CN104031166A - Tremella polysaccharide, extraction and purification method and application of tremella polysaccharide as tobacco humectant - Google Patents
Tremella polysaccharide, extraction and purification method and application of tremella polysaccharide as tobacco humectant Download PDFInfo
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- CN104031166A CN104031166A CN201410294942.5A CN201410294942A CN104031166A CN 104031166 A CN104031166 A CN 104031166A CN 201410294942 A CN201410294942 A CN 201410294942A CN 104031166 A CN104031166 A CN 104031166A
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Abstract
The invention relates to tremella polysaccharide and an extraction and purification method thereof. The method comprises the following concrete steps: treating crushed dry tremella through a Soxhlet extraction process by using diethyl ether; drying residues and then extracting by using boiling water; concentrating and centrifuging the boiling water extraction solution, precipitating the liquid supernatant by using alcohol and standing, centrifuging again and collecting precipitates, washing the precipitates, dissolving the precipitates by using distilled water, then adding 1 per mill-2per mill of papain into a water bath of which the temperature is 50+/-5 DEG C, placing for 2-4 hours, removing protein by adopting a Sevage method, decolorizing by using active carbon, decompressing, concentrating, cooling, dialyzing by using flowing water, precipitating by using the alcohol and standing again after the dialysis ends, and drying the precipitates at the temperature of 60-70 DEG C to obtain crude polysaccharide. Tests prove that the tremella polysaccharide disclosed by the invention has a quite good physical and sensorial wetting effect on cigarettes and also can be used for reducing the irritation and miscellaneous gases of the cigarettes and improving the smoothness and comfort of the smoke of the cigarettes, so the tremella polysaccharide can be applied to the cigarettes as a tobacco humectant with excellent performances.
Description
Technical field
The invention belongs to humectant technical field, be specifically related to a kind of tremella 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 tremella polysaccharide is applied in cigarette as tobacco humectant.
Summary of the invention
The object of the invention is to provide a kind of tremella 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 tremella polysaccharide, comprise the steps: the dry white fungus 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 light yellow 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 tremella polysaccharide of gained.
Utilize said extracted purification process to prepare gained tremella polysaccharide as the application of tobacco humectant.
White fungus is called again Tremella, tremella, white fungus etc., belongs to Mycophyta Tremellaceae Tremella, is a sporophore for Basidiomycota fungi white fungus, has the laudatory title of " hat in bacterium ".Tremella fructification is pure white to oyster white, and 5～10 centimetres of diameters are soft pure white, translucent, high resilience.The present invention has obtained highly purified fine silver fungus polysaccharides 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 tremella polysaccharide, utilize alcohol precipitation, protease hydrolysis, Sevage except albumen and dialysis remove the method for small molecular phase combination, effectively reduced the impurity in tremella polysaccharide, for further separation and purification lays the foundation.2) adopt gel chromatography column chromatography for separation purifying twice, obtain highly purified fine silver fungus polysaccharides.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 tremella polysaccharide of the present invention to tobacco product, therefore has significant industrial application value and wide popularizing application prospect.
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 tremella polysaccharide, comprises the steps: the dry white fungus 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), 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 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 6.7g Crude polysaccharides, yield is 2.23%.
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 352mg fine silver fungus polysaccharides solid, yield 23.5%, and LT1 sugar content is 94% after testing.
embodiment 2
An extracting and purifying method for tremella polysaccharide, comprises the steps: the dry white fungus 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 6.8g Crude polysaccharides, yield is 2.26%.
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 366mg fine silver fungus polysaccharides solid, yield 24.4%, and LT1 sugar content is 95% after testing.
application test: embodiment 1 prepares gained fine silver fungus polysaccharides as the applied research of tobacco humectant
1, physics humid keeping performance test
For investigating the physics humid keeping performance of tremella polysaccharide to cigarette, the blank cigarette shreds of Bright Yellow (great Jin circle) of take is carrier, the same glycerine of tremella polysaccharide, propylene glycol are carried out respectively together with sorbyl alcohol to physics humid keeping performance contrast test research, prepare 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 (adding the moisture content of drying base in pipe tobacco 72h after different samples changes), the results are shown in Table 1.
