CN108840845A - The method of Xanthatin is extracted from Siberian cocklebur - Google Patents
The method of Xanthatin is extracted from Siberian cocklebur Download PDFInfo
- Publication number
- CN108840845A CN108840845A CN201811172744.6A CN201811172744A CN108840845A CN 108840845 A CN108840845 A CN 108840845A CN 201811172744 A CN201811172744 A CN 201811172744A CN 108840845 A CN108840845 A CN 108840845A
- Authority
- CN
- China
- Prior art keywords
- siberian cocklebur
- xanthatin
- coarse powder
- extracted
- extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D307/93—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems condensed with a ring other than six-membered
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The method that the invention discloses a kind of to extract Xanthatin from Siberian cocklebur, which is characterized in that include the following steps：Cellulase solution, buffer solution are added into Siberian cocklebur coarse powder, 45 DEG C of heating water baths, after digesting 1.5~2.5h, inactivate 5~10min, then bionic extraction liquid is added, it is extracted under 37 DEG C, ultrasonic wave auxiliary to get the Siberian cocklebur coarse powder extract, the mass volume ratio of the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid is 1g:2.5~3.4ml:10~16.7ml:30~35ml；The Siberian cocklebur coarse powder extract is separated, purified, is concentrated to get Xanthatin.The method that Xanthatin is extracted from Siberian cocklebur of the invention, the Xanthatin purity of acquisition is up to 98%.
Description
Technical field
The present invention relates to the extracting methods of native compound, more specifically, be related to one kind extracts Xanthatin from Siberian cocklebur
Method.
Background technique
Siberian cocklebur complex chemical composition, main active are Sesquiterpene lactones compound.Currently, being related in existing research
And the achievement for extracting and separating aspect of single sesquiterpene lactone is very few.Xanthatin is in Sesquiterpene lactones compound
A kind of highest ingredient of pharmacological activity, can effectively inhibit the diseases such as the ulcer due to caused by soda acid；But due to containing in Siberian cocklebur
Some Xanthatin contents are lower, and Xanthatin content difference is larger in different Siberian cocklebur samples, thus cause in Xanthatin extraction side
The problems such as face, it is cumbersome that there are extractive techniques, low efficiency and not high purity.
Summary of the invention
It is an object of the invention to solve at least the above problems, and provide the advantages of at least will be described later.
The method that it is a still further object of the present invention to provide a kind of to extract Xanthatin from Siberian cocklebur, the Xanthatin purity of acquisition
Up to 98%.
In order to realize object of the present invention and further advantage, a kind of side that Xanthatin is extracted from Siberian cocklebur is provided
Method includes the following steps：
Cellulase solution, buffer solution are added into Siberian cocklebur coarse powder, 45 DEG C of heating water baths go out after digesting 1.5~2.5h
5~10min living, is then added bionic extraction liquid, extracts under 37 DEG C, ultrasonic wave auxiliary to get the Siberian cocklebur coarse powder extract；
The mass volume ratio of the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid is 1g:2.5~
3.4ml:10~16.7ml:30~35ml；
The Siberian cocklebur coarse powder extract is separated, purified, is concentrated to get Xanthatin.
Preferably, described from the method for extracting Xanthatin in Siberian cocklebur, the preparation of the Siberian cocklebur coarse powder includes following
Step：
S1, it rinse the Siberian cocklebur that parts by weight are 10~20 parts with clear water well after, dries to constant weight, crushes, and excessively 50~
After 100 meshes, extraction temperature is 35~40 DEG C, extracting pressure is 35~40MPa and carbon dioxide flow is 40~50L/h's
Under the conditions of carry out 1~2h of supercritical carbon dioxide extracting after, be made Siberian cocklebur extract；
S2,10~20 parts of parts by weight of water is added in the Siberian cocklebur extract in S1, in temperature be decoct 2 at 70~80 DEG C~
3h, continuous filter 23, obtains filtrate and the first filter residue, spare；
S3, the filtrate in S2 is centrifuged at revolving speed 4000rpm 20~30min through centrifuge, isolates supernatant, precipitated
Object merges with the first filter residue, obtains the second filter residue；
S4, the second filter residue in S3 is placed in the ethanol solution that volume fraction is 85~90% and impregnates 5~10h, filtering,
Obtain soak and third filter residue；
S5, the third filter residue in S4 is placed in percolating device, carries out diacolation using the soak in S4, controls diacolation stream
Speed is 300~400ml/h, obtains percolate, then mixes percolate with the supernatant in S3, is ground after vacuum concentration is dry
Mill is to get Siberian cocklebur coarse powder.
