CN104650052A - Method for preparing isoorientin extract from bamboo leaves - Google Patents
Method for preparing isoorientin extract from bamboo leaves Download PDFInfo
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- CN104650052A CN104650052A CN201510078150.9A CN201510078150A CN104650052A CN 104650052 A CN104650052 A CN 104650052A CN 201510078150 A CN201510078150 A CN 201510078150A CN 104650052 A CN104650052 A CN 104650052A
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- negative pressure
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- isorientin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
- C07D407/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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Abstract
The invention discloses a method for preparing isoorientin extract from bamboo leaves. The method is characterized by comprising the following steps: (1) processing a raw material, that is, removing impurities of bamboo leaves, and washing; (2) performing negative pressure extraction, namely, putting the raw material treated in the step (1) into a negative pressure container, adding an extraction solvent, performing digestion for 1-2 hours in the negative pressure container, filtering, collecting a filtrate, and concentrating, thereby obtaining the isoorientin extract, wherein the negative pressure extraction conditions are as follows: the negative pressure is minus 0.065-minus 0.075Mpa, the temperature is 30-35 DEG C, and the raw material and the extraction solvent are digested in a material liquid ratio of (1:10)-(1:15) (g/ml). By adopting negative pressure extraction and by controlling factors such as the material liquid ratio in negative pressure extraction, the negative pressure condition and the temperature condition, the high-purity homo-orientin extract can be prepared.
Description
Technical field
The present invention relates to effective ingredients in plant extractive technique, particularly a kind of method preparing isorientin extract from the leaf of bamboo.
Background technology
Isorientin (Isoorientin), also known as Lutonaretin, is the isomers of orientin, yellow crystal, molecular formula: C
21h
20o
11, molecular weight: 448.38, dissolves in the organic solvents such as methyl alcohol, ethanol, Virahol, n-propyl alcohol, fusing point: 230 DEG C ~ 232 DEG C.Modern study shows, isorientin has significant protective effect to myocardial ischemia, anoxic.
The common extraction source of isorientin is for Polygonum orientale (the Gan Zao Guo Miho of polygonaceae plant Polygonum knotweed (Polygonum orientale L.) and stem and branch with leaf); We study and find that in Folium Bambosae flavone, Isoorientin content is generally more than 2%, and Folium Bambosae flavone derives from the leaf of bamboo, extracts materials safety cheap, and natural resources can be utilized more fully.
Summary of the invention
Technical problem to be solved of the present invention is to provide a kind of method preparing isorientin extract from the leaf of bamboo, the present invention adopts negative pressure extraction, by controlling solid-liquid ratio, condition of negative pressure and the temperature condition etc. during negative pressure extraction, prepare highly purified isorientin extract.
The present invention solves the problems of the technologies described above adopted technical scheme:
From the leaf of bamboo, prepare a method for isorientin extract, comprise the following steps:
Step (1), Feedstock treating: by leaf of bamboo removal of impurities, clean;
Step (2), negative pressure extraction: the raw material that described step (1) processes is put into negative-pressure container, add Extraction solvent, filter after lixiviate 1-2h in described negative-pressure container and collect filtrate, concentrated, obtain isorientin extract of the present invention;
The condition of described negative pressure extraction is: negative pressure is-0.065Mpa ~-0.075Mpa, temperature 30 ~ 35 DEG C; Described raw material and Extraction solvent are according to the solid-liquid ratio lixiviate of 1:10 ~ 1:15 (g/ml).
In preferred technical scheme, in described step (1), described raw material also carries out following process after cleaning: the leaf of bamboo crushed after being dried after cleaning is crossed 20-60 mesh sieve.
In preferred technical scheme, in described step (2), described Extraction solvent is that deionized water or pH value are 1.0-3.0, massfraction is the ethanolic soln of 65-85%.
In preferred technical scheme, in described step (2), the condition of described negative pressure extraction is: negative pressure is-0.065Mpa, temperature 35 DEG C; Described Extraction solvent is described ethanolic soln be pH value is 2.0, massfraction is the ethanolic soln of 65%; Described raw material and Extraction solvent are according to the solid-liquid ratio lixiviate of 1:10 (g/ml).
In preferred technical scheme, in described step (2), described concentrated degree is: be concentrated into proportion 10 ~ 15, adopts B to measure.
Compared with prior art, tool has the following advantages in the present invention:
(1) the present invention adopts negative pressure extraction, by controlling solid-liquid ratio, condition of negative pressure and temperature condition and other factors during negative pressure extraction, prepares highly purified isorientin extract.In isorientin extract prepared by the present invention, isorientin high purity 8% ~ 10%.Described purity refers to the mass content of isorientin in isorientin extract.
(2) the inventive method is relative to prior art, and Extracting temperature is lower, can be low to moderate 30 DEG C, can guarantee that in leaching process, isorientin activeconstituents is not destroyed.
Embodiment
Below by way of specific embodiment, the present invention is described in further detail.
