CN107028996A - A kind of detection method of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its organic acid composition - Google Patents
A kind of detection method of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its organic acid composition Download PDFInfo
- Publication number
- CN107028996A CN107028996A CN201710233146.4A CN201710233146A CN107028996A CN 107028996 A CN107028996 A CN 107028996A CN 201710233146 A CN201710233146 A CN 201710233146A CN 107028996 A CN107028996 A CN 107028996A
- Authority
- CN
- China
- Prior art keywords
- extract
- chinese
- acid
- herba ixeritis
- ixeritis denticulatae
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pathology (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Engineering & Computer Science (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a kind of Chinese Herba Ixeritis Denticulatae extract, it contains caftaric acid 1.49%~3.04%, Cichoric acid 6.14%~11.43%, chlorogenic acid 2.69%~5.08%, 4,5 dicaffeoylquinic acids 1.04%~2.72%, above percentage is the purposes in the percentage by weight that each composition accounts for extract dry weight, and its medicine as preventing liver injury.The invention also discloses the preparation method of Chinese Herba Ixeritis Denticulatae extract and its detection method of content of organic acid composition.Its preparation method technique is simple, cost is low, energy consumption is small, it is easy to industrialize.
Description
Technical field
The present invention relates to a Plant Extracts and its preparation method and the wherein detection method of composition, one is specifically related to
Plant the detection method of content of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its phenols component.
Background technology
Chinese denticulate ixeris herb is composite family Ixeris Cass medicinal plant, and Chinese denticulate ixeris herb is widely distributed in China, aboundresources, with
All herbal medicine, Chinese denticulate ixeris herb has clearing heat and detoxicating, detumescence and apocenosis, cooling blood and hemostasis, effect promoted the production of body fluid to quench thirst, for acute appendicitis, lung
Purulence ulcer, cough with lung heat, enteritis, dysentery, cholecystitis, pelvic infecton, sore furuncle poison, eczema of scrotum is spitted blood, and bleeding from five sense organs or subcutaneous tissue, metrorrhagia, bruise is damaged
Wound, diabete.Chinese denticulate ixeris herb complex chemical composition, mainly containing sesquiterpenoids, triterpenes, adenosine, flavonoids, Coumarins
Deng in addition also containing compounds such as sterol, lignanoid, vitamin, amino acid, at present both at home and abroad there is not yet on Chinese bitter
The research report of organic acid composition caftaric acid, Cichoric acid, chlorogenic acid, 4,5- dicaffeoylquinic acids etc. in Mai dishes.
The domestic patent on Chinese denticulate ixeris herb has 1, is now exemplified below, Publication No. CN101564409 patent：Should
Patent is related to new application of the Chinese denticulate ixeris herb in pharmaceutical field, particular as in the medicine for preparing treatment or prevention diabetes
Application.The patent content is related to, it is demonstrated experimentally that causing rat diabetes model port to raise Chinese denticulate ixeris herb water extraction to streptozotocin
After thing, the urination amount of modeling animal is substantially reduced, and body weight is in trend is gradually increasing, and blood glucose value is substantially reduced.By Chinese denticulate ixeris herb
It is available for the maximum volume (0.4ml/10g) of gavage to be administered three times for 1st by mouse with 200 times equivalent to adult's dose,equivalent, ties
Fruit is proved without obvious toxic side effect.
The content of the invention
The technical problems to be solved by the invention overcome in existing Chinese medicine method for preparing extractive production cost it is expensive and
There is provided Chinese Herba Ixeritis Denticulatae extract and preparation method thereof for the defect of unsuitable industrialized production.
One aspect of the invention provides a kind of Chinese Herba Ixeritis Denticulatae extract, it is characterised in that：Extract is accounted for each composition
The percentage by weight meter of dry weight, it contains caftaric acid 1.49%~3.04%, Cichoric acid 6.14%~11.43%, green
Ortho acid 2.69%~5.08%, Isochlorogenic acid C 1.04%~2.72%, it is preferable that it contains caftaric acid
1.67%~2.61%, Cichoric acid 9.87%~10.89%, chlorogenic acid 3.19%~4.51%, 4,5- dicaffeoylquinic acids
1.35%~2.45%.
In the inventive solutions, Chinese Herba Ixeritis Denticulatae extract is prepared by the following method gained：
(1) extract, concentrate and centrifuge：Chinese denticulate ixeris herb medicinal material is extracted 2-3 times with more than 40% ethanol water, will be carried
Take liquid to concentrate, centrifuge, obtain supernatant；
(2) UF membrane：Supernatant obtained by step (1) is diluted with water, dilute liquid medicine passes through through organic membrane filter, filtrate
It is Chinese Herba Ixeritis Denticulatae extract to dry.
