CN104072546A - Method for extracting shephagenin A - Google Patents
Method for extracting shephagenin A Download PDFInfo
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- CN104072546A CN104072546A CN201410266205.4A CN201410266205A CN104072546A CN 104072546 A CN104072546 A CN 104072546A CN 201410266205 A CN201410266205 A CN 201410266205A CN 104072546 A CN104072546 A CN 104072546A
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Abstract
The invention discloses a method for extracting shephagenin A. The method is characterized by comprising the following steps: with leaves of shepherdia argentea as raw material, carrying out tissue smashing and cellulose enzymolysis to obtain enzymolysis raw material; carrying out flash extraction in a flash extractor, filtering to obtain an extracting solution, concentrating and drying; purifying through high pressure silica-gel column chromatography, collecting flow of the shephagenin A, concentrating at reduced pressure and carrying out ethyl acetate recrystallization to obtain the shephagenin A. The method disclosed by the invention is simple in operation, low in energy consumption, little in pollution and short in production cycle and is suitable for mass production; the method is a production process can be put into industrialization easily.
Description
Technical field
The invention belongs to Chemistry for Chinese Traditional Medicine technical field, relate to a kind of method of extracting the plain A of buffalo fruit.
Background technology
The plain A(Shephagenin A of buffalo fruit), molecular formula is C
48h
32o
32, molecular weight is 1120.75, structural formula is:
The plain A of buffalo fruit is that separation obtains a kind of active compound from the silver-colored buffalo fruit of Elaeangnaceae (Elaeagnaceae) Shepherdia argentea Nutt. leaf.Pharmacological research shows, the plain A of buffalo fruit has anti-HIV-1 reversed transcriptive enzyme, and IC50 is 0.049 μ M, is better than positive control (-)-epigallocatechin gallate (IC50 is 0.25 μ M).
In prior art, there is not yet the production technique report of the plain A of preparation of industrialization buffalo fruit.
Summary of the invention
The object of this invention is to provide a kind of method of extracting the plain A of buffalo fruit.
For achieving the above object, the present invention is by the following technical solutions:
A method of extracting the plain A of buffalo fruit, is characterized in that comprising the following steps:
(1) take the leaf of silver-colored buffalo fruit is raw material, puts into high-speed tissue mashing machine, at rotating speed, is to keep after 3-5min under 18000rpm condition, crosses 40 mesh sieves standby;
(2) raw material is according to solid-liquid ratio 1:4-6(g/ml) ratio add after aqueous ethanolic solution, then add the cellulase of 0.3-0.5%, enzymolysis 60-90min under ultrasonic power 500-700W, 45-60 ℃ condition, obtains enzymolysis raw material;
(3) enzymolysis raw material is dropped into and added in flash extracter, add extraction solvent, carry out flash extraction, filter and obtain extracting solution, concentrate drying obtains crude extract;
(4) by gained crude extract dissolve with methanol, dry method loading is to high pressure purification by silica gel column chromatography, and chloroform-methanol solvent systems gradient elution, collects the plain A flow point of buffalo fruit, concentrating under reduced pressure, and cooling crystallization, obtains coarse-grain;
(5) coarse-grain is obtained to the plain A white crystals of buffalo fruit by re-crystallizing in ethyl acetate.
In described step (2), aqueous ethanolic solution volume percent is 70-80%.
In described step (2), extract the ethanolic soln that solvent is 70-85%.
In described step (3), flash extraction voltage is 150V, and extraction time is 3-5min.
High pressure silica gel column chromatography described in described step (4), silica gel order number is 200 orders, withstand voltage 35MPa, column internal diameter is 250mm, chromatography column blade diameter length ratio is 1:10.
Described in described step (4), the volume ratio of chloroform-methanol is 1:1-1:13.
The invention has the beneficial effects as follows: the present invention is simple to operate, consuming time short, yield is high, easily large-scale production; Adopt high pressure chromatography column, applied sample amount is large, with short production cycle, and resolution is good, and product purity is high.