Preferably, described from the method for extracting Xanthatin in Siberian cocklebur, the separation, purifying, concentration include following
Step：
A1, Siberian cocklebur coarse powder extract is mixed with silica gel, obtains Siberian cocklebur silica white；
A2, by silicagel column on the Siberian cocklebur silica white in A1, carry out first time gradient elution, collect every time containing Xanthatin
Eluate, and merge, it is concentrated under reduced pressure, Xanthatin is made, study is concentrated；
A3, reverse phase silica gel column, second of gradient elution of progress, collection on the Xanthatin concentration study in A2 are contained every time
The eluate of Xanthatin, and merge, degreasing is carried out with petroleum ether, is then extracted with ethyl acetate acetic acid butyl ester, is concentrated under reduced pressure,
Up to Xanthatin.
Preferably, described from the side for extracting Xanthatin in Siberian cocklebur, the buffer solution is that NaAc-HAc buffering is molten
Liquid, solid-to-liquid ratio 1:10~1:12.5, pH 4.5；
Preferably, described from the method for extracting Xanthatin in Siberian cocklebur, the preparation method of the bionic extraction liquid is：
Sodium chloride, the pepsin of 3.2g and the hydrochloric acid of 0.02mol of 9g are added in the water of every L, and pH value is adjusted to 6.0.
Preferably, described from the method for extracting Xanthatin in Siberian cocklebur, institute in first time gradient elution described in A2
Eluant, eluent is the mixed liquor of chloroform and methanol, and the mass ratio of chloroform and methanol is followed successively by 80:1~100:1,18:1~25:
1,3:1~5:1,0.8:1~1:1.
Preferably, described from the method for extracting Xanthatin in Siberian cocklebur, institute in second of gradient elution described in A3
Eluant, eluent is methanol solution, and the mass fraction of methanol solution is followed successively by 75~85%, 85~95%, 100%.
Preferably, the second from the method for extracting Xanthatin in Siberian cocklebur, in the ethyl acetate acetic acid butyl ester
Acetoacetic ester and butyl acetate volume ratio 1.5:1~2.5:1.
The present invention is include at least the following beneficial effects：
1, the method that the present invention provides a kind of extracts Xanthatin from Siberian cocklebur, the Xanthatin purity of acquisition is up to 98%.
2, in present invention design, Xanthatin is extracted by the methods of enzymatic hydrolysis, ultrasonic wave auxiliary, the pure of Xanthatin can be improved
Degree.
3, in present invention design, a kind of method for detecting Xanthatin content in Siberian cocklebur is additionally provided, to improve Xanthatin
Purity.Based on high-efficient liquid phase chromatogram technology, efficiently quickly whether meet containing Xanthatin, content in detection Siberian cocklebur sample
It is separated it is required that carrying out purposive extraction for satisfactory sample, can be obtained the Xanthatin of high-purity.This detection method
The specific steps are：After Siberian cocklebur sample is extracted, Siberian cocklebur coarse powder extract is obtained, is detected into high performance liquid chromatograph；
The chromatographic peak peak area of retention time identical as Xanthatin reference substance is greater than 50% and (removes preceding 5 minutes chromatographic peaks after integral to use
Areas of peak normalization method calculates)；Or in the case where no reference substance, retention time has simultaneously in 9.5-11.5 minutes
The chromatographic peak peak area for having 278 nanometers of ultraviolet ends to absorb is greater than 50%, can be used as the material of Xanthatin extraction.This detection side
Method is quick and precisely, easy to operate.
4, in present invention design, a kind of preparation method of Siberian cocklebur coarse powder is additionally provided, the extraction of Siberian cocklebur coarse powder can be improved
Content, and then improve the purity of Xanthatin.In the preparation method of Siberian cocklebur coarse powder, used supercritical carbon dioxide extraction method,
Percolation extracts Siberian cocklebur, can greatly improve the yield of Siberian cocklebur coarse powder.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is the high-efficient liquid phase chromatogram of A1 group in the experiment of Xanthatin purity.
Specific embodiment
The present invention will be further described in detail below with reference to the embodiments, to enable those skilled in the art referring to specification
Text can be implemented accordingly.
It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
Embodiment 1
The method that Xanthatin is extracted from Siberian cocklebur, includes the following steps：Cellulase solution is added into Siberian cocklebur coarse powder, delays
Solution is rushed, 45 DEG C of heating water baths after digesting 1.5h, inactivate 5min, bionic extraction liquid is then added, and assist in 37 DEG C, ultrasonic wave
Lower extraction is to get the Siberian cocklebur coarse powder extract, the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid
Mass volume ratio be 1g:2.5ml:10ml:30ml；The Siberian cocklebur coarse powder extract is separated, is purified, be concentrated to get
Xanthatin.
