CN103242399A - Method for extracting apigenin-7-O-beta-D-glucuronide from broussonetia papyrifera leaves - Google Patents
Method for extracting apigenin-7-O-beta-D-glucuronide from broussonetia papyrifera leaves Download PDFInfo
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- CN103242399A CN103242399A CN 201310179358 CN201310179358A CN103242399A CN 103242399 A CN103242399 A CN 103242399A CN 201310179358 CN201310179358 CN 201310179358 CN 201310179358 A CN201310179358 A CN 201310179358A CN 103242399 A CN103242399 A CN 103242399A
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Abstract
The invention discloses a method for extracting apigenin-7-O-beta-D-glucuronide from broussonetia papyrifera leaves. The method comprises the following steps: a. pulverizing the raw material broussonetia papyrifera leaves, soaking in 5-10 times of saturated lime water 2-3 times, merging the extracting solutions, sequentially filtering with an ultrafiltration membrane and a nanofiltration membrane to carry out molecular trapping, regulating the pH value of the trapped solution to 2-3, adding into a polyamide resin for adsorption, carrying out gradient elution with an ethanol solution, and concentrating the eluate to obtain a concentrated solution; and b. purifying the concentrated solution by high-speed counter-current chromatography, carrying out on-line monitoring with an ultraviolet detector, collecting the target component according to the chromatograph, recovering the reagent, and drying under reduced pressure to obtain the apigenin-7-O-beta-D-glucuronide. When being used for producing apigenin-7-O-beta-D-glucuronide, the method has the advantages of simple technological operation and high yield, and can easily implement industrialization.
Description
Technical field
The invention belongs to biological technical field, particularly relate to a kind of method of from the paper mulberry leaf, extracting apigenin-7-O-beta-D-glucuronide.
Background technology
Paper mulberry is Moraceae paper mulberry platymiscium, is used as medicine with emulsion, root skin, bark, leaf, fruit and seed.Leaf heat-clearing, cool blood, dampness removing, desinsection are used for nosebleed epistaxis, enteritis, dysentery etc.Modern study is found, contains a large amount of flavonoid substances in the paper mulberry leaf, and wherein apigenin-7-O-beta-D-glucuronide is exactly one of composition wherein.
Apigenin-7-O-beta-D-glucuronide is to be the flavonoid glycoside of parent with the apigenin, molecular formula C21H18O11, and molecular structural formula:
。
By literature search, it is all more loaded down with trivial details to extract apigenin-7-O-beta-D-glucuronide method at present, and very difficult suitability for industrialized production.As patent (application number: 201210009876.) " extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide in the Tuberculate Speranskia Herb ", the method of this patent disclosure is to take by weighing the dry meal of Tuberculate Speranskia Herb, after adding is spent the night with 60% alcohol immersion of 10 times of amounts of meal, supersound extraction three times, united extraction liquid and concentrating under reduced pressure, concentrated solution is used sherwood oil respectively, ethyl acetate, the water-saturated n-butanol equal solvent respectively extracts three times, obtain n-butanol extract thus and carry out silica gel column chromatography, adopt different gradient chloroform-methanols=45: 1～chloroform-methanol=1: 1 pair of n-butanol extract to carry out wash-out; Detect with thin-layer chromatography TLC, merge same composition, be concentrated into small volume, obtain compound through purifying repeatedly again.
Summary of the invention
The objective of the invention is to overcome the shortcomings and deficiencies of prior art, a kind of efficient, short-cut method that extracts apigenin-7-O-beta-D-glucuronide from the paper mulberry leaf is provided.
