CN103980381A - Angelica polysaccharide, extraction and purification method thereof and application of angelica polysaccharide as tobacco humectant - Google Patents
Angelica polysaccharide, extraction and purification method thereof and application of angelica polysaccharide as tobacco humectant Download PDFInfo
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- CN103980381A CN103980381A CN201410217669.6A CN201410217669A CN103980381A CN 103980381 A CN103980381 A CN 103980381A CN 201410217669 A CN201410217669 A CN 201410217669A CN 103980381 A CN103980381 A CN 103980381A
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- radix angelicae
- angelicae sinensis
- polysaccharide
- water
- sinensis polysaccharide
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Abstract
The invention relates to an angelica polysaccharide as well as an extraction and purification method thereof. The extraction and purification method concretely comprises the steps of carrying out Soxhlet extraction on smashed dry angelica by using diethyl ether, then, volatizing residues, and next, extracting the residues by using boiling water; centrifuging and centrifuging a boiling water extracting solution, carrying out ethanol precipitation on a supernatant, and standing; centrifuging again, collecting a sediment, and dissolving by using distilled water after washing; then, adding 0.1-0.2% of papain into a water bath with the temperature of 50+/-5 DEG C for 2-4h; removing proteins by using a Sevage method, and carrying out vacuum concentration after activated carbon decoloration; dialyzing flowing water after cooling; after dialysis is ended, carrying out ethanol precipitation again and standing, and drying the sediment at the temperature of 60-70 DEG C to obtain a crude polysaccharide. A test proves that the angelica polysaccharide provided by the invention has favorable physical and sensory humectant effects, can be used for reducing the irritation and miscellaneous odor of cigarettes and improving the mellow, full and comfort levels of smoke of the cigarettes, and can be used as a performance-excellent tobacco humectant to be applied to the cigarettes.
Description
Technical field
The invention belongs to humectant technical field, be specifically related to a kind of Radix Angelicae Sinensis polysaccharide, extracting and purifying method and as the application of tobacco humectant.
Background technology
Current, in China's cigarette industry, the main polyhydroxy substances such as glycerine, propylene glycol, sorbyl alcohol that adopt are as humectant, use the main purpose of this type of humectant to be to maintain the water ratio of pipe tobacco in the course of processing, improve the resistance to processibility of pipe tobacco, but above-mentioned humectant to finished cigarettes water ratio maintain and suck comfort level to improve effect unsatisfactory.
Polysaccharide is to be polymerized by a large amount of monose, and the hydroxyl that they are a large amount of and water molecules form hydrogen bond, can make it have stronger water-holding power.At present, at home and abroad there is no the relevant report that Radix Angelicae Sinensis polysaccharide is applied in cigarette as tobacco humectant.
Summary of the invention
The object of the invention is to provide a kind of Radix Angelicae Sinensis polysaccharide, extracting and purifying method and as the application of tobacco humectant.
For achieving the above object, the present invention adopts following technical scheme:
A kind of extracting and purifying method of Radix Angelicae Sinensis polysaccharide, comprise the steps: the dry Radix Angelicae Sinensis of pulverizing to extract after (generally adding ether by solid-liquid ratio 1g:5-8ml) by ether Soxhlet, residue volatilizes and extracts with boiling water, boiling water extraction liquid is concentrated, centrifugal, get supernatant liquor alcohol precipitation standing, recentrifuge collecting precipitation thing, after throw out washing, with distilled water, dissolve, add again the papoid of weight of precipitate 1-2 ‰ to place 2-4h in 50 ± 5 ℃ of water-baths, adopt Sevage method deproteination, concentrating under reduced pressure after activated carbon decolorizing, after cooling, carry out flowing water dialysis in order to remove small molecular weight impurity, after dialysis finishes, alcohol precipitation is standing again, be deposited in 60-70 ℃ and be dried to obtain Crude polysaccharides.
Further preferably, by Crude polysaccharides chromatographic separation purifying twice, obtain refining polysaccharide; Twice separation and purification elutriant used is respectively 0.5 mol/L NaCl solution and 0.05 mol/L NH
4hCO
3solution.
Utilize said extracted purification process to prepare the Radix Angelicae Sinensis polysaccharide of gained.
Utilize said extracted purification process to prepare gained Radix Angelicae Sinensis polysaccharide as the application of tobacco humectant.
The present invention has obtained highly purified refining Radix Angelicae Sinensis polysaccharide by extracting purifying, and carried out physics humid keeping performance and tested and inherent sensory quality assessment, find that it has good physics and sense organ humectation effect to cigarette, and can reduce pungency and the assorted gas of cigarette, improve mellow and full sense and the comfortableness of cigarette smoke, the tobacco humectant that can be used as a kind of excellent property is applied in cigarette, and has determined its Optimum in cigarette.
