CN105616477A - Method for purifying polyphenol in lonicera edulis - Google Patents
Method for purifying polyphenol in lonicera edulis Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention discloses a method for purifying polyphenol in lonicera edulis. The method comprises the steps of lonicera edulis pulping, homogenizing with a high-pressure homogenizer, extraction with the ultrasonic-assisted ethanol extraction method, vacuum suction filtration and filtrate collection, filtrate concentration with a rotary evaporator, polyphenol concentrated solution collection, macroporous resin purification, tracing and checking with the thin-layer chromatography method and lonicera edulis polyphenol eluant collection, polyphenol enriched product filtration with an organic millipore filter which is 0.45 micron thick, polyphenol enriched product repurification with the Sephadex LH-20 sephadex column chromatography method, polyphenol eluant vacuum concentration and ethyl alcohol recovery, lonicera edulis polyphenol vacuum freeze drying, and purple red lonicera edulis polyphenol finished product acquisition. By the adoption of the method, the commercial value of the lonicera edulis resource is increased, the adopted macroporous resin has the advantages of being stable in physicochemical property, high in adsorptive selectivity, high in adsorption capacity, good in adsorption effect, mild in desorption condition, easy to regenerate and the like and is suitable for large-scale production of lonicera edulis polyphenol, the operation process is safe and reliable, zero pollution is achieved, cost is low, and extraction efficiency and purification efficiency are high.
Description
Technical field
The invention belongs to field of natural organic chemistry, it relates to a kind of method utilizing the polyphenol of ultrasonic-assisted extraction-in macroporous resin coupling technique purifying Lonicera edulis honeysuckle.
Background technology
Lonicera edulis Turcz. (LoniceraCaeruleaL) referred to as indigo fruit, among the people be also commonly called as Brachybotrys paridiformis, goat milk etc., its shape is oblong, and in deep blue purple color, fruit taste is similar to more climing tangerine, blueberry etc., in sour-sweet taste. Indigo fruit is a kind of emerging Wild fruits, has stronger vitality, cold tolerance, and adaptability is also fine. Containing the multiple nutritional components such as VITAMIN, mineral substance in indigo fruit, can eat raw and also can carry out process for processing. In addition, indigo fruit is also the berry of a kind of bioactive ingredients enrichment, especially the wide variety of anthocyanin class material, phenolic acid and Flavonoid substances, content are higher, also there is multiple biological activity simultaneously, such as preventing cardiovascular disease, antibacterial anti-inflammatory, preventing cancer and the physiological function such as anti-oxidant. The present invention adopts the content of ultrasonic-assisted extraction-macroporous resin coupling technique gained Lonicera edulis Turcz. polyphenol and purity all higher, it is intended that the separation of indigo fruit polyphenol and application provide new approach.
Summary of the invention
It is an object of the present invention to provide a kind of method utilizing the polyphenol of ultrasonic-assisted extraction-in macroporous resin coupling technique purifying Lonicera edulis honeysuckle, while ensureing to obtain with high-level efficiency the Lonicera edulis Turcz. polyphenol highly finished product of high-content, high purity, reduce technique cost as much as possible, reduce energy consumption and pollution on the environment.
The technical process of technical scheme provided by the present invention is as follows:
Lonicera edulis Turcz. fresh fruit �� select, clean, drain �� pull an oar �� high pressure homogenizer homogeneous �� ultrasonic assistant ethanol extraction method lixiviate �� vacuum filters and ethanolic soln wash-out �� tlc tracing detection �� collection polyphenol enriched substance �� 0.45 ��m organic filtering with microporous membrane polyphenol enriched substance �� SephadexLH-20 dextrane gel column chromatography of macroporous resin purification �� distilled water removal of impurities �� acetic acid after collecting filtrate �� Rotary Evaporators concentrated filtrate �� screening purified polyphenol enriched substance �� with ethanol-acetic acid-water (70:0.01:29.99 againv/v/v) eluant solution �� concentrating under reduced pressure elutriant �� vacuum lyophilization �� red-purple indigo fruit polyphenol highly finished product
Method steps is as follows:
(1) raw material selection: pick out fruit consistent with physical abuse and ripening degree without disease and pest in the Lonicera edulis Turcz. after harvesting, clean, drain.
