CN110376291A - A kind of coptis reference extract and its preparation method and application - Google Patents
A kind of coptis reference extract and its preparation method and application Download PDFInfo
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- CN110376291A CN110376291A CN201810330289.1A CN201810330289A CN110376291A CN 110376291 A CN110376291 A CN 110376291A CN 201810330289 A CN201810330289 A CN 201810330289A CN 110376291 A CN110376291 A CN 110376291A
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- 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
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- 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
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- 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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
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- 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
- G01N2030/042—Standards
Abstract
The invention discloses a kind of coptis reference extracts and its preparation method and application, the extract is yellow dried powder, it is made of coptis extract containing weight percent 67-84% and auxiliary material 16-33%, is wherein not less than 18% containing jateorrhizine, palmatine, jamaicin, epiberberine, coptis alkali content summation in coptis extract；Coptis reference extract consistency is good, character is stable, uniform for this, can be used for the quality control of Rhizoma Coptidis, coptis, measures to the analysis of active compound qualitative, quantitative.
Description
Technical field
The invention belongs to chemical fields, more particularly, to a kind of coptis reference extract and preparation method thereof.
Background technique
Drug inspection is an important means for verifying drug quality, more and more with being widely used for instrument analysis
Ground uses pharmaceutical standard material.Special cross-reference entry object of the Chang Huiyong reference substance as inspection drug quality in drug inspection work,
Using it as the reference standard for determining authenticity of medicament superiority and inferiority.Reference substance has highly important status in drug inspection.
In present Chinese medicine standard, the mode of single component control quality is difficult to reflect traditional Chinese medicine quality well, and use is a variety of
The method of ingredient or Characteristic chromatographic peak comprehensively control quality increasingly attracts people's attention.However, the reference substance of commercialization
Supply falls short of demand and high testing cost limits multicomponent quality control answering in actual production, scientific research and supervision area
With.Not only can solve big, the in short supply problem of monomer reference substance preparation difficulty using reference extract, but can achieve to index at
The purpose of sub-control has important meaning to the quality control and standard execution of Chinese medicine.
Existing traditional Chinese medicine quality control mode is substantially the development along Natural Medicine Chemistry, a certain or several to Chinese medicine
Effective component is the analysis method and the not only design of difinite quality but also the quality standard that can be quantified of target, referring to the matter of external botanical medicine
Amount control method is used for reference the mode of chemicals quality control, and is identified by document report foundation is simply physical and chemical accordingly, then
Develop to the identification based on spectrum, chromatography and the quality standard of assay.It is all multicomponent synthesis per Chinese medicine simply
Body also indicates that one or two using in medicinal material even several ingredients as medicinal material matter which dictates that its distinctive globality and ambiguity
There are significant limitations for the evaluation method of amount.
1990 editions " Chinese Pharmacopoeias " increase the indentification by TLC of control medicinal material, so that the identification of Chinese medicine and Chinese patent drug has
Very big progress.Chinese medicine and external herbal medicine increasingly pay attention to multicomponent or multi-component detection, such as German ginkgo at present
The quality of leaf extract controls.The analysis method of finger-print can carry out quality control to medicinal material on the whole, currently still
There is very big researching value.There are two types of " reference substances " in terms of controlling quality of medicinal material for Chinese Pharmacopoeia at present, and chemical reference substance is in
Medicine control medicinal material.Wherein chemical reference substance can be used for Qualitive test and the quantitative analysis of medicinal material, and Chinese medicine control medicinal material is then used to show
Micro- identification and thin layer identify.However, chemical reference substance and Chinese medicine control medicinal material have its limitation in traditional Chinese medicine quality control.It is first
First, chemical component diversification in Chinese medicine, single or several compounds can not reflect the overall picture of medicinal material, and existing standard is past
Past the phenomenon that having many loopholes, adulterating, happens occasionally.Chinese medicine control medicinal material is influenced by the place of production, growing environment, is difficult
Guarantee per batch of uniform quality, and be only used for Qualitive test, can not reflect the height of medicinal ingredient content.
There are four basic demand (ASCS) for Chinese medicine reference extract: Authenticity, crude drug source is reliable, has and represents
Property；Specificity, the detection method used have specificity；Con-sistency, different batches reference extract should be kept
Unanimously；Stability, character are stable, uniform, easy to use.Refer to TLC Fingerprints and high performance liquid chromatography
The method of line map, Chinese medicine reference extract are able to satisfy the needs of Qualitive test.It can also be by the reference extract of markization
One step carries out the even quantitative analysis of sxemiquantitative.Chinese medicine reference extract will have important justice to traditional Chinese medicine quality control.
The coptis is the ranunculaceae plant coptis (Coptis chinensis Franch.), the triangle leaf coptis (Coptis
Deltoidea C.Y.Cheng et Hsiao) He Yunlian (Coptis teeta Wall.) dry rhizome.Coptis rhizome contains
A variety of alkaloids, mainly jamaicin, also known as berberine (Berberine) are about 5%~8%, are secondly coptisine
(Coptisine), worenine (Worenine), palmatine (palmatine, Palmatine), jateorrhizine
(Jatrorrhizine), jateorrhizine (Columbamine) still contains obakunone (Obakunone), obakulactone
(Obakulactone), the leaf such as magnoflorine (Magnoflorine), ferulic acid (Ferulic acid) contains jamaicin 1.4%
~2.8%.In addition, also contain various trace elements in the coptis, from the triangle leaf coptis separation identify coptisine, jamaicin,
Palmatine and jateorrhizine.
In the Chinese Pharmacopoeia quality standard of Chinese Pharmacopoeia Commission's publication, one of effective component when to the coptis as medicinal material
The content of Berberine hydrochloride provides, but also just since Chinese medicine reference extract has above-mentioned specific requirement, it is existing at present
Technology does not have also or does not have mature coptis reference extract and preparation method thereof to replace the application of coptis control medicinal material.
