CN103214544A - Method for extracting ursolic acid from paulownia leaves - Google Patents
Method for extracting ursolic acid from paulownia leaves Download PDFInfo
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- CN103214544A CN103214544A CN2013101823849A CN201310182384A CN103214544A CN 103214544 A CN103214544 A CN 103214544A CN 2013101823849 A CN2013101823849 A CN 2013101823849A CN 201310182384 A CN201310182384 A CN 201310182384A CN 103214544 A CN103214544 A CN 103214544A
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- ursolic acid
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- paulownia
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Abstract
The invention relates to a method for extracting ursolic acid from paulownia leaves, which is less in pollution and low in energy consumption. The method is characterized by comprising the following steps of: weighing paulownia leaves; adding cellulase into the paulownia leaves for enzymolysis for 2-5 hours; heating to boil, and decocting for 0.5-1.5 hours; filtering; adopting a filtrate; adsorbing the filtrate through AB8 macroporous adsorption resin; eluting the filtrate with 30% ethanol; adsorbing the eluted filtrate through D101 macroporous adsorption resin; eluting the adsorbed filtrate with 70% ethanol; collecting eluant; adsorbing the eluant through strong alkali styrene cation exchange resin; eluting the eluant with a 0.5 mol/L NaOH solution; collecting the eluant; filtering the eluant; adsorbing the eluant through strong acidic styrene cation exchange resin; collecting eluate; filtering the eluate; concentrating the filtrate through a reverse osmosis membrance; adding methanol into the filtrate for crystallization; separating and washing a crystal; and drying the cystal to obtain the ursolic acid. The ursolic acid prepared by using the method is high in product purity and easy for large-scale production and operation.
Description
Technical field
The present invention relates to a kind of method of from the paulownia leaf, extracting ursolic acid, especially a kind of preparation method of ursolic acid of from plant, extracting.
Background technology
Ursolic acid, another name: urson; Ursonic acid.English name: ursolic acid.Molecular formula: C
30H
47O
3Molecular weight: 455.6929.CAS number: 77-52-1.
Structural formula is as follows:
Modern study shows, ursolic acid has anti-inflammatory, preserve moisture, promote skin cells multiple effect such as grow, whiten, and it is also as the raw material of synthetic other reactive derivative simultaneously.
Studies show that, ursolic acid mainly is present in the various plants, the abundantest with the content in the paulownia leaf, paulownia are scrophulariaceae Paulownia plant paulownia (Paulownia fortunei (Seem.) Hemsl) or royal paulownia (P.tomentosa (Thunb.) Steud).
In the prior art, prepare method that high-purity ursolic acid adopts and be traditional heating and extract, organic solvent repeated treatments such as benzene, sherwood oil, technology falls behind, and product purity is low, energy consumption is big, seriously polluted, can not adapt to the requirement of modern recycling economy far away.
Summary of the invention
Technical problem to be solved by this invention provides a kind of big production operation, preparation method of ursolic acid that product purity is high of being beneficial to.
For solving the problems of the technologies described above, the present invention adopts following technical proposal:
A kind of method of from the paulownia leaf, extracting ursolic acid, it is characterized in that described method is made up of the following step: get the paulownia leaf, the ratio that adds 5mL in every g leaf adds entry, add cellulase, at temperature 40-50 ℃, pH value 3-6, under the condition of enzyme amount 10-20U/g leaf enzymolysis 2-5 hour, be heated to boiling and decocted 0.5-1.5 hour, filter, get filtrate, by the AB8 absorption with macroporous adsorbent resin,, collect 5 times of amount column volume elutriants with 30% ethanol elution, by the D101 absorption with macroporous adsorbent resin, with 70% ethanol elution, collect 5 times of amount column volume elutriants, adsorb by strong-basicity styrene type anionite-exchange resin, NaOH eluant solution with 0.5mol/L, collect wash-out, filter, by strongly acidic styrene's type Zeo-karb, collect effluent liquid, filter, filtrate concentrates by reverse osmosis membrane, adds methanol crystallization, fractional crystallization, washing is drying to obtain.
The condition of described enzymolysis is: 45 ℃ of temperature, pH value 4, enzymolysis 3 hours.
The time of described decoction is 1 hour.
Described strong-basicity styrene type anionite-exchange resin is selected from a kind of in 201 * 7 types, 201 * 4 types, the D201 type anionite-exchange resin.
Described strongly acidic styrene's type Zeo-karb is selected from a kind of in 001 * 7 type, 001 * 4 type, the D001 type Zeo-karb.
Described reverse osmosis membrane is selected from a kind of in cellulose acetate propionate film, aromatic polyamide membrane, the triacetate fiber composite membrane.
Preparation gained ursolic acid can adopt following method to detect:
Test routine 1HPLC method and measure ursolic acid purity
Chromatographic condition
Chromatographic column: octadecylsilane bonding glue silica gel is weighting agent; Moving phase: methyl alcohol-5% phosphate aqueous solution (90: 10); Flow velocity: 1ml/ minute; Detect wavelength: 220nm; Column temperature: 30 ℃.
