CN104031162A - Ophiopogonpolysaccharide, extraction and purification method of ophiopogonpolysaccharide and application of ophiopogonpolysaccharide as tobacco humectant - Google Patents
Ophiopogonpolysaccharide, extraction and purification method of ophiopogonpolysaccharide and application of ophiopogonpolysaccharide as tobacco humectant Download PDFInfo
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- ophiopogonpolysaccharide
- radix ophiopogonis
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Abstract
The invention relates to ophiopogonpolysaccharide and an extraction and purification method of ophiopogonpolysaccharide. The method specifically comprises the following steps: carrying out Soxhlet extraction on smashed dried radix ophiopogonis by using ether; volatilizing residues and extracting by using boiled water; concentrating and centrifugalizing the boiled water-extracted liquid; carrying out alcohol precipitation on supernate and standing; centrifuging again and collecting precipitates; washing, dissolving with distilled water; then adding 0.1-0.2% of papain into a water bath with a temperature of 50+/-5 DEG C and standing for 2-4 hours; removing protein by a Sevage method; decoloring by using active carbon, and concentrating under reduced pressure; cooling and carrying out dialysis by running water; after dialysis, carrying out alcohol precipitation and standing again; and drying the precipitates at 60-70 DEG C to obtain coarse polysaccharide. The test verifies that the ophiopogonpolysaccharide provided by the invention has a good physical and sensorial moistening effect on cigarettes and can reduce irritation and miscellaneous smell of the cigarettes and improve the mellow and full feel and comfortableness of cigarette smoke. The ophiopogonpolysaccharide can be used as a tobacco humectant which is applied to cigarettes.
Description
Technical field
The invention belongs to humectant technical field, be specifically related to a kind of Radix Ophiopogonis 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 Ophiopogonis polysaccharide is applied in cigarette as tobacco humectant.
Summary of the invention
The object of the invention is to provide a kind of Radix Ophiopogonis 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 Ophiopogonis polysaccharide, comprise the steps: extract after (generally adding ether by solid-liquid ratio 1g:5-8ml) by ether Soxhlet the dry tuber of dwarf lilyturf of pulverizing, 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 khaki color 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 Ophiopogonis polysaccharide of gained.
Utilize said extracted purification process to prepare gained Radix Ophiopogonis polysaccharide as the application of tobacco humectant.
The present invention has obtained highly purified refining Radix Ophiopogonis 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 Ophiopogonis 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 Ophiopogonis polysaccharide, for further separation and purification lays the foundation.2) adopt gel chromatography column chromatography for separation purifying twice, obtain highly purified refining Radix Ophiopogonis 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 Ophiopogonis 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 Radix Ophiopogonis polysaccharide, comprises the steps: the 300g pulverizing by 1800ml ether Soxhlet, to extract 2h in order to remove degrease the dry tuber of dwarf lilyturf, 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 8.1g khaki color Crude polysaccharides, yield is 2.70%.
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 Ophiopogonis polysaccharide solid of 392mg white, yield 26.1%, and LT1 sugar content is 96.6% after testing.
embodiment 2
An extracting and purifying method for Radix Ophiopogonis polysaccharide, comprises the steps: the 300g pulverizing by 2000ml ether Soxhlet, to extract 4h in order to remove degrease the dry tuber of dwarf lilyturf, 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 7.5g khaki color Crude polysaccharides, yield is 2.50%.
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 Ophiopogonis polysaccharide solid of 383mg white, yield 25.5%, and LT1 sugar content is 96.6% after testing.
application test: embodiment 1 prepares the refining Radix Ophiopogonis polysaccharide of gained as the applied research of tobacco humectant
1, physics humid keeping performance test
For investigating the physics humid keeping performance of Radix Ophiopogonis polysaccharide to cigarette, the blank cigarette shreds of Bright Yellow (great Jin circle) of take is carrier, the same glycerine of Radix Ophiopogonis 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 Radix Ophiopogonis polysaccharide, glycerine, propylene glycol and sorbyl alcohol
The test-results of table 1 shows, adding the pipe tobacco of Radix Ophiopogonis 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 Ophiopogonis 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 Ophiopogonis polysaccharide is made into massfraction.Get respectively the above-mentioned solution containing Radix Ophiopogonis 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 Radix Ophiopogonis polysaccharide smoking result of table 2 Different adding amount
Smoking result by table 2 can find out, Radix Ophiopogonis 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 Ophiopogonis polysaccharide consumption hour, it is not clearly to the effect of cigarette; When its consumption reaches 1 ‰, occur that spine sense increases; When its consumption surpasses 1 ‰, occur that spine sense increases, concentration slightly reduces; When its consumption is 0.5 ‰, sucking quality is best.Therefore, best results while adding 0.05g Radix Ophiopogonis polysaccharide in 100g cigarette, optimum consumption is 0.5 ‰.
Claims (4)
1. the extracting and purifying method of a Radix Ophiopogonis polysaccharide, it is characterized in that, after comprising the steps: extract by ether Soxhlet the dry tuber of dwarf lilyturf of pulverizing, 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 Ophiopogonis 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 Ophiopogonis polysaccharide of gained.
Described in claim 3 Radix Ophiopogonis polysaccharide as the application of tobacco humectant.
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| CN112830977A (en) * | 2021-02-26 | 2021-05-25 | 西南科技大学 | Method for extracting and purifying polysaccharide from fibrous root residues of radix ophiopogonis |
| CN115624514A (en) * | 2022-10-12 | 2023-01-20 | 杭州时光肌生物科技有限公司 | Preparation method of composite plant extract for improving facial redness |
| CN115746154A (en) * | 2022-11-22 | 2023-03-07 | 四川农业大学 | Extraction method and application of ophiopogon japonicus fibril polysaccharide with anti-infection effect |
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