Summary of the invention
In order to solve the above-mentioned technical problem, the purpose of the present invention is to provide one kind in order to solve the above-mentioned technical problem, this
Invention is not comprehensive to examination criteria present in coptis or Rhizoma Coptidis detection, high using a variety of reference substance testing costs
Problem, the present invention provide the compound in a kind of pair of Rhizoma Coptidis, coptis extract and its coptis while being detected
Coptis reference extract, coptis reference extract consistency is good, character is stable, uniform for this, can be used for Rhizoma Coptidis, coptis prescription
Quality control, the analysis of active compound qualitative, quantitative is measured.
The present invention also provides the preparation methods of coptis reference extract.This method is easy to operate, at low cost, reproducible
And recovery rate is high.
The present invention also provides the controls for the sample that control medicinal material detection ingredient containing the coptis is replaced with coptis reference extract
Method.
The present invention is achieved through the following technical solutions:
A kind of coptis reference extract of the invention, the extract are yellow dried powder, are by containing weight percent
It is formed than coptis extract 67-84% and auxiliary material 16-33%, jateorrhizine, palmatine, barberry is wherein contained in coptis extract
Alkali, epiberberine, coptis alkali content summation are not less than 18%.
Further preferably, a kind of coptis reference extract of the invention, content >=11.0% of Berberine, palmatine
Content >=3.0%, content >=1.5% of epiberberine, content >=2.0% of coptisine；Or a kind of coptis pair of the invention
According to extract, content >=11.0% of Berberine, content >=3.0% of palmatine, content >=1.5% of epiberberine,
Content >=2.0% of coptisine, content >=1.0% of jateorrhizine.
Further preferably, in coptis reference extract of the present invention, content >=11.8% of jamaicin, palmatine
Content >=3.4%, content >=1.9% of epiberberine, content >=2.5% of coptisine.
What coptis reference extract of the present invention was prepared by the following method:
(1) coptis dry cream: the Rhizoma Coptidis powder methanol ultrasonic extraction of separate sources and batch is taken, filtrate 1 is filtered to obtain；
The dregs of a decoction again with methanol ultrasonic extraction 1-3 times, 1-3 filtrate of collection mixes with filtrate 1, is evaporated to obtain coptis dry cream；
(2) coptis extract: coptis dry cream is dissolved in methanol, add auxiliary material be evaporated be ground up, sieved up to the coptis extract
Object；
(3) coptis reference extract: separate sources or the coptis extract of batch are subjected to allotment and blent, the coptis pair is obtained
According to extract.
In above-mentioned steps 1 w/v of Rhizoma Coptidis powder and methanol be 1:8~1:50 (preferably 1:8~1:20, it is excellent
Select 1:10)；The ultrasonic time is 30~60min, and power is that (preferably ultrasonic time is 30min to 100W~3kW, and power is
500W)。
Filtering described in step 1 is with Medium speed filter paper or 2000 mesh net filtrations.
Methanol usage is 2-5 times of dry cream weight in the step 2.
10%~60% (preferably 20-40%) of dry cream weight is added in the step 2 in coptis dry cream methanol solution
Auxiliary material；
The auxiliary material includes but is not limited to: the such as superfine silica gel powder, medical starch
It is preferred that auxiliary material is 20-50% (preferably 40%) superfine silica gel powder and/or 10-30% (preferably 20%) medical starch.Institute
Stating auxiliary material and dosing can guarantee that extract character is stable, uniform and easy to use.
Being ground up, sieved after being evaporated described in step 2 was 90~200 mesh (preferably 200 mesh) the screen to filtrate.
Allotment described in step 3 refers specifically to: the coptis extract of separate sources or batch being deployed, should be made final
Coptis reference extract in correspond to crude drug ratio be 1:(2-5) (g/g), (preferably 1:3g/g), wherein (content is with wt%
Meter) jamaicin content >=11.0%, content >=3.0% of palmatine, content >=1.5% of epiberberine, coptisine contains
Amount >=2.0%.It is preferred that in coptis reference extract of the present invention, content >=11.8% of jamaicin, the content of palmatine
>=3.4%, content >=1.9% of epiberberine, content >=2.5% of coptisine.
The present invention further includes with thin-layered chromatography and/or high performance liquid chromatography between the step 2 and step 3 to not
The step of being detected with the palmatine of the coptis extract of batch, jamaicin, epiberberine and coptisine.
For reliability, content consistency, the homogeneity for guaranteeing coptis reference extract of the invention.It is preferred that in step (1)
The preceding screening step for increasing different crude drug sources: by separate sources or the raw medicinal material of batch with thin-layered chromatography and/or efficiently
Liquid chromatography is detected；Then thin-layered chromatography and/or efficient liquid are passed through to the raw medicinal material of separate sources or batch
The content of chromatogram and/or measurement that phase chromatography obtains is compared analysis, and the content for rejecting chromatogram and/or measurement is abnormal
Raw medicinal material, retain chromatogram and/or measurement the consistent raw medicinal material of content carry out next step coptis dry cream preparation.
Coptis reference extract of the invention exists in a unit, and the unit dosage form refers to the list of preparation
Position, such as every of tablet, every of capsule,
Preparation of the invention includes but is not limited to tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard
It is capsule, soft capsule, oral solution, mouth containing agent, granule, electuary, pill, powder, paste, sublimed preparation, suspension, pulvis, molten
Liquor, injection, suppository, ointment, emplastrum, creme, spray, drops, patch.
Pharmaceutical preparation of the invention, the preparation of oral administration can contain common excipient, such as adhesive, filling
Agent, diluent, tablet agent, lubricant, disintegrating agent, colorant, flavoring agent and wetting agent when necessary can be coated tablet.
Applicable filler includes cellulose, mannitol, lactose and other similar fillers.Suitable disintegrating agent include starch,
Polyvinylpyrrolidone and starch derivatives, such as sodium starch glycollate.Suitable lubricant includes, such as magnesium stearate.
Suitable pharmaceutically acceptable wetting agent includes lauryl sodium sulfate.Mixing, filling, the common method system such as tabletting can be passed through
Standby solid oral composition.Carrying out mixing repeatedly can be such that active material is distributed in entirely using those of a large amount of fillers composition
In.
