CN108362802A - A kind of root of Aucklandia lappa Decne reference extract and its preparation method and application - Google Patents
A kind of root of Aucklandia lappa Decne reference extract and its preparation method and application Download PDFInfo
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- CN108362802A CN108362802A CN201810035641.9A CN201810035641A CN108362802A CN 108362802 A CN108362802 A CN 108362802A CN 201810035641 A CN201810035641 A CN 201810035641A CN 108362802 A CN108362802 A CN 108362802A
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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
- G01N30/06—Preparation
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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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
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Abstract
The present invention provides a kind of root of Aucklandia lappa Decne reference extracts and its preparation method and application, including repeatedly being extracted to more batches of root of Aucklandia lappa Decne medicinal powders respectively, the Radix Aucklandiae extract of different batches is obtained, then the Radix Aucklandiae extract of different batches is allocated, obtains root of Aucklandia lappa Decne reference extract.The root of Aucklandia lappa Decne reference extract of the present invention preparation method is easy to operate, at low cost, reproducible and recovery rate is high；Root of Aucklandia lappa Decne reference extract is prepared with this preparation method, ensure that the consistency of the root of Aucklandia lappa Decne reference extract of different batches, and its character is stable, uniform and easy to use；Costunolide and dehydro-α-curcumene total content must not be less than 3.5% in the finally obtained reference extract of preparation method of root of Aucklandia lappa Decne reference extract using the present invention and not higher than 9.0%, water content is not higher than 5.0%；And reference extract TLC Fingerprints, HPLC finger-prints are corresponding with certified products root of Aucklandia lappa Decne medicinal material consistent.
Description
Technical field
The present invention relates to quality control technologies for traditional Chinese medicine fields, more particularly to a kind of root of Aucklandia lappa Decne reference extract and its preparation side
Method and application.
Background technology
Existing traditional Chinese medicine quality control pattern is substantially the development along Natural Medicine Chemistry, a certain or several to Chinese medicine
Active ingredient be target analysis method and not only difinite quality again can quantify quality standard design, reference foreign countries autonomic drug matter
Amount control method uses for reference the pattern of chemicals quality control, and establishes corresponding simple physics and chemistry by document report and differentiate, then
Develop to the discriminating based on spectrum, chromatography and the quality standard of assay.
All it is multicomponent synthesis per Chinese medicine simply, which dictates that its distinctive globality and ambiguity, also indicate that
One or two using in medicinal material even there are significant limitations as the evaluation method of quality of medicinal material for several ingredients.1990 editions《China
Pharmacopeia》Increase the indentification by TLC of control medicinal material so that the discriminating of Chinese medicine and Chinese patent drug has prodigious progress.In at present
Medicine and external herbal medicine increasingly pay attention to multicomponent or multi-component detection, such as the quality control of German ginkgo biloba p.e.
The analysis method of finger-print can carry out quality control to medicinal material on the whole, currently still there is prodigious researching value.
There are two types of " reference substance ", chemical reference substance and Chinese medicine comparison medicines in terms of controlling quality of medicinal material for Chinese Pharmacopoeia at present
Material.Wherein chemical reference substance can be used for Qualitive test and the quantitative analysis of medicinal material, Chinese medicine control medicinal material be then used for microscopical characters and
Thin layer differentiates.However, chemical reference substance and Chinese medicine control medicinal material have its limitation in traditional Chinese medicine quality control.This limitation
It shows, one, chemical composition diversification in Chinese medicine, single or several compounds can not reflect the overall picture of medicinal material, and existing
Standard the phenomenon that often having many loopholes, adulterating happen occasionally；Two, Chinese medicine control medicinal material is by the place of production, growing environment
Influence, it is difficult to ensure that per batch of uniform quality, and be only used for Qualitive test, can not reflect the height of medicinal ingredient content
It is low.
Chinese medicine reference extract is the character and stable components prepared with Chinese medicine, can be used for qualitative or quantitative analysis
Extract, Mr. Xie Peishan proposes four basic demands (ASCS) to Chinese medicine reference extract and reference extract is answered
Has primary condition：Authenticity, crude drug source is reliable, representative；Specificity, the detection method used
Specificity；Con-sistency, different batches reference extract should be consistent；Stability, character is stable, uniformly, just
In use.With the methods of high performance thin layer chromatography finger-print and high-efficiency liquid-phase fingerprint, it can be used for the qualitative of medicinal material
Differentiate, by the reference extract of external standard method mark, sxemiquantitative and quantitative analysis detection can be further utilized to.Chinese medicine control carries
Take object that there will be important meaning to traditional Chinese medicine quality control.
Invention content
In consideration of it, it is necessary to provide a kind of root of Aucklandia lappa Decne reference extracts and its preparation method and application.The root of Aucklandia lappa Decne pair of the present invention
According to extract preparation method is easy to operate, at low cost, reproducible and recovery rate is high, the root of Aucklandia lappa Decne pair prepared with this preparation method
According to extract consistency is good, character is stable, uniform and easy to use, it can reflect the overall picture of medicinal material.
The present invention uses following technical scheme：
A kind of root of Aucklandia lappa Decne reference extract, wherein costunolide and dehydro-α-curcumene total content must not be less than 3.5% and
Not higher than 9.0%, water content is not higher than 5.0%.
A kind of preparation method of root of Aucklandia lappa Decne reference extract, including more batches of root of Aucklandia lappa Decne medicinal powders are repeatedly extracted respectively,
The Radix Aucklandiae extract of different batches is obtained, then the Radix Aucklandiae extract of different batches is allocated, obtains root of Aucklandia lappa Decne control extraction
Object；It is described that root of Aucklandia lappa Decne medicinal powder is repeatedly extracted, it is the ingredient for making it contain costunolide and dehydro-α-curcumene
Purity and/or concentration improve, to obtain Radix Aucklandiae extract.
Further, the preparation method of the root of Aucklandia lappa Decne reference extract, includes the following steps：
Step 1: II class petroleum ether extracts：Sample is repeatedly extracted using II class petroleum ether, merges extracting solution, system
Extract I is obtained, filter residue is spare；
Step 2: ethyl alcohol extracts：The filter residue in step 1 is repeatedly extracted with ethyl alcohol, extract II is made；
Step 3: mixing：The extract II of extract I and rapid two gained obtained by step 1 is mixed, obtained root of Aucklandia lappa Decne carries
Take object；
Step 4: according to the operation of step 1 to step 3, the root of Aucklandia lappa Decne medicinal material of each batch is prepared into root of Aucklandia lappa Decne extraction respectively
Object；
Step 5: allotment：The Radix Aucklandiae extract of each batch made from step 4 is deployed into root of Aucklandia lappa Decne reference extract.
