CN102603833B - Extraction and separation process of apigenin-7-O-beta-D-glucopyranside from garden balsam stem - Google Patents
Extraction and separation process of apigenin-7-O-beta-D-glucopyranside from garden balsam stem Download PDFInfo
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- CN102603833B CN102603833B CN201210009876.3A CN201210009876A CN102603833B CN 102603833 B CN102603833 B CN 102603833B CN 201210009876 A CN201210009876 A CN 201210009876A CN 102603833 B CN102603833 B CN 102603833B
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Abstract
The invention belongs to the technical field of medicinal product research and development, and relates to an extraction and separation process of apigenin-7-O-beta-D-glucopyranside from garden balsam stem. The preparation process comprises the following steps: weighing garden balsam stem dried coarse powder, and immersing the garden balsam stem dried coarse powder in 60 percent ethanol the amount of which is 10 times of that of the coarse powder overnight; performing ultrasonic extraction for three times, combining the extracts, and concentrating at reduced pressure; extracting the concentrated solution respectively with petroleum ether, ethyl acetate, water-saturated butanol and other solvents for three times to obtain n-butyl alcohol extracts; performing silica-gel column chromatography on the n-butyl alcohol extracts, and eluting the n-butyl alcohol extracts by adopting different gradients of (chloroform-methanol of 45:1)-(chloroform-methanol of 1:1); detecting with thin-layer chromatography (TLC), combining the same component, concentrating to a small volume, and repeatedly purifying to obtain a compound; identifying the structure of the compound to be the apigenin-7-O-beta-D-glucopyranside by combining spectrum analysis. The invention aims to fulfill the quantity demand on production and scientific research, so that the yield is improved, and the cost is reduced.
Description
Technical field
Pharmaceutical prod research and development technical field under the present invention.The present invention relates to the extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide in Tuberculate Speranskia Herb.
Background technology
Tuberculate Speranskia Herb (Phryma leptostachya L.var.asiatica Hara) claims again Herba seu Radix Phrymatis, the grass etc. of delivering a child, belong to Phrymaceae (Phrymaceae) Tuberculate Speranskia Herb and belong to (Phryma) per nnial herb, grow in the dark and damp place on hillside, sylvan life, roadside, ditch bank and meadow.Tuberculate Speranskia Herb bitter, thoughts of returning home warp.There is distribution in a lot of areas of China.Tuberculate Speranskia Herb herb is all pharmaceutically acceptable, and the effect such as have clearing heat and promoting diuresis, activating blood circulation and reducing swelling, hasten parturition can be controlled the diseases such as wound, eczema, fracture, impetigo.At present, pharmacopeia is not included the relevant record that belongs to Tuberculate Speranskia Herb about Phrymaceae Tuberculate Speranskia Herb, but among the people conventional it treat diseases such as " rheumatic arthralgia ".Tuberculate Speranskia Herb is rich in abundant Lignanoids compounds.Japanese scholars Eiji Tan-iguchi and Yasuyoshi Oshima successively Fen Li phrymarolin-I (phrymalin-I), phrymarolin-II (phrymalin-II) and the acetyl Tuberculate Speranskia Herb fat element (leptostachyol acetate) of having obtained from the deep grass roots in North America.Subsequently, Eiji Taniguchi and Fumito Ishibashi have synthesized phrymarolin-I and phrymarolin-II and steric isomer thereof.Eiji Taniquchi and the also Fen Li haedoxao A of sesquilignan that obtain from the deep grass roots in North America of people such as Fumito Ishibashi, and haedoxan A is synthesized, but this synthetic method comprises 20 reactions steps, and severe reaction conditions, and combined coefficient is low.Korea S scholar SangMyung Zee etc. has isolated phrymarolin-II and ursolic acid from the ethyl acetate extract of North America Tuberculate Speranskia Herb dry aerial parts; In addition, Korea S scholar IL-kwon park etc. separates and obtains acetyl Tuberculate Speranskia Herb fat element and de-methoxy acetyl Tuberculate Speranskia Herb fat element from the chloroform extracted solution of Asia Tuberculate Speranskia Herb dry root.From Tuberculate Speranskia Herb, separate the compound obtaining, except ursolic acid is triterpene compound, all the other are Lignanoids compounds.
