CN104739902A - Preparation method of glabrous sarcandra herb injection - Google Patents
Preparation method of glabrous sarcandra herb injection Download PDFInfo
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Abstract
The invention discloses a preparation method of a glabrous sarcandra herb injection. The preparation method comprises the following steps: taking 5,000-10,000g of glabrous sarcandra herb, adding water, extracting 2-3 times, merging extract liquids, filtering, concentrating the filtrate until the relative density is 1.25-1.40 at 70 DEG C; adding ethanol to precipitate until the ethanol content is 60%-90%; refrigerating for 24-72 hours, filtering, recovering ethanol from the filtrate and concentrating until each ml of filtrate contains 5-15g of raw medicine; adjusting the pH value to 3-5, heating to 0.5-2.0 hours, refrigerating for 12-48 hours, filtering, adjusting the pH value of the filtrate to 5-7, and heating until the content ratio of chlorogenic acid to cryptochlorogenic acid in the filtrate is (3:1) to (3:2); adding a proper amount of activated carbon, filtering, adding injection water to the filtrate, and diluting into 1,000ml; and filtering, adding 3-10ml of polysorbate 80, stirring evenly, adjusting the pH value to 5.5-7.0, filtering, filling and sterilizing to obtain the glabrous sarcandra herb injection.
Description
Technical field
The present invention relates to the field of Chinese medicines, particularly a kind of preparation method of ZHONGJIEFENG ZHUSHEYE.
Background technology
ZHONGJIEFENG ZHUSHEYE is recorded in the Sanitation Ministry medicine standard Traditional Chinese medicine historical preparation the 14, and it is through extracting the sterile water solution made by Herba Sarcandrae.This product has the effect of heat-clearing and toxic substances removing, dispersing swelling and dissipating binds, and can be used for pneumonia, appendicitis caused by treatment hyperactivity of toxic heat, cellulitis, bacillary dysentery, abscess, combine with ZHONGJIEFENG PIAN and also can be used for treating the tumors such as digestive tract cancer, cancer of pancreas, hepatocarcinoma.This product antiinflammatory action is strong, but treatment function of tumor need to improve.Applicant finds in research process, and measure according to the chromatographic condition under version " Chinese Pharmacopoeia " the Herba Sarcandrae extractum characteristic pattern spectral term that records in 2010, in ZHONGJIEFENG ZHUSHEYE, the content of rosmarinic acid and chlorogenic acid is considerably less, and production process loss is serious.According to bibliographical information, rosmarinic acid and chlorogenic acid all have very strong physiologically active, but structural instability.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of ZHONGJIEFENG ZHUSHEYE, preparation method of the present invention is more scientific and reasonable, and drug quality is stablized, and curative effect is better.
ZHONGJIEFENG ZHUSHEYE provided by the invention, it is through extracting the sterile water solution made by Herba Sarcandrae, every ml soln is no less than 100 μ g containing rosmarinic acid, total amount containing chlorogenic acid and 4-dicaffeoylquinic acid is no less than 500 μ g, ratio between chlorogenic acid and 4-dicaffeoylquinic acid content is 2:1 ~ 5:4, and solution pH value is 5.0 ~ 5.5.
ZHONGJIEFENG ZHUSHEYE provided by the invention, preparation method is as follows:
Get Herba Sarcandrae 5000-10000g, extracting in water 2-3 time, merge extractive liquid, filter, when filtrate is concentrated into 70 DEG C, relative density is 1.25-1.40, add alcohol settling, alcohol content is made to be 60-90%, cold preservation 24-72 hour, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 5-15g, adjust pH value to 3-5, heating 0.5-2.0 hour, cold preservation 12-48 hour, filter, filtrate adjusts pH value to 5.5-7.0, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 3:1-3:2, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 3-10ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH value to 5.5-7.0, filter, embedding, sterilizing, obtain.
The number of times adding alcohol settling in the preparation method of ZHONGJIEFENG ZHUSHEYE provided by the invention is 2-3 time.
The number of times heating chilling treatment in the preparation method of ZHONGJIEFENG ZHUSHEYE provided by the invention is 1-3 time.
ZHONGJIEFENG ZHUSHEYE provided by the invention can also carry out filling nitrogen process before embedding.
