Patent US4361697A successfully refreshed from Google Patents.
US4361697A
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Extraction, separation and recovery of diterpene glycosides from Stevia rebaudiana plants
Assignee: F K SUZUKI INTERNATIONAL Inc
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Filing: 1981-05-21
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Granted: 1982-11-30
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Expires: 2001-05-21
Abstract
Abstract A process for recovering diterpene glycosides from the Stevia rebaudiana plant includes the steps of sequentially extracting plant material with a first solvent of intermediate polarity to extract plant substances which tend to interfere with a liquid chromatographic separation of the glycosides, and then with a second solvent of high polarity to extract glycosides, and chromatographically separating the extracted glycosides by introducing them onto a liquid chromatography column having a packing of an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support, eluting them with a solvent of polarity higher than that of the first solvent but lower than that of the second solvent, and collecting individually eluate fractions rich in respective glycosides.
Inventors
- 👤 Robert H. Dobberstein
- 👤 Mohamed S. Ahmed
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 06:54
CPC Classifications
C; C07; C07H; C07H15/00; C07H15/20; C07H15/24; C07H15/256; A; A61; A61K; A61K36/00; A61K36/18; A61K36/185; A61K36/28
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
156 words
A process for recovering diterpene glycosides from Stevia rebaudiana plant material, said process comprising: sequentially extracting Stevia rebaudiana plant material with a first solvent of intermediate polarity to remove impurities therefrom in a first extract substantially free of diterpene glycosides, and then with a second solvent of high polarity to give a second extract containing diterpene glycosides; chromatographing the resulting extracted diterpene glycosides on a liquid chromatography column having a packing comprising an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support; said extracted glycosides being introduced onto said column for chromatography thereon in solution in said second solvent or in another solvent having a high polarity not substantially greater than the polarity of the second solvent; said column being eluted with a solvent of polarity higher than that of said first solvent and lower than that of said second solvent; and collecting individually eluate fractions rich in respective diterpene glycosides.
Claim 2
Dependent Claim
156 words
A process for recovering diterpene glycosides from Stevia rebaudiana plant material, said process comprising: sequentially extracting Stevia rebaudiana plant material with a first solvent of intermediate polarity to remove impurities therefrom in a first extract substantially free of diterpene glycosides, and then with a second solvent of high polarity to give a second extract containing diterpene glycosides; chromatographing the resulting extracted diterpene glycosides on a liquid chromatography column having a packing comprising an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support; said extracted glycosides being introduced onto said column for chromatography thereon in solution in said second solvent or in another solvent having a high polarity not substantially greater than the polarity of the second solvent; said column being eluted with a solvent of polarity higher than that of said first solvent and lower than that of said second solvent; and collecting individually eluate fractions rich in respective diterpene glycosides.
Claim 3
Dependent Claim
42 words
The process of claim 1 wherein, prior to being chromatographed, said extracted diterpene glycosides are freed of said second solvent and are dissolved in a third solvent of lower polarity than that of said second solvent for said introduction onto the column.
Claim 4
Dependent Claim
30 words
The process of claim 2 wherein said third solvent is comprised of 95-100% by volume of an alkanol of from two to four carbon atoms and the balance substantially water.
Claim 5
Dependent Claim
10 words
The process of claim 3 wherein said alkanol is 1-propanol.
Claim 6
Dependent Claim
24 words
The process of claim 1, 2, 3, or 4, wherein said first solvent is a liquid haloalkane of from one to four carbon atoms.
Claim 7
Dependent Claim
10 words
The process of claim 5 wherein said haloalkane is chloroform.
Claim 8
Dependent Claim
23 words
The process of claim 1, 2, 3 or 4, wherein said second solvent is an alkanol of from one to three carbon atoms.
Claim 9
Dependent Claim
15 words
The process of claim 1, 2, 3 or 4 wherein said second solvent is methanol.
Claim 10
Dependent Claim
23 words
The process of claim 1, 2, 3 or 4 wherein said eluting solvent is an alkanol of from two to four carbon atoms.