The form of oral liquid for example can be aqueous or oily suspensions, solution, emulsion, syrup or elixir,
Or it can be a kind of dry products that can be compounded with water or other suitable carriers before use.This liquid preparation can contain
Conventional additive, such as suspending agent, such as sorbierite, syrup, methylcellulose, gelatin, hydroxyethyl cellulose, carboxymethyl are fine
Dimension element, aluminium stearate gel or hydrogenated edible fats, emulsifier, such as lecithin, anhydro sorbitol monooleate or Arab
Glue；Non-aqueous carrier (they may include edible oil), for example, apricot kernel oil, fractionated coconut oil, such as glycerol ester oily ester,
Propylene glycol or ethyl alcohol；Preservative, such as para hydroxybenzene methyl esters or propylparaben or sorbic acid, and if desired,
Contain conventional flavouring agent or colorant.
For injection, the fluid unit dosage form of preparation contains active material and sterile carrier of the invention.According to carrier
And concentration, this compound can be suspended or be dissolved.The preparation of solution is usually by the way that active material is dissolved in a kind of load
In body, disinfection is filtered before being loaded into a kind of suitable bottle or ampoule, is then sealed.For example a kind of local anaesthesia of auxiliary material
Agent, preservative and buffer are also soluble in this carrier.It, can be after being packed into bottle by this in order to improve its stability
Kind composition frost, and under vacuum remove water.
Coptis reference extract of the invention is optionally added suitable pharmaceutically acceptable when being prepared into medicament
Carrier, the pharmaceutically acceptable carrier are selected from: mannitol, sorbierite, sodium pyrosulfite, sodium hydrogensulfite, sodium thiosulfate,
Cysteine hydrochloride, thioacetic acid, methionine, injection Vitamin B_6 DTA disodium, Ethylenediaminetetraacetic Acid Calcium Salt, carbonate, the vinegar of monovalence alkali metal
Hydrochlorate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodium chloride, potassium chloride, sodium lactate, xylitol,
Maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and
Its derivative, alginates, gelatin, polyvinylpyrrolidone, glycerol, POLYSORBATE 80, agar, calcium carbonate, calcium bicarbonate, surface-active
Agent, polyethylene glycol, cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talcum powder, calcium stearate, magnesium stearate etc..
Coptis reference extract of the invention is also used as coptis standard extract, uses or for bulk pharmaceutical chemicals as medicine
The effective component of compositions, weight ratio 0.01-99.9%.
The present invention provides the preparation method of coptis reference extract, steps are as follows: seeing Fig. 4 process flow chart
1, coptis dry cream: the Rhizoma Coptidis powder methanol ultrasonic extraction of separate sources and batch is taken, filtrate 1 is filtered to obtain；
The dregs of a decoction again with methanol ultrasonic extraction 1-3 times, 1-3 filtrate of collection mixes with filtrate 1, is evaporated to obtain coptis dry cream；
2, coptis extract: coptis dry cream is dissolved in methanol, adds auxiliary material and is evaporated and is ground up, sieved up to coptis extract；
3, coptis extract detects: being extracted with thin-layered chromatography and/or high performance liquid chromatography to the coptis of different batches
The step of jateorrhizine, palmatin hydrochloride, Berberine hydrochloride, epiberberine and the coptisine of object are detected；
4, coptis reference extract: the coptis extract after will test carries out allotment and blends, and obtains coptis reference extract.
The w/v of Rhizoma Coptidis powder and methanol is 1:8~1:50 (preferably 1:10) in above-mentioned steps 1；It is described super
The sound time is 30~60min, and power is 100W~3kW (preferably ultrasonic time is 30min, power 500W).Described in step 1
Filtering be with Medium speed filter paper or 2000 mesh net filtrations.
Methanol usage is 2-5 times of dry cream weight in the step (2).
Enter 10%~60% auxiliary material of dry cream weight in the step (2) in coptis dry cream methanol solution, it is described auxiliary
Material includes but is not limited to: superfine silica gel powder, medical starch etc..Further preferred auxiliary material is 20-50% (preferably 40%) superfine silica gel powder,
And/or 10-30% (preferably 20%) medical starch.The auxiliary material, which is dosed, can guarantee that extract character is stable, uniformly and is convenient for
It uses.
Being ground up, sieved after being evaporated in the step 2 was 90~200 mesh (preferably 200 mesh) the screen to filtrate.
Detection described in above-mentioned steps 3, wherein the thin-layered chromatography of coptis extract includes the following steps:
(1) chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；It is accurate respectively
Jateorrhizine, epiberberine, coptisine and palmatine are weighed, is configured to the solution of 0.2mg/ml with methanol respectively；
(2) coptis extract solution: accurate coptis extract is ultrasonically treated with methanol, constant volume, and being made into concentration is 3mg/ml
Solution crosses 0.22 μm of filter membrane to obtain the final product；
(3) the thin-layered chromatography testing conditions are as follows:
Lamellae: G60 prefabricated board；
Point sample: 0.5 μ l, the long 10mm of ribbon point sample；
Solvent: butyl acetate: methanol: isopropanol: ammonium hydroxide=5:2:1.5:1.2；
The drying of lamellae: the lamellae after point sample is placed in phosphorus pentoxide vacuum desiccator, guarantees the drying of lamellae；
It inspects: being placed under UV366nm and inspect.
Wherein the high performance liquid chromatography of coptis extract includes the following steps:
(1) preparation of chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；Point
Precision does not weigh jateorrhizine, epiberberine, coptisine and palmatine, is configured to the solution of 0.2mg/ml with methanol respectively.
(2) preparation of coptis extract test solution: taking coptis extract that methanol-hydrochloric acid (100:1) is added, at ultrasound
Reason, lets cool, and is configured to the solution that concentration is 0.24mg/ml, and filtration takes subsequent filtrate to get coptis extract solution；
(3) chromatographic condition:
The high performance liquid chromatography testing conditions:
Chromatographic apparatus: high performance liquid chromatograph is furnished with DAD detector
Chromatographic column: C18Column (250mm × 4.6mm, 5 μm)；
Mobile phase: A- acetonitrile, the aqueous solution of B- -0.3% triethylamine Han 0.3% phosphoric acid；
Gradient elution program: 0-25min:15%A → 25%A,
25-40min:25%A → 28%A,
40-50min:28%A；
Detection wavelength 270nm (DAD detector)；Flow velocity 0.8ml/min；10 μ l of sample volume；25 DEG C of column temperature.