Innovation of the present invention is: instant invention overcomes former technique and adopt heat treated under alkali condition (PH=9) in order to make end product quality stable, cause the problem that in solution, rosmarinic acid and chlorogenic acid are seriously damaged.ZHONGJIEFENG ZHUSHEYE rosmarinic acid provided by the invention and chlorogenic acid content high, and stable system, curative effect is also better than former preparation.
Detailed description of the invention
Following embodiment for illustration of the present invention, but is not used for limiting the scope of the invention.
Embodiment 1
Get Herba Sarcandrae 5000g, add 10 times amount soak by water secondaries, each 2 hours, collecting decoction, filter, it is 1.33 (70 DEG C) that filtrate is concentrated into relative density, add alcohol settling secondary, first time makes alcohol content be 70%, second time alcohol content is 80%, each cold preservation 48 hours, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 10g, pH value to 4 is adjusted with hydrochloric acid solution, reflux 1 hour, cold preservation 24 hours, filter, filtrate reflux 1 hour again, cold preservation 24 hours, filter, filtrate adjusts pH value to 6.0 with sodium hydroxide solution, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 2:1, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 6ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH value to 5.5, filter, embedding, sterilizing, obtain.
Embodiment 2
Get Herba Sarcandrae 10000g, add 10 times amount soak by water three times, each 2 hours, collecting decoction, filter, it is 1.25 (70 DEG C) that filtrate is concentrated into relative density, add alcohol settling secondary, first time makes alcohol content be 70%, second time alcohol content is 80%, each cold preservation 72 hours, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 15g, pH value to 3 is adjusted with hydrochloric acid solution, 115 DEG C of heat treatments 0.5 hour, cold preservation 48 hours, filter, filtrate adjusts pH value to 5.5 with sodium hydroxide solution, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 3:1, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 10ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH value to 6.0, filter, fill nitrogen, embedding, sterilizing, obtain.
Embodiment 3
Get Herba Sarcandrae 5000g, add 8 times of water gaging heating and refluxing extraction three times, each 2 hours, merge extractive liquid, filter, it is 1.4 (70 DEG C) that filtrate is concentrated into relative density, add alcohol settling three times, first time makes alcohol content be 60%, second time alcohol content is 70%, second time alcohol content is 90%, each cold preservation 24 hours, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 5g, adjust pH value to 5, reflux 1 hour, cold preservation 12 hours, filter, filtrate reflux 1 hour, cold preservation 24 hours, filter, filtrate reflux 1 hour, cold preservation 48 hours, filter, filtrate adjusts pH value to 6.5 with sodium hydroxide solution, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 3:1, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 3ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH value to 6.5, filter, embedding, sterilizing, obtain.
Embodiment 4
Get Herba Sarcandrae 10000g, add 10 times amount soak by water secondaries, each 2 hours, collecting decoction, filter, it is 1.3 (70 DEG C) that filtrate is concentrated into relative density, add alcohol settling secondary, first time makes alcohol content be 70%, second time alcohol content is 80%, each cold preservation 48 hours, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 10g, pH value to 4 is adjusted with hydrochloric acid solution, 115 DEG C of heat treatments 1 hour, cold preservation 48 hours, filter, filtrate adjusts pH value to 7.0 with sodium hydroxide solution, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 3:1, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 6ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH value to 7.0, filter, fill nitrogen, embedding, sterilizing, obtain.
Embodiment 5
Get Herba Sarcandrae 5000g, add 10 times of water gaging heating and refluxing extraction three times, extraction time is respectively 3 hours, 3 hours, 2 hours, merge extractive liquid, filter, it is 1.35 (70 DEG C) that filtrate is concentrated into relative density, add alcohol settling secondary, first time makes alcohol content be 70%, second time alcohol content is 85%, each cold preservation 48 hours, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 10g, pH value to 4 is adjusted with hydrochloric acid solution, reflux 1 hour, cold preservation 24 hours, filter, filtrate reflux 1 hour again, cold preservation 24 hours, filter, filtrate adjusts pH value to 6.2 with sodium hydroxide solution, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 3:2, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 6ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH value to 5.5, filter, fill nitrogen, embedding, sterilizing, obtain.
