CN106632709B - A kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application - Google Patents
A kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application Download PDFInfo
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- CN106632709B CN106632709B CN201610313007.8A CN201610313007A CN106632709B CN 106632709 B CN106632709 B CN 106632709B CN 201610313007 A CN201610313007 A CN 201610313007A CN 106632709 B CN106632709 B CN 106632709B
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- fructus schisandrae
- carboxy methylation
- polysaccharide
- schisandrae polysaccharide
- schisandra chinensis
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 113
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 113
- 150000004676 glycans Chemical class 0.000 title claims abstract description 111
- 230000011987 methylation Effects 0.000 title claims abstract description 100
- 238000007069 methylation reaction Methods 0.000 title claims abstract description 100
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 title claims abstract description 95
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 240000006079 Schisandra chinensis Species 0.000 claims abstract description 57
- 235000008422 Schisandra chinensis Nutrition 0.000 claims abstract description 57
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- 235000009508 confectionery Nutrition 0.000 claims abstract description 44
- 238000006243 chemical reaction Methods 0.000 claims abstract description 22
- FDRCDNZGSXJAFP-UHFFFAOYSA-M sodium chloroacetate Chemical compound [Na+].[O-]C(=O)CCl FDRCDNZGSXJAFP-UHFFFAOYSA-M 0.000 claims abstract description 20
- 238000006467 substitution reaction Methods 0.000 claims abstract description 16
- 230000035484 reaction time Effects 0.000 claims abstract description 11
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims abstract description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 6
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 6
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 44
- 235000019441 ethanol Nutrition 0.000 claims description 33
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 31
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 28
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 26
- 238000000502 dialysis Methods 0.000 claims description 26
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
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- REHONNLQRWTIFF-UHFFFAOYSA-N 3,3',4,4',5-pentachlorobiphenyl Chemical compound C1=C(Cl)C(Cl)=CC=C1C1=CC(Cl)=C(Cl)C(Cl)=C1 REHONNLQRWTIFF-UHFFFAOYSA-N 0.000 abstract description 20
- 210000002966 serum Anatomy 0.000 abstract description 10
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- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 abstract description 7
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- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 6
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- 238000012360 testing method Methods 0.000 description 4
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- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 241000219991 Lythraceae Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 235000014360 Punica granatum Nutrition 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 241001506047 Tremella Species 0.000 description 2
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- 230000003078 antioxidant effect Effects 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- AYJRCSIUFZENHW-UHFFFAOYSA-L barium carbonate Chemical compound [Ba+2].[O-]C([O-])=O AYJRCSIUFZENHW-UHFFFAOYSA-L 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
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- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
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- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 241000218377 Magnoliaceae Species 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 241000221037 Pyrularia pubera Species 0.000 description 1
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- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
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- 239000002398 materia medica Substances 0.000 description 1
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- 239000010445 mica Substances 0.000 description 1
- 229910052618 mica group Inorganic materials 0.000 description 1
- 238000002715 modification method Methods 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
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- 230000000144 pharmacologic effect Effects 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
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- 235000019353 potassium silicate Nutrition 0.000 description 1
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- 238000012545 processing Methods 0.000 description 1
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- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
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- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Sustainable Development (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of carboxy methylation Fructus Schisandrae Polysaccharides and its preparation method and application, belong to polysaccharide-modified technical field.The yield of the carboxy methylation Schisandra chinensis Thick many candies is 88.37 ± 1.65%, and carboxy methylation degree of substitution is 0.724 ± 0.022, and it is 5 times of former Fructus Schisandrae Polysaccharide solubility that solubility, which is 0.5 ± 0.04 mg/mL,.The solubility of carboxy methylation Fructus Schisandrae Polysaccharide is 12.5 ± 0.26 mg/mL, and molecular weight is 3.4 × 106Da, monosaccharide composition are as follows: mannose, glucose and galactolipin, molar ratio 1:44.84:3.71.Reaction temperature of the present invention is 40-80 °C, reaction time in 1-8 h, monoxone final concentration is under conditions of 0.005-0.08 g/mL, the yield and degree of substitution of gained carboxy methylation Fructus Schisandrae Polysaccharide are higher, good water solubility, and cause mouse immune inhibition to be significantly improved effect persistence organic pollutant PCB126, mouse spleen and thymus index can be improved, increase TNF-α, the level of IFN-γ and IL-2 in serum, it can be used as immunopotentiator, be used as functional food and food additives.
Description
Technical field
The present invention relates to a kind of carboxy methylation Fructus Schisandrae Polysaccharides and its preparation method and application, belong to polysaccharide-modified technical field.
Background technique
The mellow fruit of Schisandra chinensis (Schisandra chinensis (Turcz.) Baill.) category Magnoliaceae Schisandra chinensis plant
It is real, it is main to originate in the ground such as China Heilungkiang, Liaoning, the Inner Mongol, it is a kind of common nourishing Chinese medicine, with high medicinal
And edible value, it has been put into article list (land rabbit woods, Wu Yang, the season black Fructus Schisandrae Polysaccharide extraction of moral that can be used for health food
Separation and Advance on Pharmacological Activities CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2014,39 (4): 751-754.).Research finds that Fructus Schisandrae Polysaccharide is made
For active constituent main in Schisandra chinensis, content is higher, and has a variety of lifes such as antitumor, anti-oxidant, anti-aging, antifatigue
Object activity (each ingredient latest Progress Aged in China physical-fitness medicine of Xu Bo, Shen Nan, Zhao Lijing Schisandra chinensis, 2013,11 (5):
74-74.), and it is minimum to the toxicity of organism normal cell and tissue, there are preferable development and application values.
Currently, the structural modification method of polysaccharide mainly has Phosphation, carboxy methylation, acetylation etc..Carboxy methylation modification
It as a kind of main chemical modification method, is directed to introduce carboxymethyl on polysaccharide chain, solubility of such method of modifying to polysaccharide
And electronegativity have more important influence (preparation of Jiang Leming, Fan Canmei, Nie Shaoping carboxy methylation semen plantaginis major polysaccharide and
Its bioactivity research Food Science, 2013,34 (22): 10-14.).The study found that a kind of alkali solubility tremella polysaccharides, insoluble
Yu Shui, after being modified by carboxy methylation, solubility in water is improved to 36.38mg/mL (Wu Qiong carboxy methylation alkali soluble
The research Changchun University journal of property tremella polysaccharides: natural science edition, 2009,19 (10): 66-68.).2005, Guo offered sacriffices to the gods or the spirits of the dead remote etc.
