CN100362008C - Barbat skullcap general flavone preparation method and quality control method - Google Patents

Barbat skullcap general flavone preparation method and quality control method Download PDF

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CN100362008C
CN100362008C CNB2004100888334A CN200410088833A CN100362008C CN 100362008 C CN100362008 C CN 100362008C CN B2004100888334 A CNB2004100888334 A CN B2004100888334A CN 200410088833 A CN200410088833 A CN 200410088833A CN 100362008 C CN100362008 C CN 100362008C
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herba scutellariae
scutellariae barbatae
total flavones
methyl alcohol
scutellarin
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CN1769292A (en
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王英锋
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Youcare Pharmaceutical Group Anhui Natural Pharmaceutical Co.,Ltd.
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Abstract

The present invention relates to a preparation method of barbat skullcap total flavone, a quality control method thereof and a new purpose in pharmacy. The preparation method comprises the following processes: the barbat skullcap total flavone containing more than 80 % of the total flavone content is prepared by thinning by water, regulating pH value, centrifuging, regulating pH value by hydrochloric acid, drying deposit, etc.; simultaneously, the present invention discloses an application of the barbat skullcap total flavone in the respiratory tract infection diseases, such as acute pharyngitis, acute tonsillitis, virus pneumonia, etc. The preparation of the present invention has the advantages of stabilization and controllable quality.

Description

A kind of preparation method of Herba Scutellariae Barbatae total flavones and quality controlling means
Invention field
The present invention relates to a kind of preparation method and quality controlling means and new pharmaceutical use thereof of Chinese medical extract, particularly a kind of preparation method of Herba Scutellariae Barbatae total flavones and quality controlling means and new pharmaceutical use thereof.
Background technology
Herba Scutellariae Barbatae (Scutellaria barbata D.Don) is as conventional Chinese medicine, have many-sided effects such as heat-clearing, detoxifcation, wide clinical application is in multiple infectious diseases such as treatment pharyngitis, tonsillitis, pneumonia, mazoitis, enteritis, dysentery, boils, and ephritis, hepatitis, hepatic ascites, venomous snake bite, even the treatment of multiple malignant tumour.Modern pharmacological research shows, various pharmacological activities such as that crude extract of Herba Scutellariae Barbatae and efficient part thereof, effective monomer have is antibiotic, anti-mutation, anti-inflammatory, analgesic, anti-oxidant, antitumor, immunomodulatory.In recent years, pharmaceutical use of Herba Scutellariae Barbatae and clinical application thereof more and more are subjected to paying attention to widely, and the quality standard research of the Herba Scutellariae Barbatae of being finished by national State Key Task 95 problem has been taken in the version Pharmacopoeia of the People's Republic of China in 2000.But, up to now, the Herba Scutellariae Barbatae sheet that the preparation relevant with Herba Scutellariae Barbatae only is used as medicine with herb or crude extract on the domestic medical market, ginseng lotus capsule, Reyanning sheet minority kinds such as (capsules), the deep development research work of relevant extract drugs composition is close to blank.Data-searching, at present, both at home and abroad other unit do not have as yet that the Herba Scutellariae Barbatae flavone component is antiviral, antibiotic, the research report of anti-inflammatory, analgesic, analgesia comprehensive drug and new drug development.
Summary of the invention
One object of the present invention is to disclose a kind of preparation method of Herba Scutellariae Barbatae total flavones; The object of the invention also is to disclose a kind of quality controlling means of Herba Scutellariae Barbatae total flavones; The 3rd purpose of the present invention is to disclose a kind of new pharmaceutical use of Herba Scutellariae Barbatae total flavones.
The preparation method of Herba Scutellariae Barbatae total flavones is:
Get the Herba Scutellariae Barbatae meal, adding 6-10 times of boiling water boiled 1-3 hour, filter, the dregs of a decoction are carried 1-2 hour with 6-8 times of poach, filter, merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2～6, centrifugal, get the macroporous resin of supernatant liquor, pre-treatment, dress post with same medicinal material amount, said extracted liquid is crossed post absorption, resin and crude drug weight ratio are 1: 2-8, and washing is again with 85% above ethanol elution, and the volume of eluting solvent is 4-8 a times of weight resin, reclaim ethanol, drying, promptly get Herba Scutellariae Barbatae total flavones; Add 3-5 times of water dilution, 10% sodium hydroxide is transferred pH value 8-10, and is centrifugal, and supernatant liquor is transferred pH value 1-3 with hydrochloric acid, leaves standstill 1-4 hour, and centrifugal, drying precipitate promptly gets general flavone content 80% above Herba Scutellariae Barbatae total flavones, and wherein scutellarin content is 10%.
Quality controlling means of the present invention contains one or both in the following content assaying method:
Content assaying method is:
A, total flavones
Get the scutellarin reference substance, add methyl alcohol and make the solution that every 1ml contains 0.20mg, shake up, promptly get reference substance solution; The accurate reference substance solution 0.4 of drawing, 0.6,0.8,1.0,1.2ml, adding methyl alcohol respectively to 10ml, is blank with methyl alcohol, according to spectrophotometry (an appendix VI of Chinese Pharmacopoeia version in 2000 A), wavelength place at 285nm measures optical density, with concentration is X-coordinate, and optical density is an ordinate zou, the drawing standard curve; Be taken at 80 ℃ of this Herba Scutellariae Barbatae total flavones 20mg that are dried to constant weight, put in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, supersound process 30 minutes, shake up, filter, the accurate subsequent filtrate 1ml that draws, put in the 10ml measuring bottle, add methyl alcohol, shake up to scale, the accurate 1ml that draws puts in the 10ml measuring bottle, adds methyl alcohol to scale, shaking up, is blank with methyl alcohol, according to spectrophotometry (an appendix VI of Chinese Pharmacopoeia version in 2000 A), wavelength place at 285nm measures optical density, calculates, promptly; This Herba Scutellariae Barbatae total flavones contains total flavones with scutellarin (C 21H 18O 12) meter, must not be less than 51%.
B, scutellarin
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).
With octadecylsilane chemically bonded silica is weighting agent, and methanol-water-formic acid of 38: 62: 0.05 is moving phase, and the detection wavelength is 335nm; Number of theoretical plate is pressed the scutellarin peak and is calculated, and is not less than 1500; Precision takes by weighing the scutellarin reference substance, adds moving phase and makes the solution that contains scutellarin 0.20mg among every 1ml, shakes up, and promptly gets reference substance solution; Get this Herba Scutellariae Barbatae total flavones 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, and it is an amount of to add methyl alcohol, adds methyl alcohol to scale after the dissolving in ultrasonic 20-40 minute, shakes up, and promptly gets need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, calculate, promptly; This Herba Scutellariae Barbatae total flavones contains scutellarin must not be less than 2%.
Preparation method of the present invention adopts macroporous resin adsorption technology extraction effective ingredient in Chinese to be used as medicine, and technological process is easy, does not have technical barrier, and production process is controlled easily, has guaranteed the controllability of quality product in a certain sense; Yield height of flavonoid compound in the adopting process plant, extracting solvent can recycle, energy consumption is low, and the raw medicinal material price is cheaper, guarantee that product has had lower production cost, therefore at present the medical market like product has had bigger market value adjustable space, has guaranteed that product has had the competitive edge aspect the product price in the competition of future market; The inventive method is stable, and its extract quality is controlled.
Herba Scutellariae Barbatae total flavones has been carried out antibacterial experiment in extracorporeal antivirus effect test, interior resisting virus test, in-vitro antibacterial experiment, the body, and the result shows: Herba Scutellariae Barbatae total flavones has obvious restraining effect to the cytopathic effect of parainfluenza virus (HVJ), respiratory syncytial virus (RSV), simplexvirus 1,2 types (HSV-I, II), adenovirus 3,7 types viruses such as (Adr3, Adr7); Herba Scutellariae Barbatae total flavones has obvious restraining effect to the mouse pneumonia that influenza virus causes, and it is relevant to present good dose-effect; And influenza infection is caused dead mouse significant protective effect is arranged; Herba Scutellariae Barbatae total flavones to 25 strains expressed of 10 kind of 27 strain bacterium of try bacteriostatic action in various degree, to 19 strains expressed germicidal action in various degree; Wherein, at 0.20～0.80mg/ml, minimal bactericidal concentration (MBC) can reach 0.40mg/ml to the minimal inhibitory concentration scope (MIC) of streptococcus aureus.The bacteriostatic action intensity ordering that with MIC50 is index measuring and calculating is: Pseudomonas aeruginosa, streptococcus aureus, alpha hemolytic streptococcus, block its cloth youth Durham Salmonella, staphylococcus, streptococcus pneumoniae, beta hemolytic streptococcus, colon bacillus, Klebsiella pneumonia, Bacillus proteus, the ordering of germicidal action intensity is close substantially; The Herba Scutellariae Barbatae total flavones gastric infusion has significant protective effect to the dead mouse due to the infection of staphylococcus aureus.In other related experiment, show that simultaneously 70-280mgkg-1 Herba Scutellariae Barbatae total flavones gastric infusion all can cause the exothermic reaction of rat by inhibition yeast in various degree, and be the doses dependency; 100-400mgkg-1 Herba Scutellariae Barbatae total flavones gastric infusion all can significantly suppress the mouse writhing pain reaction due to the acetic acid, and dosage correlation is obvious.
Following experimental example is used to further specify the present invention:
Experimental example 1Condition confirmed tests such as the amount of water of decocting technology, extraction time, number of times
1, determining of amount of water: three parts of Herba Scutellariae Barbataes (each 100g), add water by table 1 and extract test, boil and carry 1 hour, concentrate, drying gets dry extract, weighs, and with its general flavone content of determined by ultraviolet spectrophotometry.