(4) detection method: accurate each 10 μ l of coptis extract test solution injects liquid chromatograph, records chromatogram.
Allotment described in above-mentioned steps (4) refers specifically to: the coptis extract of separate sources or batch being deployed, should be made
It is 1:(2-5 that crude drug ratio is corresponded in final coptis reference extract), preferably 1:3 (g/g), wherein (content is in terms of wt%)
Content >=11.0% of jamaicin, content >=3.0% of palmatine, content >=1.5% of epiberberine, the content of coptisine >=
2.0%.It is preferred that in coptis reference extract of the present invention, content >=11.8% of jamaicin, the content of palmatine >=
3.4%, content >=1.9% of epiberberine, content >=2.5% of coptisine.
The range of the content of each ingredient in allotment mark of the present invention, which is also that experiment is comprehensive repeatedly by repeatedly, protects
The optimum data that the qualitative, consistency and subsequent application etc. of keeping steady obtain.
The present invention also provides the discrimination method or the coptis of a kind of Rhizoma Coptidis or Chinese materia medica preparation or effective components containing the coptis
Medicinal material or Chinese materia medica preparation method of quality control, by the above-mentioned coptis being prepared according to extract, sample to be tested is with thin
Layer chromatography and/or high performance liquid chromatography are detected, and comparison differentiates.
Beneficial effects of the present invention are explained by following test examples
The preparation method of coptis reference extract of the invention use slightly mention, refine, plus auxiliary material drying, be ground up, sieved and
The step of allotment, easy to operate, reproducible and recovery rate are high.The coptis reference extract being prepared with this preparation method because
It is multiple or different batches extracts deploy, overcomes because Chinese medicine control medicinal material is by the place of production, growing environment
It influences, it is difficult to ensure that the defect per batch of uniform quality, to ensure that the consistent of the coptis reference extract of different batches
Property；Simultaneously by adding solvent appropriate, guarantee the consistency of extract and medicinal ingredient；Representative medicinal material is chosen to make
For raw material, obtained product is representative, and the quality of medicinal material to be detected more can be intuitively reflected as reference extract；By adding
Add auxiliary material, guarantees that extract character is stable, uniform and easy to use.Using the preparation method of coptis reference extract of the invention
It is not low containing jateorrhizine, palmatine, jamaicin, epiberberine, coptis alkali content summation in finally obtained reference extract
In 18%, content >=11.0% of Berberine, content >=3.0% of palmatine, content >=1.5% of epiberberine is yellow
Connect content >=2.0% of alkali and reference extract TLC Fingerprints, HPLC finger-print are corresponding with medicinal material consistent；And with
Compared to easy to use, reference extract only needs simply to dissolve i.e. usable without cumbersome extraction control medicinal material
Journey.The present invention also provides the discrimination method or the coptis of medicinal material or Chinese materia medica preparation or containing the coptis/coptis effective component medicinal material or
The method of quality control of Chinese materia medica preparation not only can carry out Qualitive test to medicinal material or Chinese materia medica preparation, but also can be also used for half
Quantitative even quantitative analysis.
Detailed description of the invention
Fig. 1 is to carry out thin-layer chromatography to commercially available medicinal material for using chemical reference substance and coptis reference extract as reference substance
The high performance thin layer chromatography figure that method obtains.Wherein the chromatographic band of label S is corresponding in turn to from lower to upper as reference substance jateorrhizine, hydrochloric acid bar
Ma Ting, Berberine hydrochloride, epiberberine and coptisine, 1 corresponds to coptis reference extract, and 2-9 is corresponding in turn to the coptis 1, the coptis
2, the medicinal material of the coptis 3, the coptis 4, the coptis 5, the coptis 6, the coptis 7 and the coptis 8.
Fig. 2 is the finger-print common pattern map for the Rhizoma Coptidis established.Wherein No. 6 chromatographic peaks correspond to table barberry
Alkali, No. 7 chromatographic peaks correspond to jateorrhizine, and No. 8 chromatographic peaks correspond to coptisine, and No. 9 chromatographic peaks correspond to palmatin hydrochloride, and No. 10
Chromatographic peak corresponds to Berberine hydrochloride.
Fig. 3 is to carry out efficient liquid phase to commercially available medicinal material for using chemical reference substance and coptis reference extract as reference substance
The HPLC finger-print stacking chart that chromatography obtains, 1-8 respectively corresponds the coptis 1, the coptis 2, the coptis 3, the coptis 4, Huang from top to bottom
Even 5, the coptis 6, the coptis 7 and the coptis 8, ERS correspond to coptis reference extract, and R corresponds to chemical reference substance.
Fig. 4 is coptis reference extract preparation flow figure of the present invention.
Specific embodiment
With reference to the accompanying drawing, specific embodiments of the present invention will be described in detail, it is to be understood that guarantor of the invention
Shield range is not limited by the specific implementation.
Unless otherwise explicitly stated, otherwise in entire disclosure and claims, term " includes " or its change
Changing such as "comprising" or " including " etc. will be understood to comprise stated element or component, and not exclude other members
Part or other component parts.
Embodiment 1, coptis reference extract of the present invention replace the comparative test of control medicinal material
With thin-layered chromatography and high performance liquid chromatography respectively using chemical reference substance and coptis reference extract as reference
Substance analyzes commercially available medicinal material.
1 thin-layer chromatography
1.1 sample preparation
Chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；Precision weighs respectively
Jateorrhizine, epiberberine, coptisine and palmatine are configured to the solution of 0.2mg/ml with methanol respectively.
