CN102286063A - Method for preparing trillin - Google Patents
Method for preparing trillin Download PDFInfo
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- CN102286063A CN102286063A CN2011101877445A CN201110187744A CN102286063A CN 102286063 A CN102286063 A CN 102286063A CN 2011101877445 A CN2011101877445 A CN 2011101877445A CN 201110187744 A CN201110187744 A CN 201110187744A CN 102286063 A CN102286063 A CN 102286063A
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- trillenoside
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Abstract
The invention relates to a method for preparing trillin, which comprises: leaching trillium in ethanol; defatting by using petroleum ether; treating defatted medicinal liquid by alumina column chromatography; eluting with mixed solution of diethyl ether and ethanol; collecting eluent and concentrating; adding acetone to separate a yellow precipitate; centrifuging, and washing precipitate with acetone; separating and purifying by high-speed countercurrent chromatography; concentrating efflux; and drying at a low temperature to obtain the product. The method has the characteristics of simple process operation, high extraction rate and high product purity.
Description
Technical field
The invention belongs to medical technical field, be specifically related to a kind of preparation method of trillenoside.
Background technology
Trillenoside (Trillin), different name: Diosgenin glucoside, molecular formula: C
33H
52O
8Molecular weight: 576.7658, CAS accession number: 14144-06-0, molecular structural formula is as follows:
Trillenoside is to be natural compounds, is a kind of steroidal saponin, has effects such as reducing blood-fat, anti-freezing, Green Tea Extract activity, derives from liliaceous plant
Trillium tschonoskiiThe Maxim rhizome also has document to separate from Dioscorea nipponica Mak. Ningpo Yam Rhizome plants such as (Dioscorea nipponica Mak.) and obtains trillenoside.
The Chinese medicine Trillium tschonoskii Maxim is traditional rare Chinese medicine, has another name called Rhizoma Trillii Tschonoskii, contains multiple steroidal saponin, flavones and ter penoids.Have tranquilizing and allaying excitement, expelling wind and activating blood circulation, effect of prolonging life.Cure mainly have a dizzy spell, disease such as insomnia, wound, traumatic hemorrhage, neurasthenia, hypertension, cerebral concussion sequela etc.Huang Liya etc. studies show that the Rhizoma Trillii Tschonoskii anti-aging effects and strengthen cerebral tissue SOD, GSH-Px activity, reduce MDA and accumulate relevant.Left heart function after document discovery Rhizoma Trillii Tschonoskii can obviously improve ischemia reperfusion injury is also arranged, can improve SOD and GSH-Px activity in the blood, reduce the content of MDA.
In the prior art, the extracting method of trillenoside mainly is to use the 60%-70% alcohol reflux, purification process mainly adopts silicagel column, ODS mesolow column chromatography and half preparative high-performance liquid chromatographic etc., but the trillenoside content that these extracting and purifying method obtain is low, production cycle is long, cost is high, and unfavorable big production operation.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of preparation method who is beneficial to big production operation, extraction yield height, trillenoside that product purity is high.
For solving the problems of the technologies described above, the present invention adopts following technical proposal:
Get the Trillium tschonoskii Maxim rhizome, adding 4-10 doubly measures the alcoholic solution temperature of volume and soaked 2-8 hour, filters, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, the soup after the degreasing adds alumina column chromatography, ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone, adopts high-speed countercurrent chromatography to carry out separation and purification again, and effluent concentrates, cryodrying promptly gets product.
Described temperature is soaked temperature 60-85 ℃.
Described high speed adverse current chromatogram solvent systems is normal heptane-ethyl acetate-acetonitrile, and its volume ratio is (3-7): (1-3): (2-4).
Described high speed adverse current chromatogram solvent systems is petroleum ether-ethyl acetate-alcohol-water, and its volume ratio is (4-8): (3-5): (4-7): (3-6).
Adopt technique scheme to prepare trillenoside, simple to operate, extraction yield is high, sample loss is few, product purity is high, is beneficial to big production operation.
The present invention is further elaborated below in conjunction with embodiment, but the scope of protection of present invention is not limited in following embodiment.
Embodiment
Embodiment 1:
Get Trillium tschonoskii Maxim 2kg, 65% ethanol that adds 6 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 4 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get normal heptane, ethyl acetate, acetonitrile by volume 7:3:4 place separating funnel to mix, standing demix separates phase up and down, below being stationary phase mutually, is moving phase mutually down, and it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 3ml/min flow pump, rotating speed is 900rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.7%.
Embodiment 2:
Get Trillium tschonoskii Maxim 2kg, 65% ethanol that adds 4 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 2 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get normal heptane, ethyl acetate, acetonitrile by volume 5:2:3 place separating funnel to mix, standing demix separates phase up and down, below being stationary phase mutually, is moving phase mutually down, and it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 2ml/min flow pump, rotating speed is 900rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.4%.