Table 1
Sequence number Medicinal material amount (g) Amount of water (doubly) Go out cream (g) Content % To medicinal material yield %
1 2 3 100 100 100 6 8 10 11.45 12.31 12.50 17.97 17.25 17.24 2.05 2.12 2.16
2, determining of extraction time: three parts of Herba Scutellariae Barbataes (each 100g), add 10 times of water and test,, extraction time press table 2, and concentrated, drying gets dry extract, weigh, and with its general flavone content of determined by ultraviolet spectrophotometry.
Table 2
Sequence number Medicinal material amount (g) Extraction time (hour) Go out cream (g) Content % To medicinal material yield %
1 2 3 100 100 100 1 2 3 12.60 12.32 12.77 17.35 16.94 16.38 2.19 2.09 2.09
3, determining of extraction time: three parts of Herba Scutellariae Barbataes (each 100g), add 10 times of water and test,, 1 hour extraction time, extraction time press table 3, and concentrated, drying gets dry extract, weigh, and with its general flavone content of determined by ultraviolet spectrophotometry.
Table 3
Sequence number Medicinal material amount (g) Extraction time Go out cream (g) Content % To medicinal material yield %
1 2 3 100 100 100 1 2 3 12.67 14.15 14.86 17.42 16.03 15.66 2.21 2.27 2.33
4, conclusion: by above-mentioned experiment, determine that best amount of water is 10 times, the optimum extraction time is 1 hour, and the optimum extraction number of times is 2 times.
Experimental example 2The macroporous resin adsorption test
1, the orthogonal design of technology
According to the performance of macroporous resin and the character of Herba Scutellariae Barbatae total flavones, selected 3 levels (table 4) of investigating four factors (A, B, C, D) and each factor
Table 4 level of factor table
Figure C20041008883300101
The volume ratio (g/ml) of solution when A medicinal material weight and absorption
Extracting liquid pH value during B absorption
C resin and medicinal material weight ratio
D weight resin and eluent volume ratio (g/ml)
2, experimentation:
Get Herba Scutellariae Barbatae meal 900g, add 10 times of boiling water and boil and carry, filter, the dregs of a decoction add 8 times of poach again to be carried, and filters.Merge secondary filtrate, be divided into 9 parts, concentrate by orthogonal table respectively, adjust pH is crossed macroporous resin column, the absorption after washing is colourless to flowing out water liquid, resin is used 85% ethanol elution again, reclaims ethanol, drying, weighs, and ultraviolet spectrophotometry is measured optical density at the 285nm place, calculate general flavone content by made typical curve in the quality standard, and amount to into the medicinal material yield.
General flavone content=(A-0.005093) * 10 * 10 * 100%/45.41189 * sample is heavy
Table 5 test design table
Figure C20041008883300102
Test-results and variance analysis
Table 6, L9 (34) reckoner
Factor A B C D Total flavones
Row number
1 2 3 4 Receive cream rate (%) Content (%) To medicinal material yield (%)
Test number 1 2 3 4 5 6 7 8 9 1 1 1 2 2 2 3 3 3 1 2 3 1 2 3 1 2 3 1 2 3 2 3 1 3 1 2 1 2 3 3 1 2 2 3 1 1.23 0.64 0.60 0.41 0.23 2.87 0.23 1.33 1.82 44.49 51.12 55.25 40.54 44.21 55.21 38.13 53.25 55.34 0.547 0.327 0.332 0.166 0.102 1.585 0.088 0.708 1.007
I j II j IIIj I 2j II 2j III 2j Rj＝I 2j+II 2j+III 2j Sj＝RJ/3-CT 1.206 1.853 1.803 1.454 3.434 3.251 8.139 0.086 0.801 1.137 2.924 0.642 1.293 8.550 10.485 0.868 2.840 1.500 0.522 8.066 2.250 0.272 10.588 0.902 1.656 2.000 1.206 2.742 4.000 1.454 8.196 0.105 G＝4.862 CT＝G 2/9＝2.627 S E＝0.086
Table 7 analysis of variance table
The source Sum of squares Degree of freedom All the side and F ratio Significance
A B C D error e S a＝0.086 S B＝0.868 S C＝0.902 S D＝0.105 2 2 2 2 ∞ 0.043 0.434 0.451 0.053 1 10.093 10.488 1.233 ** **
F1-0.01(2，∞)＝4.61 F1-0.05(2，∞)＝3.0 F1-0.10(2，∞)＝2.3
3, result and analysis
On test-results and analysis of variance table, it is the resin demand of adsorption liquid pH and absorption medicinal material that factor B, C have the influence of highly significant to the result, D and A have certain influence to the result, influence is B＞C＞D＞A in proper order, draws optimum process condition through taking all factors into consideration: A2 B3 C1 D2 promptly: medicinal material weight is 1: 3 with the volume ratio of when absorption solution.Adsorption liquid is transferred pH2-3.Resin and crude drug weight ratio are 1: 2, for fully adsorbing available 1: 1.The volume of eluting solvent is 6 times of weight resin.
4, the macroporous resin model is preferred:
To extract total flavones with technology of the present invention available from several macroporous resins of Tianjin Chemical Plant of Nankai Univ., the 100g Herba Scutellariae Barbatae is used in every part of experiment, measure general flavone content in the cream with method in the quality standard, get its mean value, and amount to into the medicinal material yield, comparative result such as following table:
Table 8
The resin model D101 AB-8 ZTC-2 NKA-9
Paste-forming rate (%) general flavone content total flavones is to medicinal material yield (%) 1.752 40.81 0.715 2.83 55.24 1.563 1.164 41.10 0.478 1.142 39.07 0.446
As seen AB-8 type macroporous resin is fit to the extraction of Herba Scutellariae Barbatae total flavones the most.
Experimental example 3The assay of Herba Scutellariae Barbatae total flavones
1, the selection of method and condition determination
This Herba Scutellariae Barbatae total flavones is the raw material that Chinese medicine is registered 5 kind new medicines, with the total flavones is efficient part, so the object of assay is total flavones and main effective monomer thereof. according to pharmacopeia and bibliographical information, once in order to scutellarin the spectrophotometry of reference substance, measure at the 335nm place, because of inconsistent, certain error is arranged and abandon with this Herba Scutellariae Barbatae total flavones maximum absorption wavelength 285nm.Selected at last is the ultraviolet spectrophotometry of reference substance with the scutellarin, measure optical density at the 285nm place. because scutellarin has two maximum absorption wavelength 285nm and 335nm, and this Herba Scutellariae Barbatae total flavones maximum absorption wavelength also is 285nm, measures at this wavelength, sensitivity is good, and error is less.
2, methodological study
2.1 precision takes by weighing scutellarin (80 ℃ are dried to constant weight) 4.25mg, is dissolved in surely in the 25ml volumetric flask with methyl alcohol is ultrasonic, promptly gets the reference substance storing solution of 0.17mg/ml.It is fixed to get the accurate title of this Herba Scutellariae Barbatae total flavones 20mg, puts in the 10ml volumetric flask, adds methyl alcohol to scale, ultrasonic 30 minutes, shakes up, the accurate 1ml that draws puts in the 10ml volumetric flask, adds methyl alcohol to scale, shakes up, the accurate 1ml that draws, put in the 10ml volumetric flask, add methyl alcohol, shake up, as need testing solution to scale.
Accurately measure 0.40,0.60,0.80,1.00 2.2 measure, 1.20ml reference substance storing solution places the 10ml volumetric flask respectively, and methanol constant volume promptly gets serial reference substance solution to scale.With methyl alcohol is blank, measures optical density at the 285nm place according to spectrophotometry (2000 editions one appendix VI A of Chinese Pharmacopoeia), is X-coordinate with concentration, and optical density is an ordinate zou, the drawing standard working curve.
2.3 linear relationship is investigated (seeing accompanying drawing 1)
Table 9 linear relationship measurement result
Sequence number 1 2 3 4 5
Contrast solution concentration (mg/ml) optical density 0.006784 0.3173 0.010176 0.4799 0.013568 0.6192 0.016960 0.7553 0.020352 0.9096
Its regression equation is:
A＝44.401112Cx+0.011521 r＝0.9995
2.4 the mensuration of precision with the scutellarin standardized solution of same concentration, is used aforesaid method duplicate detection 5 times.
Table 10
Sequence number 1 2 3 4 5
Optical density 0.9006 0.9008 0.9010 0.9012 0.9011
On average (%) 0.9009
RSD 0.027％
2.5 reproducibility test is 2002111201 Herba Scutellariae Barbatae total flavones sample liquid with lot number with aforesaid method repetitive operation 5 times, and measures.The results are shown in following table:
Table 11 reproducibility measurement result
Sequence number Sampling amount (mg) Optical density General flavone content in the sample (%) Average content (%) RSD(％)
1 2 3 4 5 20.59 20.58 20.41 20.03 20.67 0.5087 0.5021 0.5137 0.4886 0.4990 57.15 56.44 58.23 56.43 55.85 56.82 1.61
2.6 sample stability test: get lot number and be 2002111201 sample solution, places after 3,6,9,12 hours, through spectrophotometry, its general flavone content changes little, and interpret sample solution was stablized in 12 hours.
Table 12
Time (hour) 0 3 6 9 12
General flavone content (%) 56.42 56.73 57.05 57.03 55.91
Mean value (%) 56.628
RSD(％) 0.84
2.7 average recovery test: precision takes by weighing a certain amount of scutellarin reference substance, is added to lot number and is in this Herba Scutellariae Barbatae total flavones of 2002111201, measures average recovery as stated above.
The results are shown in Table 13.
Table 13 average recovery test determination result
Sequence number Sample total flavones amount (mg) Reference substance add-on (mg) The total flavones amount of measuring (mg) The rate of recovery (%) Mean value (%) RSD (％)
1 2 3 4 5 0.00.1283 0.05808 0.05798 0.05691 0.05872 0.06688 0.06688 0.06688 0.06688 0.06688 0.1283 0.1260 0.1232 0.1213 0.1257 100.37 101.56 97.52 96.28 100.15 99.176 2.21
2.8 sample determination is got three crowdes of about 20mg of Herba Scutellariae Barbatae total flavones dry powder, the accurate title, decide, and prepares sample liquid by this current draft method, and sample liquid is measured optical density at the 285nm place, is calculated as follows general flavone content in the Herba Scutellariae Barbatae medicinal material according to regression equation.