Application patent " carboxy methyl schizophllan polysacharide preparation method and applied in cosmetics and anti-tumor drug " (application number:
CN200510101678.X in), carboxymethyl modification is carried out to schizophan using conventional reagents such as monoxone, sodium hydroxides, is changed
It has been apt to its solubility property and has maintained its good rheological property and anti-tumor activity.2011, face upward the patent of the applications such as pomegranate blueness
In " a kind of carboxy methylation Blackfungus polyhexose, Thick many candies and its preparation method and application " (application number: CN201110219047.3),
Carboxymethyl modification is carried out to Blackfungus polyhexose using isopropanol method, it is the 6 of raw sugar solubility that degree of making it dissolve, which reaches 0.6mg/mL,
Times, and its antioxidant activity in vitro is remarkably improved.
The present inventor has found in the early-stage study of the isolating and purifying of Fructus Schisandrae Polysaccharide, structural characterization and anti-tumor activity,
The water solubility of Fructus Schisandrae Polysaccharide is undesirable, constrains its further further investigation and development and application.Therefore, the present inventor is to five
Taste polysaccharide carries out carboxy methylation modification, primarily determines its structure feature, provides a kind of utilization Response Surface Method optimization carboxymethyl
The technique for changing Fructus Schisandrae Polysaccharide, and the carboxy methylation Fructus Schisandrae Polysaccharide is had studied to caused by persistence organic pollutant PCB126
The intervention effect of immunotoxicity.The research of preparation and its process optimization in relation to Fructus Schisandrae Polysaccharide carboxyl methylation derivant is so far not
See that domestic and foreign literature is reported, also has no corresponding patent disclosure.
Summary of the invention
The purpose of the present invention is to solve the water-soluble undesirable of Fructus Schisandrae Polysaccharide, constrain it and further deeply grind
Study carefully and the shortcomings that development and application, provides a kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application.
The present invention adopts the following technical scheme: a kind of carboxy methylation Fructus Schisandrae Polysaccharide, the carboxy methylation Fructus Schisandrae Polysaccharide
Molecular weight be 1.698 × 104Da, the monosaccharide composition of the carboxy methylation Fructus Schisandrae Polysaccharide are as follows: mannose, glucose and gala
Sugar, molar ratio 1:44.84:3.71, the solubility of the carboxy methylation Fructus Schisandrae Polysaccharide is 12.24-12.76mg/mL, described
Carboxy methylation Fructus Schisandrae Polysaccharide in aqueous solution be in aggregation shape formed strand, the strand with a thickness of 0.1-1nm.
The preparation method of carboxy methylation Fructus Schisandrae Polysaccharide carries out as steps described below:
(1) new fresh schisandra chinensis the preparation of Schisandra chinensis Thick many candies: is subjected to powder after 12-24h dry in 55-65 DEG C of baking oven
It is broken, reflux degreasing 2-3 times is carried out to the Schisandra chinensis after drying using petroleum ether, then by the Schisandra chinensis dregs of a decoction after degreasing with feed liquid
Distilled water progress water is added than 1:10-1:15 to mention, extracts 2-3 times and obtains Fructus Schisandrae Polysaccharide extracting solution, Fructus Schisandrae Polysaccharide extracting solution
Through being concentrated under reduced pressure, using ethyl alcohol alcohol precipitation 3-4 times of 80%, obtained alcohol hypostasis redissolves after being centrifuged, waves alcohol, through retaining molecule
Bag filter the dialysis 12-36h, vacuum drying 2-3h that amount is 3000Da obtain Schisandra chinensis Thick many candies；
(2) carboxy methylation of Fructus Schisandrae Polysaccharide: Schisandra chinensis Thick many candies are dissolved in aqueous isopropanol, and stirring keeps it sufficiently molten
Solution, Schisandra chinensis Thick many candies and isopropanol solid-liquid ratio are 1:30-1:40 (g/mL), and solution temperature is 20-25 DEG C, and mixing time is
15-20min, mixing speed 60-240r/min, it is 15- that mass fraction is added into dissolved Fructus Schisandrae Polysaccharide solution
20% NaOH solution alkalizes, and the additional amount of NaOH solution is the 20-30%, alkalization temperature 20-25 of isopropanol volume
DEG C, then alkalization time 1-2h continues that monoxone is added in the Schisandra chinensis Thick many candies reaction solution after alkalization under stirring
Carboxy methylation is carried out, monoxone additional amount is 0.005-0.08g/mL, and bath temperature is 40-80 DEG C, and the carboxymethylation reaction time is
1-8h adjusts reaction solution pH after reaction solution is cooling and is obtained alcohol precipitation product after centrifugation using ethyl alcohol alcohol precipitation to neutrality, alcohol precipitation is produced
Object is dialysed after being redissolved using a small amount of distilled water, is dried in vacuo to obtain carboxy methylation Schisandra chinensis Thick many candies after dialysis；
(3) it purifies: the carboxy methylation Schisandra chinensis Thick many candies that step (2) obtains is dissolved in the NaCl of 0.05-0.1mol/L
It in solution, is then slowly added dropwise into DEAE-52 chromatographic column, and gradient is carried out using the NaCl solution of 0.05-0.2mol/L and is washed
It is de-, elution curve is drawn, merging eluent is collected respectively according to elution curve, obtains carboxy methylation Fructus Schisandrae Polysaccharide difference group
Point, the elution fraction for choosing 0.05mol/L NaCl merges, and dialyses through the bag filter that molecular cut off is 3000Da, vacuum drying
Afterwards, carboxy methylation Fructus Schisandrae Polysaccharide is obtained.
Further, the yield of Schisandra chinensis Thick many candies is 15.9-19.8%, polyoses content 54.4- in the step (1)
65.7%, solubility 0.08-0.12mg/mL.