Further, the preparation method of the root of Aucklandia lappa Decne reference extract, specifically includes following steps：
Step 1: II class petroleum ether (with " petroleum ether (II) " hereinafter) extraction：Take root of Aucklandia lappa Decne medicinal powder cross No. 2 sieve with
Petroleum ether (II) mixes, and is filtered after ultrasonic extraction, obtains filtrate I1With filter residue I1, filter residue I1Repeat the operation, the filter residue of gained
Ⅰ2Repeat the operation；N times are recycled with this, collect each filtrate I respectively1-ⅠNWith filter residue IN；By filtrate I1- ⅠNMerge
Extracting solution I obtains extract I after extracting solution I is recycled petroleum ether (II)；
Step 2: ethyl alcohol extracts：By the filter residue I of step 1NIt is mixed with the dense ethyl alcohol for 50%-80% (preferably 60%) of volume
It closes, is filtered after homogenate extraction, obtain filtrate II1With filter residue II1, filter residue II1Repeat the operation, the filter residue II of gained2It repeats
The operation；It is recycled X times with this, collects each filtrate II respectively1-ⅡXWith filter residue IIX；By filtrate II1-ⅡXMerge to obtain filtrate
Ⅱ；It is evaporated filtrate II to obtain dry cream, dry cream is dissolved in 60% ethyl alcohol, obtain root of Aucklandia lappa Decne dry cream ethanol solution, add auxiliary material,
It is evaporated sieving and obtains extract II.
Step 3: mixing：The extract I that step 1 is obtained petroleum ether (II) dissolves, and what addition step 2 obtained carries
Object II, mixing is taken to be evaporated, sieving obtains Radix Aucklandiae extract.
Step 4: each batch root of Aucklandia lappa Decne medicinal powder in multiple batches of root of Aucklandia lappa Decne medicinal powder to be pressed to step 1 respectively to step 3
Operation be made different batches Radix Aucklandiae extract.
Step 5: allotment：The Radix Aucklandiae extract of different batches is carried out mixed in equal amounts is taken to allocate, obtains root of Aucklandia lappa Decne control extraction
Object.
Further, the w/v of the root of Aucklandia lappa Decne medicinal powder (g) in the step 1 and petroleum ether (II) (ml) is
1:5~1:50, preferably 1：5；
Further, the ultrasonic time in the step 1 is 30~60min, and power is 100W~3kW；Preferably
30min, power 500W.
Further, the filtering in the step 1 is filtered with Medium speed filter paper or 2000 mesh screens.
Further, 6 >=N ＞ 0 in the step 1, and N is integer；It is preferred that N is 3.
Further, 6 >=X ＞ 0 in the step 2, and X is integer；It is preferred that X is 5.
Further, the filter residue I in the step 2N(g) it is 1 with the w/v of ethyl alcohol (ml):5~1:50, it is excellent
It is selected as 1：5；
Further, the homogenate extraction time in the step 2 is 1~2min, and power is 100W~3kW；Preferably
1min, power 500W.
Further, the filtering in the step 2 is filtered with Medium speed filter paper or 2000 mesh screens.
Further, root of Aucklandia lappa Decne dry cream (g) and the weight of 60% ethyl alcohol (ml) and volume ratio 1 in the step 2:5-1:10；
It is preferred that 1:8.
Further, the auxiliary material weight being added in root of Aucklandia lappa Decne dry cream ethanol solution in the step 2 is the ethyl alcohol of root of Aucklandia lappa Decne dry cream
The 20%~60% of solution weight；It is preferred that 50%.
Preferably, the auxiliary material is porous silica gel.
Further, it was the filtering of 90~200 mesh screens after the step 2 is evaporated；It is preferred that 110 mesh screens.
Further, the w/v of extract I (g) and petroleum ether (II) (ml) is 1 in the step 3:8-1:
50；Preferably 1:20.
Further, it was the filtering of 90~200 mesh screens after the step 3 is evaporated；It is preferred that 110 mesh screens.
Further, step 5 further includes with thin-layered chromatography and/or high performance liquid chromatography before allotment to difference
The allotment pre-treatment that the costunolide and dehydro-α-curcumene of the Radix Aucklandiae extract of batch are detected.
Further, the standard allocated in the step 5 is：It should make in finally obtained reference extract in costene
Ester and dehydro-α-curcumene total content must not be less than 3.5% and be not higher than 9%；
Root of Aucklandia lappa Decne reference extract made from the method for the present invention root of Aucklandia lappa Decne, containing root of Aucklandia lappa Decne, the active ingredient containing root of Aucklandia lappa Decne medicinal material or in
The composition detection of medicine preparation or the application in quality control.
A kind of detection root of Aucklandia lappa Decne, containing root of Aucklandia lappa Decne, the medicinal material of the active ingredient containing root of Aucklandia lappa Decne or the compositions, method of Chinese medicine preparation, including use
Thin-layered chromatography and/or high performance liquid chromatography in root of Aucklandia lappa Decne reference extract, medicinal material or Chinese medicine preparation costunolide and
Dehydro-α-curcumene is detected, and comparison judges.
Further, it when being detected using the detection method, needs root of Aucklandia lappa Decne reference extract, product configuration Cheng Rong to be checked
Liquid carries out again.
Further, the preparation method of root of Aucklandia lappa Decne reference extract solution is in the thin-layered chromatography：Precision weighs described
Root of Aucklandia lappa Decne reference extract 0.23g-1.15g is configured to the solution of a concentration of 0.115g/mL-0.575g/mL with methanol, crosses 0.12-
0.32 μm of filter membrane is up to root of Aucklandia lappa Decne reference extract solution；It is preferred that described weigh root of Aucklandia lappa Decne reference extract 0.23g, it is configured to methanol
The solution of a concentration of 0.115g/mL crosses 0.22 μm of filter membrane to get root of Aucklandia lappa Decne reference extract solution.