In the inventive solutions, in step (1), described Chinese denticulate ixeris herb medicinal material is the medicinal material after crushing, powder
Broken mesh number is below 50 mesh；
The temperature of described extraction be 70 DEG C~100 DEG C, more preferably reflux temperature；
The concentration of more than 40% described ethanol water is that percent by volume 40%~70% (is preferably 50%-
70%) consumption, extracted every time is 8~12 times of Chinese denticulate ixeris herb medicinal material weight；
It it is respectively 2~3 times with the number of times that 40% concentrations above ethanol water is extracted, each time is 0.5~2 hour；
Described concentration be by extract solution concentration be the 1/4 of original liquid volume；
The rotating speed of described centrifugation is 4000~5000r/min.
In the inventive solutions, in step (2), described water is distilled water；
The consumption of described water is 2 times of supernatant volume；
30-60 minutes described activated carbon decolorizing time；
The pore size filter of described organic film is 300~450nm, and the material of organic film is hydrophily polyvinylidene fluoride；
The operating pressure that described process organic film is filtered is 0~0.5Mpa, preferably 0.2Mpa；
Membrane filtration temperature is 20 DEG C~35 DEG C.
Another aspect of the invention provides a kind of preparation method of Chinese Herba Ixeritis Denticulatae extract, and its feature comprises the following steps：
(1) extract, concentrate and centrifuge：Chinese denticulate ixeris herb medicinal material is extracted with more than 50% ethanol water, by extract solution
Concentration, centrifugation, obtains supernatant；
(2) UF membrane：Supernatant obtained by step (1) is diluted with water, dilute liquid medicine passes through through organic membrane filter, filtrate
It is Chinese Herba Ixeritis Denticulatae extract to dry.
In the preparation process in accordance with the present invention,
In step (1), described Chinese denticulate ixeris herb medicinal material is the medicinal material after crushing, it is preferable that the mesh number of crushing is 50 mesh
Below；
The temperature of described extraction be 70 DEG C~100 DEG C, more preferably reflux temperature；
The concentration of more than 40% described ethanol water is that percent by volume 40%~70% (is preferably 50%-
70%) consumption, extracted every time is 8~12 times of Chinese denticulate ixeris herb medicinal material weight；
The number of times that 40% concentrations above ethanol water is extracted is respectively 1~2 time, and each time is 0.5~2 hour；
Described concentration be by extract solution concentration be the 1/4 of original liquid volume；
The rotating speed of described centrifugation is 4000~5000r/min.
In the preparation process in accordance with the present invention,
In step (2), described water is distilled water；
The consumption of described water is 2 times of supernatant volume；
30-60 minutes described activated carbon decolorizing time；
The pore size filter of described organic film is 300~450nm, and the material of organic film is hydrophily polyvinylidene fluoride；
The operating pressure that described process organic film is filtered is 0~0.5Mpa, preferably 0.2Mpa；
Membrane filtration temperature is 20 DEG C~35 DEG C.
Another aspect of the present invention provides a kind of preparation method of Chinese Herba Ixeritis Denticulatae extract, it is characterised in that：It is described
Preparation method comprise the following steps：
(a) ethanol water refluxing extraction：By Chinese denticulate ixeris herb pulverizing medicinal materials to 40 mesh, add percent by volume 50%~
70% ethanol water refluxing extraction 1~2 time, each consumption is 8~12 times of amounts of Chinese denticulate ixeris herb medicinal material weight, each time
For 0.5~2 hour, merge 40%~60% ethanol extract；Merge the concentration of 40%~70% ethanol extract (b)：By step
(a) extract solution obtained by is concentrated into the 1/4 of original liquid volume；
(c) decolourize and centrifuge：Concentrate obtained by step (b) is passed through into activated carbon decolorizing, decoction is with 4000~5000r/
Min is centrifuged, and takes supernatant；
(d) UF membrane：Supernatant is taken, the distilled water for adding 2 times of supernatant volume fully dilutes, dilute liquid medicine is by organic
Film is purified, and collects filtrate；The pore size filter of organic film is 300~450nm, and the material of organic film is polyvinylidene fluoride；
(e) low temperature drying：Filtrate after membrane filtration carries out low temperature drying, and drying temperature is 50 DEG C~70 DEG C, is obtained
To Chinese Herba Ixeritis Denticulatae extract powder.
Another aspect of the present invention provides a kind of foregoing Chinese Herba Ixeritis Denticulatae extract and is preparing treatment or prevention
Purposes in hepatic injury, chemical damage, fatty liver, liver fibrosis, the medicine of hepatitis or inflammation.
Another aspect of the present invention provides a kind of content detection side of organic acid composition in Chinese Herba Ixeritis Denticulatae extract
Method, it is characterised in that it includes following condition：Detected with efficient liquid-phase chromatography method：With octadecylsilane chemically bonded silica
For filler；It is that eluent carries out gradient elution using methanol and volumetric concentration as 0.5% glacial acetic acid aqueous solution, actual conditions is shown in
Following table；Detector is UV-detector, and Detection wavelength is 330nm, and column temperature is 30 DEG C, and flow velocity is 1.0mL/min.