Measuring method
Precision takes by weighing ursolic acid 2mg, places the 50mL measuring bottle, adds methyl alcohol 20mL, and sonic oscillation makes dissolving, and methanol constant volume is drawn 10 μ L to scale, injects high performance liquid chromatograph, adopts normalization method working sample purity.
Adopt the present invention to prepare ursolic acid, be beneficial to big production operation, energy consumption is little, pollutes little.
The present invention is further elaborated below in conjunction with embodiment, but the scope of protection of present invention is not limited to following embodiment.
Embodiment
Embodiment 1
Get paulownia leaf (deriving from the leaf of scrophulariaceae Paulownia plant paulownia Paulownia fortunei (Seem.) Hemsl) 10Kg, the ratio that adds 5mL in every g leaf adds entry, add cellulase, 40 ℃ of temperature, pH value 3, enzymolysis is 2 hours under the condition of enzyme amount 10U/g leaf, be heated to the boiling and decocted 0.5 hour, filter, get filtrate, by the AB8 absorption with macroporous adsorbent resin, with 30% ethanol elution, collect 5 times of amount column volume elutriants, by the D101 absorption with macroporous adsorbent resin, with 70% ethanol elution, collect 5 times of amount column volume elutriants, by the absorption of strong basicity 201 * 7 type styrene type anionite-exchange resin,, collect wash-out with the NaOH eluant solution of 0.5mol/L, filter, by 001 * 7 type strongly acidic styrene type Zeo-karb, collect effluent liquid, filter, filtrate is by cellulose acetate propionate membrane reverse osmosis membrane concentration, add methanol crystallization, fractional crystallization, washing is drying to obtain, be drying to obtain colourless needle-ursolic acid 42.6g, detect through HPLC, purity is 98.4%, UV, IR, MS;
2HNMR,
13The data of its physical behavior of sign such as CNMR are consistent with prior art.
Embodiment 2
Get paulownia leaf (deriving from the leaf of scrophulariaceae Paulownia plant paulownia Paulownia fortunei (Seem.) Hemsl) 10Kg, the ratio that adds 5mL in every g leaf adds entry, add cellulase, 50 ℃ of temperature, pH value 6, enzymolysis is 5 hours under the condition of enzyme amount 20U/g leaf, be heated to the boiling and decocted 1.5 hours, filter, get filtrate, by the AB8 absorption with macroporous adsorbent resin, with 30% ethanol elution, collect 5 times of amount column volume elutriants, by the D101 absorption with macroporous adsorbent resin,, collect 5 times of amount column volume elutriants with 70% ethanol elution, adsorb by 201 * 4 type strong-basicity styrene type anionite-exchange resin, with the NaOH eluant solution of 0.5mol/L, collect wash-out, filter, by 001 * 4 type strongly acidic styrene type Zeo-karb, collect effluent liquid, filter, filtrate concentrates by the aromatic polyamide membrane reverse osmosis membrane, add methanol crystallization, fractional crystallization, washing is drying to obtain colourless needle-ursolic acid 35.2g, detects through HPLC, purity is 99.3%, UV, IR, MS;
2HNMR,
13The data of its physical behavior of sign such as CNMR are consistent with prior art.
Embodiment 3
Get paulownia leaf (deriving from the leaf of scrophulariaceae Paulownia plant paulownia Paulownia fortunei (Seem.) Hemsl) 10Kg, the ratio that adds 5mL in every g leaf adds entry, add cellulase, 45 ℃ of temperature, pH value 4, enzymolysis is 3 hours under the condition of enzyme amount 15U/g leaf, be heated to the boiling and decocted 1 hour, filter, get filtrate, by the AB8 absorption with macroporous adsorbent resin, with 30% ethanol elution, collect 5 times of amount column volume elutriants, by the D101 absorption with macroporous adsorbent resin,, collect 5 times of amount column volume elutriants with 70% ethanol elution, adsorb by D201 type strong-basicity styrene type anionite-exchange resin, with the NaOH eluant solution of 0.5mol/L, collect wash-out, filter, by D001 type strongly acidic styrene type Zeo-karb, collect effluent liquid, filter, filtrate concentrates by triacetate fiber composite membrane reverse osmosis membrane, add methanol crystallization, fractional crystallization, washing is drying to obtain colourless needle-ursolic acid 66.8g, detects through HPLC, purity is 99.4%, UV, IR, MS;
2HNMR,
13The data of its physical behavior of sign such as CNMR are consistent with prior art.