Further, product solution preparation method to be checked is in the thin-layered chromatography：After taking product to be checked to cross No. two sieves
Weigh 0.5g-5g, be added 10mL-100mL chloroforms, power is 100W~3kW, after 30~60min of ultrasound with Medium speed filter paper or
2000 mesh screens filter, and filter residue repeats to extract once, merges filtrate twice, chloroform is flung in 60 DEG C of -80 DEG C of water-baths, after filtrate is evaporated
Residue add methanol to dissolve surely and hold to 2-20mL, cross 0.12-0.32 μm of filter membrane up to product solution to be checked；It is preferred that weighing to be checked
Product 0.5g, is added chloroform 10mL, ultrasonic 30min, power 500w, Medium speed filter paper filtering, and chloroform, filtrate are flung in 60 DEG C of water-baths
Residue after being evaporated adds methanol to dissolve surely and is settled to 2mL, crosses 0.22 μm of filter membrane to get product solution to be checked.
Further, the thin-layered chromatography testing conditions are：
Lamellae：HPTLC G60 prefabricated boards；
Point sample：3 μ l, the long 7mm of ribbon point sample；
Solvent：Toluene, chloroform, ethyl acetate and methanol
Deployment step such as table 1：
Table 1
The drying of lamellae：Lamellae after point sample is placed in thin layer heating plate, and 50 DEG C of heating 15min ensure lamellae
Drying；
It inspects:About paradime thylaminobenzaldehyde, the ethanol solution containing 20% phosphoric acid and 20% sulfuric acid of spray 5mL 2%, 105
DEG C heating 1.5min, be placed under UV366nm and inspect.
Further, in the high performance liquid chromatography detection, the preparation method of root of Aucklandia lappa Decne reference extract solution is：It is accurate
The root of Aucklandia lappa Decne reference extract 5mg-50mg is weighed, the solution of a concentration of 2.5mg/mL-25mg/mL, mistake are configured to methanol
0.12-0.32 μm of filter membrane is up to root of Aucklandia lappa Decne reference extract solution；It is preferred that described weigh root of Aucklandia lappa Decne reference extract 5mg, matched with methanol
The solution of a concentration of 2.5mg/mL is made, crosses 0.22 μm of filter membrane to get root of Aucklandia lappa Decne reference extract solution.
Further, in the high performance liquid chromatography detection, the preparation method of product solution to be checked is：Take product to be checked
Precision weighs 0.3g-3g after crossing No. two sieves, sets in conical flask with cover, and 50mL-500mL methanol, close plug is added, and weighed weight is put
It sets overnight, 30~60min of ultrasound, power is 100W~3kW, lets cool, then weighed weight, supplies weightlessness with methanol, shake up, mistake
0.12-0.32 μm of filter membrane takes subsequent filtrate up to product solution to be checked.It is preferred that precision weighs product 0.3g to be checked, 50mL first is added
Alcohol, ultrasonic 30min, power 500W cross 0.22 μm of filter membrane.
Further, the high performance liquid chromatography testing conditions：
Chromatographic apparatus：1260 series high performance liquid chromatographs of Agilent are furnished with DAD detectors (U.S., Agilent
Technologies)
Chromatographic column：Agilent Zorbax SB C18Column (4.6mm × 250mm, 5 μm)；
Mobile phase：A- methanol, B-0.1% phosphate aqueous solutions；
Gradient elution program：0-18min：20%A → 40%A, 18-40min：40%A → 75%A, 40-65min：
75%A-100%A；65-70min:100%A；
Detection wavelength 237nm (DAD detectors)；Flow velocity 1ml/min；10 μ l of sample size；25 DEG C of column temperature；Run time：
70min。
The invention has the advantages that：
The step that the preparation method of the root of Aucklandia lappa Decne reference extract of the present invention uses extraction, prepares Radix Aucklandiae extract and allotment
Suddenly, easy to operate, at low cost, reproducible and recovery rate is high.The root of Aucklandia lappa Decne reference extract being prepared with this preparation method because
It is that the extracts of different batches allocate, overcoming is influenced because of Chinese medicine control medicinal material by the place of production, growing environment, very
Be difficult to guarantee the defect per batch of uniform quality, to ensure that different batches root of Aucklandia lappa Decne reference extract consistency, and
Its character is stable, uniform and easy to use；The finally obtained control of preparation method of root of Aucklandia lappa Decne reference extract using the present invention
Costunolide and dehydro-α-curcumene total content must not be less than 3.5% in extract and not higher than 9%, water content is not higher than
5.0%, and reference extract TLC Fingerprints, HPLC finger-prints are corresponding with certified products root of Aucklandia lappa Decne medicinal material consistent.The present invention
Additionally provide the discrimination method or root of Aucklandia lappa Decne or medicinal material or Chinese medicine preparation containing root of Aucklandia lappa Decne/root of Aucklandia lappa Decne active ingredient of medicinal material or Chinese medicine preparation
Method of quality control, can not only carry out Qualitive test to medicinal material or Chinese medicine preparation, but also can be also used for sxemiquantitative even
Quantitative analysis.
The root of Aucklandia lappa Decne reference extract made from the method for the present invention is obtained using thin-layered chromatography and/or high performance liquid chromatography
The corresponding raw medicinal material of the collection of illustrative plates that arrives is consistent；
The root of Aucklandia lappa Decne reference extract made from the method for the present invention uses in the chromatogram that thin-layered chromatography detects
The chromatogram that fluorescence spot and use high performance liquid chromatography at costunolide and dehydro-α-curcumene position detect
Chromatographic peak at costunolide and dehydro-α-curcumene position should distinguish corresponding chemical reference substance or its is corresponding
Certified products medicinal material is consistent.
Description of the drawings
Fig. 1 is detected with thin-layered chromatography to root of Aucklandia lappa Decne raw medicinal material as reference substance using chemical reference substance and control medicinal material
The TLC finger-prints arrived, S is corresponding in turn to from lower to upper to be compareed for reference substance costunolide and dehydro-α-curcumene, 1 for root of Aucklandia lappa Decne
Medicinal material, 2-7 correspond to root of Aucklandia lappa Decne raw medicinal material 1, root of Aucklandia lappa Decne raw medicinal material 2, root of Aucklandia lappa Decne raw medicinal material 3, root of Aucklandia lappa Decne raw medicinal material 4, root of Aucklandia lappa Decne respectively
Raw medicinal material 5 and root of Aucklandia lappa Decne raw medicinal material 6,8 are elecampane control medicinal material.
Fig. 2 is examined with high performance liquid chromatography to root of Aucklandia lappa Decne raw medicinal material as reference substance using chemical reference substance and control medicinal material
The HPLC finger-print stacking charts measured, S1 costunolide reference substances, S2 are dehydro-α-curcumene reference substance, from top to bottom
1 be root of Aucklandia lappa Decne medicinal material common pattern, 2 be root of Aucklandia lappa Decne control medicinal material, 3-8 correspond to respectively root of Aucklandia lappa Decne raw medicinal material 1, root of Aucklandia lappa Decne raw medicinal material 2,
Root of Aucklandia lappa Decne raw medicinal material 3, root of Aucklandia lappa Decne raw medicinal material 4, root of Aucklandia lappa Decne raw medicinal material 5 and root of Aucklandia lappa Decne raw medicinal material 6,9 are elecampane control medicinal material.