In order to solve the problems of the technologies described above, the present invention realizes by the following technical solutions:
A kind of method of extracting apigenin-7-O-beta-D-glucuronide from the paper mulberry leaf is characterized in that following steps:
A gets the paper mulberry leaf raw material and pulverizes, doubly measure saturated limewater with 5-10 and soak 2-3 time, extracting solution merges filtration and carries out molecular retention with ultra-filtration membrane, nanofiltration membrane successively, and trapped fluid is regulated in the ph2-3 adding polyamide resin and adsorbed, ethanolic soln gradient elution, elutriant concentrate concentrated solution;
The above-mentioned concentrated solution of b adopts the high speed adverse current chromatogram purifying, and the UV-detector on-line monitoring is collected target component according to collection of illustrative plates, reclaims reagent, and drying under reduced pressure namely.
Described ultra-filtration membrane is the hollow cellulose film of molecular weight cut-off 2000-6000, and nanofiltration membrane is the hollow cellulose film of molecular weight cut-off 200-400.
Described ethanolic soln gradient elution is 2-5 times of column volume 20-30% ethanol elution impurity, 3-7 times of column volume 40-55% wash-out effective constituent.
The optional chloroform of solvent systems, methyl alcohol, water solvent system that high speed adverse current chromatogram described in the step b separates, blending ratio 5-10:3-7:6-13 gets the phase that fixes mutually, does moving phase down mutually.
Utilize the present invention to extract apigenin-7-O-beta-D-glucuronide from the paper mulberry leaf, method is simple to operate, be easy to realize suitability for industrialized production, and raw material is easy to get.
Embodiment:
Further specify the present invention below in conjunction with embodiment.
Embodiment 1:
Getting the paper mulberry leaf pulverizes, take by weighing 2kg, with 7 times of amount saturated limewater solution soaking 2 times, each 3 hours, extracting solution, hollow cellulose membrane filtration with molecular weight cut-off 2000, see through liquid and use the hollow cellulose film nanofiltration of molecular weight cut-off 400 again, collect concentrated solution and regulate the saturated absorption of ph3 adding polyamide resin column, earlier with 3 times of column volumes, 30% ethanol elution impurity, use 4 times of 50% ethanol elution again, collect elutriant concentrate concentrated solution.Get chloroform, methyl alcohol, water by the 5:3:7 mixed, fully after the layering, get and inject the high speed adverse current chromatogram pipe phase that fixes mutually, start main frame, rotating speed 800rpm pumps into down and do moving phase mutually simultaneously, after the balance, flow velocity 2ml/min is set, dissolves concentrated solution with moving phase simultaneously, inject high-speed counter-current chromatograph, the online detection of UV-detector is collected flow point, drying under reduced pressure, get pale yellow powder apigenin-7-O-beta-D-glucuronide 1.2g, content 98.1%.
Embodiment 2:
Getting the paper mulberry leaf pulverizes, take by weighing 2kg, with 5 times of amount saturated limewater solution soaking 3 times, each 2 hours, extracting solution, hollow cellulose membrane filtration with molecular weight cut-off 4000, see through liquid and use the hollow cellulose film nanofiltration of molecular weight cut-off 300 again, collect concentrated solution and regulate the saturated absorption of ph2 adding polyamide resin column, earlier with 4 times of column volumes, 20% ethanol elution impurity, use 6 times of 45% ethanol elution again, collect elutriant concentrate concentrated solution.Get chloroform, methyl alcohol, water by the 8:3:5 mixed, fully after the layering, get and inject the high speed adverse current chromatogram pipe phase that fixes mutually, start main frame, rotating speed 900rpm pumps into down and do moving phase mutually simultaneously, after the balance, flow velocity 3ml/min is set, dissolves concentrated solution with moving phase simultaneously, inject high-speed counter-current chromatograph, the online detection of UV-detector is collected flow point, drying under reduced pressure, get pale yellow powder apigenin-7-O-beta-D-glucuronide 1.3g, content 98.5%.