Further, the yield of carboxy methylation Schisandra chinensis Thick many candies is 60.2-86.4%, carboxymethyl in the step (2)
Change degree of substitution is 0.41-0.71, solubility 0.46-0.54mg/mL.
Further, the temperature of reflux degreasing is 70-80 DEG C, degreasing time 1-2h in the step (1), water temperature raising degree
It is 90-100 DEG C, extraction time 4-6h.
Further, 80% ethyl alcohol alcohol precipitation 24-48h is used in the step (2), precipitation solution is with 4000-5000rpm revolving speed
It is centrifuged 5-10min.
Further, using in the step (2) through molecular cut off is the dialysis of the bag filter of 3000Da first using stream
Water dialysis finally carries out distilled water dialysis, and flowing water dialysis time is 36-48h, and distilled water dialysis time is 24-36h.
Further, in the step (2) vacuum drying time be 2-3h, temperature be 20-25 DEG C, vacuum degree be-
0.085--0.1Mpa。
Further, carboxy methylation Schisandra chinensis Thick many candies dissolve by heating in NaCl solution through concussion in the step (3),
The temperature of shaking water bath is 50-60 DEG C, and oscillation frequency is 120-160 beats/min, the time 10-12h of dissolution is shaken, then to molten
Solution after solution is centrifuged, the revolving speed of solution centrifugation are as follows: 4000-5000r/min, centrifugation time 8-10min.
Further, flowing water dialysis time is 36-48h in the step (3), and distilled water dialysis time is 24-36h, very
Empty drying time is 2-3h, and vacuum drying temperature is 20-25 DEG C, and vacuum drying vacuum degree is -0.085--0.1MPa.
Further, the carboxy methylation Fructus Schisandrae Polysaccharide can be used for functional food and food additive as immunopotentiator
Add in agent.
Preparation method of the present invention is simple, and step is easily operated, prepares the carboxy methylation five tastes using the method for molecular modification
Sub- Thick many candies and carboxy methylation Fructus Schisandrae Polysaccharide；The carboxy methylation Fructus Schisandrae Polysaccharide obtained using the preparation method in the present invention
Yield and degree of substitution are higher, and dissolubility is good, and solubility is 5 times and 125 times of former Fructus Schisandrae Polysaccharide, the carboxylic being prepared respectively
Methylation Fructus Schisandrae Polysaccharide can be effectively improved the immunosuppressive action of persistence organic pollutant PCB126, can be as immune
Reinforcing agent is applied to functional food and food additives.
Detailed description of the invention
Fig. 1 is reaction temperature, reaction time, interacting between monoxone final concentration obtains carboxy methylation Fructus Schisandrae Polysaccharide
The response surface figure (A, B, C) and contour map (D, E, F) that rate influences.
Fig. 2 is reaction temperature, reaction time, interacting between monoxone final concentration takes carboxy methylation Fructus Schisandrae Polysaccharide
The response surface figure (A, B, C) and contour map (D, E, F) influenced for degree.
Fig. 3 is the infrared spectrogram of Fructus Schisandrae Polysaccharide.
Fig. 4 is the infrared spectrogram of carboxy methylation Fructus Schisandrae Polysaccharide.
Fig. 5 is atomic force X-Y scheme (A), three-dimensional figure (B) and the strand height distribution map (C) of carboxy methylation Fructus Schisandrae Polysaccharide.
Fig. 6 is influence diagram of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice index and spleen index,aP < 0.05, with sky
White control group is compared；bP < 0.05, compared with model group；cP < 0.05, compared with CSP1 high dose group.
Fig. 7 is influence diagram of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice thymus index,aP < 0.05, with sky
White control group is compared；bP < 0.05, compared with model group；cP < 0.05, compared with CSP1 high dose group.
Fig. 8 is influence diagram of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice serum TNF-cc level,aP < 0.05,
Compared with blank control group；bP < 0.05, compared with model group.
Fig. 9 is influence diagram of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice serum I NF- γ level,ap<
0.05, compared with blank control group；bP < 0.05, compared with model group.
Figure 10 is influence diagram of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice serum IL-2 level,aP < 0.05,
Compared with blank control group；bP < 0.05, compared with model group；cP < 0.05, compared with CSP1 high dose group.
Specific embodiment
The preparation of Schisandra chinensis Thick many candies
Embodiment 1:
New fresh schisandra chinensis is crushed after 12h dry in 55 DEG C of baking ovens, using petroleum ether to the Schisandra chinensis after drying
In 70 DEG C of reflux degreasing 1h, 2 reflux degreasings are carried out, then add the Schisandra chinensis dregs of a decoction after degreasing with solid-liquid ratio 1:10 (g/mL)
Enter distilled water and carry out water to mention, mention 4h in 90 DEG C of water, extract 2 times, extracting solution merges after being concentrated under reduced pressure, 80% ethyl alcohol alcohol repeatedly
3 times heavy, after centrifugation, Schisandra chinensis is made through bag filter dialysis 12h, vacuum drying 2h that molecular cut off is 3000Da in supernatant
Thick many candies, polysaccharide yield are 15.9 ± 0.3%, and polyoses content is 54.4 ± 2.7%.
Embodiment 2
New fresh schisandra chinensis is crushed afterwards for 24 hours in drying in 65 DEG C of baking ovens, using petroleum ether to the Schisandra chinensis after drying
In 80 DEG C of reflux degreasing 2h, 3 reflux degreasings are carried out, then add the Schisandra chinensis dregs of a decoction after degreasing with solid-liquid ratio 1:15 (g/mL)
Enter distilled water progress water to mention, mentions 6h in 100 DEG C of water, extract 3 times, extracting solution merges after being concentrated under reduced pressure, and 80% ethyl alcohol is repeatedly
Alcohol precipitation 4 times, after centrifugation, the five tastes are made through bag filter dialysis 36h, vacuum drying 3h that molecular cut off is 3000Da in supernatant
Sub- Thick many candies, polysaccharide yield are 19.8 ± 0.9%, and polyoses content is 65.7 ± 2.7%, and solubility is 0.10 ± 0.02mg/mL.
The Schisandra chinensis Thick many candies prepared using embodiment 2 is the raw materials of next step.