Fig. 3 is root of Aucklandia lappa Decne reference extract preparation flow figure of the present invention.
4-1 is for being reference substance to city using chemical reference substance, root of Aucklandia lappa Decne control medicinal material and root of Aucklandia lappa Decne reference extract in Fig. 4
It sells medicinal material and carries out the high performance thin layer chromatography figure that thin-layered chromatography obtains.The wherein chromatographic band of label S be corresponding in turn to from lower to upper for
Reference substance costunolide and dehydro-α-curcumene, 1 corresponds to root of Aucklandia lappa Decne reference extract of the present invention, and 2 correspond to root of Aucklandia lappa Decne control medicinal material,
3-12 is corresponding in turn to root of Aucklandia lappa Decne 1, root of Aucklandia lappa Decne 2, root of Aucklandia lappa Decne 3, root of Aucklandia lappa Decne 4, root of Aucklandia lappa Decne 5, root of Aucklandia lappa Decne 6, root of Aucklandia lappa Decne 7, root of Aucklandia lappa Decne 8, root of Aucklandia lappa Decne 9 and root of Aucklandia lappa Decne 10,
Medicinal material, 13 correspond to elecampane control medicinal material.4-2 is that Fig. 4-1 is swept by the correspondence number that " Chempattern " software scans obtain
Tracing spectrum.
Fig. 5 is to carry out efficient liquid phase to commercially available medicinal material as reference substance for using chemical reference substance and root of Aucklandia lappa Decne reference extract
The HPLC finger-print stacking charts that chromatography obtains, S1 costunolide reference substances, S2 is dehydro-α-curcumene reference substance, from upper
Root of Aucklandia lappa Decne medicinal material common pattern is corresponded to lower 1,2 correspond to root of Aucklandia lappa Decne reference extract, and 3 correspond to root of Aucklandia lappa Decne control medicinal material, and 4-13 is corresponded to respectively
Elecampane is corresponded to for root of Aucklandia lappa Decne 1, root of Aucklandia lappa Decne 2, root of Aucklandia lappa Decne 3, root of Aucklandia lappa Decne 4, root of Aucklandia lappa Decne 5, root of Aucklandia lappa Decne 6, root of Aucklandia lappa Decne 7, root of Aucklandia lappa Decne 8, root of Aucklandia lappa Decne 9 and root of Aucklandia lappa Decne 10,14
Control medicinal material.
Fig. 6 is the finger-print common pattern collection of illustrative plates for the root of Aucklandia lappa Decne medicinal material established.Wherein No. 13 chromatographic peaks correspond to table root of Aucklandia lappa Decne
Hydrocarbon lactone, No. 14 chromatographic peaks correspond to dehydro-α-curcumene.
Specific implementation mode
In order to better illustrate technical solution of the present invention problem to be solved, the technical solution of use and reach beneficial
Effect is expanded on further in conjunction with specific implementation mode.It is worth noting that technical solution of the present invention is including but not limited to following
Embodiment.
Particular technique or condition are not specified in the embodiment of the present invention, according to technology described in document in the art or
Condition is carried out according to product description.Reagents or instruments used without specified manufacturer is that can be obtained by purchased in market
The conventional products obtained.
Through validation trial, method in the embodiment, within the scope of aforementioned parameters prepared by arbitrary selection parameter
The property of obtained root of Aucklandia lappa Decne reference extract is suitable, is used to detect the effect that the effect of sample is also enumerated with present embodiment
Quite, since the experimental record of repetition test and data volume are larger, present embodiment is only by taking some of design parameters as an example, no
It lists one by one again.
Instrument and material：
It is envisaged for preparing the medicinal material of reference extract:
Root of Aucklandia lappa Decne medicinal material is purchased from the major medicinal material market in the whole nation, is identified as composite family section plant root of Aucklandia lappa Decne Aucklandia lappa
The dry root of Decne.
The full-automatic point sample instrument of 4 thin-layer chromatographys of ATS, thin-layer chromatography double flute expansion cylinder and TLC visualizer thin-layer chromatographys
Video camera (Switzerland, CAMAG).
1260 series high performance liquid chromatographs of Agilent are furnished with DAD detectors (U.S., Agilent
Technologies)。
Methanol, ethyl alcohol are to analyze pure (Tianjin great Mao chemical reagent factories).
Toluene, ethyl acetate, chloroform (Guangzhou Chemical Reagent Factory).
Costunolide and dehydro-α-curcumene indicate purity >=98%, (Baoji time bio tech ltd)；
Test medicinal material root of Aucklandia lappa Decne：
The 11 batches of buyings of root of Aucklandia lappa Decne medicinal material are in the major pharmacy in Guangzhou and medicinal material market, root of Aucklandia lappa Decne 1-11.
The screening of 1 root of Aucklandia lappa Decne raw medicinal material of embodiment
Present embodiments provide the screening technique of the raw medicinal material of root of Aucklandia lappa Decne reference extract of the present invention.
With thin-layered chromatography and high performance liquid chromatography respectively by reference substance of chemical reference substance to root of Aucklandia lappa Decne bulk pharmaceutical chemicals
Material is analyzed and is screened.
Root of Aucklandia lappa Decne medicinal material is purchased from the major medicinal material market in the whole nation, has been identified as composite family section plant root of Aucklandia lappa Decne Aucklandia lappa
The dry root of Decne.Identified 9 batches of medicinal materials comply with standard can be spare.
1 thin-layer chromatography
1.1 sample preparation
Chemical reference substance solution：Precision weighs costunolide and dehydro-α-curcumene, is configured to 1mg/ with methanol respectively
The solution of ml.
Root of Aucklandia lappa Decne reference extract solution：Precision weighs the root of Aucklandia lappa Decne reference extract 0.23g that embodiment 1 is prepared, and sets 2mL
In volumetric flask, methanol 1.5ml is added, ultrasonic (500w) is handled 30 minutes, let cool, and with methanol constant volume to scale, crosses 0.22 μm of filter
Film is up to root of Aucklandia lappa Decne reference extract solution.