Embodiment 3:
Getting the paper mulberry leaf pulverizes, take by weighing 2kg, with 6 times of amount saturated limewater solution soaking 2 times, each 3 hours, extracting solution, hollow cellulose membrane filtration with molecular weight cut-off 3000, see through liquid and use the hollow cellulose film nanofiltration of molecular weight cut-off 300 again, collect concentrated solution and regulate the saturated absorption of ph3 adding polyamide resin column, earlier with 4 times of column volumes, 25% ethanol elution impurity, use 3 times of 60% ethanol elution again, collect elutriant concentrate concentrated solution.Get chloroform, methyl alcohol, water by the 9:4:10 mixed, fully after the layering, get and inject the high speed adverse current chromatogram pipe phase that fixes mutually, start main frame, rotating speed 850rpm pumps into down and do moving phase mutually simultaneously, after the balance, flow velocity 2ml/min is set, dissolves concentrated solution with moving phase simultaneously, inject high-speed counter-current chromatograph, the online detection of UV-detector is collected flow point, drying under reduced pressure, get pale yellow powder apigenin-7-O-beta-D-glucuronide 1.4g, content 96.5%.
Claims (4)
1. method of extracting apigenin-7-O-beta-D-glucuronide from the paper mulberry leaf is characterized in that following steps:
A gets the paper mulberry leaf raw material and pulverizes, doubly measure saturated limewater with 5-10 and soak 2-3 time, extracting solution merges filtration and carries out molecular retention with ultra-filtration membrane, nanofiltration membrane successively, and trapped fluid is regulated in the ph2-3 adding polyamide resin and adsorbed, ethanolic soln gradient elution, elutriant concentrate concentrated solution;
The above-mentioned concentrated solution of b adopts the high speed adverse current chromatogram purifying, and the UV-detector on-line monitoring is collected target component according to collection of illustrative plates, reclaims reagent, and drying under reduced pressure namely.
2. the method for from the paper mulberry leaf, extracting apigenin-7-O-beta-D-glucuronide according to claim 1, it is characterized in that the ultra-filtration membrane described in the step a is the hollow cellulose film of molecular weight cut-off 2000-6000, nanofiltration membrane is the hollow cellulose film of molecular weight cut-off 200-400.
3. the method for from the paper mulberry leaf, extracting apigenin-7-O-beta-D-glucuronide according to claim 1, it is characterized in that the ethanolic soln gradient elution described in the step a is 2-5 times of column volume 20-30% ethanol elution impurity, 3-7 times of column volume 40-55% wash-out effective constituent.
4. the method for from the paper mulberry leaf, extracting apigenin-7-O-beta-D-glucuronide according to claim 1, it is characterized in that the optional chloroform of solvent systems, methyl alcohol, water solvent system that the high speed adverse current chromatogram described in the step b separates, blending ratio 5-10:3-7:6-13, get the phase that fixes mutually, do moving phase down mutually.
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| CN 201310179358 CN103242399A (en) | 2013-05-15 | 2013-05-15 | Method for extracting apigenin-7-O-beta-D-glucuronide from broussonetia papyrifera leaves |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104997848A (en) * | 2014-04-17 | 2015-10-28 | 河北以岭医药研究院有限公司 | Applications of Broussonetia papyrifera leaf total flavone |
| CN105753918A (en) * | 2014-12-19 | 2016-07-13 | 河北以岭医药研究院有限公司 | Method for extracting apigenin-7-o-beta-D-glucuronide from paper mulberry leaves |
-
2013
- 2013-05-15 CN CN 201310179358 patent/CN103242399A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104997848A (en) * | 2014-04-17 | 2015-10-28 | 河北以岭医药研究院有限公司 | Applications of Broussonetia papyrifera leaf total flavone |
| CN105753918A (en) * | 2014-12-19 | 2016-07-13 | 河北以岭医药研究院有限公司 | Method for extracting apigenin-7-o-beta-D-glucuronide from paper mulberry leaves |
| CN105753918B (en) * | 2014-12-19 | 2020-05-19 | 河北以岭医药研究院有限公司 | method for extracting apigenin-7-o- β -D-glucuronide from broussonetia papyrifera leaves |
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Application publication date: 20130814 |