It is described from the method for extracting Xanthatin in Siberian cocklebur, the separation, purifying, concentration include the following steps：
A1, Siberian cocklebur coarse powder extract is mixed with silica gel, obtains Siberian cocklebur silica white；
A2, by silicagel column on the Siberian cocklebur silica white in A1, carry out first time gradient elution, collect every time containing Xanthatin
Eluate, and merge, it is concentrated under reduced pressure, Xanthatin is made, study is concentrated；
A3, reverse phase silica gel column, second of gradient elution of progress, collection on the Xanthatin concentration study in A2 are contained every time
The eluate of Xanthatin, and merge, degreasing is carried out with petroleum ether, is then extracted with ethyl acetate acetic acid butyl ester, is concentrated under reduced pressure,
Up to Xanthatin.
It is described that the buffer solution is NaAc-HAc buffer solution from the method for extracting Xanthatin in Siberian cocklebur, Gu
Liquor ratio is 1:10, pH 4.5；
It is described from the method for extracting Xanthatin in Siberian cocklebur, the preparation method of the bionic extraction liquid is：In the water of every L
Sodium chloride, the pepsin of 3.2g and the hydrochloric acid of 0.02mol of 9g is added, and pH value is adjusted to 6.0.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in first time gradient elution described in A2
For the mixed liquor of chloroform and methanol, the mass ratio of chloroform and methanol is followed successively by 80:1,18:1,3:1,0.8:1.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in second of gradient elution described in A3
For methanol solution, the mass fraction of methanol solution is followed successively by 75%, 85%, 100%.
It is described from the method for extracting Xanthatin in Siberian cocklebur, ethyl acetate in the ethyl acetate acetic acid butyl ester with
Butyl acetate volume ratio 1.5:1.
Embodiment 2
The method that Xanthatin is extracted from Siberian cocklebur, includes the following steps：Cellulase solution is added into Siberian cocklebur coarse powder, delays
Solution is rushed, 45 DEG C of heating water baths after digesting 2.0h, inactivate 8min, bionic extraction liquid is then added, and assist in 37 DEG C, ultrasonic wave
Lower extraction is to get the Siberian cocklebur coarse powder extract, the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid
Mass volume ratio be 1g:3.0ml:15ml:33ml；The Siberian cocklebur coarse powder extract is separated, is purified, be concentrated to get
Xanthatin.
It is described from the method for extracting Xanthatin in Siberian cocklebur, the separation, purifying, concentration include the following steps：
A1, Siberian cocklebur coarse powder extract is mixed with silica gel, obtains Siberian cocklebur silica white；
A2, by silicagel column on the Siberian cocklebur silica white in A1, carry out first time gradient elution, collect every time containing Xanthatin
Eluate, and merge, it is concentrated under reduced pressure, Xanthatin is made, study is concentrated；
A3, the Xanthatin concentration study in A2 is subjected to upper reverse phase silica gel column, carries out second of gradient elution, collects each
Eluate containing Xanthatin, and merge, degreasing is carried out with petroleum ether, is then extracted with ethyl acetate acetic acid butyl ester, is depressurized dense
Contracting is to get Xanthatin.
It is described that the buffer solution is NaAc-HAc buffer solution from the method for extracting Xanthatin in Siberian cocklebur, Gu
Liquor ratio is 1:11, pH 4.5；
It is described from the method for extracting Xanthatin in Siberian cocklebur, the preparation method of the bionic extraction liquid is：In the water of every L
Sodium chloride, the pepsin of 3.2g and the hydrochloric acid of 0.02mol of 9g is added, and pH value is adjusted to 6.0.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in first time gradient elution described in A2
For the mixed liquor of chloroform and methanol, the mass ratio of chloroform and methanol is followed successively by 90:1,21:1,4:1,0.9:1.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in second of gradient elution described in A3
For methanol solution, the mass fraction of methanol solution is followed successively by 80%, 90%, 100%.
It is described from the method for extracting Xanthatin in Siberian cocklebur, ethyl acetate in the ethyl acetate acetic acid butyl ester with
Butyl acetate volume ratio 2.0:1.
Embodiment 3
The method that Xanthatin is extracted from Siberian cocklebur, includes the following steps：Cellulase solution is added into Siberian cocklebur coarse powder, delays
Solution is rushed, 45 DEG C of heating water baths after digesting 2.5h, inactivate 10min, bionic extraction liquid is then added, and assist in 37 DEG C, ultrasonic wave
Lower extraction is to get the Siberian cocklebur coarse powder extract, the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid
Mass volume ratio be 1g:3.4ml:16.7ml:35ml；The Siberian cocklebur coarse powder extract is separated, purified, is concentrated, i.e.,
Obtain Xanthatin.