Medicinal material test solution：Pharmacy's root of Aucklandia lappa Decne medicinal powder (crossing No. two sieves) each 0.5g is taken, 10ml chloroform ultrasounds are added
30min, filtering, then add 10ml chloroform ultrasound 30min, filtering, merging filtrate, water-bath flings to chloroform, adds methanol constant volume to 2ml,
Cross 0.22 μm of filter membrane to get.
1.2 thin-layer chromatographys detect
Testing conditions are as follows：
Lamellae：HPTLC G60 prefabricated boards (Merck)；
Point sample：3 μ l, the long 7mm of ribbon point sample；
The drying of lamellae：Lamellae is set 50 DEG C of thin layer heating version and is heated 15 minutes after point sample.
Solvent：Toluene, chloroform, ethyl acetate and methanol
Expansion mode：AMD2 is unfolded, deployment step such as following table：
Table 2
It inspects：The ethanol solution that about 5mL2% paradime thylaminobenzaldehydes contain 20% sulfuric acid and 20% phosphoric acid is sprayed, thin layer is set
105 DEG C of heating 1min, chromatogram image is inspected under UV366nm in heating plate.Testing result such as Fig. 1, costunolide and dehydrogenation
Constuslactone separation asks condition fine, and each batch of medicinal material can show identical spot in same position, and similarity is high between each batch,
Preliminary screening is standard control extract raw medicinal material.
2 high performance liquid chromatography
2.1 sample preparation
Chemical reference substance solution：Precision weighs appropriate costunolide, dehydro-α-curcumene reference substance, is configured to methanol
The solution of 0.1mg/ml.
Root of Aucklandia lappa Decne reference extract solution：Precision weighs ERS 5mg, sets in 2ml volumetric flasks, adds methanol 1.5ml, is ultrasonically treated
It 30 minutes, lets cool, adds methanol constant volume scale, shake up, cross 0.22 μm of filter membrane to obtain the final product.
Medicinal material test solution：Qu Ge pharmacies Common Aucklandia Root (crossing No. four sieves) about 0.3g, it is accurately weighed, set tool plug taper
In, methanol 50ml, close plug is added in precision, and weighed weight is stood overnight, and is ultrasonically treated (power 250W, frequency 50kHz) 30min,
It lets cool, then weighed weight, supplies less loss weight with methanol, shake up, filter, take subsequent filtrate to obtain the final product.
2.2 high performance liquid chromatography detection
High performance liquid chromatography testing conditions are as follows：
Chromatographic apparatus：Agilent 1260series high performance liquid chromatographs are furnished with DAD detectors (U.S., Agilent
Technologies)
Chromatographic column：Agilent ZORBAX SB C18(4.6mm×250mm,5μm)；
Mobile phase：A- methanol, B-0.1% phosphate aqueous solutions；
Gradient elution program：0-18min：20%A → 40%A, 18-40min：40%A → 75%A, 40-65min：
75%A-100%A；65-70min:100%A；
Detection wavelength 237nm (DAD detectors)；Flow velocity 1ml/min；10 μ l of sample size；25 DEG C of column temperature；Run time：
70min。
High effective liquid chromatography for detecting：It is accurate respectively to draw chemical reference substance solution, root of Aucklandia lappa Decne reference extract solution
With each 10 μ l of medicinal material test solution, liquid chromatograph is injected.
High performance liquid chromatography testing result：
It measuring, record chromatogram is to get HPLC finger-prints stacking chart shown in Fig. 2, S1 costunolide reference substances,
S2 be dehydro-α-curcumene reference substance, from top to bottom 1 be root of Aucklandia lappa Decne medicinal material common pattern, 2 be root of Aucklandia lappa Decne control medicinal material, 3-8 difference
Corresponding root of Aucklandia lappa Decne raw medicinal material 1, root of Aucklandia lappa Decne raw medicinal material 2, root of Aucklandia lappa Decne raw medicinal material 3, root of Aucklandia lappa Decne raw medicinal material 4,5 and of root of Aucklandia lappa Decne raw medicinal material
Root of Aucklandia lappa Decne raw medicinal material 6,9 is elecampane control medicinal material.
As seen from Figure 2, the finger-print of 6 raw medicinal material of root of Aucklandia lappa Decne control medicinal material and root of Aucklandia lappa Decne 1- root of Aucklandia lappa Decne has the one of height
Thus cause property establishes the finger-print common pattern of root of Aucklandia lappa Decne medicinal material, and carries out similarity analysis to all samples.
According to set root of Aucklandia lappa Decne medicinal materials fingerprint common pattern, using included angle cosine algorithm according to each sample component and its
Peak area carries out similarity evaluation to root of Aucklandia lappa Decne control medicinal material and 6 batches of root of Aucklandia lappa Decne medicinal material samples, and the results are shown in Table 3.
Table 3
| Sample | Similarity |
| Root of Aucklandia lappa Decne control medicinal material | 0.997 |
| Root of Aucklandia lappa Decne raw medicinal material 1 | 0.998 |
| Root of Aucklandia lappa Decne raw medicinal material 2 | 0.994 |
| Root of Aucklandia lappa Decne raw medicinal material 3 | 0.993 |
| Root of Aucklandia lappa Decne raw medicinal material 4 | 0.992 |
| Root of Aucklandia lappa Decne raw medicinal material 5 | 0.999 |
| Root of Aucklandia lappa Decne raw medicinal material 6 | 0.992 |
| Common pattern | 1 |
According to above similar
Analysis result is spent, the similarity of 6 batches of root of Aucklandia lappa Decne raw medicinal materials is very high, can be selected and is extracted for root of Aucklandia lappa Decne standard control extract
Obtain raw medicinal material.
The present embodiment illustrates the selection identification method of raw material medicinal material only by taking 6 batches of raw material medicinal materials as an example.It is of the invention signified
" multiple batches of " include but not limited to the present embodiment batch.
The preparation of 2 root of Aucklandia lappa Decne reference extract of the present invention of embodiment
Root of Aucklandia lappa Decne extraction is respectively prepared in 4 batches for selecting similarity high from the raw material through screening and identification in embodiment 1
Object.
With reference to the flow chart of attached drawing 3
One：Extraction
Root of Aucklandia lappa Decne medicinal material is chosen, powder is made, medicinal powder crosses No. two sieves (850 ± 29 μm, 24 mesh), its matter is then added
(i.e. solid-to-liquid ratio is 1 to the petroleum ether (II) of 5 times of volumes of amount：5 (w/V)), ultrasonic (power 500w, room temperature) extracts middling speed after 30min
Qualitative filter paper filters, and collects filter residue I1With filtrate I1, filter residue I1It extracts three times, and will be again extracted three times again by same procedure
The filtrate of collection and filtrate I1It is extracting solution I to merge, by after I recycling design of extracting solution up to extract I.