Compared to the prior art, beneficial effect of the present invention: 1) in the leaching process of Radix Angelicae Sinensis polysaccharide, utilize alcohol precipitation, protease hydrolysis, Sevage except albumen and dialysis remove the method for small molecular phase combination, effectively reduced the impurity in Radix Angelicae Sinensis polysaccharide, for further separation and purification lays the foundation.2) adopt gel chromatography column chromatography for separation purifying twice, obtain highly purified refining Radix Angelicae Sinensis polysaccharide.Whole extraction purge process is easy and simple to handle, and raw material and the reagent of use are cheap and easy to get.3), through evidence, the humectation successful of Radix Angelicae Sinensis polysaccharide of the present invention to tobacco product, therefore has significant industrial application value and wide popularizing application prospect.
Accompanying drawing explanation
Fig. 1 is the physics humectation effect contrast figure of Radix Angelicae Sinensis polysaccharide of the present invention, glycerine, propylene glycol and sorbyl alcohol.
Embodiment
The present invention is described further by the following examples, but protection scope of the present invention is not limited to this.
test main agents and instrument used:
Ether, ethanol, acetone (Chinese Suzhou chemical reagent company limited, analytical pure), DEAE-Celluose and Sephadex G200 gel (U.S. sigma company), experimental water is distilled water.
BS200SWE1 type electronic balance (sensibility reciprocal: 0.0001g, Beijing match Doris balance company limited); R-215 type Rotary Evaporators (Switzerland BUCHI company); V-700 vacuum pump (Switzerland BUCHI company); DLSB-20/60 ℃ of subcooling recycle pump (Shanghai Mei Qiang plant and instrument company limited); KDM type regulating temp. electrothermal cover (Shandong Hua Lu electric heating Instrument Ltd.); 85-2 type constant temperature blender with magnetic force (Shanghai Si Le instrument plant), ZK-82B type electric vacunm drying case (Shanghai Shen Guang instrument company limited), CHRIST Freeze Drying Equipment (Switzerland BUCHI company), TSKGEL G2000 PW efficient gel chromatographic column (U.S. Sigma company), Agilent LC1100 type high performance liquid phase instrument and differential refraction detector (U.S. Agilent company).
embodiment 1
An extracting and purifying method for Radix Angelicae Sinensis polysaccharide, comprises the steps: the dry Radix Angelicae Sinensis of the 300g pulverizing to extract 2h in order to remove degrease by 1800ml ether Soxhlet, and residue volatilizes.Residue after volatilizing is used 750ml boiling water extraction 3 times again, merges the boiling water extraction liquid of 3 gained, concentrated, centrifugal, gets supernatant liquor, with the dehydrated alcohol of 3 times of volumes of supernatant liquor, carries out alcohol precipitation, standing over night (12h), again.Throw out dissolves with distilled water with after ethanol, washing with acetone 3 times successively, then adds the papoid of weight of precipitate 1 ‰ to place 2h in 50 ℃ of water-baths, adopts Sevage method deproteination.Add 8g gac to be placed in 70 ℃ of water-baths and heat 3h, remove by filter gac, concentrating under reduced pressure, after naturally cooling, pack flowing water in dialysis tubing into and dialyse 48h in order to remove small molecular weight impurity, after dialysis finishes, again add the dehydrated alcohol of the rear 3 times of amounts of polysaccharide soln volume of dialysis to carry out alcohol precipitation, standing over night (12h), be deposited in 70 ℃ and be dried to obtain 5.9g Crude polysaccharides, yield is 1.97%.
Get 1.5g Crude polysaccharides with after distilled water dissolving, by DEAE-Celluose gel chromatography column chromatography for separation (5.0 * 50cm), take 0.5mol/L NaCl solution as moving phase (being elutriant), flow velocity is 2.5ml/min, adopt phenol sulfuric acid method to develop the color to chromatographic solution, collect main chromatographic peak, freeze-drying.Then by freeze-drying gained solid 0.05mol/L NH
4hCO
3solution dissolves, and uses Sephadex G200 gel chromatographic columns chromatographic separation (5.0 * 50cm) again, with 0.05mol/L NH
4hCO
3solution is moving phase (being elutriant), and flow velocity is 2.0ml/min, adopts phenol sulfuric acid method to develop the color to chromatographic solution, collects main chromatographic peak, and freeze-drying obtains the refining Radix Angelicae Sinensis polysaccharide solid of 356mg, yield 23.7%, and LT1 sugar content is 94% after testing.