Comparative example 1
Get Herba Sarcandrae 5000g, add 10 times amount soak by water secondaries, each 2 hours, collecting decoction, filter, it is 1.33 (70 DEG C) that filtrate is concentrated into relative density, add alcohol settling secondary, first time makes alcohol content be 70%, second time alcohol content is 80%, each cold preservation 48 hours, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 10g, add the egg protein solution of appropriate new preparation, stir, make precipitation, cold preservation 48 hours, filter, filtrate is boiled, excessive egg protein is solidified, filter, filtrate adds ethanol, making containing amount of alcohol is 75%, place precipitation, filter, filtrate recycling ethanol, inject with water appropriate, cold preservation 24 hours, filter, filtrate is by sodium hydroxide solution adjust ph to 9.0, add active carbon make be 0.05% concentration, boil 30 minutes, filter, filtrate injecting is diluted with water to 1000ml, filter, add 6ml polyoxyethylene sorbitan monoleate, stir evenly, filter, embedding, sterilizing, obtain.
Experimental example 1: sample detection
Indices detection is carried out according to the detection method of Pharmacopoeia of the People's Republic of China version in 2010 annex.Wherein:
The content assaying method of chlorogenic acid, 4-dicaffeoylquinic acid and rosmarinic acid is as follows:
Chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filler; With acetonitrile (containing 0.1% formic acid) for mobile phase A, with 0.1% formic acid for Mobile phase B, the volume ratio adopting gradient elution: 0min → 5min → 60min → 62min → 70min, A and B is 8:92 → 8:92 → 35:65 → 100:0 → 100:0; Determined wavelength is 330nm;
It is appropriate that chlorogenic acid reference substance, 4-dicaffeoylquinic acid reference substance and rosmarinic acid reference substance are got in the preparation of reference substance solution, accurately weighed, put in brown measuring bottle, add 60% methanol and make the solution of every 1ml containing chlorogenic acid 60 μ g, 4-dicaffeoylquinic acid 35 μ g and rosmarinic acid 25 μ g, to obtain final product;
The preparation precision of need testing solution measures this product 2ml, puts in the brown measuring bottle of 10ml, adds 60% methanol dilution to scale, shakes up, and filters, gets subsequent filtrate, to obtain final product;
Algoscopy is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Fumaric content assaying method is as follows:
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler, and 0.16mol/L potassium dihydrogen phosphate (be 2.80 by phosphoric acid adjust ph) is mobile phase, and determined wavelength is 210nm; Number of theoretical plate is pressed fumaric acid and is calculated, and should be not less than 5000;
The preparation precision of reference substance solution takes the fumaric acid reference substance 10mg being dried to constant weight through 105 DEG C, put in 25ml measuring bottle, scale is diluted to water dissolution, shake up, accurate absorption 2ml, puts in 100ml measuring bottle, is diluted with water to scale, shake up, obtain (containing fumaric acid 8 μ g in every 1ml);
The preparation precision of need testing solution draws this product solution 5ml, puts in 25ml measuring bottle, is diluted with water to scale, shake up, to obtain final product;
Algoscopy precision draws reference substance solution and each 5 ~ 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Testing result is as follows:
| Sample ID | Chlorogenic acid (μ g/ml) | 4-dicaffeoylquinic acid (μ g/ml) | Rosmarinic acid (μ g/ml) | Fumaric acid (μ g/ml) | PH | Other index |
| Embodiment 1 | 381 | 192 | 143 | 72 | 5.4 | Qualified |
| Comparative example 1 | 201 | 76 | 37 | 74 | 5.8 | Qualified |
Conclusion: ZHONGJIEFENG ZHUSHEYE provided by the invention (embodiment 1) chlorogenic acid, 4-dicaffeoylquinic acid and rosmarinic acid contents are apparently higher than the ZHONGJIEFENG ZHUSHEYE (comparative example 1) obtained by ministry standard.
Experimental example 2: accelerated test
Under sample being put acceleration (40 DEG C, 75% humidity) condition, take out in time placing March, June and detect, carry out study on the stability (investigating project: pH, chlorogenic acid content, 4-dicaffeoylquinic acid content, rosmarinic acid contents and fumaric acid content).Result of the test is in table 1 and table 2.