Figure C20041008883300141
The result is as follows: table 14
Lot number Sampling amount (mg) Optical density General flavone content (%)
2002111201 20.02 0.5087 57.15
20.46 0.5423 57.82
2002111301 20.54 0.5361 56.93
20.58 0.5021 56.44
2002111401 20.17 0.5186 56.06
20.25 0.5243 56.46
According to above measurement result, limit is tentative must not remember with scutellarin and is less than 51% for this Herba Scutellariae Barbatae total flavones contains total flavones.
Experimental example 4The assay of scutellarin
In order to control the Herba Scutellariae Barbatae total flavones quality accurately, the assay project has also been drafted the content assaying method that mainly contains the effective constituent scutellarin in the total flavones except that determination of total flavonoids, uses high performance liquid chromatography as quantivative approach.
1. instrument and reagent: Japanese beam split LC-1500 type high performance liquid chromatograph, the HP-8453 of Hewlett-Packard type ultraviolet spectrophotometer, plum Teller AG-254 type electronic analytical balance, it is pure that reagent is top grade, scutellarin reference substance (Traditional Chinese Medicine Inst., Chengde Medical College's self-control, content is more than 98%).
2. chromatographic condition: chromatographic column: Discovery C18 (4.6 * 250mm)
Moving phase: methyl alcohol: water: formic acid (38: 62: 0.05)
Flow: 0.50ml/min
Column temperature: 25 ℃
Detect wavelength: 335nm
3. the preparation of need testing solution and reference substance solution; get the about 20mg of this product, the accurate title, decide, and puts in the 10ml measuring bottle; add small amount of methanol and make molten; add methyl alcohol to scale, as need testing solution, gets scutellarin reference substance 4.15mg; accurate title is fixed; put in the 25ml volumetric flask, add methyl alcohol to scale, in contrast product solution.
4. the investigation of linear relationship: precision takes by weighing scutellarin reference substance 6.40mg, puts in the 50ml measuring bottle product storing solution in contrast, (its concentration is 0.0694mg/ml).Drawing concentration is the scutellarin reference substance storing solution 2.0,4.0,6.0,8.0,10 of 0.0694mg/ml, and 12ml moves in other 5 10ml volumetric flasks successively, adds methyl alcohol to scale, respectively gets 10 μ l sample introductions, measures the scutellarin peak area under aforementioned chromatographic condition.
The results are shown in following table: table 15
Sequence number Scutellarin amount (μ g) Scutellarin peak area integrated value
1 2 3 4 5 6 0.26 0.51 0.77 1.02 1.28 1.54 16151.315 36932.570 59547.441 79285.914 99226.836 118191.281
With reference substance sample size (X) is X-coordinate, and peak area (Y) is an ordinate zou, and it is as follows that regression treatment gets straight-line equation:
Y＝80000.3*X-3455.04 r＝0.9992
Show that the scutellarin sample size has good linear relationship (typical curve is seen accompanying drawing 2) in 0.256 μ g～1.536 μ g scopes
5. precision test: above-mentioned No. 4 reference substance solution of accurate absorption, repeat sample introduction (n=6), the results are shown in following table
Table 16
Sequence number 1 2 3 4 5 6 RSD(％)
Scutellarin peak area integrated value 32689 32317 32455 32884 32855 32336 0.781％
6. to get lot number be 5 parts of 2002111201 Herba Scutellariae Barbatae total flavoness to reproducibility, prepares need testing solution by this current draft method, the results are shown in following table: table 17
Figure C20041008883300161
7. recovery test: adopt the application of sample absorption method, get lot number and be 2002111201 5 parts of Herba Scutellariae Barbatae total flavoness (2.994%), add the scutellarin reference substance respectively, the preparation need testing solution is measured in accordance with the law,
The result is as follows: table 18
Figure C20041008883300171
8. the mensuration of sample is got the about 20mg of Herba Scutellariae Barbatae total flavones dry powder, prepares sample liquid by this current draft method, draws each 10 μ l sample introduction of reference substance solution and sample solution respectively, calculates the percentage composition of scutellarin in the Herba Scutellariae Barbatae raw material with the external standard planimetry.The result is as follows: table 19
Lot number Scutellarin content (%)
2002111201 2.5
2002111301 2.2
2002111401 2.3
Standard substance concentration is 0.0694mg/ml, and peak area is 31222
According to above measurement result, the tentative this product of limit contains scutellarin and must not be less than 2%.
Experimental example 5The extracorporeal antivirus effect test
Get the 96 porocyte culture plates that grow up to monolayer cell, outwell nutrient solution, inoculate the different virus liquid 50ul/ hole of 100TCID50 respectively, put in 37 ℃, 5%CO2 incubator absorption 1 hour, outwell viral liquid, keep liquid with the Engle ' s that does not contain calf serum and wash cell face secondary, add corresponding dilution soup then, 4 multiple holes are done in 100ul/ hole, each extent of dilution.Establish virus control, normal cell contrast and positive drug contrast simultaneously.Put in 37 ℃, 5%CO2 incubator and cultivate, observation of cell pathology under the every day inverted microscope, when virus control group cytopathy is ++ ++ the time record experimental result.
Following 6 grades of criteria for classifications are pressed in the judgement of cytopathy degree (CPE):
-: the cell growth is normal, and no pathology occurs;
±: cytopathy is less than 10% of whole monolayer cell;
+ (1): cytopathy accounts for below 25% of whole monolayer cell;
++ (2): cytopathy accounts for below 50% of whole monolayer cell;
+++(3): cytopathy accounts for below 75% of whole monolayer cell;
++ ++ (4): cytopathy accounts for more than 75% of whole monolayer cell;
Calculate 50% effective concentration (IC50) by Reed-Muench.
Therapeutic index (TI)=TC50/IC50.
The result adopts rank test to carry out statistical procedures, the results are shown in Table 20,21.
Table 20,21 results show, Herba Scutellariae Barbatae total flavones has obvious restraining effect at external cytopathic effect to parainfluenza virus, RSV, HSV-I, HSV-I, A3, A7 virus, its IC50 is respectively 0.72,0.35,3.31,3.31,0.57 and 0.55mg/ml, and TI is respectively 6.32,13.10,1.38,1.38,7.96 and 8.33.CoxB virus is not demonstrated the obvious suppression effect.
Table 20 Herba Scutellariae Barbatae total flavones is to the influence of pathological changes caused by virus
Medicine Drug level Cytopathy degree (CPE)
Parainfluenza CoxB 4 CoxB 5 RSV Adr-3 Adr-7 HSV-1 HSV-2
Herba Scutellariae Barbatae 1.6 0.8 0.4 0.2 0.1 1112** 1121** 2222* 3333 3333 4334 4243 4343 4443 4444 1424 3223** 3224 3444 4444 ±111** 11±1** 1212** 2344 4443 1111** 1122** 2112** 3233 3333 1121** 1121** 2112** 3322 3333 2212** 2312* 3443 4444 4444 2221** 2311* 3344 4444 4444
Virazole 0.5 0.25 11±1** 1111** 11±1** 1112** 1111** 1111** 1111** 1111** 1121* 2321* 1111** 2223* 1111** 2121** 1112 2211
The virus control group 3333 4444 4444 4444 3333 3333 4444 4444
Annotate: compare * p＜0.05, * * p＜0.01 with the virus control group.
The IC50 and the TI of table 21 Herba Scutellariae Barbatae total flavones extracorporeal antivirus effect
Soup Virus is planted IC 50 TI
Herba Scutellariae Barbatae Parainfluenza RSV CoxB 4 CoxB 5 HSV-1 HSV-2 A 3 A 7 0.72 0.35 - - 3.31 3.31 0.57 0.55 6.32 13.10 - - 1.38 1.38 7.96 8.33
Annotate: "-" represents the viral unrestraint effect of try, unit: mg/ml.
Experimental example 6The interior resisting virus test
1, to the therapeutic action of mouse influenza virus property pneumonia
Get mouse and be divided into 6 groups at random by body weight.Be respectively three dosage groups of Herba Scutellariae Barbatae total flavones; Virazole (0.07gkg-1/d) group; Virus infection control group and intact animal control group.Except that the normal control group, mouse is slightly anaesthetized with ether, infect every 0.05ml with 15 LD50 influenza virus drop noses.Infect preceding 1 day beginning gastric infusion, every day 2 times, each 0.25ml/10g, continuous 5 days, control group under equal conditions distilled water was irritated stomach.Dissected after taking by weighing the mouse body weight on the 6th day, win full lung and weigh, calculate the lung index value, and obtain lung index inhibiting rate.
Lung index=[heavy (the g)/body weight (g) of lung] * 100
Lung index inhibiting rate=(virus control group lung index average-test group lung index average)/virus control group lung index average * 100%
Statistical procedures is carried out in the t check between employing group as a result.
Table 22 Herba Scutellariae Barbatae total flavones is to the restraining effect of mouse influenza virus property pneumonia
Group Dosage mg/kg/d Mouse number (only) Lung index value (g lung weight/100g body weight) Inhibiting rate (%)
Dosage group small dose group in the heavy dose of group of normal control papova control group virazole group - - 0.07 400 200 100 10 10 10 10 10 10 0.92±0.06 1.88±0.41## 1.04±0.10** 1.39±0.37** 1.55±0.36 1.86±0.30 - - 45 26 18 1
Annotate: compare with the normal control group: ##P＜0.01, compare with the virus control group: *P＜0.05.
Table 22 result shows that the lung index value of Herba Scutellariae Barbatae total flavones 400mg/kg/d dosage group is starkly lower than the virus control group, with the virus control group significant difference (P＜0.05) is arranged relatively; The lung index value of 200mg/kg/d dosage group is starkly lower than the virus control group, but the statistics there was no significant difference.Show that Herba Scutellariae Barbatae total flavones has obvious restraining effect to the mouse pneumonia that influenza virus causes, and it is relevant to present good dose-effect.