Coptis reference extract solution: precision weighs the coptis reference extract 60mg that embodiment 1 is prepared, and first is added
Alcohol 20ml, ultrasonic (500w) are handled 15 minutes, are shaken up, and cross 0.22 μm of filter membrane up to coptis reference extract solution).
Medicinal material test solution: taking each 0.2g of pharmacy's medicinal material, and methanol 20ml is added, and ultrasonic (500w) takes out after 15 minutes,
Centrifugation 3 minutes (revolving speed is 3200 turns per minute), supernatant is taken to cross 0.22um filter membrane up to medicinal material test solution.Pharmacy's medicinal material
It buys from Guangzhou, is respectively as follows: the coptis 1, the coptis 2, the coptis 3, the coptis 4, the coptis 5, the coptis 6, the coptis 7 and the coptis 8.
The detection of 1.2 thin-layer chromatographys
Testing conditions are as follows:
Lamellae: HPTLC G60 prefabricated board (Merck)；
Point sample: 0.5 μ l, the long 10mm of ribbon point sample；
Solvent: butyl acetate: methanol: isopropanol: ammonium hydroxide=5:2:1.5:1.2；
Expansion mode: add solvent 10ml in the side double flute expansion cylinder (20cm × 10cm), two 10ml are placed in the other side
Small beaker, each be added 10ml concentrated ammonia liquor (i.e. aqueous solution of the 25%-28% containing ammonia) pre-equilibrate 15 minutes (solvent usage amounts jointly
Volume ratio with concentrated ammonia liquor usage amount is 1:2, and the fixed sample-loading amount of the small beaker of each 10ml is that 10ml concentrated ammonia liquor (contains ammonia
The aqueous solution of 25%-28%), if the specification of the developing tank used is 10cm × 10cm, it is only necessary to place the small of a 10ml
Beaker)；
The drying of lamellae: the lamellae after point sample is placed in phosphorus pentoxide vacuum desiccator 2 hours, guarantees lamellae
It is dry；
It inspects: being placed under UV366nm and inspect.
Testing result is as shown in Figure 1, to inspect thin-layer chromatogram (T:17 DEG C, RH:37%) under UV366nm.The color of label S
Bands of a spectrum are arranged from lower to upper to be corresponding in turn to as reference substance jateorrhizine, palmatin hydrochloride, Berberine hydrochloride, epiberberine and coptisine, and 1
Coptis reference extract is corresponded to, 2-9 is corresponding in turn to the coptis 1, the coptis 2, the coptis 3, the coptis 4, the coptis 5, the coptis 6, the coptis 7 and Huang
Even 8 medicinal material.
Interpretation of result: as shown in Figure 1, five kinds of alkaloids (jateorrhizine, palmatin hydrochloride, Berberine hydrochloride, epiberberine and
Coptisine) separation situation is preferable, and coptis reference extract and medicinal material (coptis 1- coptis 8) can show identical in same position
Spot, find out coptis reference extract from map and medicinal material (coptis 1- coptis 8) have high consistency.
2 high performance liquid chromatography
2.1 sample preparation
Chemical reference substance solution: precision weighs jamaicin 5mg, and the solution of 0.1mg/ml is configured to methanol；It is accurate respectively
Jateorrhizine, epiberberine, coptisine and each 10mg of palmatine are weighed, is configured to the solution of 0.2mg/ml with methanol respectively.
Coptis reference extract solution: precision weighs the coptis control that embodiment 1 is prepared and extracts, and methanol-salt is added
Sour (volume ratio of 100:1), ultrasonic (500w) are handled 30 minutes, are let cool, and are configured to the solution that concentration is 0.24mg/ml, mistake
0.22 μm of filter membrane, take subsequent filtrate to get.
Medicinal material test solution: each pharmacy's medicinal powder is crossed No. two sieves (850 ± 29 μm, 24 mesh), accurately weighed each 0.2g,
It sets in 50ml stuffed conical flask, the mixed solution 40ml of methanol-hydrochloric acid (volume ratio of 100:1), close plug, ultrasound is added in precision
(500w) takes out after 30 minutes, lets cool, and adds at methanol to scale 50ml, shakes up, and filtration, precision measures subsequent filtrate 2ml, sets 10ml
In measuring bottle, add at methanol to scale 10ml, shake up, filter, take subsequent filtrate to get.Pharmacy's medicinal material is bought from Guangzhou, is respectively as follows:
The coptis 1, the coptis 2, the coptis 3, the coptis 4, the coptis 5, the coptis 6, the coptis 7 and the coptis 8.
2.2 high performance liquid chromatography detection
High performance liquid chromatography testing conditions are as follows:
Chromatographic apparatus: 1260 series high performance liquid chromatograph of Agilent is furnished with DAD detector (U.S., Agilent
Technologies)
Chromatographic column: Kromasil 100-5C18Column (250mm × 4.6mm, 5 μm)；
Mobile phase: A- acetonitrile, -0.3% triethylamine of B-0.3% phosphoric acid
Gradient elution program: 0-25min:15%A → 25%A, 25-40min:25%A → 28%A, 40-50min:28%
A；Detection wavelength 270nm (DAD detector)；Flow velocity 0.8ml/min；10 μ l of sample volume；25 DEG C of column temperature.
High effective liquid chromatography for detecting: accurate respectively to draw chemical reference substance solution, coptis reference extract solution
With each 10 μ l of medicinal material test solution, liquid chromatograph is injected.
High performance liquid chromatography testing result:
Measurement records chromatogram to get HPLC finger-print stacking chart shown in Fig. 3, and 1-8 respectively corresponds Huang from top to bottom
Even 1, the coptis 2, the coptis 3, the coptis 4, the coptis 5, the coptis 6, the coptis 7 and the coptis 8, ERS correspond to coptis reference extract, R correspondenceization
Learn reference substance.