It is described from the method for extracting Xanthatin in Siberian cocklebur, the separation, purifying, concentration include the following steps：
A1, Siberian cocklebur coarse powder extract is mixed with silica gel, obtains Siberian cocklebur silica white；
A2, by silicagel column on the Siberian cocklebur silica white in A1, carry out first time gradient elution, collect every time containing Xanthatin
Eluate, and merge, it is concentrated under reduced pressure, Xanthatin is made, study is concentrated；
A3, the Xanthatin concentration study in A2 is subjected to upper reverse phase silica gel column, carries out second of gradient elution, collects each
Eluate containing Xanthatin, and merge, degreasing is carried out with petroleum ether, is then extracted with ethyl acetate acetic acid butyl ester, is depressurized dense
Contracting is to get Xanthatin.
It is described that the buffer solution is NaAc-HAc buffer solution from the method for extracting Xanthatin in Siberian cocklebur, Gu
Liquor ratio is 1:12.5, pH 4.5；
It is described from the method for extracting Xanthatin in Siberian cocklebur, the preparation method of the bionic extraction liquid is：In the water of every L
Sodium chloride, the pepsin of 3.2g and the hydrochloric acid of 0.02mol of 9g is added, and pH value is adjusted to 6.0.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in first time gradient elution described in A2
For the mixed liquor of chloroform and methanol, the mass ratio of chloroform and methanol is followed successively by 100:1,25:1,5:1,1:1.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in second of gradient elution described in A3
For methanol solution, the mass fraction of methanol solution is followed successively by 85%, 95%, 100%.
It is described from the method for extracting Xanthatin in Siberian cocklebur, ethyl acetate in the ethyl acetate acetic acid butyl ester with
Butyl acetate volume ratio 2.5:1.
Embodiment 4
The method that Xanthatin is extracted from Siberian cocklebur, includes the following steps：Cellulase solution is added into Siberian cocklebur coarse powder, delays
Solution is rushed, 45 DEG C of heating water baths after digesting 2.0h, inactivate 8min, bionic extraction liquid is then added, and assist in 37 DEG C, ultrasonic wave
Lower extraction is to get the Siberian cocklebur coarse powder extract, the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid
Mass volume ratio be 1g:3.0ml:15ml:33ml；The Siberian cocklebur coarse powder extract is separated, is purified, be concentrated to get
Xanthatin.
It is described from the method for extracting Xanthatin in Siberian cocklebur, the preparation of the Siberian cocklebur coarse powder includes the following steps：
S1, it rinse the Siberian cocklebur that parts by weight are 15 parts with clear water well after, dries to constant weight, crushes, and after 60 meshes excessively,
Overcritical titanium dioxide is carried out under conditions of extraction temperature is 38 DEG C, extracting pressure is 38MPa and carbon dioxide flow is 45L/h
After carbon extracts 1.5h, Siberian cocklebur extract is made；
S2, the water that the Siberian cocklebur extract in S1 is added to 15 parts of parts by weight are to decoct 2.5h, continuous mistake at 75 DEG C in temperature
Filter 3 times, obtains filtrate and the first filter residue, spare；
S3, the filtrate in S2 is centrifuged 25min at revolving speed 4000rpm through centrifuge, isolates supernatant, sediment with
First filter residue merges, and obtains the second filter residue；
S4, the second filter residue in S3 is placed in the ethanol solution that volume fraction is 88% and impregnates 7h, filtered, impregnated
Liquid and third filter residue；
S5, the third filter residue in S4 is placed in percolating device, carries out diacolation using the soak in S4, controls diacolation stream
Speed be 350ml/h, obtain percolate, then percolate mix with the supernatant in S3, vacuum concentration drying after grind to get
Siberian cocklebur coarse powder.
It is described from the method for extracting Xanthatin in Siberian cocklebur, the separation, purifying, concentration include the following steps：
A1, Siberian cocklebur coarse powder extract is mixed with silica gel, obtains Siberian cocklebur silica white；
A2, by silicagel column on the Siberian cocklebur silica white in A1, carry out first time gradient elution, collect every time containing Xanthatin
Eluate, and merge, it is concentrated under reduced pressure, Xanthatin is made, study is concentrated；
A3, reverse phase silica gel column, second of gradient elution of progress, collection on the Xanthatin concentration study in A2 are contained every time
The eluate of Xanthatin, and merge, degreasing is carried out with petroleum ether, is then extracted with ethyl acetate acetic acid butyl ester, is concentrated under reduced pressure,
Up to Xanthatin.
