CN103980377A - Lycium barbarum polysaccharide, extracting and purifying method and application thereof as tobacco humectant - Google Patents
Lycium barbarum polysaccharide, extracting and purifying method and application thereof as tobacco humectant Download PDFInfo
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- CN103980377A CN103980377A CN201410213659.5A CN201410213659A CN103980377A CN 103980377 A CN103980377 A CN 103980377A CN 201410213659 A CN201410213659 A CN 201410213659A CN 103980377 A CN103980377 A CN 103980377A
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- barbarum polysaccharide
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Abstract
The invention relates to a lycium barbarum polysaccharide, and an extracting and purifying method thereof. The method concretely comprises the following steps: carrying out soxhlet extraction on crushed dry medlar by using a diethyl ether; drying residues and then extracting by using boiled water; concentrating and centrifuging boiling water extract solution; taking supernate to carry out alcohol precipitation and standing; centrifuging and collecting sediments; dissolving by using distilled water after washing, then adding 0.1-0.2% of nematolyt to place in a water bath at 50+/-5 DEG C for 2-4 hours; removing protein by adopting a Sevage method, carrying out vacuum concentration after decoloring activated carbon, carrying out water dialysis after cooling; carrying out alcohol precipitation and standing again after dialysis is ended, and drying the sediments at 60-70 DEG C to obtain crude polysaccharide. The test proves that the lycium barbarum polysaccharide has good physical and sensory embellishing effects on a cigarette, the thrill and offensive odor of the cigarette can be reduced, the roundness and the comfort of cigarette smoke are improved, and the lycium barbarum polysaccharide can be applied to the cigarette as a tobacco humectant with excellent performance.
Description
Technical field
The invention belongs to humectant technical field, be specifically related to a kind of lycium barbarum polysaccharide, extracting and purifying method and the application as tobacco humectant thereof.
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 lycium barbarum polysaccharide is applied in cigarette as tobacco humectant.
Summary of the invention
The object of the invention is to provide a kind of lycium barbarum polysaccharide, extracting and purifying method and the application as tobacco humectant thereof.
For achieving the above object, the present invention adopts following technical scheme:
A kind of extracting and purifying method of lycium barbarum polysaccharide, comprise the steps: the dried wolfberry of pulverizing ether Soxhlet to extract after (generally adding ether by solid-liquid ratio 1g:5-8ml), residue volatilizes and extracts with boiling water, boiling water extraction liquid is concentrated, centrifugal, getting supernatant liquor alcohol precipitation leaves standstill, recentrifuge collecting precipitation thing, after throw out washing, dissolve with distilled water, add again the papoid of weight of precipitate 1-2 ‰ to place 2-4h in 50 ± 5 DEG C 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 leaves standstill again, be deposited in 60-70 DEG C 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 lycium barbarum polysaccharide of gained.
Utilize said extracted purification process to prepare the application of gained lycium barbarum polysaccharide as tobacco humectant.
The present invention has obtained highly purified refining lycium barbarum 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 lycium barbarum polysaccharide, utilize alcohol precipitation, protease hydrolysis, Sevage except albumen and dialysis remove the method for small molecular phase combination, effectively reduced the impurity in lycium barbarum polysaccharide, for further separation and purification lays the foundation.2) adopt gel chromatography column chromatography for separation purifying twice, obtain highly purified refining lycium barbarum 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 lycium barbarum polysaccharide of the present invention to tobacco product, therefore has significant industrial application value and wide popularizing application prospect.
Brief description of the drawings
Fig. 1 is the physics humectation effect contrast figure of lycium barbarum 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 (sigma company of the U.S.), experimental water is distilled water.
BS200SWE1 type electronic balance (sensibility reciprocal: 0.0001g, Beijing match Doris balance company limited); R-215 type Rotary Evaporators (BUCHI company of Switzerland); V-700 vacuum pump (BUCHI company of Switzerland); DLSB-20/60 DEG C 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 (BUCHI company of Switzerland), TSKGEL G2000 PW efficient gel chromatographic column (Sigma company of the U.S.), Agilent LC1100 type high performance liquid phase instrument and differential refraction detector (Agilent company of the U.S.).
embodiment 1
An extracting and purifying method for lycium barbarum polysaccharide, comprises the steps: the 300g dried wolfberry of pulverizing 1800ml ether Soxhlet to extract 2h in order to remove degrease, 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, carries out alcohol precipitation with the dehydrated alcohol of 3 times of volumes of supernatant liquor, hold over night (12h), again.Throw out is successively with dissolving with distilled water after ethanol, washing with acetone 3 times, then adds the papoid of weight of precipitate 1 ‰ to place 2h in 50 DEG C of water-baths, adopts Sevage method deproteination.Add 8g gac to be placed in 70 DEG C 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, hold over night (12h), be deposited in 70 DEG C and be dried to obtain 6.7g Crude polysaccharides, yield is 2.2%.