Claim 11
Dependent Claim
15 words
The process of claim 1, 2, 3 or 4 wherein said eluting solvent is 1-propanol.
Claim 12
Dependent Claim
24 words
The process of claim 1, 2, 3 or 4 wherein said stationary phase contains an ether oxygen and said support comprises a silica gel.
Claim 13
Dependent Claim
31 words
The process of claim 11 wherein said silica gel has a mean particle size diameter of about 5-15 microns and a mean surface area of about 100-500 square meters per gram.
Claim 14
Dependent Claim
13 words
The process of claim 11 wherein said stationary phase comprises a glycerylpropyl moiety.
Claim 15
Dependent Claim
164 words
A process for recovering diterpene glycosides from Stevia rebaudiana plant material, said process comprising: sequentially extracting Stevia rebaudiana plant material with a first solvent comprising a liquid haloalkane of from one to four carbon atoms to remove impurities therefrom in a first extract substantially free of diterpene glycosides, and then with a second solvent comprising an alkanol of from one to three carbon atoms to give a second extract containing diterpene glycosides; chromatographing the resulting extracted diterpene glycosides on a liquid chromatography column having a packing comprising an ether oxygen-containing organic stationary phase covalently bonded through a silicon atom to a silica gel support; said extracted glycosides being introduced onto said column for chromatography thereon in solution in said second solvent or in another solvent having a high polarity not substantially greater than the polarity of the second solvent; said column being eluted with an alkanol of from two to four carbon atoms; and collecting individually eluate fractions rich in the respective diterpene glycosides.
Claim 16
Dependent Claim
164 words
A process for recovering diterpene glycosides from Stevia rebaudiana plant material, said process comprising: sequentially extracting Stevia rebaudiana plant material with a first solvent comprising a liquid haloalkane of from one to four carbon atoms to remove impurities therefrom in a first extract substantially free of diterpene glycosides, and then with a second solvent comprising an alkanol of from one to three carbon atoms to give a second extract containing diterpene glycosides; chromatographing the resulting extracted diterpene glycosides on a liquid chromatography column having a packing comprising an ether oxygen-containing organic stationary phase covalently bonded through a silicon atom to a silica gel support; said extracted glycosides being introduced onto said column for chromatography thereon in solution in said second solvent or in another solvent having a high polarity not substantially greater than the polarity of the second solvent; said column being eluted with an alkanol of from two to four carbon atoms; and collecting individually eluate fractions rich in the respective diterpene glycosides.
Claim 17
Dependent Claim
13 words
The process of claim 14 wherein said stationary phase comprises a glycerylpropyl moiety.
Claim 18
Dependent Claim
18 words
The process of claim 1, 2, 14 or 15 wherein said Stevia rebaudiana plant material comprises plant leaves.
Claim 19
Dependent Claim
143 words
A process for recovering diterpene glycosides from Stevia rebaudiana plant material, said process comprising: sequentially extracting Stevia rebaudiana leaves with chloroform to remove impurities therefrom in a first extract substantially free of diterpene glycosides, and then with methanol to give a second extract containing diterpene glycosides; chromatographing the resulting extracted diterpene glycosides on a liquid chromatography column having a packing comprising a glycerylpropyl moiety stationary phase covalently bonded through a silicon atom to a silica gel support having a mean particle size diameter of about 5-15 microns and a mean surface area of about 100-500 square meters per gram; said extracted glycosides being introduced onto said column for chromatography thereon in solution in a solvent comprised of 95-100% by volume of 1-propanol and the balance substantially water; said column being eluted with 1-propanol; and collecting individually eluate fractions rich in respective diterpene glycosides.