It is described that the buffer solution is NaAc-HAc buffer solution from the method for extracting Xanthatin in Siberian cocklebur, Gu
Liquor ratio is 1:11, pH 4.5；
It is described from the method for extracting Xanthatin in Siberian cocklebur, the preparation method of the bionic extraction liquid is：In the water of every L
Sodium chloride, the pepsin of 3.2g and the hydrochloric acid of 0.02mol of 9g is added, and pH value is adjusted to 6.0.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in first time gradient elution described in A2
For the mixed liquor of chloroform and methanol, the mass ratio of chloroform and methanol is followed successively by 90:1,21:1,4:1,0.9:1.
It is described from the method for extracting Xanthatin in Siberian cocklebur, eluant, eluent used in second of gradient elution described in A3
For methanol solution, the mass fraction of methanol solution is followed successively by 80%, 90%, 100%.
It is described from the method for extracting Xanthatin in Siberian cocklebur, ethyl acetate in the ethyl acetate acetic acid butyl ester with
Butyl acetate volume ratio 2.0:1.
The experiment of Xanthatin purity
1, experimental principle：First Siberian cocklebur coarse powder is detected with high performance liquid chromatograph, when Siberian cocklebur coarse powder detect it is qualified with
Afterwards, then Xanthatin extraction is carried out, the purity of Xanthatin can be improved.
2, experimental procedure：
Step 1：The detection of Siberian cocklebur coarse powder
Taking Siberian cocklebur coarse powder to number respectively is A1, A2, A3, A4 group, and cellulase solution, NaAc-HAc buffer solution is added,
After 45 degree of heating water baths digest, high-temperature inactivation, add bionic extraction liquid (by 0.9% sodium chloride, 0.32% pepsin and
0.02mol/L hydrochloric acid is formulated) ultrasonic wave assisted extraction 30min, cooled and filtered takes subsequent filtrate 20ml water bath method, adds
Methanol dissolves and is transferred to volumetric flask constant volume, and filtering is detected into high performance liquid chromatograph.If the corresponding chromatographic peak of Xanthatin
Area is greater than 50%, then illustrates that the sample can be used as the material of Xanthatin extraction.The experiment parameter of each experimental group is in a tabular form
It is presented below as：
1 Siberian cocklebur coarse powder test experience parameter of table
| A1 | A2 | A3 | A4 | |
| Siberian cocklebur meal quality (g) | 2 | 1.5 | 3 | 3 |
| Cellulase solution volume (ml) | 5 | 5 | 10 | 10 |
| Volume of buffer solution (ml) | 25 | 25 | 30 | 50 |
| Enzymolysis time (h) | 2 | 1.5 | 2.5 | 2.5 |
| Inactivation time (min) | 8 | 5 | 10 | 10 |
| Bionic extraction liquid (ml) | 70 | 50 | 90 | 100 |
| Chromatographic peak area (%) | 90 | 76 | 94 | 95 |
From the data in table 1, it can be seen that the Siberian cocklebur coarse powder in A1, A2, A3, A4 group is in the chromatographic peak face that high performance liquid chromatograph detects
Product illustrates that four groups of Siberian cocklebur coarse powder is all qualified extraction sample all greater than 50%.
Step 2：Xanthatin is extracted
It weighs respectively and detects qualified Siberian cocklebur coarse powder 500g, number B1, B2, B3, B4 group, mistake in A1, A2, A3, A4 group
After mesh, cellulase solution is added, the NaAc-HAc buffer solution that pH is 4.5, after 45 DEG C of heating water baths digest, high temperature goes out
Living, the bionic extraction liquid for being then 6.0 with pH (is prepared by 0.9% sodium chloride, 0.32% pepsin and 0.02mol/L hydrochloric acid
Into) under 37 DEG C, 500W ultrasonic wave auxiliary extract 30min, cooled and filtered, merging filtrate, 50 DEG C of heating water baths concentrations, with
The mixing of 100-200 mesh silica gel, obtains Siberian cocklebur silica white.
By silicagel column on Siberian cocklebur silica white (silica gel is 200-300 mesh), product is carried out with the mixed liquor of chloroform and methanol
Gradient elution obtains eluted product, and obtained eluted product is carried out thin-layer chromatography, colour developing is observed at ultraviolet light (278nm),
It is compared with the standard items of Xanthatin, finds out and extract the study containing Xanthatin, merging is mutually same, obtains Xanthatin study, depressurizes
Concentration obtains Xanthatin concentration study.
