CN106632709A - Carboxymethylated schisandra polysaccharide and preparation method and application thereof - Google Patents
Carboxymethylated schisandra polysaccharide and preparation method and application thereof Download PDFInfo
- Publication number
- CN106632709A CN106632709A CN201610313007.8A CN201610313007A CN106632709A CN 106632709 A CN106632709 A CN 106632709A CN 201610313007 A CN201610313007 A CN 201610313007A CN 106632709 A CN106632709 A CN 106632709A
- Authority
- CN
- China
- Prior art keywords
- carboxy methylation
- fructus schisandrae
- polysaccharide
- fruit
- schisandrae polysaccharide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 113
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 112
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 112
- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- 235000008422 Schisandra chinensis Nutrition 0.000 title claims abstract description 11
- 240000006079 Schisandra chinensis Species 0.000 title claims description 59
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 40
- 238000006467 substitution reaction Methods 0.000 claims abstract description 16
- 230000035484 reaction time Effects 0.000 claims abstract description 10
- 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
- 239000008103 glucose Substances 0.000 claims abstract description 6
- 235000013373 food additive Nutrition 0.000 claims abstract description 4
- 239000002778 food additive Substances 0.000 claims abstract description 4
- 235000013376 functional food Nutrition 0.000 claims abstract description 4
- 230000000091 immunopotentiator Effects 0.000 claims abstract description 3
- 230000011987 methylation Effects 0.000 claims description 93
- 238000007069 methylation reaction Methods 0.000 claims description 93
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 90
- 235000013399 edible fruits Nutrition 0.000 claims description 51
- 235000009508 confectionery Nutrition 0.000 claims description 44
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 33
- 238000000502 dialysis Methods 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
- 238000001556 precipitation Methods 0.000 claims description 23
- 239000012153 distilled water Substances 0.000 claims description 18
- FDRCDNZGSXJAFP-UHFFFAOYSA-M sodium chloroacetate Chemical compound [Na+].[O-]C(=O)CCl FDRCDNZGSXJAFP-UHFFFAOYSA-M 0.000 claims description 18
- 238000005119 centrifugation Methods 0.000 claims description 17
- 238000001291 vacuum drying Methods 0.000 claims description 17
- 230000004044 response Effects 0.000 claims description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 15
- 238000005238 degreasing Methods 0.000 claims description 14
- 239000011780 sodium chloride Substances 0.000 claims description 14
- 239000003480 eluent Substances 0.000 claims description 11
- 238000010828 elution Methods 0.000 claims description 11
- 239000000284 extract Substances 0.000 claims description 11
- 206010010254 Concussion Diseases 0.000 claims description 10
- 230000009514 concussion Effects 0.000 claims description 10
- AZHSSKPUVBVXLK-UHFFFAOYSA-N ethane-1,1-diol Chemical compound CC(O)O AZHSSKPUVBVXLK-UHFFFAOYSA-N 0.000 claims description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 238000003809 water extraction Methods 0.000 claims description 8
- 239000000376 reactant Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 238000013019 agitation Methods 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 claims description 4
- 238000004220 aggregation Methods 0.000 claims description 4
- 230000002776 aggregation Effects 0.000 claims description 4
- 235000011869 dried fruits Nutrition 0.000 claims description 4
- 239000003208 petroleum Substances 0.000 claims description 4
- 150000004804 polysaccharides Polymers 0.000 claims description 4
- 235000019640 taste Nutrition 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 201000002282 venous insufficiency Diseases 0.000 claims description 2
- 230000006837 decompression Effects 0.000 claims 1
- 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
- 238000006243 chemical reaction Methods 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 11
- 210000002966 serum Anatomy 0.000 abstract description 10
- 102000000588 Interleukin-2 Human genes 0.000 abstract description 8
- 108010002350 Interleukin-2 Proteins 0.000 abstract description 8
- 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
- 108060008682 Tumor Necrosis Factor Proteins 0.000 abstract description 7
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 abstract description 7
- 210000000952 spleen Anatomy 0.000 abstract description 7
- 230000004048 modification Effects 0.000 abstract description 6
- 238000012986 modification Methods 0.000 abstract description 6
- 210000001541 thymus gland Anatomy 0.000 abstract description 6
- 102100037850 Interferon gamma Human genes 0.000 abstract description 5
- 108010074328 Interferon-gamma Proteins 0.000 abstract description 5
- 230000001506 immunosuppresive effect Effects 0.000 abstract description 3
- 239000002957 persistent organic pollutant Substances 0.000 abstract description 3
- 150000002772 monosaccharides Chemical class 0.000 abstract description 2
- 241000736075 Schisandra Species 0.000 abstract 5
- 206010062016 Immunosuppression Diseases 0.000 abstract 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 abstract 1
- FOCAUTSVDIKZOP-UHFFFAOYSA-N chloroacetic acid Chemical compound OC(=O)CCl FOCAUTSVDIKZOP-UHFFFAOYSA-N 0.000 abstract 1
- 229940106681 chloroacetic acid Drugs 0.000 abstract 1
- 229930182830 galactose Natural products 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 32
- 238000000034 method Methods 0.000 description 22
