Method for extracting and purifying frangipani polysaccharide and application of frangipani polysaccharide as tobacco humectant

Technical Field

The invention belongs to the technical field of moisture, in particular relates to a polysaccharide of eggs, extracting and purifying method and its as the application of the tobacco moisture.

Background Art

The current, is mainly adopted in my cigarette industry glycerin, propylene glycol, sorbitol, multi-hydroxy material as the humectant, the use of such humectant the main purpose is to maintain the moisture content of the cut tobacco in the course of processing, improve processability of tobacco shreds, but the above-mentioned moisture keeping the moisture content of the finished cigarette smoking and to improve the comfort of the effect is not ideal.

Polysaccharide made up of a large number of monosaccharide polymerization and into, its a large amount of hydroxyl group with water molecules to form a hydrogen bond, can make its capacity with relatively strong water. At present, has not been yet both at home and abroad as a glycan of flower of the eggs in the cigarette tobacco moisture related reports of application.

Content of the invention

The purpose of this invention is to provide a polysaccharide of eggs, extracting and purifying method and its as the application of the tobacco moisture.

To achieve the above object, the invention adopts the following technical scheme:

Polysaccharide to an egg extracting and purifying method, comprises the following steps: the crushed for drying eggs of ethyl ether Soxhlet extraction (general according to the feed liquid than 1g:5-8 ml adding ethyl ether) the rear, then the dry residue extracted with boiling water, boiling water extracting solution concentration, centrifugal, is the quiet the serum is mellow , centrifugal collection depot once again, after washing the precipitate dissolved with distilled water, then adding the precipitate weight 1-2 the, the papain [...] for placing in the water bath 50 ±5 °C 2-4h, Sevage process to get rid of the protein, active carbon decolourizations after concentrating under reduced pressure, after cooling flow dialysis in order to remove small molecule impurities, is quiet once more mellow after the end of the dialysis, precipitation in the 60-70 the drying to obtain pale yellow crude polysaccharide [...].

Further preferably, the crude polysaccharide chromatography separation and purification, two, to obtain a refined polysaccharide; two separating and purifying the eluant are respectively 0.5 mol/L NaCl solution and 0.05 mol/L NH₄ HCO₃ solution.

Utilizing the above-mentioned extraction and purification method for preparing the polysaccharide of eggs.

Utilizing the above-mentioned extraction and purification spent egg prepared by the method of application of the polysaccharide as tobacco moisture.

Frangipani (Plumeria rubra l. Cv. Acutifolia), alias afzelia gardenias, yolk flower, India jasmine , large plant, family, eggs are deciduous shrub or small trees. Branchlet hypertophy multi-meat. Leaf large, thick paper, poly-born in branch top, veins in recent leaf margin place are connected while neuroptera. The top branches gatherscolored number , corolla tube-shape, diameter about 5-6cm, 5 lobed. Galochrous outside the, center fresh yellow, very aromatic. Florescence 5-10 month. Tamago flower summer flowering, fragrant concinnities. After the leaves, the trunk of the grill in the bending natural, US very its shape. Suitable for garden, meadowlands in planting, can also be potted, can be used as medicine. American origin, China has introduction and cultivation. This invention, through extraction and purification to obtain high-purity refining tamago flower polysaccharide, and the physical assuaring and moistening performance test and intrinsic sensory quality evaluation, found that the cigarette has very good physical and organoleptic assuaring and moistening function, and can reduce acrimony and impure of the cigarette, the cigarette smoke round and improving comfort, can be used as a good performance of the application in the cigarette tobacco moisture, and determines the appropriate dosage in the cigarette.

And prior art, the beneficial effect of the invention: 1) in the egg in the extraction process of the glycan of flower, using alcohol precipitation, protease hydrolysis, Sevage in addition to protein and dialysis in addition to the method of combining the small molecule, effectively reduces the tamago flower impurity in the polysaccharide, the basis for further separation and purification. 2) the gel chromatographic column chromatography separation and purification, two, to obtain high-purity refining tamago flower polysaccharide. The entire extraction and purification process is easy and simple to handle, and the raw materials used are cheap and easily obtained. 3) proved by tests, eggs of the present invention the polysaccharide is moisten effects of tobacco products, therefore it has remarkable industrial application value and broad popularization and application prospect.

Mode of execution

The following by embodiments of the present invention is used as the further description, the scope of protection of the present invention, however, is not limited to this.