As seen from Figure 3, the finger-print of 8 medicinal material of the coptis 1- coptis has high consistency, thus establishes the coptis
The finger-print common pattern of medicinal material, utilizes Chinese Pharmacopoeia Commission's similarity evaluation
(2012.130723 version) calculates, and carries out similarity analysis to all samples.Common pattern map is as shown in Fig. 2, share 10
A shared peak, wherein No. 6 chromatographic peaks are epiberberine, No. 7 chromatographic peaks are jateorrhizine, and No. 8 chromatographic peaks are coptisine, No. 9 chromatographies
Peak is palmatin hydrochloride, and No. 10 chromatographic peaks are Berberine hydrochloride.
According to set Rhizoma Coptidis finger-print common pattern, using included angle cosine algorithm according to each sample component and its
Peak area carries out similarity evaluation to 8 batches of Rhizoma Coptidis and reference extract sample, and the results are shown in Table 3.
Table 3
| Sample | Similarity |
| The coptis 1 | 0.997 |
| The coptis 2 | 0.998 |
| The coptis 3 | 0.999 |
| The coptis 4 | 0.988 |
| The coptis 5 | 0.977 |
| The coptis 6 | 0.984 |
| The coptis 7 | 0.979 |
| The coptis 8 | 0.995 |
| Coptis reference extract | 0.998 |
According to the above similarity analysis as a result, the similarity of 8 batches of medicinal materials and Rhizoma Coptidis finger-print common pattern exists
Within the scope of 0.977-0.999, similarity is very high.The similarity of coptis reference extract and Rhizoma Coptidis finger-print common pattern
It is 0.998, shows that the consistency of coptis reference extract and medicinal material of the invention is good.
According to the result of TLC and HPLC finger-print it is found that being examined to the coptis reference extract using thin-layered chromatography
The corresponding raw medicinal material of the chromatogram measured is consistent, more precisely, uses thin layer to the coptis reference extract
At the jateorrhizine in chromatogram, palmatin hydrochloride, Berberine hydrochloride, epiberberine and the coptisine position that chromatography detects
Fluorescence spot and the chromatogram that is detected using high performance liquid chromatography jateorrhizine, palmatin hydrochloride, Berberine hydrochloride,
It is consistent that epiberberine and the chromatographic peak at coptisine position distinguish corresponding chemical reference substance or its corresponding raw medicinal material,
Therefore coptis reference extract of the invention can be applied to the Qualitive test of medicinal material or Chinese materia medica preparation, such as to medicinal material or Chinese medicine system
Jateorrhizine, palmatin hydrochloride, Berberine hydrochloride, epiberberine and the coptis alkali composition of agent carry out qualitative analysis.
According to HPLC measurement result and statistical analysis, the Rhizoma Coptidis finger-print common pattern and Rhizoma Coptidis of foundation
Similarity it is high, and the similarity pole of coptis reference extract and Rhizoma Coptidis finger-print common pattern of the invention
Height, this explanation carry out jateorrhizine, palmatin hydrochloride, hydrochloric acid to medicinal material or Chinese materia medica preparation using coptis reference extract of the invention
The coptis reference extract that the assay reliability of jamaicin, epiberberine and coptisine is high therefore of the invention can be applied
In medicinal material or the sxemiquantitative discriminatory analysis of Chinese materia medica preparation, can also be applied to the coptis and containing the coptis/coptis effective component medicinal material or
In the quality control of Chinese materia medica preparation, such as the jateorrhizine to medicinal material or Chinese materia medica preparation, palmatin hydrochloride, Berberine hydrochloride, table are small
Bark of a cork tree alkali and coptis alkali composition carry out sxemiquantitative discriminatory analysis, or to the coptis and containing the coptis/coptis effective component medicinal material or in
Medicine preparation carries out half-quantitative detection and then controls its quality.
The character analysis of embodiment 2, coptis reference extract
1. apparent state: the coptis reference extract that embodiment 3 obtains is yellow powder.
2. determination of moisture: being carried out according to 2010 editions Chinese Pharmacopoeia annex IX G (hypobaric drying method).Testing result is the coptis
Reference extract water content is 6.45%.
3. uniformity test: 10 batch coptis reference extracts are prepared according to the method for embodiment 1, after measured between each batch
For epiberberine, coptisine, palmatine and jamaicin thin-layer chromatography testing result difference very little, epiberberine, the coptis
The fluorescence spot of alkali, palmatine and jamaicin has apparent display, and is distributed very consistent；Through high performance liquid chromatography detection its
Epiberberine, coptisine, palmatine and content of berberine are within allotment critical field.Therefore the coptis pair of the invention is utilized
The coptis reference extract consistency being prepared according to the preparation method of extract is very good.
4. stability test:
The test sample for taking the coptis reference extract prepared according to the method for embodiment 1 of 10 different batches, according to state
The relevant regulations of " requirements of national drug standards substance Development Techniques " that pharmacopoeia commission, family works out, inspection target include character with
Dissolubility.
Test sample opening is set in suitable clean container, is placed 10 days at a temperature of 60 DEG C, is sampled in the 5th day and the 10th day,
It is detected by stability high spot reviews project.
The results show that before and after hot test test sample character without significant change, before and after hot test thin-layer chromatogram also without
Significant change, dissolution rate measurement result carry out Independent samples t-test with SPSS, and calculated result P > 0.05 illustrates no conspicuousness
Difference.
The preparation of embodiment 3, coptis reference extract
Instrument and reagent: the semi-automatic point sample instrument of 5 thin-layer chromatography of Linomat, the full-automatic point sample instrument of 4 thin-layer chromatography of ATS, thin
Layer chromatography double flute expansion cylinder, the colour developing of Chromatogram Immersion Device III chromatography dipping tank and TLC
Visualizer thin-layer chromatography video camera (Switzerland, CAMAG).
Agilent 1260series high performance liquid chromatograph is furnished with DAD detector (U.S., Agilent
Technologies)。
Methanol, ethyl alcohol, isopropanol are to analyze pure (Tianjin great Mao chemical reagent factory).Toluene, ethyl acetate, acetic acid fourth
Ester, concentrated ammonia liquor and triethylamine are to analyze pure (Guangzhou Chemical Reagent Factory).