Claim 20
Dependent Claim
143 words
A process for recovering diterpene glycosides from Stevia rebaudiana plant material, said process comprising: sequentially extracting Stevia rebaudiana leaves with chloroform to remove impurities therefrom in a first extract substantially free of diterpene glycosides, and then with methanol to give a second extract containing diterpene glycosides; chromatographing the resulting extracted diterpene glycosides on a liquid chromatography column having a packing comprising a glycerylpropyl moiety stationary phase covalently bonded through a silicon atom to a silica gel support having a mean particle size diameter of about 5-15 microns and a mean surface area of about 100-500 square meters per gram; said extracted glycosides being introduced onto said column for chromatography thereon in solution in a solvent comprised of 95-100% by volume of 1-propanol and the balance substantially water; said column being eluted with 1-propanol; and collecting individually eluate fractions rich in respective diterpene glycosides.
Claim 21
Dependent Claim
46 words
In a process for separating a mixture of diterpene glycosides of the Stevia rebaudiana plant by liquid chromatography on a packed column, the improvement which comprises employing as the column packing an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support.
Claim 22
Dependent Claim
46 words
In a process for separating a mixture of diterpene glycosides of the Stevia rebaudiana plant by liquid chromatography on a packed column, the improvement which comprises employing as the column packing an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support.
Claim 23
Dependent Claim
20 words
The process of claim 18 wherein said stationary phase contains an ether oxygen and said inorganic support is silica gel.
Claim 24
Dependent Claim
13 words
The process of claim 19 wherein said stationary phase comprises a glycerylpropyl moiety.
Claim 25
Dependent Claim
31 words
The process of claim 19 wherein said silica gel has a mean particle size diameter of about 5-15 microns and a mean surface area of about 100-500 square meters per gram.
Claim 26
Dependent Claim
13 words
The process of claim 21 wherein said stationary phase comprises a glycerylpropyl moiety.
Claim 27
Dependent Claim
25 words
The process of claim 18, 19, 20, 21 or 22 wherein said column is eluted with an alkanol of from two to four carbon atoms.
Claim 28
Dependent Claim
17 words
The process of claim 18, 19, 20, 21 or 22 wherein said column is eluted with 1-propanol.
Claim 29
Dependent Claim
99 words
A process for separating a mixture of diterpene glycosides of the Stevia rebaudiana plant, said process comprising: introducing a solution of said mixture of diterpene glycosides in a solvent onto a liquid chromatography column, said column having a packing comprising an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support; said solvent comprising 95-100% by volume of an alkanol of from one to four carbon atoms and the balance substantially water; eluting said column with an alkanol of from two to four carbon atoms; and collecting individually eluate fractions rich in respective diterpene glycosides.
Claim 30
Dependent Claim
99 words
A process for separating a mixture of diterpene glycosides of the Stevia rebaudiana plant, said process comprising: introducing a solution of said mixture of diterpene glycosides in a solvent onto a liquid chromatography column, said column having a packing comprising an oxygen-containing organic stationary phase covalently bonded through a silicon atom to an inorganic support; said solvent comprising 95-100% by volume of an alkanol of from one to four carbon atoms and the balance substantially water; eluting said column with an alkanol of from two to four carbon atoms; and collecting individually eluate fractions rich in respective diterpene glycosides.
Claim 31
Dependent Claim
13 words
The process of claim 25 wherein said stationary phase contains an ether oxygen.
Claim 32
Dependent Claim
13 words
The process of claim 25 wherein said stationary phase comprises a glycerylpropyl moiety.
Claim 33
Dependent Claim
12 words
The process of claim 26 wherein said inorganic support is silica gel.
Claim 34
Dependent Claim
13 words
The process of claim 28 wherein said stationary phase comprises a glycerylpropyl moiety.
Claim 35
Dependent Claim
42 words
The process of claim 25 wherein said stationary phase comprises a glycerylpropyl moiety and said support comprises a silica gel having a mean particle size diameter of from 5-15 microns and a mean surface area of from 100-500 square meters per gram.
Claim 36
Dependent Claim
18 words
The process of claim 25, 26, 27, 28, 29 or 30 wherein each of said alkanols is 1-propanol.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1985-11-29 | 1986-05-31 | 1986-11-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-11-29 | 1990-05-31 | 1990-11-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-11-29 | 1994-05-31 | 1994-11-30 | $7700.00 | Window Closed / Lapsed |