Below in conjunction with embodiment, further illustrate the present invention, but the scope of protection of present invention is not limited to following embodiment.
embodiment:
Embodiment 1:
The silver-colored buffalo leaf really of take is raw material, be cut into 1cm left and right section shape, put into high-speed tissue mashing machine, at rotating speed, be to keep after 5min under 18000rpm condition, cross 40 mesh sieves standby, take 1kg, according to solid-liquid ratio 1:4(g/ml) ratio add after 80% aqueous ethanolic solution, add again 0.5% cellulase, at ultrasonic power 700W, enzymolysis 70min under 50 ℃ of conditions, obtain enzymolysis raw material, add in flash extracter, add the ethanolic soln of 6L80% as extracting solvent, extraction voltage is 150V, extraction time is 3min, carry out flash extraction 2 times, filtration obtains extracting solution, after extracting solution concentrate drying, obtain crude extract, with dissolve with methanol, dry method loading is to high pressure silica gel (silica gel order number is 200 orders) column chromatography purification, withstand voltage 35MPa, column internal diameter is 250mm, chromatography column blade diameter length ratio is 1:10, chloroform-methanol is 1:1 by volume, 1:6, 1:13 gradient elution, collect the plain A flow point of buffalo fruit, concentrating under reduced pressure, cooling crystallization, obtain coarse-grain, coarse-grain obtains the plain A white crystals of buffalo fruit by re-crystallizing in ethyl acetate, content is 95.8%.
Embodiment 2:
The silver-colored buffalo leaf really of take is raw material, be cut into 1cm left and right section shape, put into high-speed tissue mashing machine, at rotating speed, be to keep after 3min under 18000rpm condition, cross 40 mesh sieves standby, take 1kg, according to solid-liquid ratio 1:6(g/ml) ratio add after 70% aqueous ethanolic solution, add again 0.4% cellulase, at ultrasonic power 700W, enzymolysis 90min under 45 ℃ of conditions, obtain enzymolysis raw material, add in flash extracter, add the ethanolic soln of 4.5L75% as extracting solvent, extraction voltage is 150V, extraction time is 5min, carry out flash extraction 2 times, filtration obtains extracting solution, after extracting solution concentrate drying, obtain crude extract, with dissolve with methanol, dry method loading is to high pressure silica gel (silica gel order number is 200 orders) column chromatography purification, withstand voltage 35MPa, column internal diameter is 250mm, chromatography column blade diameter length ratio is 1:10, chloroform-methanol is 1:1 by volume, 1:7, 1:13 gradient elution, collect the plain A flow point of buffalo fruit, concentrating under reduced pressure, cooling crystallization, obtain coarse-grain, coarse-grain obtains the plain A white crystals of buffalo fruit by re-crystallizing in ethyl acetate, content is 94.4%.
Embodiment 3:
The silver-colored buffalo leaf really of take is raw material, be cut into 1cm left and right section shape, put into high-speed tissue mashing machine, at rotating speed, be to keep after 5min under 18000rpm condition, cross 40 mesh sieves standby, take 1kg, according to solid-liquid ratio 1:5(g/ml) ratio add after 75% aqueous ethanolic solution, add again 0.5% cellulase, at ultrasonic power 600W, enzymolysis 75min under 55 ℃ of conditions, obtain enzymolysis raw material, add in flash extracter, add the ethanolic soln of 4L70% as extracting solvent, extraction voltage is 150V, extraction time is 4min, carry out flash extraction 2 times, filtration obtains extracting solution, after extracting solution concentrate drying, obtain crude extract, with dissolve with methanol, dry method loading is to high pressure silica gel (silica gel order number is 200 orders) column chromatography purification, withstand voltage 35MPa, column internal diameter is 250mm, chromatography column blade diameter length ratio is 1:10, chloroform-methanol is 1:1 by volume, 1:5, 1:12 gradient elution, collect the plain A flow point of buffalo fruit, concentrating under reduced pressure, cooling crystallization, obtain coarse-grain, coarse-grain obtains the plain A white crystals of buffalo fruit by re-crystallizing in ethyl acetate, content is 95.4%.