Table 1 accelerates result of the test in March
| Title | Chlorogenic acid (μ g/ml) | 4-dicaffeoylquinic acid (μ g/ml) | Rosmarinic acid (μ g/ml) | Fumaric acid (μ g/ml) | PH | Conclusion |
| Embodiment 1 | 336 | 227 | 137 | 71 | 5.3 | Qualified |
| Comparative example 1 | 158 | 94 | 33 | 72 | 5.2 | Qualified |
| Regulation promulgated by the ministries or commissions of the Central Government | Be not less than 60 | 5.0~6.0 |
Table 2 accelerates result of the test in June
| Title | Chlorogenic acid (μ g/ml) | 4-dicaffeoylquinic acid (μ g/ml) | Rosmarinic acid (μ g/ml) | Fumaric acid (μ g/ml) | PH | Conclusion |
| Embodiment 1 | 312 | 245 | 132 | 70 | 5.2 | Qualified |
| Comparative example 1 | 137 | 112 | 29 | 71 | 5.1 | Qualified |
| Regulation promulgated by the ministries or commissions of the Central Government | Be not less than 60 | 5.0~6.0 |
Conclusion: after ZHONGJIEFENG ZHUSHEYE provided by the invention (embodiment 1) was accelerated through 6 months, check item meets ministry standard regulation, the change of solution pH value is little, and chlorogenic acid, 4-dicaffeoylquinic acid and rosmarinic acid contents are apparently higher than the ZHONGJIEFENG ZHUSHEYE (comparative example 1) obtained by ministry standard.
Experimental example 3: the experimentation of antitumor action
1, to rat liver cancer H
22the inhibitory action of solid tumor
Select the inoculation tumor kind animal of latter 8 days, extract seroperitoneum, with 0.9% sodium chloride injection dilution, counting, adjustment cell concentration to 2.0 × 10
7individual/mL, in mice right fore axillary fossa subcutaneous vaccination H
22cell suspension 0.2 mL.Be divided into 4 groups at random after within 2nd, weighing with inoculation is rear, often organize 10.Group is: 1. embodiment 1 medicine group: by 2 g crude drug/kg intravenous administrations; 2. comparative example 1 medicine group: by 2 g crude drug/kg intravenous administrations; 3. 5 one fluorouracil groups: 25mg/kg intravenous administration, in administration on the the 1st, 4,7,10, totally 4 times; 4. model control group: intravenous injection every day isopyknic aseptic 0.9% sodium chloride injection of gavage.Administration from inoculating next day, administration every day 1 time, continuous 10 d, every day records body weight, and within after drug withdrawal the 2nd day, weighed and marrow execution of breaking by each treated animal, peel off tumor tissue, scales/electronic balance weighing, the results are shown in Table 3.
Table 3 couple rat liver cancer H
22the inhibitory action of solid tumor
| Group | Dosage | Body weight/g before administration | Body weight/g after administration | Tumor weight/g |
| Model control group | — | 20.5±0.7 | 35.2±1.2 | 2.72±0.53 |
| 5-fluorouracil group | 25mg／kg | 20.6±0.1.0 | 27.6±0.9 | 1.29±0.41** |
| Embodiment 1 medicine group | 2 g crude drug/kg | 20.5±0.8 | 29.3±0.7 | 1.43±0.56** |
| Comparative example 1 medicine group | 2 g crude drug/kg | 20.6±0.9 | 34.9±1.0 | 1.91±0.44* |
Note: compare with model control group, * * P < 0.01, * P < 0.05.
2, to rat liver cancer H
22the inhibitory action of ascites tumor
Kunming mouse 40, gets H
22cell suspension 1.0 × 10
6individual/mL, in mouse peritoneal inoculation H
22cell suspension 0.2 mL.Inoculate after weighing next day and be divided into 4 groups at random, only often organize l0.Group is: 1. model control group, every day gavage isopyknic aseptic 0.9% sodium chloride injection of lumbar injection; 2. cyclophosphamide group: 80 mg/kg, in l, intraperitoneal injection on the 5th, twice totally; 3. embodiment 1 medicine group: by 2.5 g crude drug/kg intravenously administrables; 4. comparative example 1 medicine group: by 2.5 g crude drug/kg intravenously administrables.Administration from inoculating next day, administration every day 1 time, continuous 10 d, record each treated animal death time, the results are shown in Table 4.