Reverse phase silica gel column on study is concentrated in Xanthatin, study gradient elution is concentrated to the Xanthatin with methanol solution,
Continue thin-layer chromatography, colour developing is observed at ultraviolet light (278nm), is compared with the standard specimen of Xanthatin, by the mark with Xanthatin
The identical spot of sample and observed under fluorescent light without other impurities sample merge, obtain the first of the purified product containing the Xanthatin
Alcoholic solution after the solution petroleum ether degreasing, then is extracted with ethyl acetate acetic acid butyl ester, is concentrated under reduced pressure and removes solvent to get grey
The chemical structure of ear pavilion, Xanthatin is as follows：
In this experiment, the quality of chloroform and methanol is as follows than data in each experimental group：
B1 group：The mass ratio of chloroform and methanol is followed successively by 100：1,25：1,5：1,1：1；B2 group：The quality of chloroform and methanol
Than being followed successively by 80：1,20：1,4：1,1：1；B3 group：The mass ratio of chloroform and methanol is followed successively by 90：1,18：1,3：1,0.8：1；B4
Group:The mass ratio of chloroform and methanol is followed successively by 100：1,25：1,5：1,1：1.
The data of the mass fraction of methanol solution are as follows in each experimental group：
B1 group：The mass fraction of methanol solution is followed successively by 75%, 85% and 100%；B2 group：The mass fraction of methanol solution
It is followed successively by 80%, 90% and 100%；B3 group：The mass fraction of methanol solution is followed successively by 85%, 95% and 100%；B4 group:First
The mass fraction of alcoholic solution is followed successively by 85%, 95% and 100%.
Other experimental datas in each group are presented below as in a tabular form：
2 Xanthatin of table extracts experiment parameter
| B1 | B2 | B3 | B4 | |
| Mesh (mesh) | 40 | 24 | 60 | 70 |
| Cellulase solution volume (L) | 1.25 | 1.7 | 1.7 | 1.7 |
| Cellulase activity (U/g) | 20 | 30 | 35 | 35 |
| Volume of buffer solution (L) | 6.25 | 8.3 | 5 | 8.3 |
| The solid-to-liquid ratio of buffer solution | 1:12.5 | 1:15 | 1:10 | 1:12 |
| Enzymolysis time (h) | 2 | 1.5 | 2.5 | 2.5 |
| Inactivation time (min) | 8 | 5 | 10 | 10 |
| Bionic extraction liquid (L) | 17.5 | 16.7 | 15 | 16.7 |
| Ethyl acetate acetic acid butyl ester volume ratio | 2:1 | 1.5:1 | 2.5:1 | 2.5:1 |
After testing, every group of obtained Xanthatin purity is identified, it is as a result as follows：
3 Xanthatin purity of table
| B1 | B2 | B3 | B4 | |
| Purity (%) | 95 | 88 | 96 | 98 |
From the data in table 3, it can be seen that every group of Xanthatin purity has all reached higher degree, the purity of B4 group is had reached
98%；The method for illustrating to extract Xanthatin in Siberian cocklebur of the invention, can obtain the Xanthatin of high-purity, purity is up to
98%.
The preparation of Siberian cocklebur coarse powder is tested
Method one
S1, it rinse the Siberian cocklebur that parts by weight are 15g with clear water well after, dries to constant weight, crushes, and after 60 meshes excessively,
Overcritical titanium dioxide is carried out under conditions of extraction temperature is 38 DEG C, extracting pressure is 38MPa and carbon dioxide flow is 45L/h
After carbon extracts 1.5h, Siberian cocklebur extract is made；
S2, the water that the Siberian cocklebur extract in S1 is added to parts by weight 15g are to decoct 2.5h, continuous mistake at 75 DEG C in temperature
Filter 3 times, obtains filtrate and the first filter residue, spare；
S3, the filtrate in S2 is centrifuged 25min at revolving speed 4000rpm through centrifuge, isolates supernatant, sediment with
First filter residue merges, and obtains the second filter residue；
S4, the second filter residue in S3 is placed in the ethanol solution that volume fraction is 88% and impregnates 7h, filtered, impregnated
Liquid and third filter residue；
S5, the third filter residue in S4 is placed in percolating device, carries out diacolation using the soak in S4, controls diacolation stream
Speed is 350ml/h, obtains percolate, then mixes percolate with the supernatant in S3, is ground after vacuum concentration is dry, finally
Obtain the Siberian cocklebur coarse powder of 2.15mg.
Method two
S1, it rinse the Siberian cocklebur that parts by weight are 15g with clear water well after, dries to constant weight, crushes, and after 60 meshes excessively,
Obtain Siberian cocklebur crushed material；
The water of parts by weight 15g is added in S2, the Siberian cocklebur crushed material into S1, is to decoct 2.5h at 75 DEG C in temperature, continuously
Filter 23, obtains filtrate, after filter vacuum is concentrated and dried, finally obtains the Siberian cocklebur coarse powder of 1.35mg.
Method one is the extracting method of Siberian cocklebur coarse powder of the invention, and method two is traditional extracting method, from result above
Know that the yield of the extracting method of Siberian cocklebur coarse powder of the invention is higher than traditional extraction process.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details.