Test the main reagent and instrument:

Ethyl ether, ethanol, acetone (China Suzhou chemical reagent Company limited, analytical pure), and DEAE-Celluose Sephadex G200 gel (sigma Company of the United States), experimental water is distilled water.

BS200SWE1 type electronic balance (identical: 0.0001g, Beijing [...] balance Company limited); R-215-shaped rotary evaporimeter (BUCHI Company of Switzerland); V-700 vacuum pump (BUCHI Company of Switzerland); DLSB-20/60 °C low-temperature cooling circulating pump (Shanghai-US strong instrument and equipment Company limited); KDM-type temperature regulating electric heating sleeve (Shandong hua Lu electric instrument Company limited); 85-2-type constant-temperature magnetic stirrer (Shanghai si Leinstrument plant), ZK-82B-type electric vacuum drying oven (Shanghai shen Guang instrument instrument Company limited), freeze dryer CHRIST (BUCHI Company of Switzerland), TSKGEL G2000 PW efficient gel chromatographic column (the United States Company Sigma), Agilent LC1100-type efficient liquid phase meter and the differential refraction detector (Agilent Company of the United States).

Embodiment 1

Polysaccharide to an egg extracting and purifying method, comprises the following steps: of the comminuted 300g for drying eggs 1800 ml ethyl ether Soxhlet extraction 2h used for removing the oil, the dry residue. For the dry residue after 750 ml boiling water extracting 3 times, the merger 3 extract as the boiling water, concentration, centrifugation, the supernatant, the supernatant fluid 3 times with ethanol anhydrous ethanol volume, static sleepovers (12h), sediment centrifugally collected again. Precipitate with ethanol, acetone washing 3 times with distilled water, then adding the precipitate weight of the 1, the papain [...] for the 50 is placed in the water bath [...] 2h, Sevage process to get rid of the protein. Adding 8g activated carbon in the 70 the heating in water bath [...] 3h, is filtered to remove the activated carbon, concentrated under reduced pressure, a natural cooling after dialysis in dialysis bag 48h in order to remove small molecule impurities for, after the end of the dialysis, dialysis to join again the volume the post-sugar solution 3 times with ethanol anhydrous ethanol, standing sleepovers (12h), precipitate in the 70 the drying to obtain [...] 6.7g crude polysaccharide, the yield is 2.2%.

Taking 1.5g crude polysaccharide dissolved with distilled water, using DEAE-Celluose gel chromatographic column chromatography separation (5.0 × 50 cm), to 0.5mol/L NaCl solution as mobile phase (in other words eluant), flow rate is 2.5 ml/min, the phenol-sulfuric acid method for color rendering chromatographic analysis fluid , collecting main chromatographic peaks, freeze-dried. Then the resulting solid for the lyophilized 0.05mol/L NH₄ HCO₃ solution dissolving, using Sephadex G200 gel chromatographic column again chromatography separation (5.0 × 50 cm), to 0.05mol/L NH₄ HCO₃ solution as mobile phase (in other words eluant), flow rate is 2.0 ml/min, the phenol-sulfuric acid method for color rendering chromatographic analysis fluid , collecting main chromatographic peaks, freeze-dried, to obtain 356 mg refining tamago flower polysaccharide solid, yield 23.7%, through measuring LT1 the sugar content is 94%.

Embodiment 2

Polysaccharide to an egg extracting and purifying method, comprises the following steps: of the comminuted 300g for drying eggs 2000 ml ethyl ether Soxhlet extraction 4h is used to remove the grease, the dry residue. For the dry residue after 750 ml boiling water extracting 3 times, the merger 3 extract as the boiling water, concentration, centrifugation, the supernatant, the supernatant fluid 3 times with ethanol anhydrous ethanol volume, static 12h, the sediment centrifugally collected again. Precipitate with ethanol, acetone washing 3 times with distilled water, then adding the precipitate weight of the 2, the papain [...] for the 50 is placed in the water bath [...] 4h, Sevage process to get rid of the protein. Adding 8g activated carbon in the 70 the heating in water bath [...] 4h, filtering to remove the activated carbon, concentrated under reduced pressure, a natural cooling after dialysis in dialysis bag 48h in order to remove small molecule impurities for, after the end of the dialysis, dialysis to join again the volume the post-sugar solution 3 times with ethanol anhydrous ethanol, standing sleepovers, precipitate in the 70 the drying to obtain [...] 6.4g crude polysaccharide, the yield is 2.1%.