With caftaric acid, Cichoric acid, chlorogenic acid, 4,5- DCQs for reference substance, above percentage
It is percent by volume.
In the present invention, described Chinese denticulate ixeris herb medicinal material is preferably selected from the Chinese denticulate ixeris herb of composite family Ixeris Cass plant
Ixeris chinensis(Thunb.)Nakai)。
The invention further relates to the detection method of content of organic acid composition in above-mentioned Chinese Herba Ixeritis Denticulatae extract, it is wrapped
Containing following condition：Detected with efficient liquid-phase chromatography method；Using octadecylsilane chemically bonded silica as filler；With methanol and
The glacial acetic acid solution that volumetric concentration is 0.5% is that eluent carries out gradient elution, and actual conditions is shown in Table 1；Detection wavelength is
330nm, column temperature is 30 DEG C, and flow velocity is 1.0mL/min；
Table 1
Using caftaric acid, Cichoric acid, chlorogenic acid and 4,5- dicaffeoylquinic acids as reference substance；Above percentage is equal
For percent by volume.
In this detection method, the preparation of reference substance solution, the preparation of need testing solution, Specification Curve of Increasing and containing measurement
Fixed can be existing conventional method and condition, preferably following methods and condition：
The preparation of reference substance solution：
Accurately weighed reference substance chlorogenic acid 4.12mg, Cichoric acid 11.20mg, galuteolin 4.01mg, 4,5-Dicaffeoylquinic acid
1.20mg is in 10mL volumetric flasks, plus 70% ethanol constant volume, and mixed reference substance solution is made.
The preparation of need testing solution：Chinese Herba Ixeritis Denticulatae extract powder (crossing 40 mesh sieves) about 1g is weighed, it is accurately weighed, put circle
In the flask of bottom, precision adds 70% ethanol 50mL, weighs, heating and refluxing extraction 1h is put to room temperature, supplies weightlessness, centrifuges
(4000r/min, 10min), supernatant is filtered with 0.45 μm of miillpore filter, takes filtrate, you can；
Specification Curve of Increasing：Precision draw hybrid standard product solution 0.1mL, 0.2mL, 0.4mL, 0.8mL, 1.2mL,
1.6mL, 2.0mL, put in 2mL volumetric flasks respectively, with methanol dilution to scale, obtain various concentrations reference substance solution.Take above-mentioned molten
Liquid sample introduction successively, the μ L of sample size 5 determine each chromatographic peak peak area, with reference substance sample size (μ g) for abscissa, chromatographic peak peak face
Product is ordinate, draws standard curve；
Assay：Precision draws each 10 μ L of 5 batches of need testing solutions, injects liquid chromatograph, determines 50 minutes chromatograms
Peak area, calculates content.
Unless otherwise specified, raw material of the present invention is collection or commercially available, and reagent is commercially available.
The positive effect of the present invention is：
(1) in Chinese Herba Ixeritis Denticulatae extract of the invention the content of organic acid composition at home and in foreign standard at present
Have no report.The content of organic acid composition is 11.36%~22.27%, and its preparation method technique is simple, cost is low, energy
Consumption is small, it is easy to industrialize.
(2) in the preparation method of Chinese Herba Ixeritis Denticulatae extract of the invention, removed with membrane separation technique in extract
Large biological molecule such as protein, tannin etc., effect is preferable.
Brief description of the drawings
Fig. 1 is the high-efficient liquid phase chromatogram of caftaric acid reference substance
Fig. 2 is the high-efficient liquid phase chromatogram of Cichoric acid reference substance.
Fig. 3 is the high-efficient liquid phase chromatogram of chlorogenic acid reference substance.
Fig. 4 is the high-efficient liquid phase chromatogram of 4,5- dicaffeoylquinic acid reference substances.
Fig. 5 is the high-efficient liquid phase chromatogram of Chinese Herba Ixeritis Denticulatae extract test sample made from embodiment 3.
Embodiment
The present invention is further illustrated with embodiment below, but the present invention is not intended to be limited thereto.
In following examples, the concentration of ethanol water is volumetric concentration.