Take filter residue I3, be added 5 times of volumes of its quality 60% ethyl alcohol (i.e. solid-to-liquid ratio be 1：5 (w/V), during sudden strain of a muscle carries after 1min
Fast qualitative filter paper filtering, collects filter residue II1With filtrate II1, filter residue II1It extracts five times, and will be extracted again again by same procedure
The filtrate of five collections and filtrate II1It is extracting solution II to merge, and extracting solution II is evaporated, dry cream is obtained.
Dry cream is dissolved with 60% ethyl alcohol of 8 times of dry cream (w/v), then adds 50% porous silica gel of dry cream weight, is used
Rotary Evaporators are evaporated, and be crushed 110 mesh sieve, are obtained extract II.
Two：Prepare Radix Aucklandiae extract
Extract I is dissolved with the petroleum ether (II) of extract obtained above I 20 times (w/v), is added
Extract II obtained is stated, is evaporated with Rotary Evaporators, 110 mesh sieve is crushed, obtains Radix Aucklandiae extract.
According to the method described above, 4 batches of Radix Aucklandiae extracts are prepared, the detection of high effective liquid chromatography for measuring is utilized by reference substance
The results are shown in Table 4.
Table 4
Three：Allotment
The Radix Aucklandiae extract of 4 above-mentioned batches is pressed 1:1:1:The mixing of 1 ratio blends to obtain root of Aucklandia lappa Decne reference extract, most
It is about 1 that finished product, which corresponds to crude drug ratio,：2 (g/g), wherein the content of each ingredient utilizes high performance liquid chromatography by reference substance
Method measures, and testing result is as shown in table 5.
Table 5
Allotment standard is：It should make in final root of Aucklandia lappa Decne reference extract costunolide and dehydro-α-curcumene total content not
It obtains less than 3.5% and is not higher than 9% (content is in terms of wt%).The range of the content of each ingredient in this allotment standard is also process
The optimum data that repeatedly the comprehensive holding stability of experiment, consistency and subsequent application etc. obtain repeatedly.
The character analysis of 3 root of Aucklandia lappa Decne reference extract of embodiment
1. apparent state：The root of Aucklandia lappa Decne reference extract that embodiment 2 obtains is yellow powder.
2. determination of moisture：It is carried out according to 2015 editions Chinese Pharmacopoeia annex IX G (hypobaric drying method).Testing result is root of Aucklandia lappa Decne
Reference extract water content is 1.51%.
3. uniformity test：4 batch root of Aucklandia lappa Decne reference extracts are prepared according to the method for embodiment 2, after measured between each batch
The thin-layer chromatography testing result difference very little for costunolide and dehydro-α-curcumene, costunolide and dehydrogenation root of Aucklandia lappa Decne
The fluorescence spot of lactone has apparent display, and is distributed very consistent；Through its costunolide of high performance liquid chromatography detection and
Dehydro-α-curcumene content is within allotment critical field.Therefore the preparation method of the root of Aucklandia lappa Decne reference extract of the present invention is utilized
The root of Aucklandia lappa Decne reference extract consistency being prepared is very good.
4. stability test：
The test sample for taking the root of Aucklandia lappa Decne reference extract of the method preparation according to embodiment 2 of 4 different batches, according to country
The relevant regulations for " requirement of national drug standards substance Development Techniques " that pharmacopoeia commission works out, inspection target include character and molten
Xie Xing.
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 sample t-inspection with SPSS, and result of calculation P ＞ 0.05 illustrate no conspicuousness
Difference.
The application for the root of Aucklandia lappa Decne reference extract that 4 embodiment of the present invention 2 of embodiment is prepared
With thin-layered chromatography and high performance liquid chromatography respectively using chemical reference substance and root of Aucklandia lappa Decne reference extract as reference
Substance analyzes commercially available medicinal material.
1 thin-layer chromatography
1.1 sample preparation
Chemical reference substance solution：Precision weighs costunolide and dehydro-α-curcumene, is configured to 1mg/ with methanol respectively
The solution of ml.
Root of Aucklandia lappa Decne reference extract solution：Precision weighs the root of Aucklandia lappa Decne reference extract 0.23g that embodiment 1 is prepared, and sets 2mL
In volumetric flask, methanol 1.5ml is added, ultrasonic (500w) is handled 30 minutes, let cool, and with methanol constant volume to scale, crosses 0.22 μm of filter
Film is up to root of Aucklandia lappa Decne reference extract solution.
Medicinal material test solution：Pharmacy's root of Aucklandia lappa Decne medicinal powder (crossing No. two sieves) each 0.5g is taken, 10ml chloroform ultrasounds are added
30min, filtering, then add 10ml chloroform ultrasound 30min, filtering, merging filtrate, water-bath flings to chloroform, adds methanol constant volume to 2ml,
Cross 0.22 μm of filter membrane to get.
1.2 thin-layer chromatographys detect
Testing conditions are as follows：
Lamellae：HPTLC G60 prefabricated boards (Merck)；
Point sample：3 μ l, the long 7mm of ribbon point sample；
The drying of lamellae：Lamellae is set 50 DEG C of thin layer heating version and is heated 15 minutes after point sample.
Solvent：Toluene, chloroform, ethyl acetate and methanol
Expansion mode：AMD2 is unfolded, deployment step such as following table：
Table 6
It inspects：The ethanol solution that about 5mL2% paradime thylaminobenzaldehydes contain 20% sulfuric acid and 20% phosphoric acid is sprayed, thin layer is set
105 DEG C of heating 1min, inspect and take pictures under UV366nm in heating plate.
Testing result is as shown in Fig. 4-1, to inspect thin-layer chromatogram (T under UV366nm：20℃).The chromatographic band of label S by
Under up row be corresponding in turn to and correspond to root of Aucklandia lappa Decne reference extract of the present invention for reference substance costunolide and dehydro-α-curcumene, 1,
2 correspond to root of Aucklandia lappa Decne control medicinal material, and 3-12 is corresponding in turn to root of Aucklandia lappa Decne 1, root of Aucklandia lappa Decne 2, root of Aucklandia lappa Decne 3, root of Aucklandia lappa Decne 4, root of Aucklandia lappa Decne 5, root of Aucklandia lappa Decne 6, root of Aucklandia lappa Decne 7, wood
Perfume 8,10 medicinal material of root of Aucklandia lappa Decne 9 and root of Aucklandia lappa Decne, 13 correspond to elecampane control medicinal material.