Table 4 couple rat liver cancer H
22the inhibitory action of ascites tumor
| Group | Dosage | Survival day/sky |
| Model control group | — | 15.4±0.8 |
| Cyclophosphamide group | 80mg／kg | 21.7±0.6* |
| Embodiment 1 medicine group | 2.5 g crude drug/kg | 20.9±0.9* |
| Comparative example 1 medicine group | 2.5 g crude drug/kg | 17.6±1.1 |
Note: compare with model control group, * P < 0.05.
Conclusion: ZHONGJIEFENG ZHUSHEYE provided by the invention (embodiment 1) is to rat liver cancer H
22the inhibitory action of solid tumor and ascites tumor is better than by the obtained ZHONGJIEFENG ZHUSHEYE (comparative example 1) of ministry standard.
Claims (5)
1. the preparation method of a ZHONGJIEFENG ZHUSHEYE, described ZHONGJIEFENG ZHUSHEYE is through extracting the sterile water solution made by Herba Sarcandrae, it is characterized in that preparation method is as follows: get Herba Sarcandrae 5000-10000g, extracting in water 2-3 time, merge extractive liquid, filter, when filtrate is concentrated into 70 DEG C, relative density is 1.25-1.40, add alcohol settling, alcohol content is made to be 60%-90%, cold preservation 24-72 hour, filter, filtrate recycling ethanol is also concentrated into every 1ml containing crude drug 5-15g, adjust pH is to 3-5, heating 0.5-2.0 hour, cold preservation 12-48 hour, filter, filtrate adjust pH is to 5-7, be heated to filtrate Content of Chlorogenic Acid and 4-dicaffeoylquinic acid content ratio is 3:1-3:2, add active carbon appropriate, filter, filtrate injecting is diluted with water to 1000ml, filter, add 3-10ml polyoxyethylene sorbitan monoleate, stir evenly, adjust pH is to 5.5-7.0, filter, embedding, sterilizing, obtain.
2. preparation method as claimed in claim 1, is characterized in that: the every ml soln of described ZHONGJIEFENG ZHUSHEYE is no less than 100 μ g containing rosmarinic acid; Every ml soln is no less than 500 μ g containing the total amount of chlorogenic acid and 4-dicaffeoylquinic acid, and the ratio between chlorogenic acid and 4-dicaffeoylquinic acid is 2:1 ~ 5:4; Solution ph is 5.0 ~ 5.5.
3. preparation method as claimed in claim 1 or 2, is characterized in that: the number of times adding alcohol settling is 2-3 time.
4. preparation method as claimed in claim 1 or 2, is characterized in that: the number of times of heating chilling treatment is 1-3 time.
5. preparation method as claimed in claim 1 or 2, is characterized in that: carry out before embedding filling nitrogen process.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108159092A (en) * | 2018-02-05 | 2018-06-15 | 张立峰 | A kind of acupoint injection therapy drug injection and its preparation method for being used to treat heel string disease |
| CN111721880A (en) * | 2020-07-02 | 2020-09-29 | 江中药业股份有限公司 | Method for establishing fingerprint of sarcandra glabra by using double-column tandem HPLC-MS |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105878922A (en) * | 2016-02-26 | 2016-08-24 | 青岛市市立医院 | Traditional Chinese medicinal injection for treating ischemic stroke with syndromes of stagnation and obstruction of heat-toxin and blood stasis |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679873A (en) * | 2004-03-18 | 2005-10-12 | 诺氏制药（吉林）有限公司 | Sarcadra injection, its making method and venous injection |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679841A (en) * | 2005-01-21 | 2005-10-12 | 山东恒瑞医药科技发展有限公司 | Powder injection of sarcandra and its making method |
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2011
- 2011-09-19 CN CN201510140108.5A patent/CN104739902A/en active Pending
- 2011-09-19 CN CN2011102766736A patent/CN102462711A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679873A (en) * | 2004-03-18 | 2005-10-12 | 诺氏制药（吉林）有限公司 | Sarcadra injection, its making method and venous injection |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108159092A (en) * | 2018-02-05 | 2018-06-15 | 张立峰 | A kind of acupoint injection therapy drug injection and its preparation method for being used to treat heel string disease |
| CN111721880A (en) * | 2020-07-02 | 2020-09-29 | 江中药业股份有限公司 | Method for establishing fingerprint of sarcandra glabra by using double-column tandem HPLC-MS |
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Application publication date: 20150701 |