The preparation of the Chinese Herba Ixeritis Denticulatae extract of embodiment 1
Take Chinese denticulate ixeris herb herb to dry medicinal material 1kg, crush as 40 mesh, add 8 times of amount 50% ethanol waters backflows and carries
Take 1 time, 1.0 hours every time, pour out extract solution, residue adds 10 times to measure 50% ethanol water refluxing extraction 1 hour, merges all
Extract solution, is concentrated under reduced pressure into the 1/4 of original volume, and with activated carbon decolorizing 30 minutes, 4000r/min centrifugation 10min poured out supernatant
Liquid, adds 2 times of supernatant volume distilled water dilutings, and dilution is filtered by organic film, membrane aperture 450nm, operating pressure
0.2MPa, 25 DEG C of operation temperature, obtains organic film permeate, and the filtrate after membrane filtration carries out low temperature drying, temperature 65
DEG C, dried object is collected, the Chinese Herba Ixeritis Denticulatae extract of the present invention is produced.
The preparation of the Chinese Herba Ixeritis Denticulatae extract of embodiment 2
Take Chinese denticulate ixeris herb to dry medicinal material 1kg, crush as 40 mesh, 10 times of 70% ethanol water refluxing extractions 2 of amount of addition
It is secondary, 1.0 hours every time, extract solution is poured out, residue adds 10 times to measure 70% ethanol water refluxing extraction 1 hour, merges and all carries
Liquid is taken, the 1/4 of original volume is concentrated under reduced pressure into, with activated carbon decolorizing 30 minutes, 4000r/min centrifugation 10min poured out supernatant,
2 times of supernatant volume distilled water dilutings are added, dilution is filtered by organic film, membrane aperture 450nm, operation pressure 0.2MPa, behaviour
Make 28 DEG C of temperature, obtain organic film permeate, the filtrate after membrane filtration carries out low temperature drying, 70 DEG C of drying temperature is received
Collect dried object, produce the Chinese Herba Ixeritis Denticulatae extract of the present invention.
The preparation of the Chinese Herba Ixeritis Denticulatae extract of embodiment 3
Take Chinese denticulate ixeris herb to dry medicinal material 1kg, crush as 40 mesh, 12 times of 60% ethanol water refluxing extractions 1 of amount of addition
It is secondary, 1.5 hours every time, extract solution is poured out, residue adds 10 times to measure 60% ethanol water refluxing extraction 1 hour, merges and all carries
Liquid is taken, the 1/4 of original volume is concentrated under reduced pressure into, with activated carbon decolorizing 30 minutes, 4500r/min centrifugation 10min poured out supernatant,
2 times of supernatant volume distilled water dilutings are added, dilution is filtered by organic film, membrane aperture 450nm, operating pressure 0.2MPa,
23 DEG C of operation temperature, obtains organic film permeate, the filtrate progress low temperature drying after membrane filtration, 65 DEG C of drying temperature,
Dried object is collected, the Chinese Herba Ixeritis Denticulatae extract of the present invention is produced.
Equipment used in the present invention is the common apparatus used in pharmaceutical industry, such as membrane separation plant, centrifuge.
The preparation of the Chinese Herba Ixeritis Denticulatae extract of embodiment 4
Take Chinese denticulate ixeris herb to dry medicinal material 1kg, crush as 40 mesh, 8 times of 50% ethanol water refluxing extractions 1 of amount of addition
It is secondary, 1.0 hours every time, extract solution is poured out, residue adds 10 times to measure 50% ethanol water refluxing extraction 1 hour, merges and all carries
Liquid is taken, the 1/4 of original volume is concentrated under reduced pressure into, with activated carbon decolorizing 30 minutes, 4500r/min centrifugation 10min poured out supernatant, led to
Organic film filtration is crossed, membrane aperture 450nm, operating pressure 0.2MPa, 23 DEG C of operation temperature obtain organic film permeate, through filter membrane
Filtrate after filtering carries out low temperature drying, and 65 DEG C of drying temperature collects dried object, produces the Chinese Herba Ixeritis Denticulatae extract of comparative example 1.
The preparation of the Chinese Herba Ixeritis Denticulatae extract of embodiment 5
Take Chinese denticulate ixeris herb to dry medicinal material 1kg, crush as 40 mesh, 8 times of 10% ethanol water refluxing extractions 1 of amount of addition
It is secondary, 1.0 hours every time, extract solution is poured out, residue adds 10 times to measure 10% ethanol water refluxing extraction 1 hour, merges and all carries
Liquid is taken, the 1/4 of original volume is concentrated under reduced pressure into, with activated carbon decolorizing 30 minutes, 4500r/min centrifugation 10min poured out supernatant, led to
Organic film filtration is crossed, membrane aperture 450nm, operating pressure 0.2MPa, 23 DEG C of operation temperature obtain organic film permeate, through filter membrane
Filtrate after filtering carries out low temperature drying, and 65 DEG C of drying temperature collects dried object, produces the Chinese Herba Ixeritis Denticulatae extract of comparative example 2.