The physics humectation effect comparison of table 1, tremella polysaccharide, glycerine, propylene glycol and sorbyl alcohol
The test-results of table 1 shows, adding the pipe tobacco of tremella 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 tremella 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 tremella polysaccharide is made into massfraction.Get respectively the above-mentioned solution containing tremella 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 2.
The tremella polysaccharide smoking result of table 2 Different adding amount
Smoking result by table 2 can find out, tremella polysaccharide shows as and makes flue gas obviously become mellow and full, soft the Main Function of cigarette, and comfortable taste promotes, and sugariness increases, and assorted gas and pungency reduce.When tremella 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 tremella polysaccharide in 100g cigarette, optimum consumption is 1 ‰.
Claims (4)
1. the extracting and purifying method of a tremella polysaccharide, it is characterized in that, after comprising the steps: the dry white fungus 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 tremella 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 tremella polysaccharide of gained.
Described in claim 3 tremella polysaccharide as the application of tobacco humectant.
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| CN104434949A (en) * | 2014-11-25 | 2015-03-25 | 郭霞 | Medicine for conditioning internal thermal and Yang hyperactivity, hyperactivity of heart-liver fire and swollen sore throat |
| CN104497157A (en) * | 2014-12-22 | 2015-04-08 | 绿雅（江苏）食用菌有限公司 | Method for separating and purifying tremella intercellular polysaccharide |
| CN104585866A (en) * | 2015-01-20 | 2015-05-06 | 川渝中烟工业有限责任公司 | Ultrasonic atomized electronic cigarette liquid rich in biological active polysaccharide and preparation method thereof |
| CN105481993A (en) * | 2016-02-03 | 2016-04-13 | 赵志鑫 | Tremella polysaccharide extracting method |
| CN109957040A (en) * | 2019-04-08 | 2019-07-02 | 上海应用技术大学 | A kind of method that tremella polysaccharides are extracted in the mashing of enzyme process Aided Wet |
| CN111454375A (en) * | 2020-05-12 | 2020-07-28 | 西安绿天生物技术有限公司 | Extraction method of tremella polysaccharide |
| CN113383987A (en) * | 2021-07-26 | 2021-09-14 | 云南中烟工业有限责任公司 | Essence formula containing tremella polysaccharide extract and application of essence formula in tobacco flavoring |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104211829A (en) * | 2014-09-17 | 2014-12-17 | 上海交通大学 | Preparation and application of white fungus functional polysaccharide |
| CN104434949A (en) * | 2014-11-25 | 2015-03-25 | 郭霞 | Medicine for conditioning internal thermal and Yang hyperactivity, hyperactivity of heart-liver fire and swollen sore throat |
| CN104497157A (en) * | 2014-12-22 | 2015-04-08 | 绿雅（江苏）食用菌有限公司 | Method for separating and purifying tremella intercellular polysaccharide |
| CN104585866A (en) * | 2015-01-20 | 2015-05-06 | 川渝中烟工业有限责任公司 | Ultrasonic atomized electronic cigarette liquid rich in biological active polysaccharide and preparation method thereof |
| CN104585866B (en) * | 2015-01-20 | 2016-03-30 | 川渝中烟工业有限责任公司 | Ultrasonic atomizatio electronic cigarette liquid being rich in bioactive polysaccharide and preparation method thereof |
| CN105481993A (en) * | 2016-02-03 | 2016-04-13 | 赵志鑫 | Tremella polysaccharide extracting method |
| CN109957040A (en) * | 2019-04-08 | 2019-07-02 | 上海应用技术大学 | A kind of method that tremella polysaccharides are extracted in the mashing of enzyme process Aided Wet |
| CN111454375A (en) * | 2020-05-12 | 2020-07-28 | 西安绿天生物技术有限公司 | Extraction method of tremella polysaccharide |
| CN113383987A (en) * | 2021-07-26 | 2021-09-14 | 云南中烟工业有限责任公司 | Essence formula containing tremella polysaccharide extract and application of essence formula in tobacco flavoring |
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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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Application publication date: 20140910 |