Summary of the invention
1, the extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide in Tuberculate Speranskia Herb, is characterized in that this technique comprises following process:
Step (1): take the dry meal 4.1kg of Tuberculate Speranskia Herb, after adding and spending the night with 60% alcohol immersion of 10 times of amounts of meal, supersound extraction 30min-35min, ultrasonic temperature is 30 ℃-32 ℃, and ultrasonic power is 80W, supersound extraction three times, united extraction liquid concentrating under reduced pressure, concentrated solution is placed in 4 ℃ of refrigerator 48h-60h, suction filtration, the concentrated solution of the partial pigment that is removed precipitation;
Step (2): the concentrated solution in step (1) is respectively extracted three times with sherwood oil, ethyl acetate, water-saturated n-butanol equal solvent respectively, each extraction partial concentration is to thick paste shape medicinal extract, reclaim solvent, obtain thus n-butanol extract 45g;
Step (3): n-butanol extract 45g in step (2) is dissolved in 200ml chloroform, mixes 100～200 order silica gel with 100g, volatilize solvent, select 450g100～200 order silica gel chloroform to mix thoroughly, wet method dress post, dry method loading;
Step (4): adopt different gradients chloroform-methanol=45: 1～chloroform-methanol=1: the n-butanol extract in 1 pair of step (3) carries out elution chromatography, with every 50ml elutriant, collect one bottle, detect with thin-layer chromatography TLC, merge same composition, be concentrated into small volume, then obtain compound 42mg through purifying repeatedly;
Step (5): identify that in conjunction with spectroscopic analysis in (4), compound structure is apigenin-7-O-beta-D-phranoglucuronide.
According to the present invention, " % " in the present invention is weight percent.
Embodiment
In Tuberculate Speranskia Herb of the present invention, the extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide comprises following examples, and the following examples can further illustrate the present invention, but do not limit the present invention in any way.
Embodiment 1:
1, the extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide in Tuberculate Speranskia Herb, is characterized in that this technique comprises following process:
Step (1): take the dry meal 4.1kg of Tuberculate Speranskia Herb, after adding and spending the night with 60% alcohol immersion of 10 times of amounts of meal, supersound extraction 30min, ultrasonic temperature is 30 ℃, and ultrasonic power is 80W, supersound extraction three times, united extraction liquid concentrating under reduced pressure, concentrated solution is placed in 4 ℃ of refrigerator 48h, suction filtration, the concentrated solution of the partial pigment that is removed precipitation;
Step (2): the concentrated solution in step (1) is respectively extracted three times with sherwood oil, ethyl acetate, water-saturated n-butanol equal solvent respectively, each extraction partial concentration is to thick paste shape medicinal extract, reclaim solvent, obtain thus n-butanol extract 45g;
Step (3): n-butanol extract 45g in step (2) is dissolved in 200ml chloroform, mixes 100～200 order silica gel with 100g, volatilize solvent, select 450g100～200 order silica gel chloroform to mix thoroughly, wet method dress post, dry method loading;
Step (4): adopt different gradients chloroform-methanol=45: 1～chloroform-methanol=1: the n-butanol extract in 1 pair of step (3) carries out elution chromatography, with every 50ml elutriant, collect one bottle, detect with thin-layer chromatography TLC, merge same composition, be concentrated into small volume, then obtain compound 42mg through purifying repeatedly;
Step (5): identify that in conjunction with spectroscopic analysis in (4), compound structure is apigenin-7-O-beta-D-phranoglucuronide.