2, to the provide protection of influenza virus induced mice death
(1) virus causes the mensuration of the minimum virulence of dead mouse: virus is diluted to the different virus liquid of 0.5-4LD50 with physiological saline, gets 50 of mouse, and 10 of each extent of dilution, 0.05ml/ collunarium infects.Observe the death condition of animal, get and infect the 7-9 days animal dead rates in back, the infective dose during for experiment in the concentration more than 90%.Experimental result shows: when 2LD50 infected, mortality of mice was 90%, so it is decided to be infective dose when testing.
(2) provide protection of medicine: get 100 of mouse, be divided into 5 groups at random by body weight.Be respectively three dosage groups of Herba Scutellariae Barbatae total flavones, virazole (0.07gkg-1/d) group and virus infection control group.Each administration group gastric infusion, every day 2 times, each 0.25ml/10g, continuous 7 days, the virus control group under equal conditions gave distilled water.Administration respectively organized in second day mouse with the slight anesthesia of ether after, infect every 0.05ml with 2 LD50 influenza virus drop noses.Record infects the death toll of back mouse, calculates mortality ratio, dead protection ratio and increase in life span.
Mortality ratio %=dead animal number/animal sum
Dead protection ratio=(control group mortality ratio-experimental group mortality ratio)/control group mortality ratio
Increase in life span=(experimental group on average survive fate-infected group on average survive fate)/infected group fate of on average surviving
The results are shown in Table 23,24.
Table 23,24 results show, in the mouse infection virus 15 days, the dead number average of three dosage treated animals of Herba Scutellariae Barbatae total flavones is less than the infection control group; Dead protection ratio is respectively 27.78%, 33.33% and 11.1 1% (but statistics is meaningless); And the survival fate of mouse obviously prolongs than infected group, with control group significant difference (P＜0.01) is arranged relatively.Increase in life span is respectively 18.97% and 17.94% and 8.20%.The expression Herba Scutellariae Barbatae total flavones causes dead mouse to influenza infection when this dosage have significant protective effect.
Table 23 Herba Scutellariae Barbatae total flavones causes the provide protection of dead mouse to influenza virus
Group Dosage mg/kg/d Number of animals (only) Death toll Mortality ratio % Protection ratio %
Dosage group small dose group in the heavy dose of group of infection group and viral infection group virazole -- 0.07 400 200 100 20 20 20 20 20 18 8 13 12 16 90 40 65 60 80 55.56** 27.78 33.33 11.11
Annotate: compare with control group *P＜0.05.
Table 24 Herba Scutellariae Barbatae total flavones causes the provide protection of dead mouse to influenza virus
Group Dosage g/kg/d Number of animals (only) Average survival fate Increase in life span (%)
Dosage group small dose group in the heavy dose of group of infection group and viral infection group virazole -- 0.07 400 200 100 20 20 20 20 20 9.75±1.92 12.85±1.49** 11.60±2.13** 11.50±2.13** 10.55±1.96 31.79 18.97 17.94 8.20
Annotate: compare with control group *P＜0.01.
Experimental example 7The in-vitro antibacterial test
1, the preparation of test organisms liquid
Alpha hemolytic streptococcus, beta hemolytic streptococcus, streptococcus pneumoniae increased bacterium in 18 hours with 37 ℃ of 10%FCS-nutrient broth mediums respectively to be cultivated, and same medium 10-1 dilution is test organisms liquid.Other bacterium increased bacterium in 18 hours with 37 ℃ of nutrient broth mediums respectively to be cultivated, and same medium 10-3 dilution is test organisms liquid.
2, the mensuration of minimal inhibitory concentration (MIC)
Sterilization Herba Scutellariae Barbatae total flavones 25.64mg/ml (raw material 50mg/ml) concentration is initial, the 10%FCS-nutrient broth medium is adopted in alpha hemolytic streptococcus, beta hemolytic streptococcus, streptococcus pneumoniae test, nutrient broth medium doubling dilution soup to 10 concentration is adopted in other bacteria test, every hole 1ml adds 24 porocyte culture plates, every extent of dilution soup is established 2 multiple holes, add above test organisms liquid 50 μ l respectively, and establish the control wells that each bacterium liquid does not add medicine, cultivated 18 hours for 37 ℃, naked eyes and inverted microscope are observed down and are had or not bacterial growth.
Sterilization SHUANGHUANGLIAN KOUFUYE two-fold dilution's concentration (40mg/ml) concentration is initial, corresponding substratum doubling dilution soup to 10 concentration, and above similarity condition adds test organisms liquid, observes the bacterial growth phenomenon.
With no bacterial growth the high dilution of medicine be minimal inhibitory concentration (MIC).
The mensuration of 3 minimal bactericidal concentration (MBC)
More than observing no bacterial growth culture 50 μ l is inoculated in the plain agar plate culture medium (alpha hemolytic streptococcus, beta hemolytic streptococcus, pneumococcal vaccination is in the blood agar plate culture medium, Bacillus proteus is inoculated in the SS Agar Plating), cultivated 18 hours the counting bacterium colony for 37 ℃.
The high dilution of medicine that is less than 5 (2 multiple hole mean values) with colony number is the minimum bactericidal concentration (MBC) of medicine to this bacterium.Experimental result sees Table 25,26,27.
The vitro antibacterial activity of table 25 Herba Scutellariae Barbatae total flavones (1) extent of dilution concentration (mg/ml)
Bacterial classification Bacterium lot number number Herba Scutellariae Barbatae total flavones SHUANGHUANGLIAN KOUFUYE Carefully bacterium is shone
Extent of dilution/MIC Extent of dilution/MBC Extent of dilution/MIC Extent of dilution/MBC
Its cloth youth Durham Salmonella Klebsiella pneumonia of streptococcus aureus staphylococcal pneumonia suis alpha hemolytic streptococcus beta hemolytic streptococcus colon bacillus Pseudomonas aeruginosa Bacillus proteus card 26112-6 26003-20 26010-18 faces 25923 and faces 2-18 and face 2-20 26101-18 and face 11 and face 14 and face 22 32213-6 32209-13 32210-17 32210-18 and face 10 and face 11 and face 12 and face 23 and face 24 and face 25 and face 5 and face 55 49109 and face 1 and face 12 29,108 46101 1∶256(0.20) 1∶64(0.80) 1∶64(0.80) 1∶256(0.20) 1∶256(0.20) 1∶256(0.20) 1∶32(1.60) 1∶4(12.82) 1∶128(0.40) 1∶4(12.82) 1∶64(0.80) 1∶64(0.80) 1∶16(3.21) 1∶4(12.82) 1∶16(3.21) 1∶64(0.80) 1∶16(3.21) 1∶8(6.41) 1∶4(12.82) 1∶4(12.82) 1∶256(0.20) 1∶256(0.20) 1∶4(12.82) ＞1∶2(25.64) ＞1∶2(25.64) 1∶64(0.80) 1∶4(12.82) 1∶64(0.80) 1∶16(3.21) 1∶16(3.21) 1∶64(3.21) 1∶128(0.40) 1∶128(0.40) 1∶16(3.21) ＞1∶2(25.64) 1∶4(12.82) ＞1∶2(25.64) 1∶16(3.21) 1∶4(12.82) 1∶2(25.64) 1∶4(12.82) 1∶2(25.64) 1∶4(12.82) 1∶4(12.82) 1∶4(12.82) ＞1∶2(25.64) ＞1∶2(25.64) 1∶128(0.40) 1∶128(0.40) 1∶2(25.64) ＞1∶2(25.65) ＞1∶2(25.64) 1∶16(3.21) ＞1∶2(25.64) 1∶256(0.31) 1∶64(1.25) 1∶64(1.25) 1∶128(0.63) 1∶256(0.31) 1∶128(0.63) 1∶128(0.63) 1∶4(20.0) 1∶16(5.00) 1∶16(5.00) 1∶16(5.00) 1∶8(10.0) 1∶16(5.00) 1∶8(10.0) 1∶8(10.0) 1∶8(10.0) 1∶16(5.00) 1∶8(10.0) 1∶4(20.0) 1∶4(20.0) 1∶128(0.63) 1∶64(1.25) 1∶32(2.50) 1∶16(5.00) 1∶16(5.00) 1∶16(5.00) 1∶4(2.50) 1∶32(2.50) 1∶32(2.50) 1∶32(2.50) 1∶16(5.00) 1∶32(2.50) 1∶32(2.50) 1∶32(2.50) ＞1∶2(40.0) 1∶4(20.0) 1∶4(20.0) 1∶2(40.0) 1∶2(40.0) 1∶16(5.0 0) ＞1∶2(40.0) ＞1∶2(40.0) ＞1∶2(40.0) 1∶2(40.0) ＞1∶2(40.0) 1∶2(40.0) ＞1∶2(40.0) 1∶16(5.00) 1∶16(5.00) 1∶16(5.00) 1∶4(20.0) 1∶16(5.00) ＞1∶2(40.0) ＞1∶2(40.0) + + + + + + + + + + + + + + + + + + + + + + + + + + + +
Annotate: "+" has bacterial growth for liquid nutrient medium, and Agar Plating is grown to lawn.