(2) indigo fruit polyphenol extracts: adopt refiner to be pulled an oar by pretreated Lonicera edulis Turcz., and by high pressure homogenizer to indigo fruit pulp homogeneous 5min, then additional proportion is 40��55:0.1:59.9��44.9(v/v/v) ethanol-acetic acid-aqueous solution, be 1:15��25(g/mL according to solid-liquid ratio) ratio mix, to be mixed be evenly placed on lixiviate 75��100min in 35��50 DEG C of Ultrasonic Cleanerss (power is 500W).
(3) vacuum filtration: the polyphenolic extract after lixiviate is carried out vacuum and filters and collect filtrate.
(4) concentrated: indigo fruit polyphenol filtrate to be placed in round-bottomed flask and rotary evaporation in vacuo instrument process 30��35min under 35��40 DEG C of conditions, reclaim ethanol and also collect polyphenol concentrated solution.
(5) screening of macroporous resin: to collect polyphenol concentrated solution again vacuum filter, and the adsorptive capacity of macroporous resin to polyphenol filtrate of XAD-7, X-5, S-8, D101, AB-8, D1400, adsorption rate and desorption efficiency it is respectively by comparative analysis model, therefrom filter out the suitableeest macroporous resin of purifying Lonicera edulis honeysuckle polyphenol.
(6) macroporous resin purification: the concentrated filtrate being 2.03mg/mL by concentration with constant flow pump carries out loading with the flow velocity of 0.5��2BV/h, and applied sample amount is 1.5��2.5 times of resin volume, to macroporous resin in absorption state of saturation. Be first the distilled water flushing pillar of 2.5��4BV with elution volume, remove the impurity such as polysaccharide, protein in indigo fruit polyphenol, then with the 65��95% of 4 times of column volumes (v/v) ethanol eluate control elution flow rate be that 1��2BV/h carries out gradient elution, and with tlc follow the tracks of and detection, collect indigo fruit polyphenol enriched substance.
(7) organic membrane filtration: by 0.45 ��m of organic filtering with microporous membrane indigo fruit polyphenol enriched substance and collect filtrate.
(8) sephadex column layer analysis: the polyphenol enriched substance after filtering is carried out secondarily purified after the absorption of SephadexLH-20 sephadex column is saturated, with the ethanol-acetic acid-water of 4��5BV (70:0.01:29.99,v/v/v) solution carries out wash-out, collects secondarily purified polyphenol elutriant.
(9) concentrated and vacuum lyophilization: the polyphenol elutriant collected is carried out concentrating under reduced pressure under 35��45 DEG C of conditions and reclaims ethanol, then the vacuum freeze drier being placed in by polyphenol concentrated solution under 30��40 DEG C of conditions is dried process, make indigo fruit polyphenol highly finished product powder, in red-purple.
It is an advantage of the invention that: 1. macroporous resin have stable in physicochemical property, loading capacity effective greatly, regenerate the advantages such as simple, be applicable to indigo fruit polyphenol large-scale production; 2. whole operating procedure is safe and reliable, and operative technique is with low cost; 3. the indigo fruit polyphenol content and the purity that are obtained by this technique are all higher; 4. this technology improves the commercial value of indigo fruit resource.