- 241000699670 Mus sp. Species 0.000 description 12
- 238000011160 research Methods 0.000 description 12
- 239000000523 sample Substances 0.000 description 9
- 238000001035 drying Methods 0.000 description 8
- 238000003304 gavage Methods 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 6
- 238000012512 characterization method Methods 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 206010001497 Agitation Diseases 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- BZHJMEDXRYGGRV-UHFFFAOYSA-N Vinyl chloride Chemical compound ClC=C BZHJMEDXRYGGRV-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000003651 drinking water Substances 0.000 description 4
- 235000020188 drinking water Nutrition 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000001900 immune effect Effects 0.000 description 4
- 238000005342 ion exchange Methods 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 238000009776 industrial production Methods 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- 238000000611 regression analysis Methods 0.000 description 3
- 210000002784 stomach Anatomy 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 244000025254 Cannabis sativa Species 0.000 description 2
- 241000219991 Lythraceae 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
- 230000021736 acetylation Effects 0.000 description 2
- 238000006640 acetylation reaction Methods 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000013401 experimental design Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- -1 has Phosphation Chemical class 0.000 description 2
- 230000007233 immunological mechanism Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 230000016507 interphase Effects 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000012821 model calculation Methods 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000002688 persistence Effects 0.000 description 2
- 238000010926 purge Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 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
- 241000196324 Embryophyta Species 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 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
- 238000010162 Tukey test Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 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
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000007688 immunotoxicity Effects 0.000 description 1
- 231100000386 immunotoxicity Toxicity 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 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
- 238000000569 multi-angle light scattering Methods 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 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
- 235000019353 potassium silicate Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000012744 reinforcing agent Substances 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 238000004621 scanning probe microscopy Methods 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 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
- 238000012109 statistical procedure Methods 0.000 description 1
- 238000005211 surface analysis Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
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
Landscapes
- 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 carboxymethylated schisandra polysaccharide and a preparation method and application thereof and belongs to the technical field of polysaccharide modification. Yield of carboxymethylated schisandra coarse polysaccharide is 88.37+/-1.65%, carboxymethylation substitution degree is 0.724+/-0.022, and solubility is 0.5+/-0.04mg/mL, which is 5 times of that of original schisandra polysaccharide. Solubility of the carboxymethylated schisandra polysaccharide is 12.5+/-0.26mg/mL, molecular weight is 3.4x106 Da, and monosaccharide is composed of mannose, glucose and galactose according to a molar ratio of 1:44.84:3.71. When reaction temperature is 40-80 DEG C, reaction time is 1-8h and final concentration of chloroacetic acid is 0.005-0.08g/mL, the carboxymethylated schisandra polysaccharide is high in yield, substitution degree and water solubility, has remarkable effect on improving mouse immunosuppression caused by persistent organic pollutant PCB126, can improve mouse spleen and thymus index and increase content of TNF-alpha, IFN-gamma and IL-2 in serum, can serve as an immunopotentiator and can be used as functional food and a food additive.
Description
Technical field
The present invention relates to a kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application, belongs to polysaccharide-modified technology neck
Domain.
Background technology
The fruit of Chinese magnoliavine (Schisandra chinensis (Turcz.) Baill.) belongs to the mellow fruit of Magnoliaceae fruit of Chinese magnoliavine plant
Real, main originates in the ground such as China Heilungkiang, Liaoning, the Inner Mongol, is a kind of common nourishing Chinese medicine, with high medicinal
And edibility, be put into can be used for health food article list (land rabbit woods, Wu Yang, season moral it is black. Fructus Schisandrae Polysaccharide extract
Separate 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 component main in the fruit of Chinese magnoliavine, its content is higher, and with various lifes such as antitumor, anti-oxidant, anti-aging, antifatigue
Thing activity (Xu Bo, Shen Nan, Zhao Lijing. each composition latest Progress of the fruit of Chinese magnoliavine. Aged in China physical-fitness medicine, 2013,11 (5)：
74-74.), it is and minimum to the toxicity of organism normal cell and tissue, with preferable development and application values.