Embodiment 1:
From the leaf of bamboo, prepare a method for isorientin extract, comprise the following steps:
Step (1), Feedstock treating: the leaf of bamboo removed foreign material, clean;
Step (2), negative pressure extraction: the leaf of bamboo that described step (1) processes is put into negative-pressure container, add the ethanolic soln (the described leaf of bamboo and ethanolic soln, according to the solid-liquid ratio lixiviate of 1:10 (g/ml), namely add the ratio of 10ml ethanolic soln according to the 1g leaf of bamboo) that pH value is 2.0, massfraction is 65%; Filter after lixiviate 2h in negative-pressure container and collect filtrate, being concentrated into proportion 10 (B measurement), obtaining isorientin extract of the present invention.The condition of described negative pressure extraction is: negative pressure-0.065Mpa, temperature 35 DEG C.
Measure through HPLC, in final isorientin extract, the purity of isorientin is 8.3%.
Embodiment 2:
From the leaf of bamboo, prepare a method for isorientin extract, comprise the following steps:
Step (1), Feedstock treating: the leaf of bamboo removed foreign material, clean; Crushed after being dried crosses 50 mesh sieves, obtains bamboo leaf powder;
Step (2), negative pressure extraction: in the bamboo leaf powder of described step (1), add the ethanolic soln (described bamboo leaf powder and ethanolic soln, according to the solid-liquid ratio lixiviate of 1:10 (g/ml), namely add the ratio of 10ml ethanolic soln according to 1g bamboo leaf powder) that pH value is 2.0, massfraction is 70%; Filter after lixiviate 1.5h in negative-pressure container and collect filtrate, concentrated, obtain isorientin extract of the present invention.The condition of described negative pressure extraction is: negative pressure-0.065Mpa, temperature 30 DEG C.
Measure through HPLC, in final isorientin extract, the purity of isorientin is 11.8%.
Embodiment 3:
From the leaf of bamboo, prepare a method for isorientin extract, comprise the following steps:
Step (1), Feedstock treating: the leaf of bamboo removed foreign material, clean;
Step (2), negative pressure extraction: in the bamboo leaf powder of described step (1), add the ethanolic soln (the described leaf of bamboo and ethanolic soln, according to the solid-liquid ratio lixiviate of 1:12 (g/ml), namely add the ratio of 12ml ethanolic soln according to the 1g leaf of bamboo) that pH value is 2.0, massfraction is 75%; Filter after lixiviate 1.5h in negative-pressure container and collect filtrate, being concentrated into proportion 10 (B measurement), obtaining isorientin extract of the present invention.The condition of described negative pressure extraction is: negative pressure-0.075Mpa, temperature 30 DEG C.
Measure through HPLC, in final isorientin extract, the purity of isorientin is 8.5%.
Embodiment 4:
From the leaf of bamboo, prepare a method for isorientin extract, comprise the following steps:
Step (1), Feedstock treating: the leaf of bamboo removed foreign material, clean; Crushed after being dried crosses 20 mesh sieves, obtains bamboo leaf powder;
Step (2), negative pressure extraction: in the bamboo leaf powder of described step (1), add deionized water (described bamboo leaf powder and deionized water, according to the solid-liquid ratio lixiviate of 1:15 (g/ml), namely add the ratio of 15ml deionized water according to 1g bamboo leaf powder); Filter after lixiviate 1.5h in negative-pressure container and collect filtrate, concentrated, obtain isorientin extract of the present invention.The condition of described negative pressure extraction is: negative pressure-0.065Mpa, temperature 30 DEG C.
Measure through HPLC, in final isorientin extract, the purity of isorientin is 8.6%.
The test of embodiment 5 eosinophilic apoptosis-inducing and the experiment of MBP kinase activity
Eosinophilic's apoptosis-inducing is tested: the method (Journal ofImmunological Methods 184 volume 39-51 page nineteen ninety-five) of experimental evidence Vermes etc. is carried out.Make eosinophilic granulocyte sexual cell HL-60 cell suspension in RPMI1640 substratum, cell concn is 1-3 × 10
6cell/ml.Tested substance (finally collecting the isorientin extract obtained in above-described embodiment 1-4) or substratum is added in this cell suspending liquid 500 μ l, the concentration of isorientin extract in cell suspending liquid is made to be 100 μ g/ml, at 37 DEG C, 5%CO
2condition under cultivate 6 hours or 24 hours.Centrifugal clean after, add phosphatide phthalein associated proteins (Annexin) V damping fluid, then add 5u1 phosphatide phthalein associated proteins V-FITC.By flow cytometry method, using phosphatide phthalein associated proteins V positive cell as apoptosis-inducing cell, with the per-cent evaluation of relatively total cell count.