Embodiment
Embodiment 1
After picking out fruit consistent with physical abuse and ripening degree without disease and pest in the Lonicera edulis Turcz. after harvesting, clean, draining, pull an oar with refiner, and by high pressure homogenizer to indigo fruit pulp homogeneous 5min, then additional proportion is 40:0.1:59.9(v/v/v) ethanol-acetic acid-aqueous solution, be 1:15(g/mL according to solid-liquid ratio) ratio mix, to be mixed be evenly placed on lixiviate 75min in 35 DEG C of Ultrasonic Cleanerss (power is 500W); Then polyphenol vat liquor being carried out vacuum to filter, collect indigo fruit polyphenol filtrate and be placed in round-bottomed flask rotary evaporation 30min under 35 DEG C of conditions, reclaim ethanol and collect polyphenol concentrated solution, vacuum filters polyphenol concentrated solution again; Be respectively the adsorptive capacity of macroporous resin to polyphenol filtrate of XAD-7, X-5, S-8, D101, AB-8, D1400, adsorption rate and desorption efficiency by comparative analysis model, the selection result is the best results of XAD-7 type macroporous resin purification Lonicera edulis Turcz. polyphenol. The concentrated filtrate being 2.03mg/mL by concentration with constant flow pump carries out loading with the flow velocity of 0.5BV/h, applied sample amount is 1.5 times of resin volume, to macroporous resin in absorption state of saturation, it it is first the distilled water flushing pillar of 3BV with elution volume, remove the impurity such as polysaccharide, protein in indigo fruit polyphenol, then with the 65��95% of 4 times of column volumes (v/v) ethanol eluate control elution flow rate be that 1BV/h carries out gradient elution, and follow the tracks of by tlc and detection, collect indigo fruit polyphenol enriched substance, then by 0.45 ��m of organic filtering with microporous membrane indigo fruit polyphenol enriched substance and collect filtrate, polyphenol filtrate is carried out secondarily purified after the absorption of SephadexLH-20 sephadex column is saturated, with the ethanol-acetic acid-water (70:0.01:29.99 of 4BVv/v/v) solution carries out wash-out, collects secondarily purified polyphenol elutriant. Under 35 DEG C of conditions, polyphenol elutriant being carried out concentrating under reduced pressure and reclaim ethanol, the vacuum freeze drier being then placed in by polyphenol concentrated solution under 30 DEG C of conditions is dried process, makes indigo fruit polyphenol highly finished product powder, in red-purple.
Embodiment 2
After picking out fruit consistent with physical abuse and ripening degree without disease and pest in the Lonicera edulis Turcz. after harvesting, clean, draining, pull an oar with refiner, and by high pressure homogenizer to indigo fruit pulp homogeneous 5min, then additional proportion is 50:0.1:49.9(v/v/v) ethanol-acetic acid-aqueous solution, be 1:20(g/mL according to solid-liquid ratio) ratio mix, to be mixed be evenly placed on lixiviate 90min in 40 DEG C of Ultrasonic Cleanerss (power is 500W); Then polyphenol vat liquor being carried out vacuum to filter, collect indigo fruit polyphenol filtrate and be placed in round-bottomed flask rotary evaporation 35min under 40 DEG C of conditions, reclaim ethanol and collect polyphenol concentrated solution, vacuum filters polyphenol concentrated solution again; Be respectively the adsorptive capacity of macroporous resin to polyphenol filtrate of XAD-7, X-5, S-8, D101, AB-8, D1400, adsorption rate and desorption efficiency by comparative analysis model, the selection result is the best results of XAD-7 type macroporous resin purification Lonicera edulis Turcz. polyphenol. The concentrated filtrate being 2.03mg/mL by concentration with constant flow pump carries out loading with the flow velocity of 1BV/h, applied sample amount is 2 times of resin volume, to macroporous resin in absorption state of saturation, it it is first the distilled water flushing pillar of 3.5BV with elution volume, remove the impurity such as polysaccharide, protein in indigo fruit polyphenol, then with the 65��95% of 4 times of column volumes (v/v) ethanol eluate control elution flow rate be that 1.5BV/h carries out gradient elution, and follow the tracks of by tlc and detection, collect indigo fruit polyphenol enriched substance, then by 0.45 ��m of organic filtering with microporous membrane indigo fruit polyphenol enriched substance and collect filtrate, polyphenol filtrate is carried out secondarily purified after the absorption of SephadexLH-20 sephadex column is saturated, with the ethanol-acetic acid-water (70:0.01:29.99 of 4.5BVv/v/v) solution carries out wash-out, collects secondarily purified polyphenol elutriant. Under 40 DEG C of conditions, polyphenol elutriant being carried out concentrating under reduced pressure and reclaim ethanol, the vacuum freeze drier being then placed in by polyphenol concentrated solution under 35 DEG C of conditions is dried process, makes indigo fruit polyphenol highly finished product powder, in red-purple.