(the isolating and purifying of Zhao Ting Fructus Schisandrae Polysaccharide, structural characterization compared with Fructus Schisandrae Polysaccharide preparation process in existing literature
And its activity research [D] Jiangsu University, 2009.), the preparation process of the Thick many candies uses alcohol precipitation 3-4 times repeatedly of 80% ethyl alcohol,
The polyoses content of Schisandra chinensis Thick many candies is significantly improved, polyoses content improves about 21% or so；And use boulton process can
Effectively shorten drying time, drying time reduces about 93% or so, which can be energy saving, is conducive to industrial production
In application.
The preparation of carboxy methylation Schisandra chinensis Thick many candies
Embodiment 3:
It weighs Schisandra chinensis Thick many candies sample and is mixed with isopropanol with solid-liquid ratio 1:30 (g/mL), at 20 DEG C, be with revolving speed
60r/min mechanical stirring 15min, is sufficiently dissolved；Mass fraction is added into dissolved Fructus Schisandrae Polysaccharide solution is
15% NaOH solution alkalizes, and the additional amount of NaOH solution is the 20% of isopropanol volume, continues at 20 DEG C of temperature mechanical
Stir 1h；Monoxone is added in above-mentioned reaction solution, chloroethene acid concentration is made to reach 0.005g/mL, is placed in 40 DEG C of waters bath with thermostatic control
In pot, continues mechanic whirl-nett reaction 1h, reaction product is adjusted to neutrality with hydrochloric acid after cooling, for 24 hours through 80% ethyl alcohol alcohol precipitation, alcohol precipitation
After liquid is centrifuged 5min with 4000rpm revolving speed, precipitating is redissolved, and is the bag filter flowing water dialysis 36h of 3000Da through molecular cut off,
After distilled water dialysis for 24 hours, 2h is dried in vacuo under 20 DEG C, vacuum degree -0.085MPa to get carboxy methylation Schisandra chinensis Thick many candies
(CSP), polysaccharide yield is 75.9 ± 1.1%, and carboxy methylation degree of substitution is 0.41 ± 0.02.
Embodiment 4:
It weighs Schisandra chinensis Thick many candies sample and is mixed with isopropanol with solid-liquid ratio 1:35 (g/mL), at 25 DEG C, be with revolving speed
240r/min mechanical stirring 20min, is sufficiently dissolved；Mass fraction is added into dissolved Fructus Schisandrae Polysaccharide solution is
20% NaOH solution alkalizes, and the additional amount of NaOH solution is the 25% of isopropanol volume, continues at 25 DEG C of temperature mechanical
Stir 2h；Monoxone is added in above-mentioned reaction solution, chloroethene acid concentration is made to reach 0.02g/mL, is placed in 60 DEG C of thermostat water baths
In, continue mechanic whirl-nett reaction 4h, reaction product is adjusted to neutrality with hydrochloric acid after cooling, through 80% ethyl alcohol alcohol precipitation 36h, precipitation solution
After being centrifuged 10min with 4000rpm revolving speed, precipitating is redissolved, and is the bag filter flowing water dialysis 48h of 3000Da through molecular cut off,
Distilled water is dialysed after 36h, is dried in vacuo 3h under 25 DEG C, vacuum degree -0.1MPa to get carboxy methylation Schisandra chinensis Thick many candies
(CSP), it is 86.4 ± 1.5% that polysaccharide yield, which is polysaccharide yield, and carboxy methylation degree of substitution is 0.71 ± 0.03, solubility 0.50
±0.04mg/mL。
Embodiment 5:
It weighs Schisandra chinensis Thick many candies sample and is mixed with isopropanol with solid-liquid ratio 1:40 (g/mL), at 25 DEG C, be with revolving speed
240r/min mechanical stirring 20min, is sufficiently dissolved；Mass fraction is added into dissolved Fructus Schisandrae Polysaccharide solution is
20% NaOH solution alkalizes, and the additional amount of NaOH solution is the 30% of isopropanol volume, continues at 25 DEG C of temperature mechanical
Stir 2h；Monoxone is added in above-mentioned reaction solution, chloroethene acid concentration is made to reach 0.08g/mL, is placed in 80 DEG C of thermostat water baths
In, continue mechanic whirl-nett reaction 8h, reaction product is adjusted to neutrality with hydrochloric acid after cooling, through 80% ethyl alcohol alcohol precipitation 48h, precipitation solution
After being centrifuged 10min with 5000rpm revolving speed, precipitating is redissolved, and is the bag filter flowing water dialysis 48h of 3000Da through molecular cut off,
Distilled water is dialysed after 36h, is dried in vacuo 3h under 25 DEG C, vacuum degree -0.1MPa to get carboxy methylation Schisandra chinensis Thick many candies
(CSP), polysaccharide yield is 60.2 ± 2.2%, and carboxy methylation degree of substitution is 0.68 ± 0.02.
Response phase method optimizes carboxy methylation Schisandra chinensis Thick many candies synthesis technology
Based on experiment of single factor, reaction temperature chooses 50 DEG C, 60 DEG C and 70 DEG C three levels, and the reaction time is chosen
Tri- levels of 2h, 4h and 6h, monoxone final concentration choose tri- levels of 0.01g/mL, 0.02g/mL and 0.04g/mL, utilize
8.0 software of Design Expert carries out experimental design according to Box-Benhnken center combination design principle, with reaction temperature
(A), reaction time (B) and monoxone final concentration (C) are independent variable, with the yield (Y of CSP1) and degree of substitution (Y2) it is response,
Optimize Schisandra chinensis Thick many candies carboxy methylation process conditions using Response Surface Method.Response surface empirical factor level is shown in Table 1, response
Surface analysis the results are shown in Table 2, table 3 and table 4.
Table 1: empirical factor water-glass
Table 2: response surface experimental design and response result
The analysis of variance table of 3 yield of table
Note: P < 0.0001 be it is extremely significant, P < 0.05 is significant.
Regression analysis is carried out to experimental data using 8 software of Design-Expert, yield is tested according to each factor and is tied
The influence of fruit carries out quadratic equation fitting, and fitting obtains following formula:
Y1=-84.48711+2.53102A+28.50936B+67.94377C-0.036938AB+0.21919 AC -
1.66175BC–0.020539A2–2.94075B2–36.32919C2
Y in formula1For yield.