MBP Kinase activity assays: the method (Blood 99 volume 3432-3438 page 2002) of experimental evidence De Souza etc. is carried out.By eosinophilic granulocyte sexual cell HL-60 cell suspension in RPMI1640 substratum, cell concn is 1-3 × 10
6cell/ml.Add tested substance or substratum in this cell suspending liquid 500 μ l, make concentration be 100 μ g/ml, at 37 DEG C, 5%CO
2condition under cultivate 6 hours or 24 hours.Centrifugal clean after, add solubilized damping fluid, dissolved cell, use the gel containing MBP 5mg/ml to carry out electrophoresis.Gel, after de-sex change, again denaturing treatment, is used
32p-ATP marks, and implements the phosphorylation reaction of 3 hours.After gel drying, Quantimet is used to resolve radioactivity.When 36kDa grows indicia band, be denoted as the MBP kinase activation positive (+).
Isorientin extract observed the apoptosis-inducing of concentration dependent in HL-60 cell.When carrying out gel MBP kinase assays (In-gel MBP kinase assay) in addition, isorientin extract activates 36kDa MBP kinases.Above result prompting.Isorientin extract is to eosinophilic granulocyte energy cell death inducing, and this apoptosis is relevant with 36kDa MBP kinases.Infer in isorientin extract of the present invention containing the composition that may be used for anti-eosinophilic's inflammatory effect medicine thus.
Acquired results is as shown in table 1 below:
Confirm that embodiment 2 has strong apoptosis-inducing activity, and with the kinase whose activation of MBP.
Table 1
As mentioned above, just the present invention can be realized preferably.
Claims (4)
1. from the leaf of bamboo, prepare a method for isorientin extract, it is characterized in that, comprise the following steps:
Step (1), Feedstock treating: by leaf of bamboo removal of impurities, clean;
Step (2), negative pressure extraction: the raw material that described step (1) processes is put into negative-pressure container, add Extraction solvent, filter after lixiviate 1-2h in described negative-pressure container and collect filtrate, concentrated, obtain isorientin extract of the present invention;
The condition of described negative pressure extraction is: negative pressure is-0.065Mpa ~-0.075Mpa, temperature 30 ~ 35 DEG C; Described raw material and Extraction solvent are according to the solid-liquid ratio lixiviate of 1:10 ~ 1:15 (g/ml).
2. a kind of method preparing isorientin extract from the leaf of bamboo as claimed in claim 1, is characterized in that, in described step (1), after described impurity removing, also carry out following process: the leaf of bamboo crushed after being dried after cleaning is crossed 20-60 mesh sieve.
3. a kind of method preparing isorientin extract from the leaf of bamboo as claimed in claim 1, it is characterized in that, in described step (2), described Extraction solvent is that deionized water or pH value are 1.0-3.0, massfraction is the ethanolic soln of 65-85%.
4. a kind of method preparing isorientin extract from the leaf of bamboo as claimed in claim 1, is characterized in that, in described step (2),
The condition of described negative pressure extraction is: negative pressure is-0.065Mpa, temperature 35 DEG C;
Described Extraction solvent is described ethanolic soln be pH value is 2.0, massfraction is the ethanolic soln of 65%;
Described raw material and Extraction solvent are according to the solid-liquid ratio lixiviate of 1:10 (g/ml).
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| CN201510078150.9A CN104650052B (en) | 2015-02-13 | 2015-02-13 | A kind of method that homo-orientin extract is prepared from the leaf of bamboo |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008201725A (en) * | 2007-02-20 | 2008-09-04 | Univ Of Ryukyus | Anticaries agent |
| CN102285976A (en) * | 2011-09-27 | 2011-12-21 | 天津市尖峰天然产物研究开发有限公司 | Method for extracting isoorientin from bamboo leaf flavones |
| CN103263514A (en) * | 2013-05-23 | 2013-08-28 | 浙江省柑桔研究所 | Method for extracting flavones, low-molecule pectin and cellulose from orange peels in combined way |
| CN104587274A (en) * | 2015-02-13 | 2015-05-06 | 宁波杰顺生物科技有限公司 | Bamboo leaf homo-orientin extract |
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2015
- 2015-02-13 CN CN201510078150.9A patent/CN104650052B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008201725A (en) * | 2007-02-20 | 2008-09-04 | Univ Of Ryukyus | Anticaries agent |
| CN102285976A (en) * | 2011-09-27 | 2011-12-21 | 天津市尖峰天然产物研究开发有限公司 | Method for extracting isoorientin from bamboo leaf flavones |
| CN103263514A (en) * | 2013-05-23 | 2013-08-28 | 浙江省柑桔研究所 | Method for extracting flavones, low-molecule pectin and cellulose from orange peels in combined way |
| CN104587274A (en) * | 2015-02-13 | 2015-05-06 | 宁波杰顺生物科技有限公司 | Bamboo leaf homo-orientin extract |
Non-Patent Citations (2)
| Title |
|---|
| HAE-SUK PARK，等: "Antioxidant Flavone Glycosides from the Leaves of Sasa borealis", 《ARCH. PHARM. RES.》 * |
| 刘威: "木豆叶具有雌激素样活性的黄酮类成分提取分离及其抗肿瘤活性研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
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