The vitro antibacterial activity of table 26 Herba Scutellariae Barbatae total flavones (2)-Mlc scope (mg/ml)
Bacterial classification (strain number) Herba Scutellariae Barbatae total flavones SHUANGHUANGLIAN KOUFUYE
The MIC scope MIC 50 The MIC scope MIC 50
Its cloth youth Durham Salmonella Klebsiella pneumonia of streptococcus aureus staphylococcal pneumonia suis alpha hemolytic streptococcus beta hemolytic streptococcus colon bacillus Pseudomonas aeruginosa Bacillus proteus card (6) (1) (4) (2) (5) (3) (2) (3) (1) (1) 0.20-0.80 1.60 0.40-12.82 0.80 0.80-12.82 6.41-12.82 0.20 12.82-＞25.64 0.80 12.82 0.34 1.60 1.60 0.80 1.80 7.14 0.20 0.80 12.82 0.31-1.25 0.63 5.00-20.00 5.00-10.00 5.00-10.00 10.00-20.00 0.63-1.25 1.25-5.00 5.00 2.50 0.54 0.63 8.56 5.00 5.34 11.13 0.63 1.78 5.00 2.50
The vitro antibacterial activity of table 27 Herba Scutellariae Barbatae total flavones (3)-germicidal action strength ratio is (MBC value)
MBC(mg/ml) Extent of dilution Bacterial classification Bacterium number-lot number Ratio
0.40 0.80 3.21 12.82 25.64 ＞25.64 1∶128 1∶64 1∶16 1∶4 1∶2 ＞1∶2 Its cloth youth Durham Salmonella streptococcus pneumoniae alpha hemolytic streptococcus beta hemolytic streptococcus colon bacillus beta hemolytic streptococcus proteus pneumonia suis colon bacillus proteus pneumonia klebsiella of streptococcus aureus Pseudomonas aeruginosa streptococcus aureus streptococcus aureus staphylococcus alpha hemolytic streptococcus card Face 18, face 20 face 5, face 55 26112-6, face 25923 26003-20,26010-18 26101-18 32213-6 29108 faces 14 32209-13 32210-18, face and 11,12 face 23 32210-17, face and 10 49109 face 11, face and 22 face 24, face and 25 face 1, face 12 46101 2/6 2/2 2/6 2/6 1/1 1/2 1/1 1/3 1/2 3/5 1/3 2/5 1/3 2/3 2/3 2/3 1/1
Experimental result, 10 kind of 27 strain bacterium of this experiment control wells all has bacterial growth, and Agar Plating is grown to lawn.Herba Scutellariae Barbatae total flavones to try in the 27 strain bacteriums 25 strains expressed anti-microbial effect in various degree.
Try under the bacterial strain condition in this experiment, calculate respectively with MIC50 and drug dilution degree, Herba Scutellariae Barbatae total flavones is to the bacteriostatic action intensity of examination bacterium, and except that minority bacterial strains such as Klebsiella pneumonia, Bacillus proteus, staphylococcus, majority is higher than SHUANGHUANGLIAN KOUFUYE; The germicidal action intensity of close method measuring and calculating is higher than SHUANGHUANGLIAN KOUFUYE.Try under the bacterial strain condition in this experiment, the ordering of the bacteriostatic action intensity of Herba Scutellariae Barbatae total flavones is: Pseudomonas aeruginosa, streptococcus aureus, alpha hemolytic streptococcus, block its cloth youth Durham Salmonella, staphylococcus, streptococcus pneumoniae, beta hemolytic streptococcus, colon bacillus, Klebsiella pneumonia, Bacillus proteus.Try under the bacterial strain condition in this experiment, the ordering of the germicidal action intensity of Herba Scutellariae Barbatae total flavones is: Pseudomonas aeruginosa, streptococcus aureus, staphylococcus, block its cloth youth Durham Salmonella, alpha hemolytic streptococcus, beta hemolytic streptococcus, streptococcus pneumoniae, colon bacillus, Bacillus proteus.
Above experimental result shows, its cloth youth Durham Salmonella of gram positive bacterium such as the Herba Scutellariae Barbatae total flavones pair streptococcus aureus relevant with respiratory tract infection clinically, staphylococcus, alpha hemolytic streptococcus, beta hemolytic streptococcus and gram-negative card etc. has stronger anti-microbial effect.
Experimental example 8Herba Scutellariae Barbatae total flavones is to the dead provide protection of lethality infection of staphylococcus aureus
1, infectation of bacteria minimum lethal dose (concentration) determines
37 ℃ of streptococcus aureuses (clinical strain 2-18) increased bacterium in 18 hours and cultivate, prepare 5% sterilised yeast suspension 10-1～10-3 dilution with 0.5%MC, to mouse peritoneal injection 0.5ml/ only, each extent of dilution infects 5 mouse, observe the death condition of respectively organizing mouse, observed 5 days continuously, determine to cause in 3 days that the bacterial concentration of mouse 80～100% death is the infectation of bacteria minimum lethal concentration.
Experimental result, 5% yeast MC suspension 10-1 dilution streptococcus aureus (clinical strain 2-18) infecting mouse, mortality ratio reaches 80% in 3 days, is defined as this experiment infectation of bacteria minimum lethal concentration.
2, to the dead provide protection of lethality infection of staphylococcus aureus
110 of Kunming mouses are divided into 6 groups at random.Experimental mice (each 20) give Herba Scutellariae Barbatae total flavones 400,200,100mgkg-1 respectively, positive drug and infect that control group mice (each 20) gives SHUANGHUANGLIAN JIAONANG 624mgkg-1 respectively and with volume distilled water, 0.2ml10g-1w volume gastric infusion, 12 hours (± 2 hours) 1 time, for three days on end.Mouse makes lethal hit with 0.5ml/ abdominal injection of minimum lethal concentration streptococcus aureus more than the 4th day, infects the back and continues administration 4 times (2 days).Remain 10 mouse peritoneal bacterial injection diluents (no bacterium adds) in contrast.The dead mouse situation is respectively organized in 6 hours 1 time observation behind the infecting mouse, adds up dead mouse number in 48 hours, calculates mortality ratio (%).
Mortality ratio (%)=dead mouse number/total mice * 100%
The significance of animal dead rate difference between X2 check comparative group.
Experimental result sees Table 28.
Experimental result, behind the mouse infection streptococcus aureus 24,48 hours, infecting control group had 85% and 95% death respectively.The mouse death rate of 3 dosage groups of Herba Scutellariae Barbatae total flavones and 24,48 hours 2 time points of SHUANGHUANGLIAN KOUFUYE all is lower than the infection control group.Wherein, 24 hours mortality ratio of SHUANGHUANGLIAN KOUFUYE is minimum, but does not have significance (P＞0.05) with Herba Scutellariae Barbatae total flavones difference.Heavy dose of Herba Scutellariae Barbatae total flavones has significance (P＜0.05) with infection control group comparing difference; middle dosage group 24 hours and infection control group comparing difference have significance (P＜0.05); show that it has significant protective effect to the dead mouse that the lethality infection of staphylococcus aureus causes, and the doses dependency is arranged.5% yeast MC suspension intraperitoneal injection of mice does not have death.
Table 28 Herba Scutellariae Barbatae total flavones is to the dead provide protection of lethality infection of staphylococcus aureus
Group Dosage (mgkg -1) Example number (n) 24 hours death condition 48 hours death condition
The example number Mortality ratio (%) The example number Mortality ratio (%)
Infect dosage group flavones small dose group diluent control group in the heavy dose of group of the control group SHUANGHUANLIAN control group flavones flavones - 624 400 200 100 - 20 20 20 20 20 10 17 9 10 10 14 0 85 45** 50* 50* 70 0 19 14 13 16 17 0 95 70* 65* 80 85 0
Annotate: compare with the infection control group *P＜0.01; *P＜0.05.
Experimental example 9The anti-inflammatory action test
1, the Herba Scutellariae Barbatae total flavones p-Xylol causes the influence of mice auricle swelling reaction
60 of 26～30g male mice in kunming, random packet, experimental mice gives Herba Scutellariae Barbatae total flavones 400,200,100mgkg-1 respectively, positive drug contrast and the solvent control group mice gives Asprin 100mgkg-1, SHUANGHUANGLIAN JIAONANG 624mgkg-1 respectively and with volume distilled water, 0.2ml10g-1w volume gastric infusion, 12 hours (± 2 hours) 1 time, continuous 3 times.
After the last administration 60 minutes, dip in cotton balls and to get dimethylbenzene 50 μ l contact test mouse right ears two-sided 5 seconds, took off cervical vertebra in 15 minutes to put to death mouse, ears are downcut with the position homalographic with diameter 6mm punch tool, take by weighing left and right sides auricle weight, calculate auricle edema rate (%) and inhibitory rate of intumesce (%).
Auricle edema rate (%)=(auris dextra sheet weight-left auricle weight)/left auricle weight * 100%
Inhibitory rate of intumesce (%)=(control group swelling rate-experimental group swelling rate)/control group swelling rate * 100%
The t check is the significance of auricle edema rate (%) group difference relatively.Experimental result sees Table 29.
Table 29 Herba Scutellariae Barbatae total flavones p-Xylol causes the acutely inflamed restraining effect of Mice Auricle (X ± s)
Group Dosage (mgkg -1) Example number (n) Swelling rate (%) Inflammation inhibiting rate (%)
Flavones group small dose group in the heavy dose of group of solvent control group Asprin control group SHUANGHUANLIAN control group - 100 624 400 200 100 10 10 10 10 10 10 177.42±40.91 104.31±48.63** 122.86±42.94** 111.11±27.29** 120.65±64.24* 136.67±40.91*△ - 41.42 30.75 37.37 32.00 22.97
Annotate: compare with solvent *P＜0.01, *P＜0.05; △ and Asprin group be P＜0.05 relatively.
Experimental result, 100～400mgkg-1 Herba Scutellariae Barbatae total flavones gastric infusion p-Xylol induced mice auricle acute inflammatory reaction has restraining effect in various degree, with solvent control group comparing difference significance or highly significant (P＜0.05 or 0.01) are arranged, dose-dependence is more obvious.Wherein, heavy dose of group action intensity is higher than institute's amount of reagent SHUANGHUANGLIAN JIAONANG, and is close with Asprin, but difference does not have significance; Middle dosage group action intensity is close with institute's amount of reagent SHUANGHUANGLIAN JIAONANG.
2, Herba Scutellariae Barbatae total flavones causes the influence that the rat skin capillary permeability increases reaction to histamine
60 of Wistar rats, random packet, the experimental group rat gives Herba Scutellariae Barbatae total flavones 280,140,70mgkg-1 respectively, positive drug contrast and the solvent control rats gives Asprin 100mgkg-1, SHUANGHUANGLIAN JIAONANG 436.8mgkg-1 respectively and with volume distilled water, 1.0ml100g-1w volume gastric infusion, 12 hours (± 2 hours) 1 time, continuous 3 times.