Embodiment 3
After picking out fruit consistent with physical abuse and ripening degree without disease and pest in the Lonicera edulis Turcz. after harvesting, clean, draining, pull an oar with refiner, and by high pressure homogenizer to indigo fruit pulp homogeneous 5min, then additional proportion is 55:0.1:44.9(v/v/v) ethanol-acetic acid-aqueous solution, be 1:25(g/mL according to solid-liquid ratio) ratio mix, to be mixed be evenly placed on lixiviate 100min in 50 DEG C of Ultrasonic Cleanerss (power is 500W); Then polyphenol vat liquor being carried out vacuum to filter, collect indigo fruit polyphenol filtrate and be placed in round-bottomed flask rotary evaporation 35min under 40 DEG C of conditions, reclaim ethanol and collect polyphenol concentrated solution, vacuum filters polyphenol concentrated solution again; Be respectively the adsorptive capacity of macroporous resin to polyphenol filtrate of XAD-7, X-5, S-8, D101, AB-8, D1400, adsorption rate and desorption efficiency by comparative analysis model, the selection result is the best results of XAD-7 type macroporous resin purification Lonicera edulis Turcz. polyphenol. The concentrated filtrate being 2.03mg/mL by concentration with constant flow pump carries out loading with the flow velocity of 1.5BV/h, applied sample amount is 2.5 times of resin volume, to macroporous resin in absorption state of saturation, it it is first the distilled water flushing pillar of 4BV with elution volume, remove the impurity such as polysaccharide, protein in indigo fruit polyphenol, then with the 65��95% of 4 times of column volumes (v/v) ethanol eluate control elution flow rate be that 2BV/h carries out gradient elution, and follow the tracks of by tlc and detection, collect indigo fruit polyphenol enriched substance, then by 0.45 ��m of organic filtering with microporous membrane indigo fruit polyphenol enriched substance and collect filtrate, polyphenol filtrate is carried out secondarily purified after the absorption of SephadexLH-20 sephadex column is saturated, with the ethanol-acetic acid-water (70:0.01:29.99 of 5BVv/v/v) solution carries out wash-out, collects secondarily purified polyphenol elutriant. Under 45 DEG C of conditions, polyphenol elutriant being carried out concentrating under reduced pressure and reclaim ethanol, the vacuum freeze drier being then placed in by polyphenol concentrated solution under 40 DEG C of conditions is dried process, makes indigo fruit polyphenol highly finished product powder, in red-purple.