The preparation of the Chinese Herba Ixeritis Denticulatae extract of embodiment 6
Take Chinese denticulate ixeris herb to dry medicinal material 1kg, crush as 40 mesh, add 8 times of amount water and extract 1 time, 1.0 hours every time, incline
Go out extract solution, residue adds 10 times of amount water to extract 1 hour, merges whole extract solutions, the 1/4 of original volume is concentrated under reduced pressure into, with activity
Carbon decoloring 30 minutes, 4500r/min centrifugation 10min, pours out supernatant, is filtered by organic film, membrane aperture 450nm, operation pressure
Power 0.2MPa, 23 DEG C of operation temperature, obtains organic film permeate, and the filtrate after membrane filtration carries out low temperature drying, dries
65 DEG C of temperature, collects dried object, produces the Chinese Herba Ixeritis Denticulatae extract of comparative example 3.
The Chinese Herba Ixeritis Denticulatae extract liver protection experiment of embodiment 7
CCl is set up in conventional manner4Liver damage animal model is caused, mouse is randomly divided into 9 groups, every group 10, is respectively
Normal group, model group, positive controls, Chinese Herba Ixeritis Denticulatae extract group, Chinese Herba Ixeritis Denticulatae extract reference examples group.Positive control
Group gives DDB 0.20g/kg, Chinese Herba Ixeritis Denticulatae extract group and Chinese Herba Ixeritis Denticulatae extract reference examples group, every kind of extraction
Thing or reference examples give 1.95g/kg or 1.00g/kg respectively, and normal and model group gives isometric physiological saline respectively.Continuously
Gastric infusion 8 days.Go out after last dose 2h outside normal group, remaining each group intraperitoneal injection 0.2%CCl4Vegetable oil 10ml/kg,
Isometric vegetable oil is injected intraperitoneally in normal group, and fasting be can't help taking blood after water, 16h, and 5000r/min is centrifuged 20 minutes, determine AST and
ALT enzyme activities, by MDA contents in thio barbital colorimetric method for determining serum.Experimental result is referring to table 1 below.
The Chinese Herba Ixeritis Denticulatae extract liver protection result of the test of table 1 (N=10)
Verified by embodiment, it is known that Chinese Herba Ixeritis Denticulatae extract has certain protective effect for hepatic injury.
Had greatly improved relative to model group, but Different Extraction Method and various dose have for ALT, AST and MDA
Different Effects, inventor compared for different extracting methods in extraction process, and different solvents and different step orders are right
The product obtained in extraction has considerable influence.It is not desired to be constrained by theory, can be guessed according to experimental result, may be due to supplement
The product obtained by insoluble matter is removed after the aqueous solution and not only fully extracts that obtained being capable of liver-protective caftaric acid, witloof
The organic acids such as acid, chlorogenic acid, Isochlorogenic acid C, bear larger while eliminating in extract for liver metabolism
Compound, so as to cause to generate preferable hepatoprotective effect.
The Chinese Herba Ixeritis Denticulatae extract Carrageenan of embodiment 8 causes the influence of rat paw edema
200~220g SD rats are taken to be randomly divided into 6 groups, every group 10, be model group, positive controls, denticulate ixeris herb respectively
Extract high and low dose group, each group medicine is made into the suspension of respective concentration with distilled water, and rat oral gavage volume presses 10mL
kg-1Volume calculations are administered.Positive controls give metacortandracin 0.015gkg-1, denticulate ixeris herb high and low dose group is given respectively to be carried
Take thing 1.35kg-1、0.54g·kg-1, model group gives isometric physiological saline daily.Continuous gavage is administered 4 days.In last
1.5h is in the carrageenan of Rat Right metapedes plantar subcutaneous injection 1% after administration, and 1h after preceding and injection is injected in every 0.1mL, measurement,
2h, 4h, 6h, 8h vola pedis volume, calculate swelling percentage, are shown in Table 2.
Volume × 100% before swelling percentage=(volume before volume-injection after injection)/injection
The Chinese Herba Ixeritis Denticulatae extract of table 2 to rat paw edema influence (N=10)
Note：Chinese denticulate ixeris herb and model group compare Δ P<0.01
The Chinese Herba Ixeritis Denticulatae extract of embodiment 9 causes the increased influence of mouse peritoneal capillary permeability to glacial acetic acid
(20 ± 5) g Kunming mouses 60 are taken, male and female half and half are randomly divided into 6 groups, packet and medication are with 2.2
Under.Successive administration 7d, in the Evans blue normal saline solution 0.1mL10g of 1.5h tail vein injections after last dose 0.5%-1,
10min pneumoretroperitoneums inject 0.7% glacial acetic acid 0.1mL10g-1, cervical vertebra is taken off after 20min and puts to death animal, every intraperitoneal injection physiology
Salt solution 5mL, and gently rub belly 3min, then from intraperitoneal sucking-off peritoneal fluid 3mL or so (flushing liquor is such as contaminated by the blood,
Then it is discarded), 1000r/min centrifugation 5min determine absorbance with spectrophotometer at 590nm wavelength, are shown in Table 3.