The yield of each factor pair CSP all has more significant influence (P < 0.05).Influence of three factors for yield is suitable
Sequence are as follows: monoxone final concentration > reaction temperature > reaction time.Value < 0.0001 test model P, it was demonstrated that model is significant.Lose quasi- error
Not significant (value=2.14 F of value；Value=0.2374 P), illustrate that inspection result is not significantly different with the model calculation, can be used for
The theoretical prediction of Fructus Schisandrae Polysaccharide carboxy methylation compound experiment.
Table 4: the variance analysis of carboxy methylation Fructus Schisandrae Polysaccharide degree of substitution
Note: P < 0.0001 be it is extremely significant, P < 0.05 is significant
Regression analysis is carried out to experimental data using 8 software of Design-Expert, degree of substitution is tested according to each factor
As a result influence carries out quadratic equation fitting, and fitting obtains following formula:
Y2=-0.68299+0.026417A+0.14229B+0.45287C+4.6875 × 10-3AB+4.00000 × 10-
3AC–4.84211×10-3BC–2.26187×10-3A2–0.018494B2–0.18924C2
Y in formula2For degree of substitution.
The degree of substitution of each factor pair CSP has more significant influence (P < 0.05).Influence of three factors for degree of substitution
Sequentially are as follows: monoxone final concentration > reaction time > reaction temperature.Value < 0.0001 P of test model, it was demonstrated that model is significant.It loses quasi-
Not significant (value=6.37 F of error amount；Value=0.0528 P), illustrate that inspection result is not significantly different with the model calculation, table
Bright regression model and actual conditions fitting very well, can be used for the theoretical prediction of Fructus Schisandrae Polysaccharide carboxy methylation compound experiment.
Plot analysis is carried out according to multiple regression equation using 8.0 software of Design Expert, obtains regression equation
Three factors of response surface figure (see Fig. 1 and Fig. 2) all have significant impact for the yield and degree of substitution of CSP, wherein monoxone
The influence of final concentration is most significant.
(5) analysis of experimental results and optimization:
Regression analysis calculating is carried out to experimental data using 8 software of Design-Expert, obtains the best work of synthesis CSP
Skill condition are as follows: reaction temperature is 63 DEG C, reaction time 4.5h, the final concentration of 0.022g/mL of monoxone.Under this condition, it puts down
Row experiment 5 times, the yield for obtaining CSP is 88.3 ± 1.65%, and degree of substitution is 0.72 ± 0.022.Actual measured value and response surface
Model prediction result is closer to, and the model for illustrating that the response phase method optimizes is accurate and reliable, is suitable for Schisandra chinensis Thick many candies
Carboxymethylated modification.With 2011, face upward the applications such as pomegranate blueness patent " a kind of carboxy methylation Blackfungus polyhexose, Thick many candies and its
Preparation method and application " (application number: CN201110219047.3) compare, and monoxone dosage reduces in the process conditions
46%, the reduction of monoxone dosage can reduce the generation of polysaccharide degradation side reaction in reaction process, so that carboxymethylated polysaccharides obtain
Rate significantly improves, and improves about 30% or so；Secondly, mechanical stirring is selected to replace magnetic agitation in the process conditions, stirring is strong
Degree is big, and good mixing effect can easily be accommodated and operate, and be more suitable for a large amount of preparations of carboxy methylation Fructus Schisandrae Polysaccharide；And it uses true
Empty seasoning can effectively shorten drying time, energy saving, be conducive to the application in industrial production.
Embodiment 6:
For the optimal processing parameter reaction temperature for using response surface to obtain for 63 DEG C, reaction time 4.5h, monoxone is dense eventually
Degree is 0.022g/mL, and carboxy methylation Schisandra chinensis Thick many candies are prepared.
The purifying and structure feature of carboxy methylation Schisandra chinensis Thick many candies
Preparation carboxy methylation Schisandra chinensis Thick many candies (CSP) 0.5g in above-described embodiment 6 is weighed, carboxy methylation Schisandra chinensis is carried out
The purifying of Thick many candies.
Embodiment 7:
It is dissolved in 0.05mol/LNaCl solution, is 160 beats/min, filled in the concussion water bath with thermostatic control of temperature 50 C in oscillation frequency
Divide dissolution 10h, lysate is centrifuged 10min through 4000r/min, and after removing insoluble matter, supernatant passes through DEAE-52 ion exchange column
(2.0 × 50cm) is separated；It is eluted with the NaCl solution of 0.05,0.1,0.15 and 2mol/L, is received with automatic collector by pipe
Collect eluent, by the flow control of constant flow pump in 1mL/min, every pipe collects 5mL.Collection liquid sulfuric acid-phynol method is detected by pipe,
Collect merging eluent respectively according to elution curve, wherein the eluent of 0.05mol/L NaCl is concentrated under reduced pressure, and retains molecule
The bag filter flowing water that amount is 3000Da is dialysed after 48h, distilled water dialysis for 24 hours, and 20 DEG C of Yu Wendu, vacuum degree are -0.085MPa's
Dry 2h, obtains carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) in vacuum oven.
Embodiment 8:
It is dissolved in 0.08mol/LNaCl solution, is 150 beats/min, filled in 55 DEG C of temperature of concussion water bath with thermostatic control in oscillation frequency
Divide dissolution 11h, lysate is centrifuged 9min through 4500r/min, and after removing insoluble matter, supernatant passes through DEAE-52 ion exchange column
(2.0 × 50cm) is separated；It is eluted with the NaCl solution of 0.05,0.1,0.15 and 2mol/L, is received with automatic collector by pipe
Collect eluent, by the flow control of constant flow pump in 1mL/min, every pipe collects 5mL.Collection liquid sulfuric acid-phynol method is detected by pipe,
Collect merging eluent respectively according to elution curve, wherein the eluent of 0.05mol/L NaCl is concentrated under reduced pressure, and retains molecule
The bag filter flowing water that amount is 3000Da is dialysed after 40h, distilled water dialysis 30h, and 21 DEG C of Yu Wendu, vacuum degree are -0.090MPa's
Dry 2.5h, obtains carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) in vacuum oven.