Claims (1)
1. the method for polyphenol in the purifying Lonicera edulis honeysuckle, it is characterised in that concrete operation steps is as follows:
(1) fruit consistent with physical abuse and ripening degree without disease and pest in the Lonicera edulis Turcz. after harvesting is picked out, after cleaning drains, employing refiner is pulled an oar, and by high pressure homogenizer to indigo fruit pulp homogeneous 5min, then additional proportion is 40��55:0.1:59.9��44.9(v/v/v) ethanol-acetic acid-aqueous solution, be 1:15��25(g/mL according to solid-liquid ratio) ratio mix, to be mixed be evenly placed on lixiviate 75��100min in 35��50 DEG C of Ultrasonic Cleanerss, gained vat liquor carried out vacuum and filters and collect filtrate;
(2) indigo fruit polyphenol filtrate is placed in round-bottomed flask and rotary evaporation in vacuo instrument process 30��35min under 35��40 DEG C of conditions, reclaim ethanol and also collect polyphenol concentrated solution;
(3) to collect polyphenol concentrated solution again vacuum filter, and the adsorptive capacity of macroporous resin to polyphenol filtrate of XAD-7, X-5, S-8, D101, AB-8, D1400, adsorption rate and desorption efficiency it is respectively by comparative analysis model, therefrom filter out the suitableeest macroporous resin of purifying Lonicera edulis honeysuckle polyphenol;
(4) concentrated filtrate being 2.03mg/mL by concentration with constant flow pump carries out loading with the flow velocity of 0.5��2BV/h, and applied sample amount is 1.5��2.5 times of resin volume, to macroporous resin in absorption state of saturation;
Be first the distilled water flushing pillar of 2.5��4BV with elution volume, remove polysaccharide, protein impurities in indigo fruit polyphenol, then with the 65��95% of 4 times of column volumes (v/v) ethanol eluate control elution flow rate be that 1��2BV/h carries out gradient elution, and with tlc follow the tracks of and detection, collect indigo fruit polyphenol enriched substance;
(5) indigo fruit polyphenol enriched substance is collected also with 0.45 ��m of organic filtering with microporous membrane, then polyphenol enriched substance is carried out secondarily purified after the absorption of SephadexLH-20 sephadex column is saturated, wash-out is carried out, the proportions 70:0.01:29.99(of ethanol-acetic acid-aqueous solution with the ethanol-acetic acid-aqueous solution of 4��5BVv/v/v), collect secondarily purified polyphenol elutriant;
(6) the polyphenol elutriant collected is carried out concentrating under reduced pressure under 35��45 DEG C of conditions and reclaim ethanol, then the vacuum freeze drier being placed in by polyphenol concentrated solution under 30��40 DEG C of conditions is dried process, make indigo fruit polyphenol highly finished product powder, in red-purple.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107647399A (en) * | 2017-06-08 | 2018-02-02 | 湖北理工学院 | A kind of fruit of glossy privet polyphenol and its production method |
| CN112493482A (en) * | 2020-09-22 | 2021-03-16 | 沈阳农业大学 | Composite nutritional powder with functions of regulating intestinal flora and barrier and preparation method thereof |
| CN113754703A (en) * | 2021-09-10 | 2021-12-07 | 黑龙江格爱自动化科技有限公司 | Method for extracting berry polyphenol, flavonoid and flavonoid compounds |
Citations (2)
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| CN101463056A (en) * | 2007-12-20 | 2009-06-24 | 东北林业大学 | Preparation technique for purifying Lonicera edulis anthocyanin by using macroporous resin |
| CN102342952A (en) * | 2011-07-28 | 2012-02-08 | 哈尔滨工业大学 | Application of lonicera edulis anthocyanin in anti-radiation medicament |
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2016
- 2016-03-02 CN CN201610114628.3A patent/CN105616477A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101463056A (en) * | 2007-12-20 | 2009-06-24 | 东北林业大学 | Preparation technique for purifying Lonicera edulis anthocyanin by using macroporous resin |
| CN102342952A (en) * | 2011-07-28 | 2012-02-08 | 哈尔滨工业大学 | Application of lonicera edulis anthocyanin in anti-radiation medicament |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107647399A (en) * | 2017-06-08 | 2018-02-02 | 湖北理工学院 | A kind of fruit of glossy privet polyphenol and its production method |
| CN112493482A (en) * | 2020-09-22 | 2021-03-16 | 沈阳农业大学 | Composite nutritional powder with functions of regulating intestinal flora and barrier and preparation method thereof |
| CN113754703A (en) * | 2021-09-10 | 2021-12-07 | 黑龙江格爱自动化科技有限公司 | Method for extracting berry polyphenol, flavonoid and flavonoid compounds |
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Application publication date: 20160601 |