embodiment 2
An extracting and purifying method for Radix Angelicae Sinensis polysaccharide, comprises the steps: the dry Radix Angelicae Sinensis of the 300g pulverizing to extract 4h in order to remove degrease by 2000ml ether Soxhlet, and residue volatilizes.Residue after volatilizing is used 750ml boiling water extraction 3 times again, merges the boiling water extraction liquid of 3 gained, concentrated, centrifugal, gets supernatant liquor, with the dehydrated alcohol of 3 times of volumes of supernatant liquor, carries out alcohol precipitation, standing 12h, recentrifuge collecting precipitation thing.Throw out dissolves with distilled water with after ethanol, washing with acetone 3 times successively, then adds the papoid of weight of precipitate 2 ‰ to place 4h in 50 ℃ of water-baths, adopts Sevage method deproteination.Add 8g gac to be placed in 70 ℃ of water-baths and heat 4h, remove by filter gac, concentrating under reduced pressure, after naturally cooling, pack flowing water in dialysis tubing into and dialyse 48h in order to remove small molecular weight impurity, after dialysis finishes, again add the dehydrated alcohol of the rear 3 times of amounts of polysaccharide soln volume of dialysis to carry out alcohol precipitation, standing over night, be deposited in 70 ℃ and be dried to obtain 6.2g Crude polysaccharides, yield is 2.46%.
Get 1.5g Crude polysaccharides with after distilled water dissolving, by DEAE-Celluose gel chromatography column chromatography for separation (5.0 * 50cm), take 0.5mol/L NaCl solution as moving phase (being elutriant), flow velocity is 2.5ml/min, adopt phenol sulfuric acid method to develop the color to chromatographic solution, collect main chromatographic peak, freeze-drying.Then by freeze-drying gained solid 0.05mol/L NH
4hCO
3solution dissolves, and uses Sephadex G200 gel chromatographic columns chromatographic separation (5.0 * 50cm) again, with 0.05mol/L NH
4hCO
3solution is moving phase (being elutriant), and flow velocity is 2.0ml/min, adopts phenol sulfuric acid method to develop the color to chromatographic solution, collects main chromatographic peak, and freeze-drying obtains the refining Radix Angelicae Sinensis polysaccharide solid of 370mg, yield 24.7%, and LT1 sugar content is 95% after testing.
application test: Radix Angelicae Sinensis polysaccharide is as the applied research of tobacco humectant
1, physics humid keeping performance test
For investigating the physics humid keeping performance of Radix Angelicae Sinensis polysaccharide to cigarette, the blank cigarette shreds of Bright Yellow (great Jin circle) of take is carrier, by embodiment 1 prepare the refining same glycerine of Radix Angelicae Sinensis polysaccharide of gained, propylene glycol carries out respectively the research of physics humid keeping performance contrast test together with sorbyl alcohol, prepares respectively massfraction and be 5% the above-claimed cpd aqueous solution.The blank cigarette shreds first 1000g being mixed is placed in 22 ± 1 ℃ of temperature, relative humidity 60 ± 2% environment balance 48h.Then equivalent takes five parts of pipe tobaccos, and every part of 100g evenly applies the 4g above-claimed cpd aqueous solution respectively in pipe tobacco, and contrasting blank sample is that pipe tobacco evenly applies 4g water.Then five parts of pipe tobaccos are placed in respectively and under 22 ± 1 ℃, the environment of relative humidity 40 ± 2%, investigate it and separate wet process, and the variation of instant water ratio is analyzed, investigate its physics humectation effect, the results are shown in Figure 1.
Test-results shows, adding the pipe tobacco of Radix Angelicae Sinensis polysaccharide, glycerine, four kinds of compounds of propylene glycol and sorbyl alcohol compares with blank pipe tobacco, moisture loss speed is slower, this is because the contained hydrophilic radical hydroxyl of polyol can form hydrogen bond with water and water be fettered, delay scattering and disappearing of moisture, improved the physics humid keeping performance of pipe tobacco; Wherein the physics humid keeping performance of Radix Angelicae Sinensis polysaccharide is best, is secondly propylene glycol and glycerine, is finally sorbyl alcohol.
2, sense organ humid keeping performance test
With 20v% aqueous ethanolic solution, make solvent, it is 1% solution that Radix Angelicae Sinensis polysaccharide is made into massfraction.Get respectively the above-mentioned solution containing Radix Angelicae Sinensis polysaccharide of 0.100g, 0.500g, 1.000g, 5.000g, 10.000g and 20.000g, then evenly spray is added in five parts of blank cigarette shreds of not spiced 100g Bright Yellow respectively, roll, respectively pick out 100 same weight cigarette, the climatic chamber balance 48h that is placed in 22 ℃ ± 1 ℃ of temperature, humidity 60% ± 2%, smokes panel test.Control sample is the blank cigarette of Bright Yellow, and control sample is balance 48h under uniform temp, humidity condition equally.Smoking result is in Table 1.