Embodiment 2:
1, the extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide in Tuberculate Speranskia Herb, is characterized in that this technique comprises following process:
Step (1): take the dry meal 4.1kg of Tuberculate Speranskia Herb, after adding and spending the night with 60% alcohol immersion of 10 times of amounts of meal, supersound extraction 35min, ultrasonic temperature is 32 ℃, and ultrasonic power is 80W, supersound extraction three times, united extraction liquid concentrating under reduced pressure, concentrated solution is placed in 4 ℃ of refrigerator 60h, suction filtration, the concentrated solution of the partial pigment that is removed precipitation;
Step (2): the concentrated solution in step (1) is respectively extracted three times with sherwood oil, ethyl acetate, water-saturated n-butanol equal solvent respectively, each extraction partial concentration is to thick paste shape medicinal extract, reclaim solvent, obtain thus n-butanol extract 45g;
Step (3): n-butanol extract 45g in step (2) is dissolved in 200ml chloroform, mixes 100～200 order silica gel with 100g, volatilize solvent, select 450g100～200 order silica gel chloroform to mix thoroughly, wet method dress post, dry method loading;
Step (4): adopt different gradients chloroform-methanol=45: 1～chloroform-methanol=1: the n-butanol extract in 1 pair of step (3) carries out elution chromatography, with every 50ml elutriant, collect one bottle, detect with thin-layer chromatography TLC, merge same composition, be concentrated into small volume, then obtain compound 42mg through purifying repeatedly;
Step (5): identify that in conjunction with spectroscopic analysis in (4), compound structure is apigenin-7-O-beta-D-phranoglucuronide.The Structural Identification of compound
Yellow amorphous powder, is dissolved in hot methanol, hot water, hot ethanol.The reaction of hydrochloric acid-magnesium powder is positive, and Molish reaction is positive, and the reaction of iron trichloride-Tripotassium iron hexacyanide is aobvious blue, and pointing out this compound is flavonoid glycoside compounds.Aubepine-sulfate spray displaing yellow (105 ℃).With 2% sulphuric acid hydrolysis, the existence to glucuronic acid is known in thin layer inspection.
1h-NMR has 7 hydrogen proton signal: δ 7.95 (2H, d, J=8.3Hz, H-2 ' in fragrant district, 6 ') and δ 6.94 (2H, d, J=8.3Hz, H-3 ', 5 ') be respectively that dihydro is bimodal, coupling constant J=8.3Hz, illustrates that Flavone aglycone B ring is for AA ' BB ' system; δ 6.83 (1H, br.s, H-8) and δ 6.44 (1H, br.s, H-6) have respectively a single hydrogen wide unimodal, the two hydrogen protons that close for A interannular digit pair; It is unimodal that δ 6.85 (1H, s, H-3) has located a sharp-pointed single hydrogen, is 3 hydrogen proton signal peaks of Flavone aglycone; The δ 12.97 (1H, br.s) of low place is the characteristic signal peak of flavones 5-OH, illustrates that 7 have oxygen-containing substituents; δ 10～12 (1H, br.s) is the characteristic signal peak of flavones 4-OH.(1H, d, J=7.1Hz, H-1 ") locate as glucuronic acid terminal hydrogen fignal center δ 5.16, by its coupling constant J=7.1Hz, learn that its glycosidic bond is beta comfiguration.At δ 3.2～δ 3.8 places, there is proton signal on 4 sugar in addition.
13c-NMR has provided 21 carbon signals, and wherein δ 170.9 (C-6 "), δ 99.4 (C-1 "), δ 76.0 (C-5 "), δ 74.5 (C-3 "), δ 72.8 (C-2 "), δ 71.6 (C-4 ") are one group of glucal acidic group fignal center.The carbon signal that also has in addition 15 Flavone aglycones, wherein δ 99.5 and δ 94.6 are respectively the characteristic signal peaks of 6,8 carbon on 5,7-dioxo flavone A ring; δ 181.9 is the characteristic signal peak of 4 C=O of flavonoid compound; δ 164.2 and δ 102.9 are respectively the characteristic signal peak of flavonoid compound C-2 and C-3; δ 128.5 and δ 115.9 are respectively the C-2 ' in AA ' BB ' system on flavonoid compound B ring, 6 ', C-3 ', 5 ' characteristic signal peak; δ 162.7 illustrates that flavonoid compound C-7 and glucuronic acid are connected to form oxygen glycosides.By NMR spectrum, data are belonged to.Basically identical through By consulting literatures and its spectral data.Do not decline with reference substance contrast exhibition distance, all consistent and mixed melting points that develops the color.According to above parsing, inferring this compound structure is apigenin-7-O-beta-D-phranoglucuronide.