Claims (8)
1. extracting the method for Xanthatin from Siberian cocklebur, which is characterized in that include the following steps：
Cellulase solution, buffer solution are added into Siberian cocklebur coarse powder, 45 DEG C of heating water baths after digesting 1.5~2.5h, inactivate 5
Then bionic extraction liquid is added in~10min, extract under 37 DEG C, ultrasonic wave auxiliary to get the Siberian cocklebur coarse powder extract；
The mass volume ratio of the Siberian cocklebur coarse powder and cellulase solution, buffer solution, bionic extraction liquid is 1g:2.5~
3.4ml:10~16.7ml:30~35ml；
The Siberian cocklebur coarse powder extract is separated, purified, is concentrated to get Xanthatin.
2. the method for Xanthatin is extracted from Siberian cocklebur as described in claim 1, which is characterized in that the preparation of the Siberian cocklebur coarse powder
Include the following steps：
S1, it rinse the Siberian cocklebur that parts by weight are 10~20 parts with clear water well after, dries to constant weight, crushes, and cross 50~100 mesh
After sieve, extraction temperature be 35~40 DEG C, the condition that extracting pressure is 35~40MPa and carbon dioxide flow is 40~50L/h
After 1~2h of lower progress supercritical carbon dioxide extracting, Siberian cocklebur extract is made；
S2, the water that the Siberian cocklebur extract in S1 is added to 10~20 parts of parts by weight are that 2~3h is decocted at 70~80 DEG C in temperature,
Continuous filter 23, obtains filtrate and the first filter residue, spare；
S3, the filtrate in S2 is centrifuged at revolving speed 4000rpm 20~30min through centrifuge, isolates supernatant, sediment with
First filter residue merges, and obtains the second filter residue；
S4, the second filter residue in S3 is placed in the ethanol solution that volume fraction is 85~90% and impregnates 5~10h, filtering obtains
Soak and third filter residue；
S5, the third filter residue in S4 is placed in percolating device, carries out diacolation using the soak in S4, control diacolation flow velocity is
300~400ml/h obtains percolate, then mixes percolate with the supernatant in S3, grinds after vacuum concentration is dry, i.e.,
Obtain Siberian cocklebur coarse powder.
3. the method for Xanthatin is extracted from Siberian cocklebur as described in claim 1, which is characterized in that the separation, purifying, concentration
Include the following steps：
A1, Siberian cocklebur coarse powder extract is mixed with silica gel, obtains Siberian cocklebur silica white；
A2, by silicagel column on the Siberian cocklebur silica white in A1, carry out first time gradient elution, collect every time containing the elution of Xanthatin
Object, and merge, it is concentrated under reduced pressure, Xanthatin is made, study is concentrated；
A3, reverse phase silica gel column, second of gradient elution of progress on the Xanthatin concentration study in A2 are collected and contains Siberian cocklebur every time
The eluate of pavilion, and merging, carries out degreasing with petroleum ether, is then extracted with ethyl acetate acetic acid butyl ester, be concentrated under reduced pressure to get
Xanthatin.
4. the method for Xanthatin is extracted from Siberian cocklebur as described in claim 1, which is characterized in that the buffer solution is
NaAc-HAc buffer solution, solid-to-liquid ratio 1:10~1:12.5 pH 4.5.
5. the method for Xanthatin is extracted from Siberian cocklebur as described in claim 1, which is characterized in that the bionic extraction liquid is matched
Method processed is：Sodium chloride, the pepsin of 3.2g and the hydrochloric acid of 0.02mol of 9g are added in the water of every L, and pH value is adjusted to
6.0。
6. the method for Xanthatin is extracted from Siberian cocklebur as claimed in claim 3, which is characterized in that the first subgradient described in A2
Eluant, eluent used in elution is the mixed liquor of chloroform and methanol, and the mass ratio of chloroform and methanol is followed successively by 80:1~100:1,
18:1~25:1,3:1~5:1,0.8:1~1:1.
7. the method for Xanthatin is extracted from Siberian cocklebur as claimed in claim 3, which is characterized in that the second subgradient described in A3
Eluant, eluent used in elution is methanol solution, and the mass fraction of methanol solution is followed successively by 75~85%, 85~95%, 100%.
