonizing Decoction standard soup | |
| CN104215698B (en) | The fingerprint atlas detection method of Callicarpa nudiflora medical material, intermediate and preparation thereof and standard finger-print | |
| CN107727753B (en) | Method for establishing baile sleeping capsule fingerprint | |
| CN104076099B (en) | A kind of detection method of pharmaceutical composition | |
| CN105758949B (en) | Qizhi weitong granules based on dose-effect colour atla promotees gastroenteritic power method of quality control | |
| CN114689774B (en) | Preparation process and quality control method of standard decoction of radix angelicae sinensis blood replenishing decoction | |
| CN103207255A (en) | Content detection method for Naoxintong capsule | |
| Liu et al. | Determination of the active ingredients and biopotency in Polygala tenuifolia Willd. and the ecological factors that influence them | |
| CN105738519A (en) | Qi stagnation stomachache particle anti-ulcer quality control method based on dose-effect color card | |
| CN103105444A (en) | Method for detecting quality of Xianing Gubao capsules | |
| CN104237444A (en) | Construction method of HPLC (High Performance Liquid Chromatography) fingerprint spectrum of salvia miltiorrhiza and radix puerariae depression relieving drug | |
| CN106706786A (en) | Method for determining content of six ginsenoside ingredients of folium ginseng | |
| CN103169864B (en) | Zengye Tang granule and preparation method thereof, purposes and detection method | |
| Cai et al. | Xianlian Jiedu Decoction alleviates colorectal cancer by regulating metabolic profiles, intestinal microbiota and metabolites | |
| CN109633003A (en) | Radix Polygalae mouth mountain ketone III and 3,6 in a kind of measurement Radix Polygalae, the method for-two mustard acyl cane sugar contents | |
| CN104007198B (en) | A kind of glossy ganoderma emperor's preparation HPLC standard finger-print and construction method thereof and application | |
| CN106918673B (en) | A kind of method for building up of the finger-print of Chinese medicine composition | |
| CN104950068B (en) | The kidney invigorating is dispeled the wind the quality determining method of climacteric mixture | |
| CN103604898B (en) | The benefit heart relaxes the fingerprint atlas detection method of preparation | |
| CN114689776B (en) | Construction method of high-efficiency liquid-phase characteristic spectrum of lindera root decoction | |
| CN101816749A (en) | Medicament for curing dysuria, preparation method and quality control method thereof | |
| CN106526000B (en) | The method for building up and finger-print of Shengma Gegen Tang composition finger-print | |
| CN105044260B (en) | The construction method of Yupingfeng preparation finger and detection method | |
| CN104666771A (en) | Traditional Chinese medicine capsule for treating depressive disorder and preparation technology and detection and analysis method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190710 Address after: 117000 Medical Park of Benxi Economic and Technological Development Zone, Liaoning Province Patentee after: Liaoning Huarun Benxi Sanyao Co., Ltd. Address before: 110847 No. 79, Chong Shan Road, Huanggu District, Shenyang, Liaoning. Patentee before: Liaoning University of TCM |
|
| TR01 | Transfer of patent right |