After the last administration 40 minutes, each mouse abdominal injection 3% vetanarcol 0.133ml100g-1w anesthesia, back part intradermal injection in 20 minutes 0.01% histamine normal saline solution 0.1ml, sublingual vein is injected 1% Evans Blue normal saline solution 0.4ml100g-1w immediately, take off cervical vertebra after 30 minutes and put to death rat, measure the locus coeruleus diameter, downcut back indigo plant with diameter 1.5cm punch tool and dye skin graft, shred rearmounted 7: 3 acetone normal saline solution 5ml, 45 ℃ are soaked 96 hours until the blue completely dissolve of skin, centrifugal 15 minutes of 1500rpm/min gets supernatant liquor and measures the A620nm value, calculates inhibiting rate (%).
Locus coeruleus diameter inhibiting rate (%)=(experimental group diameter-control group diameter)/control group diameter * 100%
Ooze out inhibiting rate (%)=(control group A 620nm value-experimental group A620nm)/control group A 620nm * 100%
Locus coeruleus diameter and the different significance of skin graft immersion liquid A620nm value difference between t check comparative group; The uneven person of homoscedasticity analysis and judgement variance between laboratories, t, the significance of check comparative group differences.
Experimental result sees Table 30.
Table 30 Herba Scutellariae Barbatae total flavones to histamine cause the rat skin capillary permeability increase reaction influence ( )
Group Dosage (mgkg -1) Example number (n) Locus coeruleus diameter (cm) Inhibiting rate (%) Locus coeruleus skin graft immersion liquid A 620The nm value Inhibiting rate (%)
Dosage group small dose group in the heavy dose of group of solvent control group Asprin group SHUANGHUANGLIAN JIAONANG group - 100 436.8 280 140 70 10 10 10 10 10 10 1.23±0.07 1.08±0.13 ** 1.07±0.07 ** 1.05±0.04 ** 1.01±0.11 ** 1.07±0.08 ** - 12.20 13.41 15.04 17.89 13.01 0.78±0.26 0.50±0.16 ** 0.53±0.19 * 0.47±0.15 ** 0.50±0.19 * 0.71±0.17 - 36.40 31.71 39.66 35.92 8.97
Annotate: compare with solvent *P＜0.05, *P＜0.01.
Experimental result, the Herba Scutellariae Barbatae total flavones gastric infusion of 3 dosage causes the rat skin capillary permeability to histamine and increases reaction in various degree restraining effect is all arranged, with solvent control group comparing difference significance or highly significant (P＜0.05 or 0.01) are arranged respectively except that small dose group locus coeruleus skin graft immersion liquid A value, locus coeruleus skin graft immersion liquid A value has dosage correlation.Wherein, the heavy dose of group of flavones action intensity is higher than institute's amount of reagent Asprin and SHUANGHUANGLIAN JIAONANG, but difference does not have significance (P＞0.05); Middle dosage group action intensity is close with institute's amount of reagent Asprin, is higher than SHUANGHUANGLIAN JIAONANG.
Experimental example 10The refrigeration function test
80 of Wistar rats, 20 ± 1 ℃ of room temperatures adapt to 7 days; Test preceding 3 days adaptability and measure the rat rectal temperature, select that basal body temperature is normal, 60 of the temperature difference＜0.3 ℃ qualified rats, random packet, every group 12, the experimental group rat gives Herba Scutellariae Barbatae total flavones 70,140,280mgkg-1 respectively, positive drug control group and solvent control rats give Asprin 100mgkg-1 respectively or with volume distilled water, 1.0mlkg-1w gastric infusion, 12 hours (± 2 hours) No. 1 meter 3 times (the 3rd administration time be after the pyrogenicity 6 hours) at interval; Experiment is when 1 hour thermometric of day interval 2 times, and averaging is (basis) body temperature before the pyrogenicity; Every mouse back subcutaneous injection 10% yeast MC suspension 5.0mlkg-1; Thermometric 6 hours time the after the pyrogenicity is rejected temperature and is raise behind＜0.8 ℃ of rat, and each organizes rat (the 3rd time) the same dosage, volume administration once more, behind the medicine every No. 1 meter of 1 hour thermometric 3 times.
Each organizes data after test for normality, respectively organizes behind the medicine each time point and the significance of 6 hours body temperature differences of pyrogenicity difference on the same group with t check, and the uneven person of homoscedasticity analysis and judgement variance between laboratories uses t instead, check.
Experimental result sees Table 31.
Experimental result, 70～280mgkg-1 Herba Scutellariae Barbatae total flavones gastric infusion has in various degree restraining effect to the exothermic reaction of rat due to the yeast, and certain dosage correlation arranged, with solvent control group comparing difference significance (P＜0.05) was arranged in 1,3 hour behind wherein heavy dose of each time point of group and the middle dosage group medicine; Heavy dose of group action intensity is higher than the Asprin of institute's amount of reagent, but there was no significant difference (P＞0.0
Table 31 Herba Scutellariae Barbatae total flavones to the refrigeration function of rat yeast pyrogenicity (
Figure C20041008883300281
)
Group Dosage (mgkg -1) Example number (n) Basal body temperature (℃) 6h after the pyrogenicity (℃) Body temperature behind the medicine (℃)/with pyrogenicity after 6 little time difference value (℃)
1h behind the medicine 2h behind the medicine 3h behind the medicine
Dosage group flavones small dose group in the heavy dose of group of the solvent control group Asprin contrast flavones flavones -group 100 280 140 70 10 9 10 8 8 38.09±0.40 37.87±0.41 37.94±0.24 37.83±0.44 38.03±0.94 39.22±0.42 38.94±0.36 39.34±0.36 39.05±0.51 39.51±0.63 39.42±0.50 0.20±0.66 38.22±0.37 -0.71±0.38 ** 38.39±1.16 -0.95±1.22 * 38.48±0.68 -0.58±0.72 * 39.20±0.63 -0.31±0.56 39.25±0.49 0.03±0.65 38.47±0.49 -0.48±0.66 38.34±0.96 -1.00±1.03 *38.53±0.68 -0.53±0.60 39.56±0.82 0.05±0.84 39.28±0.30 0.06±0.37 38.22±0.25 -0.72±0.38 **38.52±1.11 -0.82±1.21 *38.76±0.54 -0.30±0.30 *39.56±0.40 0.05±0.46
Annotate: compare with the solvent control group *P＜0.05, *P＜0.01.
Experimental result, 3 dosage groups of Herba Scutellariae Barbatae total flavones all have refrigeration function in various degree, and it is relevant to be dosage, and heavy dose of group intensity is higher than institute's amount of reagent Asprin, and acting duration is behind the medicine more than 3 hours.
Description of drawings
Fig. 1 linear relationship is investigated
Fig. 2 typical curve
Embodiment 1:
Get 10 kilograms of Herba Scutellariae Barbatae meal, adding 10 times of boiling water boiled 1 hour, filter, the dregs of a decoction are carried 1 hour with 8 times of poach, filter, merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2, centrifugal, get the macroporous resin of supernatant liquor, pre-treatment, dress post with same medicinal material amount, said extracted liquid is crossed post absorption, resin and crude drug weight ratio are 1: 2, and washing is again with 85% above ethanol elution, and the volume of eluting solvent is 6 times of weight resin, reclaim ethanol, drying, promptly get Herba Scutellariae Barbatae total flavones; Make 1000 of injections according to ordinary method, every 10ml, once a day, each one.Be applicable to upper respiratory tract infection diseases such as the acute pharyngitis that causes by viral cold, acute tonsillitis.
Embodiment 2:
Get 10 kilograms of Herba Scutellariae Barbatae meal, adding 8 times of boiling water boiled 2 hours, filter, the dregs of a decoction are carried 2 hours with 6 times of poach, filter, merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH3, centrifugal, get the AB-8 type macroporous resin of supernatant liquor, pre-treatment, dress post with same medicinal material amount, said extracted liquid is crossed post absorption, resin and crude drug weight ratio are 1: 8, and washing is again with 85% above ethanol elution, and the volume of eluting solvent is 8 times of weight resin, reclaim ethanol, drying, promptly get Herba Scutellariae Barbatae total flavones; Make oral liquid 10000ml according to ordinary method, each 10ml, every day 2-3 time.Be applicable to influenza.
Embodiment 3
Get 10 kilograms of Herba Scutellariae Barbatae meal, add 9 times of boiling water and boiled 2.5 hours, filter, the dregs of a decoction are carried 1.5 hours with 7 times of poach, filter, and merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH5, centrifugal, get the macroporous resin of supernatant liquor, pre-treatment with same medicinal material amount, the dress post is crossed post absorption with said extracted liquid, and resin and crude drug weight ratio are 1: 6, washing is again with 85% above ethanol elution, the volume of eluting solvent is 5 times of weight resin, reclaims ethanol, add 3 times of water dilutions, 10% sodium hydroxide is transferred pH value 9, centrifugal, supernatant liquor is transferred pH value 2 with hydrochloric acid, leaves standstill 2 hours, and is centrifugal, drying precipitate promptly gets Herba Scutellariae Barbatae total flavones.Make granule 1kg according to ordinary method, each 10 grams, every day 3 times.Be applicable to influenza.
Embodiment 4
Get 10 kilograms of Herba Scutellariae Barbatae meal, add 7 times of boiling water and boiled 3 hours, filter, the dregs of a decoction are carried 1 hour with 8 times of poach, filter, and merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH6, centrifugal, get the macroporous resin of supernatant liquor, pre-treatment with same medicinal material amount, the dress post is crossed post absorption with said extracted liquid, and resin and crude drug weight ratio are 1: 4, washing is again with 85% above ethanol elution, the volume of eluting solvent is 6 times of weight resin, reclaims ethanol, add 3 times of water dilutions, 10% sodium hydroxide is transferred pH value 9, centrifugal, supernatant liquor is transferred pH value 2 with hydrochloric acid, leaves standstill 3 hours, and is centrifugal, drying precipitate promptly gets general flavone content 80% above Herba Scutellariae Barbatae total flavones.Make 4000 in tablet according to ordinary method, each 4, every day 3 times.Be applicable to upper respiratory tract infection diseases such as the acute pharyngitis that causes by viral cold, acute tonsillitis.