Embodiment 3:
Get Trillium tschonoskii Maxim 2kg, 70% ethanol that adds 5 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 8 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get normal heptane, ethyl acetate, acetonitrile by volume 3:2:2 place separating funnel to mix, standing demix separates phase up and down, below being stationary phase mutually, is moving phase mutually down, and it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 3ml/min flow pump, rotating speed is 900rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.5%.
Embodiment 4:
Get Trillium tschonoskii Maxim 2kg, 65% ethanol that adds 6 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 5 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get normal heptane, ethyl acetate, acetonitrile by volume 4:1:4 place separating funnel to mix, standing demix separates phase up and down, below being stationary phase mutually, is moving phase mutually down, and it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 3ml/min flow pump, rotating speed is 850rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.1%.
Embodiment 5:
Get Trillium tschonoskii Maxim 2kg, 65% ethanol that adds 5 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 3 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get sherwood oil, ethyl acetate, ethanol, water by volume 4:4:5:3 place separating funnel to mix, standing demix, separate phase up and down, below be stationary phase mutually, be moving phase mutually down, it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 3ml/min flow pump, rotating speed is 850rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.3%.
Embodiment 6:
Get Trillium tschonoskii Maxim 3kg, 70% ethanol that adds 8 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 4 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get sherwood oil, ethyl acetate, ethanol, water by volume 8:3:4:6 place separating funnel to mix, standing demix, separate phase up and down, below be stationary phase mutually, be moving phase mutually down, it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 2ml/min flow pump, rotating speed is 900rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.8%.
Embodiment 7:
Get Trillium tschonoskii Maxim 5kg, 70% ethanol that adds 4 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 7 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get sherwood oil, ethyl acetate, ethanol, water by volume 5:5:7:4 place separating funnel to mix, standing demix, separate phase up and down, below be stationary phase mutually, be moving phase mutually down, it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 3ml/min flow pump, rotating speed is 900rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.4%.
Embodiment 8:
Get Trillium tschonoskii Maxim 5kg, 65% ethanol that adds 7 times of amounts of its weight volume is heated to 80 ℃, and temperature was soaked 6 hours, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing is added the neutral alumina column chromatography, with ether-ethanol (65:35) wash-out, collect elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, precipitation is used washing with acetone again, and cryodrying gets the trillenoside crude product.Get sherwood oil, ethyl acetate, ethanol, water by volume 7:4:6:5 place separating funnel to mix, standing demix, separate phase up and down, below be stationary phase mutually, be moving phase mutually down, it is stand-by in being dissolved under the 10ml mutually to get above-mentioned trillenoside crude product 150mg, open high-speed counter-current chromatograph, go into moving phase, begin continuous sample introduction behind the ready to balance with the 3ml/min flow pump, rotating speed is 900rpm, collect the trillenoside flow point, concentrate, cryodrying promptly gets trillenoside, measure through HPLC, purity is 98.2%.
Claims (4)
1. the preparation method of a trillenoside, it is characterized in that described method comprises the following steps: to get the Trillium tschonoskii Maxim rhizome, adding 4-10 doubly measures the alcoholic solution temperature of volume and soaked 2-8 hour, filter, get decompression filtrate recycling ethanol, add the equal-volume petroleum ether degreasing, soup after the degreasing adds alumina column chromatography, and ether-ethanol (65:35) wash-out is collected elutriant, concentrate, add acetone and separate out yellow mercury oxide, centrifugal, the precipitation washing with acetone, adopt high-speed countercurrent chromatography to carry out separation and purification again, effluent concentrates, cryodrying promptly gets product.
2. according to the preparation method of the described trillenoside of claim 1, it is characterized in that described temperature soaks temperature 60-85 ℃.
3. according to the preparation method of the described trillenoside of claim 1, it is characterized in that described high speed adverse current chromatogram solvent systems is normal heptane-ethyl acetate-acetonitrile, its volume ratio is (3-7): (1-3): (2-4).
4. according to the preparation method of the described trillenoside of claim 1, it is characterized in that described high speed adverse current chromatogram solvent systems is petroleum ether-ethyl acetate-alcohol-water, its volume ratio is (4-8): (3-5): (4-7): (3-6).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101877445A CN102286063A (en) | 2011-07-06 | 2011-07-06 | Method for preparing trillin |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101877445A CN102286063A (en) | 2011-07-06 | 2011-07-06 | Method for preparing trillin |
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|---|---|
| CN102286063A true CN102286063A (en) | 2011-12-21 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105263945A (en) * | 2013-12-06 | 2016-01-20 | 于跃 | Method for synthesizing saponin |
| CN110604736A (en) * | 2019-08-22 | 2019-12-24 | 西南医科大学 | Application of trillium saponin in preparing nerve protection medicine |
-
2011
- 2011-07-06 CN CN2011101877445A patent/CN102286063A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105263945A (en) * | 2013-12-06 | 2016-01-20 | 于跃 | Method for synthesizing saponin |
| CN110604736A (en) * | 2019-08-22 | 2019-12-24 | 西南医科大学 | Application of trillium saponin in preparing nerve protection medicine |
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Application publication date: 20111221 |