Taking 1.5g crude polysaccharide dissolved with distilled water, using DEAE-Celluose gel chromatographic column chromatography separation (5.0 × 50 cm), to 0.5mol/L NaCl solution as mobile phase (in other words eluant), flow rate is 2.5 ml/min, the phenol-sulfuric acid method for color rendering chromatographic analysis fluid , collecting main chromatographic peaks, freeze-dried. Then the resulting solid for the lyophilized 0.05mol/L NH₄ HCO₃ solution dissolving, using Sephadex G200 gel chromatographic column again chromatography separation (5.0 × 50 cm), to 0.05mol/L NH₄ HCO₃ solution as mobile phase (in other words eluant), flow rate is 2.0 ml/min, the phenol-sulfuric acid method for color rendering chromatographic analysis fluid , collecting main chromatographic peaks, freeze-dried, to obtain 360 mg refining tamago flower polysaccharide solid, yield 24%, through measuring LT1 the sugar content is 95%.

Application test: embodiment 1 preparation of the polysaccharide of eggs as tobacco humectant applied research

1, physical assuaring and moistening performance test

The polysaccharide of eggs for inspection physical assuaring and moistening performance of the cigarette, to gold leaf (large jin Yuan) blank cigarette tobacco as a carrier, the refined tamago flower polysaccharide with glycerol, propylene glycol and sorbitiol together the respective physical assuaring and moistening performance than testing and research, separately prepared for the mass fraction of 5% aqueous solution of the above-mentioned compound. Firstly the 1000g mixing the temperature of the blank of the cigarette tobacco 22 ±1 °C, the relative humidity of the environment 60 ±2% 48h. Then equivalent weighing five cut tobacco, each 100g, the cut tobacco is evenly applied 4g aqueous solution of the above-mentioned compound, compared with the blank sample to the uniform application of 4g water. Then cut tobacco are respectively arranged in five portions 22 ±1 °C, relative humidity of 40 ±2% inspection its wet process under the environment, and to the change of the immediate analysis of the water content, investigate its physical assuaring and moistening effect (that is, adding different samples tobacco after the cut tobacco 72h dry basis of the change in the water content), the results shown in table 1.

Table 1, tamago flower polysaccharide, glycerin, propylene glycol and sorbierite physical assuaring and moistening effect contrast

Table 1 test result indicates that, adding tamago flower polysaccharide, glycerin, propylene glycol and sorbitiol four kinds of compound compared with the blank and the cut tobacco, the moisture loss rate is relatively slow, this is because multi-hydroxy compound containing hydrophilic groups of the hydroxy can be and water to form hydrogen bond to the bound water, delaying the moisture loss, improves the cut tobacco physical assuaring and moistening performance; wherein the egg polysaccharide the best physical assuaring and moistening performance, followed by propylene glycol and glycerol, sorbitol is the last.

2, sensory assuaring and moistening performance test

To 20v % ethanol aqueous solution as the solvent, the eggs into polysaccharide the mass fraction of 1% of the solution. Separately taking 0.100g, 0 . 500g, 1.000g, 5.000g, 10 . 000g and 20.000g the above-mentioned polysaccharide solution containing eggs, and then respectively applied to the can without perfume five portions 100g gold leaf blank tobacco shreds, rolling, all selected 100 supporting the same weight cigarette, a temperature the 22 the   ± 1 [...] , humidity 60% ± 2% balance in the case of constant temperature and humidity 48h, smoking. Cigarette leaf blank for Golden the control sample, the control sample at the same temperature the same, humidity conditions balance 48h. Evaluating smoking results are shown in table 2.

Table 2 different quantity tamago flower polysaccharide evaluating smoking results

From table 2 it can be seen of the results caused, tamago polysaccharide the main role of the cigarettes is shown as obviously the flue gas becomes smooth, soft, taste improving comfort, impurity gas and acrimony are reduced. When the egg is relatively hours polysaccharide, the role of the cigarette is not very obvious; when its consumption exceed the 1 when [...] , spines [...] occur, the phenomenon of concentration are reduced; when its consumption is 1 the best quality when [...] use. Therefore, in the 100g added in cigarette 0.1g tamago polysaccharide the best effect when, that is, the most suitable for the amount of the 1 [...].