Berberine hydrochloride (also referred to as jamaicin), palmatin hydrochloride (also referred to as palmatine in text), coptisine, epiberberine, mark
Show purity >=98%, (Sichuan Wei Keqi Biotechnology Co., Ltd)；
It is envisaged for preparing the medicinal material of reference extract: the coptis.
Rhizoma Coptidis 8 batches buyings are in the major pharmacy in Guangzhou and medicinal material market, the coptis 1, the coptis 2, the coptis 3 and the coptis 4: Guangzhou
The Qingping Free Market；5: great Can woods pharmacy of the coptis；The shop coptis 6:1；The coptis 7: the pharmacy of anterior approach；The coptis 8: collection and hall pharmacy.
The preparation method of coptis reference extract of the present invention is present embodiments provided, method flow diagram is as shown in Figure 4.
One, it extracts
It chooses Rhizoma Coptidis and powder is made, medicinal powder crosses No. two sieves (850 ± 29 μm, 24 mesh), its quality is then added
The methanol (i.e. solid-to-liquid ratio is 1:10 (w/V)) of 10 times of volumes, middling speed is qualitative after ultrasonic (power 500w, room temperature) extracts 30min
Filter paper filtering, collects filter residue 1 and filtrate 1, and filter residue 1 is extracted secondary again by same procedure, and will extract secondary collection again
Filtrate merges with filtrate 1 Ji Wei extracting solution, and extracting solution solvent evaporated is got dry extract；
Two, coptis extract is prepared
After dry cream is dissolved completely with the methanol of dry cream 2-5 times (w/v), then plus 40% micro mist silicon of dry cream weight
20% medical starch of glue and dry cream weight, is evaporated with Rotary Evaporators, crushed 200 meshes, obtain coptis extract.
Three, extract analyte detection: 6 batches of coptis extracts of preparation are detected according to the detection method of test example two, and testing result is such as
Shown in table 4.
The component content table of 4 coptis extract of table
Four, it deploys
The coptis extract of 6 batches of step 3 is mixed in a certain ratio and blends coptis reference extract, is deployed
Standard are as follows: it is about 1:3 (g/g) that final products, which correspond to crude drug ratio, should make final coptis reference extract Berberine
Content >=11.0%, content >=3.0% of palmatine, content >=1.5% of epiberberine, content >=2.0% of coptisine
(content is in terms of wt%) wherein the measurement result of the content of each ingredient is as shown in table 5.
Component content in 5 coptis reference extract of table
The aforementioned description to specific exemplary embodiment of the invention is in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to the above instruction, can much be changed
And variation.The purpose of selecting and describing the exemplary embodiment is that explaining specific principle of the invention and its actually answering
With so that those skilled in the art can be realized and utilize a variety of different exemplary implementation schemes of the invention and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (10)
1. a kind of coptis reference extract, it is characterised in that the extract is yellow dried powder, is by containing weight percent
It is formed than coptis extract 67-84% and auxiliary material 16-33%, jateorrhizine, palmatine, barberry is wherein contained in coptis extract
Alkali, epiberberine, coptis alkali content summation are not less than 18%.
2. coptis reference extract as described in claim 1, which is characterized in that jamaicin described in coptis reference extract contains
Amount >=11.0%, content >=3.0% of palmatine, content >=1.5% of epiberberine, content >=2.0% of coptisine；Or it is yellow
Even content >=11.0% of jamaicin described in reference extract, content >=3.0% of palmatine, the content of epiberberine >=
1.5%, content >=2.0% of coptisine, content >=1.0% of jateorrhizine.
3. coptis reference extract as claimed in claim 2, which is characterized in that in the coptis reference extract, barberry
Content >=11.8% of alkali, content >=3.4% of palmatine, content >=1.9% of epiberberine, the content of coptisine >=
2.5%.
4. coptis reference extract as described in claim 1, which is characterized in that the auxiliary material is superfine silica gel powder, medical starch
Middle one or two.
5. a kind of coptis reference extract as described in claim 1, which is characterized in that be to be prepared by the following method
:
(1) coptis dry cream: the Rhizoma Coptidis powder methanol ultrasonic extraction of separate sources and batch is taken, filtrate 1 is filtered to obtain；The dregs of a decoction
Again with methanol ultrasonic extraction 1-3 times, 1-3 filtrate of collection mix with filtrate 1, are evaporated to obtain coptis dry cream；
(2) coptis extract: coptis dry cream is dissolved in methanol, adds auxiliary material and is evaporated and is ground up, sieved up to coptis extract；
(3) coptis reference extract: carrying out allotment for separate sources or the coptis extract of batch and blend, and obtains coptis control and mentions
Take object.
6. a kind of coptis reference extract as claimed in claim 5, which is characterized in that increase different coptis medicines before step (1)
The screening step in material source: by separate sources or the raw medicinal material of batch with thin-layered chromatography and/or high performance liquid chromatography into
Row detection；Then the raw medicinal material of separate sources or batch is obtained by thin-layered chromatography and/or high performance liquid chromatography
Chromatogram and/or palmatine, jamaicin, epiberberine and the coptis alkali content of measurement be compared analysis, reject chromatogram
And/or the raw medicinal material of the content exception of measurement, the consistent raw medicinal material progress of content for retaining chromatogram and/or measurement are next
The preparation of the coptis dry cream of step.
7. a kind of coptis reference extract as claimed in claim 5, which is characterized in that further include between step 2 and step 3 with
Thin-layered chromatography and/or high performance liquid chromatography to the palmatine of the coptis extract of different batches, jamaicin, epiberberine and
The step of coptisine is detected.
8. a kind of coptis reference extract as described in claim 1, which is characterized in that Rhizoma Coptidis powder and first in step 1
The w/v of alcohol is 1:8~1:50；The ultrasonic time is 30~60min, and power is 100W~3kW；Methanol in step 2
Dosage is 2-5 times of dry cream weight；Allotment described in step 3 refers specifically to: separate sources or the coptis extract of batch are carried out
Allotment, should make to correspond to crude drug ratio 1:(2-5 in final coptis reference extract) g/g, wherein content is small in terms of wt%
Content >=11.0% of bark of a cork tree alkali, content >=3.0% of palmatine, content >=1.5% of epiberberine, the content of coptisine >=
2.0%.