Embodiment 5
Get 10 kilograms of Herba Scutellariae Barbatae meal, add 8 times of boiling water and boiled 1 hour, filter, the dregs of a decoction are carried 2 hours with 6 times of poach, filter, and merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2, centrifugal, get the macroporous resin of supernatant liquor, pre-treatment with same medicinal material amount, the dress post is crossed post absorption with said extracted liquid, and resin and crude drug weight ratio are 1: 4, washing is again with 85% above ethanol elution, the volume of eluting solvent is 8 times of weight resin, reclaims ethanol, add 4 times of water dilutions, 10% sodium hydroxide is transferred pH value 8, centrifugal, supernatant liquor is transferred pH value 2 with hydrochloric acid, leaves standstill 2 hours, and is centrifugal, drying precipitate promptly gets general flavone content 80% above Herba Scutellariae Barbatae total flavones.Make 3000 of capsules according to ordinary method, each 3, every day 3 times.Be applicable to upper respiratory tract infection diseases such as the acute pharyngitis that causes by viral cold, acute tonsillitis.
Embodiment 6:The assay of Herba Scutellariae Barbatae total flavones
Get the scutellarin reference substance, add methyl alcohol and make the solution that every 1ml contains 0.20mg, shake up, promptly get reference substance solution; The accurate reference substance solution 0.4 of drawing, 0.6,0.8,1.0,1.2ml, adding methyl alcohol respectively to 10ml, is blank with methyl alcohol, according to spectrophotometry (an appendix VI of Chinese Pharmacopoeia version in 2000 A), wavelength place at 285nm measures optical density, with concentration is X-coordinate, and optical density is an ordinate zou, the drawing standard curve; Be taken at 80 ℃ of this Herba Scutellariae Barbatae total flavones 20mg that are dried to constant weight, put in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, supersound process 30 minutes, shake up, filter, the accurate subsequent filtrate 1ml that draws, put in the 10ml measuring bottle, add methyl alcohol, shake up to scale, the accurate 1ml that draws puts in the 10ml measuring bottle, adds methyl alcohol to scale, shaking up, is blank with methyl alcohol, according to spectrophotometry (an appendix VI of Chinese Pharmacopoeia version in 2000 A), wavelength place at 285nm measures optical density, calculates, promptly; This Herba Scutellariae Barbatae total flavones contains total flavones with scutellarin (C 21H 18O 12) meter, must not be less than 51%.
Embodiment 7The content assaying method of scutellarin in the Herba Scutellariae Barbatae total flavones:
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).
With octadecylsilane chemically bonded silica is weighting agent, and methanol-water-formic acid of 38: 62: 0.05 is moving phase, and the detection wavelength is 335nm; Number of theoretical plate is pressed the scutellarin peak and is calculated, and is not less than 1500; Precision takes by weighing the scutellarin reference substance, adds moving phase and makes the solution that contains scutellarin 0.20mg among every 1ml, shakes up, and promptly gets reference substance solution; Get this Herba Scutellariae Barbatae total flavones 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, and it is an amount of to add methyl alcohol, adds methyl alcohol to scale after the dissolving in ultrasonic 30 minutes, shakes up, and promptly gets need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, calculate, promptly; This Herba Scutellariae Barbatae total flavones contains scutellarin must not be less than 2%.
Embodiment 8The quality controlling means of Herba Scutellariae Barbatae total flavones:
1. the assay of Herba Scutellariae Barbatae total flavones:
Get the scutellarin reference substance, add methyl alcohol and make the solution that every 1ml contains 0.20mg, shake up, promptly get reference substance solution; The accurate reference substance solution 0.4 of drawing, 0.6,0.8,1.0,1.2ml, adding methyl alcohol respectively to 10ml, is blank with methyl alcohol, according to spectrophotometry (an appendix VI of Chinese Pharmacopoeia version in 2000 A), wavelength place at 285nm measures optical density, with concentration is X-coordinate, and optical density is an ordinate zou, the drawing standard curve; Be taken at 80 ℃ of this Herba Scutellariae Barbatae total flavones 20mg that are dried to constant weight, put in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, supersound process 30 minutes, shake up, filter, the accurate subsequent filtrate 1ml that draws, put in the 10ml measuring bottle, add methyl alcohol, shake up to scale, the accurate 1ml that draws puts in the 10ml measuring bottle, adds methyl alcohol to scale, shaking up, is blank with methyl alcohol, according to spectrophotometry (an appendix VI of Chinese Pharmacopoeia version in 2000 A), wavelength place at 285nm measures optical density, calculates, promptly; This Herba Scutellariae Barbatae total flavones contains total flavones with scutellarin (C 21H 18O 12) meter, must not be less than 51%.
2. the content assaying method of scutellarin in the Herba Scutellariae Barbatae total flavones:
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).
With octadecylsilane chemically bonded silica is weighting agent, and methanol-water-formic acid of 38: 62: 0.05 is moving phase, and the detection wavelength is 335nm; Number of theoretical plate is pressed the scutellarin peak and is calculated, and is not less than 1500; Precision takes by weighing the scutellarin reference substance, adds moving phase and makes the solution that contains scutellarin 0.20mg among every 1ml, shakes up, and promptly gets reference substance solution; Get this Herba Scutellariae Barbatae total flavones 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, and it is an amount of to add methyl alcohol, adds methyl alcohol to scale after the dissolving in ultrasonic 35 minutes, shakes up, and promptly gets need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, calculate, promptly; This Herba Scutellariae Barbatae total flavones contains scutellarin must not be less than 2%.

Claims (13)

1. the preparation method of a Herba Scutellariae Barbatae total flavones is characterized in that this method is:
Get the Herba Scutellariae Barbatae meal, adding 6-10 times of boiling water boiled 1-3 hour, filter, the dregs of a decoction are carried 1-2 hour with 6-8 times of poach, filter, merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2～6, centrifugal, get supernatant liquor, with the macroporous resin pre-treatment of same medicinal material amount, the dress post, said extracted liquid is crossed post absorption, resin and crude drug weight ratio are 1: 2-8, and washing is again with 85% above ethanol elution, and the volume of eluting solvent is 4-8 a times of weight resin, reclaim ethanol, drying, promptly get Herba Scutellariae Barbatae total flavones.
2. the preparation method of Herba Scutellariae Barbatae total flavones as claimed in claim 1 is characterized in that this method is:
Get the Herba Scutellariae Barbatae meal, add 10 times of boiling water and boiled 1 hour, filter, the dregs of a decoction are carried 1 hour with 8 times of poach, filter, merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2, centrifugal, get supernatant liquor, with the macroporous resin pre-treatment of same medicinal material amount, the dress post, said extracted liquid is crossed post absorption, and resin and crude drug weight ratio are 1: 2, and washing is again with 85% above ethanol elution, the volume of eluting solvent is 6 times of weight resin, reclaims ethanol, drying, promptly gets Herba Scutellariae Barbatae total flavones.
3. the preparation method of Herba Scutellariae Barbatae total flavones as claimed in claim 1 is characterized in that this method is:
Get the Herba Scutellariae Barbatae meal, adding 8 times of boiling water boiled 2 hours, filter, the dregs of a decoction are carried 2 hours with 6 times of poach, filter, merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH3, centrifugal, get supernatant liquor, with the AB-8 type macroporous resin pre-treatment of same medicinal material amount, the dress post, said extracted liquid is crossed post absorption, resin and crude drug weight ratio are 1: 8, and washing is again with 85% above ethanol elution, and the volume of eluting solvent is 8 times of weight resin, reclaim ethanol, drying, promptly get Herba Scutellariae Barbatae total flavones.
4. the preparation method of a Herba Scutellariae Barbatae total flavones is characterized in that this method is:
Get the Herba Scutellariae Barbatae meal, add 6-10 times of boiling water and boiled 1-3 hour, filter, the dregs of a decoction are carried 1-2 hour with 6-8 times of poach, filter, and merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2～6, centrifugal, get supernatant liquor, with the macroporous resin pre-treatment of same medicinal material amount, the dress post is crossed post absorption with said extracted liquid, and resin and crude drug weight ratio are 1: 2-8, washing is again with 85% above ethanol elution, and the volume of eluting solvent is 4-8 a times of weight resin, reclaims ethanol, drying; Add 3-5 times of water dilution, 10% sodium hydroxide adjust pH 8-10, centrifugal, supernatant liquor leaves standstill 1-4 hour with hydrochloric acid adjust pH 1-3, and centrifugal, drying precipitate promptly gets general flavone content 80% above Herba Scutellariae Barbatae total flavones, and wherein scutellarin content is 10%.
5. the preparation method of Herba Scutellariae Barbatae total flavones as claimed in claim 4 is characterized in that this method is:
Get the Herba Scutellariae Barbatae meal, add 10 times of boiling water and boiled 1 hour, filter, the dregs of a decoction are carried 1 hour with 8 times of poach, filter, and merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH2, centrifugal, get supernatant liquor, with the macroporous resin pre-treatment of same medicinal material amount, the dress post, said extracted liquid is crossed post absorption, and resin and crude drug weight ratio are 1: 2, and washing is again with 85% above ethanol elution, the volume of eluting solvent is 6 times of weight resin, reclaim ethanol, add 3 times of water dilutions, 10% sodium hydroxide adjust pH 9, centrifugal, supernatant liquor hydrochloric acid adjust pH 2, leave standstill 2 hours, centrifugal, drying precipitate; Add 3 times of water dilutions, 10% sodium hydroxide adjust pH 9, centrifugal, supernatant liquor leaves standstill 3 hours with hydrochloric acid adjust pH 2, and centrifugal, drying precipitate promptly gets general flavone content 80% above Herba Scutellariae Barbatae total flavones, and wherein scutellarin content is 10%.