Embodiment 9:
It is dissolved in 0.1mol/LNaCl solution, is 120 beats/min, filled in the concussion water bath with thermostatic control of temperature 60 C in oscillation frequency
Divide dissolution 12h, lysate is centrifuged 8min through 5000r/min, and after removing insoluble matter, supernatant passes through DEAE-52 ion exchange column
(2.0 × 50cm) is separated；It is eluted with the NaCl solution of 0.05,0.1,0.15 and 2mol/L, is received with automatic collector by pipe
Collect eluent, by the flow control of constant flow pump in 1mL/min, every pipe collects 5mL.Collection liquid sulfuric acid-phynol method is detected by pipe,
Collect merging eluent respectively according to elution curve, wherein the eluent of 0.05mol/L NaCl is concentrated under reduced pressure, and retains molecule
The bag filter flowing water that amount is 3000Da is dialysed after 36h, distilled water dialysis 36h, and 25 DEG C of Yu Wendu, vacuum degree are the true of -0.1MPa
Dry 3h, obtains carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) in empty drying box.
(the isolating and purifying of Zhao Ting Fructus Schisandrae Polysaccharide, structural characterization and its Jiangsu activity research [D] are big with existing literature
Learn, 2009.) it is compared with patent (application number: CN201110219047.3 and CN201210132340.0), it is more in the purification process
Sugar is sufficiently dissolved through isothermal vibration, reduces the waste of carboxy methylation Fructus Schisandrae Polysaccharide raw material；And column purification process uses
The ion exchange column being relatively large in diameter is dramatically speeded up on the basis of not changing purification effect so that applied sample amount increases one times
Purify speed；And drying time can be effectively shortened using boulton process, and it is energy saving, be conducive to the application in industrial production.
Carboxy methylation Fructus Schisandrae Polysaccharide (CSP1), solubility 12.50mg/mL is prepared in Example 8.
The measurement of solubility is carried out according to 2005 editions pharmacopeia, the solubility of Schisandra chinensis Thick many candies is 0.10 ± 0.03mg/mL, carboxylic
The solubility of methylation Schisandra chinensis Thick many candies is 0.50 ± 0.04mg/mL, the solubility of carboxy methylation Fructus Schisandrae Polysaccharide (CSP1)
For 12.50 ± 0.26mg/mL.
The measurement of molecular weight: efficient gel size exclusion chromatography-multi-angle laser light scattering instrument-differential pulse polarograpll is used
Instrument combination analysis method (HPSEC-MALLS-RI System) is measured polysaccharide relative molecular mass.Chromatographic condition is with TSK-
GEL series G6000PWXL and G4000PWXL chromatographic column (300mm, 7.8mm, TOSOH, Japan) are analytical column, with 0.15mol/L
NaNO3 and 0.05mol/L NaH2PO4 (pH=7) is mobile phase, under conditions of column temperature is 35 DEG C, with the stream of 0.5mL/min
Speed carries out elution analysis.The solution for being 5mg/mL at mass concentration by CSP1 sample preparation, the high speed centrifugation through 12000r/min
100 μ L of sample introduction after taking supernatant to cross film after machine centrifugation 10min.CSP1 molecular weight is 1.698 × 104Da.With existing literature (Zhao
The the isolating and purifying of graceful Fructus Schisandrae Polysaccharide, structural characterization and its activity research [D] Jiangsu University, 2009.) and patent (application number:
CN201110219047.3 and CN201210132340.0) it compares, CSP1 molecular weight is smaller, and the lesser polysaccharide dissolution of molecular weight
Degree is high and is easier to the performance of its active function.
Monosaccharide composition analysis: precision weighs polysaccharide sample 5mg in ampoule bottle, is dissolved with the sulfuric acid solution of 2mol/L, envelope
Mouthful, in 100 DEG C of baking ovens after heating hydrolysis 8h, hydrolyzate adds barium carbonate to be neutralized to neutrality, is centrifuged off BaSO with 4000r/min4
Precipitating, supernatant freeze-drying.Hydrolysate is analyzed after acetylation with 2010 gas chromatography system of Shimadzu GC.The list of CSP1
Sugar composition are as follows: mannose, glucose and galactolipin, molar ratio 1:44.84:3.71.It can be seen that CSP1 is made of three kinds of monosaccharide,
Respectively mannose, glucose and galactolipin, and existing literature (the isolating and purifying of Zhao Ting Fructus Schisandrae Polysaccharide, structural characterization and its
Activity research [D] Jiangsu University, 2009.) and in patent (CN201210132340.0), Fructus Schisandrae Polysaccharide is by six kinds of monosaccharide structures
At (rhamnose, arabinose, xylose, mannose, glucose and galactolipin).
Infrared analysis: with potassium bromide (KBr) pressed disc method, the polysaccharide sample of a certain amount of drying is taken, using Fourier transform
Infrared spectrometer (ATAVAR360) scans 4000~400cm-1Wavelength period carries out structural characterization.Fructus Schisandrae Polysaccharide is through carboxy methylation
Polysaccharide afterwards is in 1600cm-1、1420cm-1There is strong absworption peak in place, (for the characteristic absorption peak of-COO-, wherein 1600cm-1It is left
Right place is the asymmetric stretching vibration absworption peak of-C-O-, 1420cm-1It is the symmetrical stretching vibration absorption peak of-C-O- at left and right),
Show carboxy methylation success.
Polysaccharide sample: being configured to the solution of 1mg/mL by atomic force Electronic Speculum, is crossed film and is collected filtrate, 20 μ L filtrates is taken to make it
It is uniformly attached on clean mica sheet, anti-gray is dry at room temperature.Sample is seen with scanning probe microscopy (MFP-3D)
It surveys.In aqueous solution in aggregation shape, the thickness of strand is in 0.1-1nm.With existing literature (structure of Zhao Ting Fructus Schisandrae Polysaccharide,
Bioactivity and its immunologic mechanism research [D] Jiangsu University, 2013.) it compares, carboxy methylation Fructus Schisandrae Polysaccharide is in aqueous solution
Aggregation reduce, and polysaccharide chain thickness compared with Fructus Schisandrae Polysaccharide component (1-5nm) significantly reduce.