9. a kind of coptis reference extract as claimed in claim 6, which is characterized in that the thin-layer chromatography of the coptis extract
Method and/or high effective liquid chromatography for detecting, include the following steps:
Wherein thin-layered chromatography includes the following steps:
(1) chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；Precision weighs respectively
Jateorrhizine, epiberberine, coptisine and palmatine are configured to the solution of 0.2mg/ml with methanol respectively；
(2) coptis extract solution: accurate coptis extract is ultrasonically treated with methanol, constant volume, and being made into concentration is 3mg/ml solution,
Cross 0.22 μm of filter membrane to obtain the final product；
(3) the thin-layered chromatography testing conditions are as follows:
Lamellae: G60 prefabricated board；
Point sample: 0.5 μ l, the long 10mm of ribbon point sample；
Solvent: butyl acetate: methanol: isopropanol: ammonium hydroxide=5:2:1.5:1.2；
The drying of lamellae: the lamellae after point sample is placed in phosphorus pentoxide vacuum desiccator, guarantees the drying of lamellae；
It inspects: being placed under UV366nm and inspect；
Wherein high performance liquid chromatography includes the following steps:
(1) preparation of chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；Essence respectively
It is close to weigh jateorrhizine, epiberberine, coptisine and palmatine, the solution of 0.2mg/ml is configured to methanol respectively；
(2) preparation of coptis extract test solution: taking coptis extract that methanol-hydrochloric acid (100:1) is added, ultrasonic treatment,
It lets cool, is configured to the solution that concentration is 0.24mg/ml, filtration takes subsequent filtrate to get coptis extract solution；
(3) chromatographic condition:
The high performance liquid chromatography testing conditions:
Chromatographic apparatus: high performance liquid chromatograph is furnished with DAD detector
Chromatographic column: C18Column (250mm × 4.6mm, 5 μm)；
Mobile phase: A- acetonitrile, the aqueous solution of B- -0.3% triethylamine Han 0.3% phosphoric acid；
Gradient elution program: 0-25min:15%A → 25%A,
25-40min:25%A → 28%A,
40-50min:28%A；
Detection wavelength 270nm, DAD detector；Flow velocity 0.8ml/min；10 μ l of sample volume；25 DEG C of column temperature；
(4) detection method: precision draws each 10 μ l of test solution, injects liquid chromatograph, records chromatogram and calculates medicine root
Alkali, epiberberine, coptisine and palmatine, content of berberine.
10. the discrimination method or the coptis of a kind of Rhizoma Coptidis or Chinese materia medica preparation or the medicinal material or Chinese medicine system of the effective component containing the coptis
The method of quality control of agent, which is characterized in that by coptis reference extract described in claim 1, sample to be tested with thin layer
Chromatography and/or high performance liquid chromatography are detected, and comparison differentiates,
Wherein thin-layered chromatography includes the following steps:
(1) chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；Precision weighs respectively
Jateorrhizine, epiberberine, coptisine and palmatine are configured to the solution of 0.2mg/ml with methanol respectively；
(2) coptis reference extract solution: accurate coptis reference extract is ultrasonically treated with methanol, constant volume, is made into concentration and is
3mg/ml solution crosses 0.22 μm of filter membrane to obtain the final product；
(3) sample to be tested: medicinal material test solution contains coptis: taking sample 0.2g, methanol 20ml is added, takes after ultrasound
Out, it is centrifuged, supernatant is taken to cross 0.22um filter membrane up to medicinal material test solution；
(4) the thin-layered chromatography testing conditions are as follows:
Lamellae: G60 prefabricated board；
Point sample: 0.5 μ l, the long 10mm of ribbon point sample；
Solvent: butyl acetate: methanol: isopropanol: ammonium hydroxide=5:2:1.5:1.2；
The drying of lamellae: the lamellae after point sample is placed in phosphorus pentoxide vacuum desiccator, guarantees the drying of lamellae；
It inspects: being placed under UV366nm and inspect；
Wherein high performance liquid chromatography includes the following steps:
(1) preparation of chemical reference substance solution: precision weighs jamaicin, and the solution of 0.1mg/ml is configured to methanol；Essence respectively
It is close to weigh jateorrhizine, epiberberine, coptisine and palmatine, the solution of 0.2mg/ml is configured to methanol respectively；
(2) preparation of coptis reference extract solution: taking coptis reference extract that methanol-hydrochloric acid (100:1) is added, at ultrasound
Reason, lets cool, and is configured to the solution that concentration is 0.24mg/ml, and filtration takes subsequent filtrate to get coptis reference extract solution；
(3) sample to be tested test solution: Rhizoma Coptidis powder contains coptis, accurately weighed each 0.2g, sets 50ml tool plug
In conical flask, the mixed solution 40ml of methanol-hydrochloric acid (volume ratio of 100:1), close plug is added in precision, and ultrasound is taken out, lets cool, add
It at methanol to scale 50ml, shaking up, filters, precision measures subsequent filtrate 2ml, and it sets in 10ml measuring bottle, adds at methanol to scale 10ml,
Shake up, filter, take subsequent filtrate to get；
(4) chromatographic condition:
The high performance liquid chromatography testing conditions:
Chromatographic apparatus: high performance liquid chromatograph is furnished with DAD detector
Chromatographic column: C18Column (250mm × 4.6mm, 5 μm)；
Mobile phase: A- acetonitrile, the aqueous solution of B- -0.3% triethylamine Han 0.3% phosphoric acid；
Gradient elution program: 0-25min:15%A → 25%A,
25-40min:25%A → 28%A,
40-50min:28%A；
Detection wavelength 270nm, DAD detector；Flow velocity 0.8ml/min；10 μ l of sample volume；25 DEG C of column temperature；
(5) detection method: precision draws each 10 μ l of medicinal material test solution, injects liquid chromatograph, records chromatogram and calculating
Jateorrhizine, epiberberine, coptisine and palmatine, content of berberine.
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Application publication date: 20191025 |
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