6. the preparation method of Herba Scutellariae Barbatae total flavones as claimed in claim 4 is characterized in that this method is:
Get the Herba Scutellariae Barbatae meal, add 8 times of boiling water and boiled 2 hours, filter, the dregs of a decoction are carried 2 hours with 6 times of poach, filter, and merge filtrate twice, be concentrated into 3 times of medicinal material amounts, centrifugal, get supernatant liquor and add hydrochloric acid accent pH3, centrifugal, get supernatant liquor, with the AB-8 type macroporous resin pre-treatment of same medicinal material amount, the dress post is crossed post absorption with said extracted liquid, and resin and crude drug weight ratio are 1: 8, washing is again with 85% above ethanol elution, and the volume of eluting solvent is 8 times of weight resin, reclaims ethanol; Add 4 times of water dilutions, 10% sodium hydroxide adjust pH 8, centrifugal, supernatant liquor leaves standstill 2 hours with hydrochloric acid adjust pH 2, and centrifugal, drying precipitate promptly gets general flavone content 80% above Herba Scutellariae Barbatae total flavones, and wherein scutellarin content is 10%.
7. the quality controlling means of a Herba Scutellariae Barbatae total flavones is characterized in that comprising in this method following content assaying method:
A, total flavones
Get the scutellarin reference substance, add methyl alcohol and make the solution that every 1ml contains 0.20mg, shake up, promptly get reference substance solution; The accurate reference substance solution 0.4,0.6,0.8,1.0 of drawing, 1.2ml adds methyl alcohol respectively to 10ml, is blank with methyl alcohol, according to spectrophotometry, measures optical density at the wavelength place of 285nm, is X-coordinate with concentration, and optical density is an ordinate zou, the drawing standard curve; Be taken at 80 ℃ of Herba Scutellariae Barbatae total flavones 20mg that are dried to constant weight, put in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, supersound process 30 minutes, shake up, filter, the accurate subsequent filtrate 1ml that draws, put in the 10ml measuring bottle, add methyl alcohol, shake up to scale, the accurate 1ml that draws puts in the 10ml measuring bottle, adds methyl alcohol to scale, shaking up, is blank with methyl alcohol, according to spectrophotometry, wavelength place at 285nm measures optical density, calculates, promptly; Herba Scutellariae Barbatae total flavones contains total flavones with scutellarin C 21H 18O 12Meter must not be less than 51%;
B, scutellarin
According to high effective liquid chromatography for measuring; With octadecylsilane chemically bonded silica is weighting agent, and methanol-water-formic acid of 38: 62: 0.05 is moving phase, and the detection wavelength is 335nm; Number of theoretical plate is pressed the scutellarin peak and is calculated, and is not less than 1500; Precision takes by weighing the scutellarin reference substance, adds moving phase and makes the solution that contains scutellarin 0.20mg among every 1ml, shakes up, and promptly gets reference substance solution; Get Herba Scutellariae Barbatae total flavones 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, and it is an amount of to add methyl alcohol, adds methyl alcohol to scale after the dissolving in ultrasonic 20-40 minute, shakes up, and promptly gets need testing solution; Draw each 10 μ 1 of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, calculate, promptly; Herba Scutellariae Barbatae total flavones contains scutellarin must not be less than 2%.
8. quality controlling means as claimed in claim 7 is characterized in that the content of total flavone measuring method is in this method:
Get the scutellarin reference substance, add methyl alcohol and make the solution that every 1ml contains 0.20mg, shake up, promptly get reference substance solution; The accurate reference substance solution 0.4,0.6,0.8,1.0 of drawing, 1.2ml adds methyl alcohol respectively to 10ml, is blank with methyl alcohol, according to spectrophotometry, measures optical density at the wavelength place of 285nm, is X-coordinate with concentration, and optical density is an ordinate zou, the drawing standard curve; Be taken at 80 ℃ of Herba Scutellariae Barbatae total flavones 20mg that are dried to constant weight, put in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, supersound process 30 minutes, shake up, filter, the accurate subsequent filtrate 1ml that draws, put in the 10ml measuring bottle, add methyl alcohol, shake up to scale, the accurate 1ml that draws puts in the 10ml measuring bottle, adds methyl alcohol to scale, shaking up, is blank with methyl alcohol, according to spectrophotometry, wavelength place at 285nm measures optical density, calculates, promptly; Herba Scutellariae Barbatae total flavones contains total flavones with scutellarin C 21H 18O 12Meter must not be less than 51%.
9. quality controlling means as claimed in claim 7 is characterized in that the content assaying method of scutellarin in this method is:
According to high effective liquid chromatography for measuring; With octadecylsilane chemically bonded silica is weighting agent, and methanol-water-formic acid of 38: 62: 0.05 is moving phase, and the detection wavelength is 335nm; Number of theoretical plate is pressed the scutellarin peak and is calculated, and is not less than 1500; Precision takes by weighing the scutellarin reference substance, adds moving phase and makes the solution that contains scutellarin 0.20mg among every 1ml, shakes up, and promptly gets reference substance solution; Get Herba Scutellariae Barbatae total flavones 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, and it is an amount of to add methyl alcohol, adds methyl alcohol to scale after the dissolving in ultrasonic 30 minutes, shakes up, and promptly gets need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, calculate, promptly; Herba Scutellariae Barbatae total flavones contains scutellarin must not be less than 2%.
10. quality controlling means as claimed in claim 7 is characterized in that comprising in this method following content assaying method and is:
A. the assay of Herba Scutellariae Barbatae total flavones:
Get the scutellarin reference substance, add methyl alcohol and make the solution that every 1ml contains 0.20mg, shake up, promptly get reference substance solution; The accurate reference substance solution 0.4,0.6,0.8,1.0 of drawing, 1.2ml adds methyl alcohol respectively to 10ml, is blank with methyl alcohol, according to spectrophotometry, measures optical density at the wavelength place of 285nm, is X-coordinate with concentration, and optical density is an ordinate zou, the drawing standard curve; Be taken at 80 ℃ of Herba Scutellariae Barbatae total flavones 20mg that are dried to constant weight, put in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, supersound process 30 minutes, shake up, filter, the accurate subsequent filtrate 1ml that draws, put in the 10ml measuring bottle, add methyl alcohol, shake up to scale, the accurate 1ml that draws puts in the 10ml measuring bottle, adds methyl alcohol to scale, shaking up, is blank with methyl alcohol, according to spectrophotometry, wavelength place at 285nm measures optical density, calculates, promptly; Herba Scutellariae Barbatae total flavones contains total flavones with scutellarin C 21H 18O 12Meter must not be less than 51%;
The content assaying method of scutellarin in b, the Herba Scutellariae Barbatae total flavones:
According to high effective liquid chromatography for measuring; With octadecylsilane chemically bonded silica is weighting agent, and methanol-water-formic acid of 38: 62: 0.05 is moving phase, and the detection wavelength is 335nm; Number of theoretical plate is pressed the scutellarin peak and is calculated, and is not less than 1500; Precision takes by weighing the scutellarin reference substance, adds moving phase and makes the solution that contains scutellarin 0.20mg among every 1ml, shakes up, and promptly gets reference substance solution; Get Herba Scutellariae Barbatae total flavones 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, and it is an amount of to add methyl alcohol, adds methyl alcohol to scale after the dissolving in ultrasonic 35 minutes, shakes up, and promptly gets need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, calculate, promptly; Herba Scutellariae Barbatae total flavones contains scutellarin must not be less than 2%.
11. Herba Scutellariae Barbatae total flavones has application in antivirus action, anti-microbial effect, anti-inflammatory action, refrigeration function or the analgesic medicine in preparation.
12. the application of Herba Scutellariae Barbatae total flavones in preparation treatment respiratory tract infection medicine.
13. the application of Herba Scutellariae Barbatae total flavones as claimed in claim 12 in preparation treatment respiratory tract infection medicine is characterized in that described respiratory tract infection is meant acute pharyngitis, acute tonsillitis or virus pneumonia.
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Publication number Priority date Publication date Assignee Title
CN101167799B (en) * 2007-10-19 2012-09-05 复旦大学 Scutellaria barbata total flavone and its application in preparing influenza virus resisting medicine
CN102552359A (en) * 2008-06-27 2012-07-11 四川大学 Preparation method of Erigeron multiradiatus extractive
CN101623326B (en) * 2009-07-30 2011-07-27 扬州大学 Sculellaria barbata formulation and preparation method thereof
CN102516218A (en) * 2011-12-20 2012-06-27 苏州宝泽堂医药科技有限公司 Method for preparing herba scutellariae barbatae with high purity
CN102688291B (en) * 2012-06-01 2014-07-09 康阳润和（北京）医药科技有限公司 Sculellaria barbata general flavone compound, oral preparations prepared from Sculellaria barbata general flavone compound and preparation methods respectively for Sculellaria barbata general flavone compound and oral preparations
CN104277086B (en) * 2013-07-12 2018-08-28 河北以岭医药研究院有限公司 Scutellarin extracting method in a kind of Sculellaria barbata
CN104644744A (en) * 2014-10-21 2015-05-27 中山大学 Scutellaria barbata extract as well as preparation method and application thereof
CN113310979A (en) * 2021-05-27 2021-08-27 南京益唯森生物科技有限公司 Method for accurately and rapidly determining total flavone content in scutellaria baicalensis medicinal material and product thereof
CN113599416A (en) * 2021-06-11 2021-11-05 宋坪 New application of sculellaria barbata extract

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
中国中药杂志. 王文蜀等，957-959，半枝莲中黄酮类化学成分研究. 2004 *
中成药. 邸多隆等，530-532，半枝莲提取工艺的研究. 2003 *
华西药学杂志. 袁崇均等，112-113，正交试验法优选半枝莲提取工艺的研究. 2002 *

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