Carboxy methylation Fructus Schisandrae Polysaccharide causes the intervention effect of immunity of organism toxicity to PCB126
Experimental animal
ICR mouse (cleaning grade), half male and half female, weight are 20 ± 2g, are purchased from Jiangsu University's animal experimental center, test it
Preceding quarantine 3 days, raising temperature range are 21 ± 1 DEG C, and raising humidity range is 60 ± 5%.
Material and reagent
Example 8 purifies obtained carboxy methylation Fructus Schisandrae Polysaccharide；
PCB126, physiological saline, picric acid；
TNF-α enzyme linked immunological kit, IL-2 enzyme linked immunological kit, IFN-γ enzyme linked immunological kit.
Experimental group and administration
It adapts to feed 70 before experiment after ICR mouse 3 days, is randomly divided into 5 groups, every group 14, half male and half female simultaneously separates
It feeds, blank control group, PCB126 model group, the high, medium and low three kinds of concentration of carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) is set
Dosage group.It is administered once daily, continuous 14 days.And in administration the 7th day, PCB126 model group, CSP1 high, in CSP1, CSP1 low three
50 μ g/kg PCB126 (stomach-filling amount is every 0.1mL) is given in the stomach-filling simultaneously of kind concentration dose group, persistently to poison 5 days.
Grouping, dosage and administration route are as follows:
Blank control group: the physiological saline (0.1mL/20g/d) of daily stomach-filling same dose, feed and drinking-water are freely；
PCB126 model group: the physiological saline (0.1mL/20g/d) of daily stomach-filling same dose, in beginning stomach-filling in the 7th day
PCB126 is given, is given 5 days, feed and drinking-water are freely；
CSP1 administration group: point high, medium and low three kinds of concentration dose groups,
CSP1 high dose group: the CSP1 (400mg/kg/d) of difference stomach-filling same dose daily, in administration, the 7th day starts to fill
Stomach gives PCB126, gives 5 days, and feed and drinking-water are freely；
CSP1 middle dose group: the CSP1 (200mg/kg/d) of difference stomach-filling same dose daily, in administration, the 7th day starts to fill
Stomach gives PCB126, gives 5 days, and feed and drinking-water are freely；
CSP1 low dose group: the CSP1 (100mg/kg/d) of difference stomach-filling same dose daily, in administration, the 7th day starts to fill
Stomach gives PCB126, gives 5 days, and feed and drinking-water are freely.
(1) Immune Organs Index
After the last administration, Yu Ci fasting 8h weighs the weight of animals and records.It extracts after eyeball takes blood and puts to death, take out chest
Gland, spleen are blotted after physiological saline cleans with filter paper, are weighed and are recorded.Each organ index is calculated as follows:
(2) physiochemical indice
With ELISA enzyme linked immunological kit measurement mice serum in IL-2, INF- γ, TNF-α content.
Statistical procedures
Data are analyzed with SPSS16.0 statistical software.As a result it is expressed asGroup difference uses single factor test variance
It analyzes (One-way ANOVA), the significance of difference between each group is compared using Tukey method of inspection, P ﹤ 0.05 indicates that conspicuousness is poor
It is different.Experimental result
CSP1 is shown in Fig. 6 and Fig. 7 to the index and spleen index of exposed Mice, thymus index respectively.Compared with blank control group, model
The mouse spleen and thymus index of group are remarkably decreased (P < 0.05), illustrate that PCB126 causes mouse immune to inhibit model foundation success；
And the high, medium and low dosage of CSP1 has restored the Thymus and spleen index of mouse in varying degrees, wherein CSP1 high dose group can be same
When significantly improve mouse spleen index and thymus index.CSP1 is to TNF-α in exposed Mice serum, IFN-γ, IL-2 horizontal
Fig. 8, Fig. 9 and Figure 10 are shown in influence respectively.Compared with blank control group, TNF-α, IFN-γ and IL-2 in the mice serum of model group
Level is remarkably decreased (P < 0.05), and TNF-α, IFN-γ and IL-2 content are equal in the high, medium and low dosage mice serum of CSP1
There is different degrees of rising, wherein the content of the TNF-α in CSP1 high dose group mice serum significantly rises (P < compared with model group
0.05), and in CSP1 high dose group and middle dosage mice serum the content of IL-2 significantly rises (P < 0.05) compared with model group.Mesh
Before, existing literature report, (structure, bioactivity and its immunologic mechanism of Zhao Ting Fructus Schisandrae Polysaccharide study [D] to Fructus Schisandrae Polysaccharide
Jiangsu University, 2013.) and Blackfungus polyhexose (the separation preparation of Song Guanglei Blackfungus polyhexose and the Zhejiang bioactivity research [D]
Industrial and commercial university, 2011.) there is immunoregulatory activity, the immunosuppressive action of caused by cyclophosphamide can be significantly improved, but for holding
The intervention effect of immunity of organism toxicity caused by long property organic pollutant PCB126 has no relevant report.