Claims (1)
1. the extraction and separation process of apigenin-7-O-beta-D-phranoglucuronide in Tuberculate Speranskia Herb, is characterized in that this technique comprises following process:
Step (1): take the dry meal 4.1kg of Tuberculate Speranskia Herb, after adding and spending the night with 60% alcohol immersion of 10 times of amounts of meal, supersound extraction 30min-35min, ultrasonic temperature is 30 ℃-32 ℃, and ultrasonic power is 80W, supersound extraction three times, united extraction liquid concentrating under reduced pressure, concentrated solution is placed in 4 ℃ of refrigerator 48h-60h, suction filtration, the concentrated solution of the partial pigment that is removed precipitation;
Step (2): the concentrated solution in step (1) is respectively extracted three times with sherwood oil, ethyl acetate, water-saturated n-butanol solvent respectively, each extraction partial concentration is to thick paste shape medicinal extract, reclaim solvent, obtain thus n-butanol extract 45g;
Step (3): n-butanol extract 45g in step (2) is dissolved in 200ml chloroform, mixes with 100g100～200 order silica gel, volatilize solvent, select 450g100～200 order silica gel chloroform to mix thoroughly, wet method dress post, dry method loading;
Step (4): adopt different gradient chloroform-methanol=45:1～chloroform-methanol=1:1 to carry out elution chromatography to the n-butanol extract in step (3), with every 50ml elutriant, collect one bottle, detect with thin-layer chromatography TLC, merge same composition, be concentrated into small volume, then obtain compound 42mg through purifying repeatedly;
Step (5): identify that in conjunction with spectroscopic analysis in (4), compound structure is apigenin-7-O-beta-D-phranoglucuronide.
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| CN107812010A (en) * | 2017-10-30 | 2018-03-20 | 成都中医药大学 | Leonurus extract adjusts the new application of uterine smooth muscle |
| CN110731978A (en) * | 2019-12-09 | 2020-01-31 | 张宇静 | preparation method of garden balsam stem extract and effervescent tablet thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1682746A (en) * | 2005-03-02 | 2005-10-19 | 姜瑞芝 | Use of clasping gutweed flavonoid in preparing medicine for treating coronary heart disease |
| CN101292988A (en) * | 2008-06-25 | 2008-10-29 | 通化华夏药业有限责任公司 | Use of apigenin -7-O-beta-D glycosides glucuronic acid in preparing medicaments for treating dementia diseases |
| WO2010026666A1 (en) * | 2008-09-04 | 2010-03-11 | サントリーホールディングス株式会社 | Glucuronyltransferase and polynucleotide encoding the same |
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| JP2011213699A (en) * | 2010-04-02 | 2011-10-27 | Kinki Univ | Skin care external preparation having collagen production promotion action obtained from daisy inflorescence |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1682746A (en) * | 2005-03-02 | 2005-10-19 | 姜瑞芝 | Use of clasping gutweed flavonoid in preparing medicine for treating coronary heart disease |
| CN101292988A (en) * | 2008-06-25 | 2008-10-29 | 通化华夏药业有限责任公司 | Use of apigenin -7-O-beta-D glycosides glucuronic acid in preparing medicaments for treating dementia diseases |
| WO2010026666A1 (en) * | 2008-09-04 | 2010-03-11 | サントリーホールディングス株式会社 | Glucuronyltransferase and polynucleotide encoding the same |
Non-Patent Citations (5)
| Title |
|---|
| JP特开2011-213699A 2011.10.27 |
| Ufuk Ozgen，等."A New Sulfated α-Ionone Glycoside from Sonchus erzincanicus Matthews".《Molecules》.2010,第15卷第2593-2599页. |
| Ufuk Ozgen,等."A New Sulfated α-Ionone Glycoside from Sonchus erzincanicus Matthews".《Molecules》.2010,第15卷第2593-2599页. * |
| 关键，等."透骨草不同极性成分抗氧化活性研究".《包装与食品机械》.2011,第29卷(第2期),第18-21页. |
| 关键,等."透骨草不同极性成分抗氧化活性研究".《包装与食品机械》.2011,第29卷(第2期),第18-21页. * |
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