8. the method for Xanthatin is extracted from Siberian cocklebur as claimed in claim 3, which is characterized in that the ethyl acetate acetic acid
Ethyl acetate and butyl acetate volume ratio 1.5 in butyl ester:1~2.5:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811172744.6A CN108840845A (en) | 2018-10-09 | 2018-10-09 | The method of Xanthatin is extracted from Siberian cocklebur |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811172744.6A CN108840845A (en) | 2018-10-09 | 2018-10-09 | The method of Xanthatin is extracted from Siberian cocklebur |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108840845A true CN108840845A (en) | 2018-11-20 |
Family
ID=64188058
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811172744.6A Pending CN108840845A (en) | 2018-10-09 | 2018-10-09 | The method of Xanthatin is extracted from Siberian cocklebur |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108840845A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109528796A (en) * | 2018-12-11 | 2019-03-29 | 江西赣隆药业有限公司 | A kind of Siberian cocklebur grass granule and preparation method thereof |
| CN110373490A (en) * | 2019-08-05 | 2019-10-25 | 上海出入境检验检疫局动植物与食品检验检疫技术中心 | A kind of primer and kit of Rapid identification spiny cocklebur |
| CN110452204A (en) * | 2019-08-16 | 2019-11-15 | 山西农业大学 | Xanthatin thioether class and imines analog derivative and its preparation method and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101830874A (en) * | 2010-05-13 | 2010-09-15 | 苏州派腾生物医药科技有限公司 | Preparation method of Xanthatin |
-
2018
- 2018-10-09 CN CN201811172744.6A patent/CN108840845A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101830874A (en) * | 2010-05-13 | 2010-09-15 | 苏州派腾生物医药科技有限公司 | Preparation method of Xanthatin |
Non-Patent Citations (2)
| Title |
|---|
| SCHERER, R.等: "Biological activity and chemical composition of hydrodistilled and supercritical extracts of Xanthium strumarium L.leaves", 《JOURNAL OF ESSENTIAL OIL RESEARCH》 * |
| 胡冬燕等: "苍耳化学成分的分离与鉴定", 《中草药》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109528796A (en) * | 2018-12-11 | 2019-03-29 | 江西赣隆药业有限公司 | A kind of Siberian cocklebur grass granule and preparation method thereof |
| CN110373490A (en) * | 2019-08-05 | 2019-10-25 | 上海出入境检验检疫局动植物与食品检验检疫技术中心 | A kind of primer and kit of Rapid identification spiny cocklebur |
| CN110452204A (en) * | 2019-08-16 | 2019-11-15 | 山西农业大学 | Xanthatin thioether class and imines analog derivative and its preparation method and application |
| CN110452204B (en) * | 2019-08-16 | 2021-03-12 | 山西农业大学 | Xanthatin thioether and imine derivatives, and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101723996B (en) | Method for separating high-purity anthocyanin monomers from black rice | |
| CN102276679B (en) | Method for extracting high-purity tea saponin from oil-tea-cake by decompression boiling | |
| CN108840845A (en) | The method of Xanthatin is extracted from Siberian cocklebur | |
| CN101863935B (en) | Preparation method of 1,4-di-[4-(glucosyloxy) benzyl]-2-isobutyl malate comparison product | |
| CN101721516B (en) | Preparation method of gardenia extract | |
| CN110437059B (en) | Method for extracting pachymic acid A and pachymic acid B from Poria peel | |
| CN106317148A (en) | Method for extracting cordycepin from cordyceps militaris | |
| CN101401897A (en) | Edulcoration purification process for rhizoma paris saponin extract solution | |
| CN106632546A (en) | Method for preparing two chemical reference substances of Rhoifolin and naringin simultaneously | |
| CN106596426A (en) | Detection method applied in gynostemma total saponin extraction and separation process | |
| CN106432167A (en) | Method for extracting theaflavin from black tea | |
| CN107573255A (en) | A kind of method that capsaicine and dihydrocapsaicin are isolated and purified from pepper fruit | |
| CN108003211A (en) | A kind of method of quick separating ganoderic acid in accessory substance from ganoderma lucidum | |
| CN111675741A (en) | Separation method for simultaneously obtaining four kinds of epimedium rare flavone by using preparative liquid phase method | |
| CN101387586B (en) | Method for producing orientin standard substance | |
| CN106420871A (en) | Preparation method of Panax notoginseng saponins rich in Fe and Fd and monomers Fe and Fd thereof | |
| CN113583077B (en) | Process for preparing sterols | |
| CN113354697B (en) | Method for separating and purifying pectolinarin and linarin | |
| CN102924545A (en) | Enrichment and purifying method of akebiaquinata saponin D | |
| CN104926904B (en) | A kind of method of extraction purification cordycepin in mycoderma from Cordyceps militaris | |
| CN114920795A (en) | Preparation method of dried toad skin bufogenin lactone component | |
| CN108341845A (en) | The method that cornel extractive prepares high-purity morroniside | |
| CN107353296B (en) | A method of extracting activated protein and eurycomanone from Tongkat Ali | |
| CN109232674B (en) | Method for extracting chrysin-8-C-beta-D-glucoside from Nanshan tea | |
| CN105085498B (en) | The method and system of extraction separation isovitexin from Desmodium styracifolium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181120 |