Claims (9)
1. a kind of carboxy methylation Fructus Schisandrae Polysaccharide, it is characterised in that: the molecular weight of the carboxy methylation Fructus Schisandrae Polysaccharide is 1.698
×104Da, the monosaccharide composition of the carboxy methylation Fructus Schisandrae Polysaccharide are as follows: mannose, glucose and galactolipin, molar ratio 1:
44.84:3.71, the solubility of the carboxy methylation Fructus Schisandrae Polysaccharide are 12.24-12.76 mg/mL, the carboxy methylation five tastes
Sub- polysaccharide in aqueous solution be in aggregation shape formed strand, the strand with a thickness of 0.1-1 nm；
It carries out as steps described below:
(1) preparation of Schisandra chinensis Thick many candies: new fresh schisandra chinensis is crushed after 12-24h dry in 55-65 DEG C of baking oven, is adopted
Reflux degreasing 2-3 times is carried out to the Schisandra chinensis after drying with petroleum ether, then by the Schisandra chinensis dregs of a decoction after degreasing with solid-liquid ratio 1:
10-1:15 is added distilled water progress water and mentions, and takes 2-3 times and obtains Fructus Schisandrae Polysaccharide extracting solution, Fructus Schisandrae Polysaccharide extracting solution is through depressurizing
Concentration, using ethyl alcohol alcohol precipitation 3-4 times of 80%, obtained alcohol hypostasis redissolves after being centrifuged, waves alcohol, is 3000 through molecular cut off
Bag filter the dialysis 12-36h, vacuum drying 2-3h of Da obtains Schisandra chinensis Thick many candies；
(2) carboxy methylation of Fructus Schisandrae Polysaccharide: Schisandra chinensis Thick many candies are dissolved in aqueous isopropanol, and mechanical stirring keeps it sufficiently molten
Solution, Schisandra chinensis Thick many candies and isopropanol solid-liquid ratio are 1:30-1:40(g/mL), solution temperature is 20-25 DEG C, and mixing time is
15-20 min, mixing speed are 60-240 r/min, and it is 15- that mass fraction is added into dissolved Fructus Schisandrae Polysaccharide solution
20% NaOH solution alkalizes, and the additional amount of NaOH solution is the 20-30% of isopropanol volume, and alkalization temperature is 20-25 DEG C,
Alkalization time is 1-2 h, then continues that monoxone is added in the Schisandra chinensis Thick many candies reaction solution after alkalization under stirring
Carboxy methylation is carried out, monoxone additional amount is 0.005-0.08 g/mL, and bath temperature is 40-80 DEG C, the carboxymethylation reaction time
For 1-8 h, reaction solution pH is adjusted after reaction solution is cooling and to neutrality alcohol precipitation product is obtained using ethyl alcohol alcohol precipitation after centrifugation, by alcohol
Heavy product is dialysed after being redissolved using a small amount of distilled water, is dried in vacuo to obtain carboxy methylation Schisandra chinensis after dialysis thick
Polysaccharide；
(3) it purifies: the carboxy methylation Schisandra chinensis Thick many candies that step (2) obtains is dissolved in the NaCl solution of 0.05-0.1mol/L
In, it is then slowly added dropwise into DEAE-52 chromatographic column, and gradient elution is carried out using the NaCl solution of 0.05-0.2 mol/L,
Elution curve is drawn, merging eluent is collected respectively according to elution curve, obtains carboxy methylation Fructus Schisandrae Polysaccharide different component, is selected
It takes the elution fraction of 0.05mol/L NaCl to merge, dialyses, after vacuum drying through the bag filter that molecular cut off is 3000 Da,
Obtain carboxy methylation Fructus Schisandrae Polysaccharide.
2. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: Schisandra chinensis is thick in the step (1)
The yield of polysaccharide is 15.9-19.8%, and polyoses content 54.4-65.7%, solubility is 0.08-0.12 mg/mL.
3. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: carboxy methylation in the step (2)
The yield of Schisandra chinensis Thick many candies is 60.2-86.4%, and carboxy methylation degree of substitution is 0.41-0.71, solubility 0.46-0.54
mg/mL。
4. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: flow back degreasing in the step (1)
Temperature be 70-80 DEG C, degreasing time be 1-2 h, water temperature raising degree be 90-100 DEG C, extraction time be 4-6 h.
5. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: use 80% second in the step (2)
Alcohol alcohol precipitation 24-48h, precipitation solution are centrifuged 5-10 min with 4000-5000rpm revolving speed.
6. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: using through cutting in the step (2)
It stays the bag filter dialysis that molecular weight is 3000 Da to dialyse first using flowing water, finally carries out distilled water dialysis, flowing water dialysis time
For 36-48h, distilled water dialysis time is 24-36h.
7. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: vacuum drying in the step (2)
Time is 2-3h, and temperature is 20-25 DEG C, and vacuum degree is-0.085-- 0.1Mpa.
8. carboxy methylation Fructus Schisandrae Polysaccharide according to claim 1, it is characterised in that: carboxy methylation in the step (3)
Schisandra chinensis Thick many candies dissolve by heating in NaCl solution through concussion, and the temperature of shaking water bath is 50-60 DEG C, oscillation frequency 120-
160 beats/min, the time 10-12h of dissolution is shaken, then dissolved solution is centrifuged, the revolving speed of solution centrifugation
Are as follows: 4000-5000 r/min, centrifugation time are 8-10 min；
Flowing water dialysis time is 36-48h in the step (3), and distilled water dialysis time is 24-36h, vacuum drying time 2-
3h, vacuum drying temperature are 20-25 DEG C, and vacuum drying vacuum degree is-0.085-- 0.1MPa.
9. the application of carboxy methylation Fructus Schisandrae Polysaccharide described in claim 1, it is characterised in that: the carboxy methylation Fructus Schisandrae Polysaccharide
It can be as immunopotentiator in functional food and food additives.
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| CN102321186A (en) * | 2011-08-02 | 2012-01-18 | 江苏大学 | Carboxymethylated auricularia auricula polysaccharide and crude polysaccharide, and preparation method and application thereof |
| CN102492050A (en) * | 2011-12-01 | 2012-06-13 | 通化师范学院 | Method for preparing active ingredient polysaccharide from schisandra chinensis herb residues |
| CN102757510A (en) * | 2012-04-28 | 2012-10-31 | 江苏大学 | Low-molecular weight schisandra chinensis polysaccharides and preparation method and application thereof |
| CN102805762A (en) * | 2011-06-02 | 2012-12-05 | 上海华拓医药科技发展股份有限公司 | Chinese magnoliavine fruit polysaccharide extract and preparation method and application thereof |
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| CN102321186A (en) * | 2011-08-02 | 2012-01-18 | 江苏大学 | Carboxymethylated auricularia auricula polysaccharide and crude polysaccharide, and preparation method and application thereof |
| CN102492050A (en) * | 2011-12-01 | 2012-06-13 | 通化师范学院 | Method for preparing active ingredient polysaccharide from schisandra chinensis herb residues |
| CN102757510A (en) * | 2012-04-28 | 2012-10-31 | 江苏大学 | Low-molecular weight schisandra chinensis polysaccharides and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 五味子多糖的分离纯化及清除自由基研究;孟宪军等;《食品科学》;20080115(第01期);第91-94页 |
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