Compared with model group, Chinese denticulate ixeris herb can effectively suppress the increase of glacial acetic acid induced mice capillary permeability.
The Chinese Herba Ixeritis Denticulatae extract of table 3 to mouse peritoneal permeability influence (N=10)
Note：Chinese denticulate ixeris herb is compared △ P with sowthistle and model group<0.01
The detection of organic acid composition in the Chinese Herba Ixeritis Denticulatae extract of embodiment 10
The detection method of organic acid composition：
Efficient liquid-phase chromatography method is detected that chromatographic condition is：Using octadecylsilane chemically bonded silica as filler, with
Methanol-volumetric concentration is 0.5% glacial acetic acid solution gradient elution, and actual conditions is shown in Table 4, and Detection wavelength is 330nm, and column temperature is 30
DEG C, flow velocity is that percentage is percent by volume in 1.0ml/min, table.
Table 4
The preparation of reference substance solution：Accurately weighed reference substance chlorogenic acid 4.12mg, Cichoric acid 11.20mg, galuteolin
4.01mg, 4,5-Dicaffeoylquinic acid 1.20mg are in 10mL volumetric flasks, plus 70% ethanol constant volume, and mixed reference substance solution is made.
Leave out the preparation of need testing solution：Chinese Herba Ixeritis Denticulatae extract powder (crossing 40 mesh sieves) about 1g is weighed, it is accurately weighed,
Put in round-bottomed flask, precision adds 70% ethanol 50mL, weighs, heating and refluxing extraction 1h is put to room temperature, supplies weightlessness, centrifuge
(4000r/min, 10min), supernatant is filtered with 0.45 μm of miillpore filter, is taken subsequent filtrate, is produced；
Specification Curve of Increasing：Precision draw hybrid standard product solution 0.1mL, 0.2mL, 0.4mL, 0.8mL, 1.2mL,
1.6mL, 2.0mL, put in 2mL volumetric flasks respectively, with methanol dilution to scale, obtain various concentrations reference substance solution.
Above-mentioned solution sample introduction successively is taken, the μ L of sample size 5 determine each chromatographic peak peak area, are with reference substance sample size (μ g)
Abscissa, chromatographic peak peak area is ordinate, draws standard curve；
Determination method：Precision draws each 10 μ L of 3 batches of need testing solutions, injects liquid chromatograph, determines 50 minutes chromatogram peak outs
Area, calculates content, while by the corresponding chromatographic peak gained mobile phase of testing sample by Mass Spectrometer Method, respectively at caffeoyl winestone
Acid, Cichoric acid, chlorogenic acid are corresponding with 4,5- dicaffeoylquinic acid molecular weight.
After testing, the ratio of each composition is in embodiment 1-3：
Caftaric acid, Cichoric acid, chlorogenic acid and 4,5- dicaffeoylquinic acid content sums in the extract of embodiment 1
For 17.02%, it contains caftaric acid 1.67%, Cichoric acid 10.81%, chlorogenic acid 3.19%, 4, the caffeoyl Kuis of 5- bis-
Peaceful acid 1.35%.
Caftaric acid, Cichoric acid, chlorogenic acid, 4,5- dicaffeoylquinic acid content sums in the extract of embodiment 2
For 19.21%, it contains caftaric acid 2.61%, Cichoric acid 9.87%, chlorogenic acid 4.28%, 4, the caffeoyl quinines of 5- bis-
Acid 2.45%.
Caftaric acid, Cichoric acid, chlorogenic acid, 4,5- dicaffeoylquinic acid content sums in the extract of embodiment 3
For 20.23%, it contains caftaric acid 2.46%, Cichoric acid 10.89%, chlorogenic acid 4.51%, 4, the caffeoyl Kuis of 5- bis-
Peaceful acid 2.37%.
Claims (10)
1. a kind of Chinese Herba Ixeritis Denticulatae extract, it is characterised in that：Accounted for by each composition in terms of the percentage by weight of extract dry weight, it contains
There are caftaric acid 1.49%~3.04%, Cichoric acid 6.14%~11.43%, chlorogenic acid 2.69%~5.08%, 4,5-
Dicaffeoylquinic acid 1.04%~2.72%, it is preferable that it contains caftaric acid 1.67%~2.61%, Cichoric acid
9.87%~10.89%, chlorogenic acid 3.19%~4.51%, 4,5- dicaffeoylquinic acids 1.35%~2.45%.
2. China's Herba Ixeritis Denticulatae extract as claimed in claim 1, it is prepared by the following method gained：
(1) extract, concentrate and centrifuge：Chinese denticulate ixeris herb medicinal material is extracted 2-3 times with more than 40% ethanol water, by extract solution
Concentration, centrifugation, obtains supernatant；
(2) UF membrane：Supernatant obtained by step (1) is diluted with water, dilute liquid medicine passes through drying through organic membrane filter, filtrate
As Chinese Herba Ixeritis Denticulatae extract.