Embodiment 4
Get paulownia leaf (deriving from the leaf of scrophulariaceae Paulownia vegetable wool paulownia P.tomentosa) 10Kg, the ratio that adds 5mL in every g leaf adds entry, add cellulase, 45 ℃ of temperature, pH value 4, enzymolysis is 3 hours under the condition of enzyme amount 15U/g leaf, be heated to the boiling and decocted 1 hour, filter, get filtrate, by the AB8 absorption with macroporous adsorbent resin, with 30% ethanol elution, collect 5 times of amount column volume elutriants, by the D101 absorption with macroporous adsorbent resin,, collect 5 times of amount column volume elutriants with 70% ethanol elution, adsorb by D201 type strong-basicity styrene type anionite-exchange resin, with the NaOH eluant solution of 0.5mol/L, collect wash-out, filter, by D001 type strongly acidic styrene type Zeo-karb, collect effluent liquid, filter, filtrate concentrates by triacetate fiber composite membrane reverse osmosis membrane, add methanol crystallization, fractional crystallization, washing is drying to obtain colourless needle-ursolic acid 62.2g, detects through HPLC, purity is 99.1%, UV, IR, MS;
2HNMR,
13The data of its physical behavior of sign such as CNMR are consistent with prior art.
Claims (6)
1. method of from the paulownia leaf, extracting ursolic acid, it is characterized in that described method is made up of the following step: get the paulownia leaf, the ratio that adds 5mL in every g leaf adds entry, add cellulase, at temperature 40-50 ℃, pH value 3-6, under the condition of enzyme amount 15U/g leaf enzymolysis 2-5 hour, be heated to boiling and decocted 0.5-1.5 hour, filter, get filtrate, by the AB8 absorption with macroporous adsorbent resin,, collect 5 times of amount column volume elutriants with 30% ethanol elution, by the D101 absorption with macroporous adsorbent resin, with 70% ethanol elution, collect 5 times of amount column volume elutriants, adsorb by strong-basicity styrene type anionite-exchange resin, NaOH eluant solution with 0.5mol/L, collect wash-out, filter, by strongly acidic styrene's type Zeo-karb, collect effluent liquid, filter, filtrate concentrates by reverse osmosis membrane, adds methanol crystallization, fractional crystallization, washing is drying to obtain.
2. according to the described a kind of method of from the paulownia leaf, extracting ursolic acid of claim 1, it is characterized in that the condition of described enzymolysis is: 45 ℃ of temperature, pH value 4, enzymolysis 3 hours.
3. according to the described a kind of method of extracting ursolic acid from the paulownia leaf of claim 1, the time that it is characterized in that described decoction is 1 hour.
4. according to the preparation method of the described a kind of bryonia alkyd of claim 1, it is characterized in that described strong-basicity styrene type anionite-exchange resin is selected from a kind of in 201 * 7 types, 201 * 4 types, the D201 type anionite-exchange resin.
5. according to the preparation method of the described a kind of bryonia alkyd of claim 1, it is characterized in that described strongly acidic styrene's type Zeo-karb is selected from a kind of in 001 * 7 type, 001 * 4 type, the D001 type Zeo-karb.
6. according to the preparation method of the described a kind of bryonia alkyd of claim 1, it is characterized in that described reverse osmosis membrane is selected from a kind of in cellulose acetate propionate film, aromatic polyamide membrane, the triacetate fiber composite membrane.
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| CN2013101823849A CN103214544A (en) | 2013-05-17 | 2013-05-17 | Method for extracting ursolic acid from paulownia leaves |
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| CN2013101823849A CN103214544A (en) | 2013-05-17 | 2013-05-17 | Method for extracting ursolic acid from paulownia leaves |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497094A (en) * | 2014-12-29 | 2015-04-08 | 江苏千药堂国医研究院有限公司 | Processing method based on comprehensive utilization of stems and roots of cucurbitaceae melon plants |
| CN106983803A (en) * | 2017-05-22 | 2017-07-28 | 贵州大学 | Logical application of the leaf volatile oil in anti-inflammatory anti-oxidation medicine is prepared in sea |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1453289A (en) * | 2003-05-19 | 2003-11-05 | 西安交通大学 | Process of extracting and separating ursolic acid from plant |
| CN101205248A (en) * | 2007-11-28 | 2008-06-25 | 付建明 | Method for preparing ursolic acid by using paulownia leaves as raw material |
-
2013
- 2013-05-17 CN CN2013101823849A patent/CN103214544A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1453289A (en) * | 2003-05-19 | 2003-11-05 | 西安交通大学 | Process of extracting and separating ursolic acid from plant |
| CN101205248A (en) * | 2007-11-28 | 2008-06-25 | 付建明 | Method for preparing ursolic acid by using paulownia leaves as raw material |
Non-Patent Citations (1)
| Title |
|---|
| 张利等: "酶法提取车前草中熊果酸的工艺研究", 《安徽农业科学》, vol. 38, no. 11, 31 December 2010 (2010-12-31), pages 5846 - 5847 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497094A (en) * | 2014-12-29 | 2015-04-08 | 江苏千药堂国医研究院有限公司 | Processing method based on comprehensive utilization of stems and roots of cucurbitaceae melon plants |
| CN104497094B (en) * | 2014-12-29 | 2017-01-11 | 江苏千药堂国医研究院有限公司 | Processing method based on comprehensive utilization of stems and roots of cucurbitaceae melon plants |
| CN106983803A (en) * | 2017-05-22 | 2017-07-28 | 贵州大学 | Logical application of the leaf volatile oil in anti-inflammatory anti-oxidation medicine is prepared in sea |
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Application publication date: 20130724 |