The Radix Angelicae Sinensis polysaccharide smoking result of table 1 Different adding amount
Smoking result by table 1 can find out, Radix Angelicae Sinensis polysaccharide shows as and makes flue gas obviously become mellow and full, soft the Main Function of cigarette, and comfortable taste promotes, and assorted gas and pungency reduce.When Radix Angelicae Sinensis polysaccharide consumption hour, it is not clearly to the effect of cigarette; When its consumption surpasses 1 ‰, there is the phenomenon that spine sense increase, concentration reduce; When its consumption is 1 ‰, sucking quality is best.Therefore, best results while adding 0.1g Radix Angelicae Sinensis polysaccharide in 100g cigarette, optimum consumption is 1 ‰.
Claims (4)
1. the extracting and purifying method of a Radix Angelicae Sinensis polysaccharide, it is characterized in that, after comprising the steps: the dry Radix Angelicae Sinensis of pulverizing to extract by ether Soxhlet, residue volatilizes and extracts with boiling water, boiling water extraction liquid is concentrated, centrifugal, get supernatant liquor alcohol precipitation standing, recentrifuge collecting precipitation thing, after throw out washing, with distilled water, dissolve, add again the papoid of 1-2 ‰ to place 2-4h in 50 ± 5 ℃ of water-baths, adopt Sevage method deproteination, concentrating under reduced pressure after activated carbon decolorizing, after cooling, carry out flowing water dialysis, after dialysis finishes, alcohol precipitation is standing again, be deposited in 60-70 ℃ and be dried to obtain Crude polysaccharides.
2. the extracting and purifying method of Radix Angelicae Sinensis polysaccharide as claimed in claim 1, is characterized in that, by Crude polysaccharides chromatographic separation purifying twice, obtains refining polysaccharide; Twice separation and purification elutriant used is respectively 0.5 mol/L NaCl solution and 0.05 mol/L NH
4hCO
3solution.
3. utilize extracting and purifying method described in claim 1 or 2 to prepare the Radix Angelicae Sinensis polysaccharide of gained.
Described in claim 3 Radix Angelicae Sinensis polysaccharide as the application of tobacco humectant.
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Cited By (5)
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| CN104522683A (en) * | 2014-12-19 | 2015-04-22 | 无限极（中国）有限公司 | Application of Chinese angelica polypeptide with effects of resisting oxidization and delaying ageing in preparation of food |
| CN105985451A (en) * | 2015-02-15 | 2016-10-05 | 中国人民解放军第四军医大学 | Chinese angelica acidic polysaccharide component, and preparation method and application thereof |
| CN107723092A (en) * | 2017-11-29 | 2018-02-23 | 湖北中烟工业有限责任公司 | A kind of preparation method and applications with fragrant and sweet fermentation angelica extract |
| CN107714720A (en) * | 2017-10-30 | 2018-02-23 | 兰州大学 | A kind of preparation method for the Radix Angelicae Sinensis astragalus polyose iron complexes for treating hypoferric anemia |
| CN108771005A (en) * | 2018-06-19 | 2018-11-09 | 佛山推启农业研究院（普通合伙） | A kind of lactagogue forage and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104522683A (en) * | 2014-12-19 | 2015-04-22 | 无限极（中国）有限公司 | Application of Chinese angelica polypeptide with effects of resisting oxidization and delaying ageing in preparation of food |
| CN105985451A (en) * | 2015-02-15 | 2016-10-05 | 中国人民解放军第四军医大学 | Chinese angelica acidic polysaccharide component, and preparation method and application thereof |
| CN105985451B (en) * | 2015-02-15 | 2018-04-03 | 中国人民解放军第四军医大学 | A kind of Radix Angelicae Sinensis acidity polysaccharide component and its extracting method and application |
| CN107714720A (en) * | 2017-10-30 | 2018-02-23 | 兰州大学 | A kind of preparation method for the Radix Angelicae Sinensis astragalus polyose iron complexes for treating hypoferric anemia |
| CN107723092A (en) * | 2017-11-29 | 2018-02-23 | 湖北中烟工业有限责任公司 | A kind of preparation method and applications with fragrant and sweet fermentation angelica extract |
| CN108771005A (en) * | 2018-06-19 | 2018-11-09 | 佛山推启农业研究院（普通合伙） | A kind of lactagogue forage and preparation method thereof |
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Address after: 450000 Yulin South Road, Henan, Zheng Dong, No. 16 South Road, Zhengzhou Applicant after: China Tobacco Henan Industrial Co., Ltd. Address before: 450000 Zhengzhou agricultural road, Henan, No. 29 Applicant before: China Tobacco Henan Industrial Co., Ltd. |
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