Get after the dissolving of 1.5g Crude polysaccharides distilled water, by DEAE-Celluose gel chromatography column chromatography for separation (5.0 × 50cm), taking 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, by 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 lycium barbarum polysaccharide solid of 356mg, yield 23.7%, and LT1 sugar content is 94% after testing.
embodiment 2
An extracting and purifying method for lycium barbarum polysaccharide, comprises the steps: the 300g dried wolfberry of pulverizing 2000ml ether Soxhlet to extract 4h in order to remove degrease, 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, carries out alcohol precipitation with the dehydrated alcohol of 3 times of volumes of supernatant liquor, leaves standstill 12h, recentrifuge collecting precipitation thing.Throw out is successively with dissolving with distilled water after ethanol, washing with acetone 3 times, then adds the papoid of weight of precipitate 2 ‰ to place 4h in 50 DEG C of water-baths, adopts Sevage method deproteination.Add 8g gac to be placed in 70 DEG C 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, hold over night, be deposited in 70 DEG C and be dried to obtain 6.4g Crude polysaccharides, yield is 2.1%.
Get after the dissolving of 1.5g Crude polysaccharides distilled water, by DEAE-Celluose gel chromatography column chromatography for separation (5.0 × 50cm), taking 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, by 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 lycium barbarum polysaccharide solid of 360mg, yield 24%, and LT1 sugar content is 95% after testing.
application test: lycium barbarum polysaccharide is as the applied research of tobacco humectant
1, physics humid keeping performance test
For investigating the physics humid keeping performance of lycium barbarum polysaccharide to cigarette, taking the blank cigarette shreds of Bright Yellow (great Jin circle) as carrier, by embodiment 1 prepare the refining same glycerine of lycium barbarum 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 DEG C 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 DEG C, 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, add the pipe tobacco of lycium barbarum polysaccharide, glycerine, four kinds of compounds of propylene glycol and sorbyl alcohol compared 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 lycium barbarum polysaccharide is best, is secondly propylene glycol and glycerine, is finally sorbyl alcohol.
2, sense organ humid keeping performance test
Make solvent with 20v% aqueous ethanolic solution, it is 1% solution that lycium barbarum polysaccharide is made into massfraction.Get respectively the above-mentioned solution containing lycium barbarum 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 DEG C ± 1 DEG C 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 lycium barbarum polysaccharide smoking result of table 1 Different adding amount
Smoking result by table 1 can find out, lycium barbarum 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 lycium barbarum polysaccharide consumption hour, it is not clearly to the effect of cigarette; In the time that its consumption exceedes 1 ‰, there is the phenomenon that spine sense increase, concentration reduce; In the time that its consumption is 1 ‰, sucking quality is best.Therefore, best results add 0.1g lycium barbarum polysaccharide in 100g cigarette time, optimum consumption is 1 ‰.
Claims (4)
1. the extracting and purifying method of a lycium barbarum polysaccharide, it is characterized in that, after comprising the steps: the dried wolfberry of pulverizing ether Soxhlet to extract, residue volatilizes and extracts with boiling water, boiling water extraction liquid is concentrated, centrifugal, getting supernatant liquor alcohol precipitation leaves standstill, recentrifuge collecting precipitation thing, after throw out washing, dissolve with distilled water, add again the papoid of 1-2 ‰ to place 2-4h in 50 ± 5 DEG C 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 leaves standstill again, be deposited in 60-70 DEG C and be dried to obtain Crude polysaccharides.
2. the extracting and purifying method of lycium barbarum 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 lycium barbarum polysaccharide of gained.
Described in claim 3 lycium barbarum polysaccharide as the application of tobacco humectant.
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| CN109942723A (en) * | 2019-03-15 | 2019-06-28 | 苏州大学 | A kind of separation method of polysaccharides |
| CN111944072A (en) * | 2020-08-20 | 2020-11-17 | 宋现伟 | Wolfberry polysaccharide extraction method, wolfberry polysaccharide tablet and preparation method thereof |
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Cited By (3)
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|---|---|---|---|---|
| CN105861152A (en) * | 2016-05-27 | 2016-08-17 | 山东中烟工业有限责任公司 | Tobacco flavor with sweet smoke and application |
| CN109942723A (en) * | 2019-03-15 | 2019-06-28 | 苏州大学 | A kind of separation method of polysaccharides |
| CN111944072A (en) * | 2020-08-20 | 2020-11-17 | 宋现伟 | Wolfberry polysaccharide extraction method, wolfberry polysaccharide tablet 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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Application publication date: 20140813 |