Interpretation of result：By Fig. 4-1 it is found that costunolide and dehydro-α-curcumene separation situation are preferable, root of Aucklandia lappa Decne control carries
Take object, root of Aucklandia lappa Decne control medicinal material and root of Aucklandia lappa Decne medicinal material medicinal material (root of Aucklandia lappa Decne 1- root of Aucklandia lappa Decne 10) that can show identical spot in same position,
Find out that root of Aucklandia lappa Decne reference extract has high consistency with root of Aucklandia lappa Decne control medicinal material and root of Aucklandia lappa Decne medicinal material (root of Aucklandia lappa Decne 1- root of Aucklandia lappa Decne 10) from collection of illustrative plates.
Fig. 4-1 can be obtained into its scanning spectra (Fig. 4-2) by self-programmed software " Digiscan " scanning.
2 high performance liquid chromatography
2.1 sample preparation
Chemical reference substance solution：Precision weighs appropriate costunolide, dehydro-α-curcumene reference substance, is configured to methanol
The solution of 0.1mg/ml.
Root of Aucklandia lappa Decne reference extract solution：Precision weighs ERS 5mg, sets in 2ml volumetric flasks, adds methanol 1.5ml, is ultrasonically treated
It 30 minutes, lets cool, adds methanol constant volume scale, shake up, cross 0.22 μm of filter membrane to obtain the final product.
Medicinal material test solution：Qu Ge pharmacies Common Aucklandia Root (crossing No. four sieves) about 0.3g, it is accurately weighed, set tool plug taper
In, methanol 50ml, close plug is added in precision, and weighed weight is stood overnight, and is ultrasonically treated (power 250W, frequency 50kHz) 30min,
It lets cool, then weighed weight, supplies less loss weight with methanol, shake up, filter, take subsequent filtrate to obtain the final product.
2.2 high performance liquid chromatography detection
High performance liquid chromatography testing conditions are as follows：
Chromatographic apparatus：1260 series high performance liquid chromatographs of Agilent are furnished with DAD detectors (U.S., Agilent
Technologies)
Chromatographic column：Agilent ZORBAX SB C18(4.6mm×250mm,5μm)；
Mobile phase：A- methanol, B-0.1% phosphate aqueous solutions；
Gradient elution program：0-18min：20%A → 40%A, 18-40min：40%A → 75%A, 40-65min：
75%A-100%A；65-70min:100%A；
Detection wavelength 237nm (DAD detectors)；Flow velocity 1ml/min；10 μ l of sample size；25 DEG C of column temperature；Run time：
70min。
High effective liquid chromatography for detecting：It is accurate respectively to draw chemical reference substance solution, root of Aucklandia lappa Decne reference extract solution
With each 10 μ l of medicinal material test solution, liquid chromatograph is injected.
High performance liquid chromatography testing result：
It measuring, record chromatogram is to get HPLC finger-prints stacking chart shown in fig. 5, S1 costunolide reference substances,
S2 is dehydro-α-curcumene reference substance, from top to bottom the common pattern of 1 correspondence root of Aucklandia lappa Decne medicinal material, 2 correspondence root of Aucklandia lappa Decne reference extracts, 3
Corresponding root of Aucklandia lappa Decne control medicinal material, 4-13 correspond to root of Aucklandia lappa Decne 1, root of Aucklandia lappa Decne 2, root of Aucklandia lappa Decne 3, root of Aucklandia lappa Decne 4, root of Aucklandia lappa Decne 5, root of Aucklandia lappa Decne 6, root of Aucklandia lappa Decne 7, wood respectively
Perfume 8, root of Aucklandia lappa Decne 9 and root of Aucklandia lappa Decne 10,14 correspond to elecampane control medicinal material.
As seen from Figure 5, root of Aucklandia lappa Decne control medicinal material and the finger-print of 10 medicinal material of root of Aucklandia lappa Decne 1- root of Aucklandia lappa Decne have the consistent of height
Property, the finger-print common pattern of root of Aucklandia lappa Decne medicinal material is thus established, and similarity analysis is carried out to all samples.Common pattern collection of illustrative plates
As shown in fig. 6, wherein No. 13 chromatographic peaks are costunolide, No. 14 color dehydro-α-curcumenes.According to ERS finger-prints, use
Included angle cosine algorithm is according to each sample component and its peak area to root of Aucklandia lappa Decne control medicinal material, 10 batches of root of Aucklandia lappa Decne medicinal materials and set root of Aucklandia lappa Decne medicine
Material finger-print common pattern carries out similarity evaluation, and the results are shown in Table 7.
Table 7
According to the above similarity analysis as a result, root of Aucklandia lappa Decne control medicinal material, 10 batches of medicinal materials and root of Aucklandia lappa Decne reference extract finger-print
Similarity within the scope of 0.953-0.985, similarity is very high.Root of Aucklandia lappa Decne reference extract and root of Aucklandia lappa Decne medicinal materials fingerprint share mould
The similarity of formula is 0.981, and the consistency of the root of Aucklandia lappa Decne reference extract and medicinal material that show the present invention is good.
According to the result of TLC and HPLC finger-prints it is found that being examined using thin-layered chromatography to the root of Aucklandia lappa Decne reference extract
The corresponding raw medicinal material of the chromatogram that measures is consistent, more precisely, thin layer is used to the root of Aucklandia lappa Decne reference extract
Fluorescence spot at costunolide and dehydro-α-curcumene position in the chromatogram that chromatography detects and using efficient
Chromatographic peak of the chromatogram that liquid chromatography detects at costunolide and dehydro-α-curcumene position is right with it respectively
The chemical reference substance answered or its corresponding raw medicinal material are consistent, thus the present invention root of Aucklandia lappa Decne reference extract can be applied to medicinal material or
The Qualitive test of Chinese medicine preparation, such as the costunolide and dehydro-α-curcumene ingredient of medicinal material or Chinese medicine preparation are carried out qualitative
Analysis.