3. China's Herba Ixeritis Denticulatae extract as claimed in claim 2, it is characterised in that：
In step (1), described Chinese denticulate ixeris herb medicinal material is the medicinal material after crushing, and the mesh number of crushing is below 50 mesh；
The temperature of described extraction be 70 DEG C~100 DEG C, more preferably reflux temperature；
The concentration of more than 40% described ethanol water is percent by volume 40%~70% (being preferably 50%-70%), often
The consumption of secondary extraction is 8~12 times of Chinese denticulate ixeris herb medicinal material weight；
It it is respectively 2~3 times with the number of times that 40% concentrations above ethanol water is extracted, each time is 0.5~2 hour；
Described concentration be by extract solution concentration be the 1/4 of original liquid volume；
The rotating speed of described centrifugation is 4000~5000r/min.
4. China's Herba Ixeritis Denticulatae extract as claimed in claim 2, it is characterised in that：
In step (2), described water is distilled water；
The consumption of described water is 2 times of supernatant volume；
30-60 minutes described activated carbon decolorizing time；
The pore size filter of described organic film is 300~450nm, and the material of organic film is hydrophily polyvinylidene fluoride；
The operating pressure that described process organic film is filtered is 0~0.5Mpa, preferably 0.2Mpa；
Membrane filtration temperature is 20 DEG C~35 DEG C.
5. the preparation method of the Chinese Herba Ixeritis Denticulatae extract as described in claim any one of 1-4, its feature comprises the following steps：
(1) extract, concentrate and centrifuge：Chinese denticulate ixeris herb medicinal material is extracted with more than 40% ethanol water, extract solution is concentrated,
Centrifugation, obtains supernatant；
(2) UF membrane：Supernatant obtained by step (1) is diluted with water, dilute liquid medicine passes through drying through organic membrane filter, filtrate
As Chinese Herba Ixeritis Denticulatae extract.
6. preparation method as claimed in claim 5, it is characterised in that：
In step (1), described Chinese denticulate ixeris herb medicinal material is the medicinal material after crushing, it is preferable that the mesh number of crushing is below 50 mesh；
The temperature of described extraction be 70 DEG C~100 DEG C, more preferably reflux temperature；
The concentration of more than 40% described ethanol water is percent by volume 40%~70% (being preferably 50%-70%), often
The consumption of secondary extraction is 8~12 times of Chinese denticulate ixeris herb medicinal material weight；
The number of times that ethanol water is extracted is respectively 1~2 time, and each time is 0.5~2 hour；
Described concentration be by extract solution concentration be the 1/4 of original liquid volume；
The rotating speed of described centrifugation is 4000~5000r/min.
7. the preparation method as described in claim 5 or 6, it is characterised in that：
In step (2), described water is distilled water；
The consumption of described water is 2 times of supernatant volume；
30-60 minutes described activated carbon decolorizing time；
The pore size filter of described organic film is 300~450nm, and the material of organic film is hydrophily polyvinylidene fluoride；
The operating pressure that described process organic film is filtered is 0~0.5Mpa, preferably 0.2Mpa；
Membrane filtration temperature is 20 DEG C~35 DEG C.
8. the preparation method of Chinese Herba Ixeritis Denticulatae extract as described in claim any one of 1-4, it is characterised in that：Described preparation
Method comprises the following steps：
(a) ethanol water refluxing extraction：By Chinese denticulate ixeris herb pulverizing medicinal materials to 40 mesh, percent by volume 40%~70% is added
Ethanol water refluxing extraction 1~2 time, each consumption is 8~12 times of amounts of Chinese denticulate ixeris herb medicinal material weight, and each time is
0.5~2 hour, merge 40%~60% ethanol extract；Merge the concentration of 40%~70% ethanol extract (b)：By step (a)
The extract solution of gained is concentrated into the 1/4 of original liquid volume；
(c) decolourize and centrifuge：Concentrate obtained by step (b) is passed through into activated carbon decolorizing, decoction with 4000~5000r/min from
The heart, takes supernatant；
(d) UF membrane：Supernatant is taken, the distilled water for adding 2 times of supernatant volume fully dilutes, and dilute liquid medicine enters by organic film
Row purifying, collects filtrate；The pore size filter of organic film is 300~450nm, and the material of organic film is polyvinylidene fluoride；
(e) low temperature drying：Filtrate after membrane filtration carries out low temperature drying, and drying temperature is 50 DEG C~70 DEG C, in obtaining
Magnificent Herba Ixeritis Denticulatae extract powder.