Embodiment 4:
The silver-colored buffalo leaf really of take is raw material, be cut into 1cm left and right section shape, put into high-speed tissue mashing machine, at rotating speed, be to keep after 5min under 18000rpm condition, cross 40 mesh sieves standby, take 1kg, according to solid-liquid ratio 1:6(g/ml) ratio add after 75% aqueous ethanolic solution, add again 0.3% cellulase, at ultrasonic power 500W, enzymolysis 60min under 60 ℃ of conditions, obtain enzymolysis raw material, add in flash extracter, add the ethanolic soln of 5L85% as extracting solvent, extraction voltage is 150V, extraction time is 3min, carry out flash extraction 2 times, filtration obtains extracting solution, after extracting solution concentrate drying, obtain crude extract, with dissolve with methanol, dry method loading is to high pressure silica gel (silica gel order number is 200 orders) column chromatography purification, withstand voltage 35MPa, column internal diameter is 250mm, chromatography column blade diameter length ratio is 1:10, chloroform-methanol is 1:1 by volume, 1:7, 1:13 gradient elution, collect the plain A flow point of buffalo fruit, concentrating under reduced pressure, cooling crystallization, obtain coarse-grain, coarse-grain obtains the plain A white crystals of buffalo fruit by re-crystallizing in ethyl acetate, content is 94.9%.
Claims (6)
1. a method of extracting the plain A of buffalo fruit, is characterized in that comprising the following steps:
(1) take the leaf of silver-colored buffalo fruit is raw material, puts into high-speed tissue mashing machine, at rotating speed, is to keep after 3-5min under 18000rpm condition, crosses 40 mesh sieves standby;
(2) raw material is according to solid-liquid ratio 1:4-6(g/ml) ratio add after aqueous ethanolic solution, then add the cellulase of 0.3-0.5%, enzymolysis 60-90min under ultrasonic power 500-700W, 45-60 ℃ condition, obtains enzymolysis raw material;
(3) enzymolysis raw material is dropped into and added in flash extracter, carry out flash extraction, filter and obtain extracting solution, concentrate drying obtains crude extract;
(4) by gained crude extract dissolve with methanol, dry method loading is to high pressure purification by silica gel column chromatography, and chloroform-methanol solvent systems gradient elution, collects the plain A flow point of buffalo fruit, concentrating under reduced pressure, and cooling crystallization, obtains coarse-grain;
(5) coarse-grain is obtained to the plain A crystallization of buffalo fruit by re-crystallizing in ethyl acetate.
2. a kind of method of extracting the plain A of buffalo fruit as claimed in claim 1, is characterized in that in described step (2), aqueous ethanolic solution volume percent is 70-80%.
3. a kind of method of extracting the plain A of buffalo fruit as claimed in claim 1, is characterized in that the ethanolic soln that in described step (2), extraction solvent is 70-85%.
4. a kind of method of extracting the plain A of buffalo fruit as claimed in claim 1, is characterized in that in described step (3), flash extraction voltage is 150V, and extraction time is 3-5min.
5. a kind of method of extracting the plain A of buffalo fruit as claimed in claim 1, is characterized in that high pressure silica gel column chromatography described in described step (4), and silica gel order number is 200 orders, withstand voltage 35MPa, and column internal diameter is 250mm, chromatography column blade diameter length ratio is 1:10.
6. a kind of method of extracting the plain A of buffalo fruit as claimed in claim 1, the volume ratio that it is characterized in that chloroform-methanol described in described step (4) is 1:1-1:13.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410266205.4A CN104072546A (en) | 2014-06-16 | 2014-06-16 | Method for extracting shephagenin A |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410266205.4A CN104072546A (en) | 2014-06-16 | 2014-06-16 | Method for extracting shephagenin A |
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| CN104072546A true CN104072546A (en) | 2014-10-01 |
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| CN201410266205.4A Pending CN104072546A (en) | 2014-06-16 | 2014-06-16 | Method for extracting shephagenin A |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104833560A (en) * | 2015-04-27 | 2015-08-12 | 宁夏森淼种业生物工程有限公司 | Quickly-shearing nondestructive method for extracting mineral elements from wolfberry leaf |
-
2014
- 2014-06-16 CN CN201410266205.4A patent/CN104072546A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104833560A (en) * | 2015-04-27 | 2015-08-12 | 宁夏森淼种业生物工程有限公司 | Quickly-shearing nondestructive method for extracting mineral elements from wolfberry leaf |
| CN104833560B (en) * | 2015-04-27 | 2017-09-19 | 宁夏森淼种业生物工程有限公司 | A kind of method that flash shearing non-destructive extracts folium lycii mineral element |
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Application publication date: 20141001 |