According to HPLC measurement results and statistical analysis, root of Aucklandia lappa Decne medicinal materials fingerprint common pattern and the root of Aucklandia lappa Decne medicinal material of foundation
Similarity it is high, and the present invention root of Aucklandia lappa Decne reference extract and root of Aucklandia lappa Decne medicinal materials fingerprint common pattern similarity pole
Height, this explanation carry out in costunolide and dehydrogenation root of Aucklandia lappa Decne medicinal material or Chinese medicine preparation using the root of Aucklandia lappa Decne reference extract of the present invention
The assay reliability of ester is high, therefore the root of Aucklandia lappa Decne reference extract of the present invention can be applied to the semidefinite of medicinal material or Chinese medicine preparation
Discriminatory analysis is measured, can also be applied in the quality control of root of Aucklandia lappa Decne and the medicinal material or Chinese medicine preparation containing root of Aucklandia lappa Decne/root of Aucklandia lappa Decne active ingredient, example
Medicine costunolide and dehydro-α-curcumene ingredient such as to medicinal material or Chinese medicine preparation carry out sxemiquantitative discriminatory analysis, or to wood
The fragrant and medicinal material containing root of Aucklandia lappa Decne/root of Aucklandia lappa Decne active ingredient or Chinese medicine preparation carry out half-quantitative detection and then control its quality.
Several embodiments of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
Cannot the limitation to the scope of the claims of the present invention therefore be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention
Protect range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
1. a kind of root of Aucklandia lappa Decne reference extract, which is characterized in that described its costunolide of root of Aucklandia lappa Decne reference extract and dehydrogenation root of Aucklandia lappa Decne
Lactone total content must not be less than 3.5% and not higher than 9.0%, and water content is not higher than 5.0%.
2. a kind of preparation method of root of Aucklandia lappa Decne reference extract, which is characterized in that including being carried out respectively to more batches of root of Aucklandia lappa Decne medicinal powders
Repeatedly extraction, obtains the Radix Aucklandiae extract of different batches, is then allocated to the Radix Aucklandiae extract of different batches, obtain root of Aucklandia lappa Decne
Reference extract.
3. the preparation method of root of Aucklandia lappa Decne reference extract according to claim 2, which is characterized in that include the following steps：
Step 1: II class petroleum ether extracts：Sample is repeatedly extracted using II class petroleum ether, merges extracting solution, is made and carries
Object I is taken, filter residue is spare；
Step 2: ethyl alcohol extracts：The filter residue in step 1 is repeatedly extracted with ethyl alcohol, extract II is made；
Step 3: mixing：The extract II of extract I and rapid two gained obtained by step 1 is mixed, Radix Aucklandiae extract is made；
Step 4: according to the operation of step 1 to step 3, the root of Aucklandia lappa Decne medicinal material of each batch is prepared into Radix Aucklandiae extract respectively；
Step 5: allotment：The Radix Aucklandiae extract of each batch made from step 4 is deployed into root of Aucklandia lappa Decne reference extract.
4. the preparation method of root of Aucklandia lappa Decne reference extract according to claim 3, which is characterized in that include the following steps：
Step 1: II class petroleum ether extracts：It takes root of Aucklandia lappa Decne medicinal powder to be mixed with II class petroleum ether, filters, obtain after ultrasonic extraction
Filtrate I1With filter residue I1；Filter residue I1Repeat the operation；The filter residue I of gained2Repeat the operation；N times are recycled with this, respectively
Collect each filtrate I1-ⅠNWith filter residue IN；By filtrate I1-ⅠNMerge to obtain extracting solution I, after extracting solution I is recycled petroleum ether (II)
Obtain extract I；
Step 2: ethyl alcohol extracts：By the filter residue I of step 1NIt mixes with ethyl alcohol, is filtered after homogenate extraction, obtain filtrate II1And filter
Slag II1；Filter residue II1Repeat the operation；The filter residue II of gained2Repeat the operation；It is recycled X times, is collected respectively each with this
Filtrate II1-ⅡXWith filter residue IIX；By filtrate II1-ⅡXMerge to obtain filtrate II；It is evaporated filtrate II to obtain dry cream, dry cream is molten
Solution obtains root of Aucklandia lappa Decne dry cream ethanol solution, adds auxiliary material in ethyl alcohol, is evaporated sieving and obtains extract II；
Step 3: mixing：The extract I of step 1 petroleum ether (II) is dissolved, is added the extract II of step 2, mixing,
It is evaporated, sieving obtains Radix Aucklandiae extract；
Step 4: according to the operation of step 1 to step 3, the root of Aucklandia lappa Decne medicinal material of each batch is prepared into Radix Aucklandiae extract respectively；
Step 5: allotment：The Radix Aucklandiae extract of different batches obtained by step 4 is allocated, root of Aucklandia lappa Decne control extraction is obtained
Object.
5. the preparation method of root of Aucklandia lappa Decne reference extract according to claim 4, which is characterized in that the wood in the step 1
The w/v of fragrant medicinal powder (g) and petroleum ether (II) (ml) is 1:5~1:50；Ultrasonic time is 30~60min, power
For 100W~3kW.
6. the preparation method of root of Aucklandia lappa Decne reference extract according to claim 4, which is characterized in that the filter in the step 2
Slag IN(g) it is 1 with the w/v of ethyl alcohol (ml):5~1:50；The homogenate extraction time is 1~2min, power 100W
~3kW.
7. the preparation method of root of Aucklandia lappa Decne reference extract according to claim 4, which is characterized in that root of Aucklandia lappa Decne in the step 2
The weight and volume ratio 1 of dry cream (g) and ethyl alcohol (ml):5-1:10；The auxiliary material weight that the root of Aucklandia lappa Decne dry cream ethanol solution is added is
The 20%~60% of the ethanol solution weight of root of Aucklandia lappa Decne dry cream.
8. the preparation method of root of Aucklandia lappa Decne reference extract according to claim 4, which is characterized in that wherein 6 >=N ＞ 0；6≥X
＞ 0；And N, X are integer.
9. root of Aucklandia lappa Decne made from the method described in root of Aucklandia lappa Decne reference extract described in claim 1 or claim 2-8 any one
Reference extract in root of Aucklandia lappa Decne, containing in root of Aucklandia lappa Decne, the composition detection of the medicinal material of the active ingredient containing root of Aucklandia lappa Decne or Chinese medicine preparation or quality control
Application.
10. a kind of detection root of Aucklandia lappa Decne, containing root of Aucklandia lappa Decne, the medicinal material of the active ingredient containing root of Aucklandia lappa Decne or the compositions, method of Chinese medicine preparation, feature exists
In, including using thin-layered chromatography and/or high performance liquid chromatography to root of Aucklandia lappa Decne reference extract described in claim 1 or right
It is required that the costunolide in root of Aucklandia lappa Decne reference extract, medicinal material and/or Chinese medicine preparation made from method described in 2-8 any one
It is detected with dehydro-α-curcumene, comparison judges.
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