9. Chinese Herba Ixeritis Denticulatae extract as described in claim any one of 1-4 treats or prevents hepatic injury, chemically preparing
Purposes in hepatic injury, fatty liver, liver fibrosis, the medicine of hepatitis or inflammation.
10. the detection method of organic acid component content in a kind of Chinese Herba Ixeritis Denticulatae extract, it is characterised in that it is comprising following
Condition：Detected with efficient liquid-phase chromatography method：Using octadecylsilane chemically bonded silica as filler；It is dense with methanol and volume
The glacial acetic acid aqueous solution that degree is 0.5% is that eluent carries out gradient elution, and actual conditions see the table below 1；Detector is ultraviolet detection
Device, Detection wavelength is 330nm, and column temperature is 30 DEG C, and flow velocity is 1.0mL/min；
With caftaric acid, Cichoric acid, chlorogenic acid, 4,5- DCQs for reference substance, above percentage is
Percent by volume.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710233146.4A CN107028996A (en) | 2017-04-11 | 2017-04-11 | A kind of detection method of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its organic acid composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710233146.4A CN107028996A (en) | 2017-04-11 | 2017-04-11 | A kind of detection method of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its organic acid composition |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107028996A true CN107028996A (en) | 2017-08-11 |
Family
ID=59534965
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710233146.4A Pending CN107028996A (en) | 2017-04-11 | 2017-04-11 | A kind of detection method of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its organic acid composition |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107028996A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109030678A (en) * | 2018-07-18 | 2018-12-18 | 天津中医药大学 | The content assaying method of 5 kinds of chemical components in a kind of northeast Chinese Ixeris |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101766667A (en) * | 2008-12-30 | 2010-07-07 | 上海雷允上科技发展有限公司 | Echinacea purpurea extract and preparing method and phenols ingredient detection method thereof |
-
2017
- 2017-04-11 CN CN201710233146.4A patent/CN107028996A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101766667A (en) * | 2008-12-30 | 2010-07-07 | 上海雷允上科技发展有限公司 | Echinacea purpurea extract and preparing method and phenols ingredient detection method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 刘海霞: "中华苦荬菜与苣荬菜主要化学成分及保肝抗炎药理作用的比较研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109030678A (en) * | 2018-07-18 | 2018-12-18 | 天津中医药大学 | The content assaying method of 5 kinds of chemical components in a kind of northeast Chinese Ixeris |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101810793B (en) | Application of corn stigma flavone in preparing medicament for curing hyperlipidemia | |
| CN101396384B (en) | Asiatic centella extract and preparation methode thereof | |
| CN101336987A (en) | Preparation method of total flavone of Hovenia dulcisThunb | |
| CN111187159A (en) | Separation process and application of natural free radical scavenger in saxifraga tangutica | |
| CN107998166A (en) | The preparation method of dandelion extract | |
| CN102731586A (en) | Preparation technology of stilbene glucoside in polygonum multiflorum | |
| CN109528803A (en) | A kind of preparation method of Radix Pseudostellariae extract and its Radix Pseudostellariae extract and detection method of preparation | |
| CN103923043B (en) | A method of effectively preparing tanshin polyphenolic acid B extract | |
| CN107028996A (en) | A kind of detection method of Chinese Herba Ixeritis Denticulatae extract and its preparation method and its organic acid composition | |
| CN103923042B (en) | The preparation method of tanshin polyphenolic acid B extract | |
| CN104224867B (en) | A kind of desmodium sugar-free particle and preparation method thereof | |
| CN101269182A (en) | Pharmaceutical composition for treating children's wind-heat cold, preparation method and quality control method | |
| CN106943354B (en) | A kind of preparation method of novel biochemical particles | |
| CN1969963A (en) | Method for extracting medicament effective component for treating hepatitis and medicinal composition thereof | |
| CN101422522B (en) | Preparation method of traditional Chinese medicine preparation | |
| CN107582593A (en) | The method that general flavone is extracted from beggar-ticks | |
| CN107669751A (en) | A kind of clarification process and method of quality control of the pharmaceutical composition that there is treatment red blood trace on face to act on | |
| CN103977354B (en) | Traditional Chinese medicine extract with anti-depression effect and preparation method and application thereof | |
| CN105037311A (en) | Apigenin on-line two-dimensional composite liquid-phase extraction purification method | |
| CN102078400B (en) | Preparation method of extract of total triterpene acid of loquat leaf | |
| CN100411653C (en) | Chinese medicine composition for treating diabetes mellitus | |
| CN105017235B (en) | The method for isolating and purifying Puerarin | |
| CN101244220A (en) | Black thorn chinaroot greenbrier total saponin extract and preparation thereof | |
| CN108042682A (en) | A kind of Long Lu Zishuiqinggan ball and preparation method thereof | |
| CN108030813A (en) | A kind of clarification process and method of quality control of the pharmaceutical composition with delay skin aging |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170811 |