At present, the structural modification method of polysaccharide mainly has Phosphation, carboxy methylation, acetylation etc..Carboxy methylation is modified
As a kind of main chemical modification method, it is directed to introduce carboxymethyl on polysaccharide chain, solubility of this kind of method of modifying to polysaccharide
And electronegativity have more important impact (Jiang Leming, Fan Canmei, Nie Shaoping. the preparation of carboxy methylation semen plantaginis major polysaccharide and
Its bioactivity research. Food Science, 2013,34 (22)：10-14.).Research finds that a kind of alkali solubility tremella polysaccharides is insoluble
Yu Shui, by carboxy methylation modify after, its solubility in water improve to 36.38mg/mL (Wu Qiong. carboxy methylation alkali soluble
The research of property tremella polysaccharides. Changchun University's journal：Natural science edition, 2009,19 (10)：66-68.).2005, Guo offered sacriffices to the gods or the spirits of the dead remote etc.
Patent " carboxy methyl schizophllan polysacharide preparation method and apply in cosmetics and the antineoplastic " (application number of application：
CN200510101678.X in), carboxymethyl modification is carried out to schizophan using conventional reagents such as monoxone, NaOH, is changed
It has been apt to its solubility property and has maintained its good rheological property and antitumor activity.2011, face upward the patent of the applications such as pomegranate green grass or young crops
A kind of " carboxy methylation Blackfungus polyhexose, Thick many candies and its preparation method and application " (application number：CN201110219047.3 in),
Carboxymethyl modification is carried out to Blackfungus polyhexose using isopropanol method so as to which solubility reaches 0.6mg/mL, be the 6 of raw sugar solubility
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 antitumor activity,
Fructus Schisandrae Polysaccharide it is water-soluble undesirable, constrain its further further investigation and development and application.Therefore, the present inventor is to five
Taste polysaccharide carries out carboxy methylation modification, primarily determines that its architectural feature, there is provided one kind optimizes carboxymethyl using Response Surface Method
The technique for changing Fructus Schisandrae Polysaccharide, and caused by have studied the carboxy methylation Fructus Schisandrae Polysaccharide to persistence organic pollutant PCB126
The intervention effect of immunotoxicity.The research of preparation and its process optimization about Fructus Schisandrae Polysaccharide carboxyl methylation derivant is so far not
See that domestic and foreign literature is reported, also have no that corresponding patent is disclosed.
The content of the invention
The invention aims to solve the water-soluble undesirable of Fructus Schisandrae Polysaccharide, constrain it and further deeply grind
Study carefully and development and application shortcoming, there is provided a kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application.
The present invention is adopted the following technical scheme that：A kind of carboxy methylation Fructus Schisandrae Polysaccharide, the carboxy methylation Fructus Schisandrae Polysaccharide
Molecular weight be 1.698 × 104Da, the monose of the carboxy methylation Fructus Schisandrae Polysaccharide is consisted of：Mannose, glucose and gala
Sugar, mol ratio is 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 forms strand in aggregation shape in aqueous, and the thickness of the strand is 0.1-1nm.
The preparation method of carboxy methylation Fructus Schisandrae Polysaccharide, is carried out as steps described below：
(1) preparation of fruit of Chinese magnoliavine Thick many candies：New fresh schisandra chinensis are dried after 12-24h in 55-65 DEG C of baking oven carries out powder
It is broken, degreasing 2-3 time that flows back is carried out to the dried fruit of Chinese magnoliavine using petroleum ether, then by the fruit of Chinese magnoliavine dregs of a decoction after degreasing with feed liquid
Than 1:10-1:15 addition distilled water carry out water extraction, extract 2-3 time and obtain Fructus Schisandrae Polysaccharide extract, Fructus Schisandrae Polysaccharide extract
Jing reduced pressure concentrations, using 80% ethanol alcohol precipitation 3-4 time, the alcohol hypostasis for obtaining redissolves, waves alcohol, Jing retention molecules Jing after centrifugation
The bag filter dialysis 12-36h for 3000Da is measured, vacuum drying 2-3h obtains fruit of Chinese magnoliavine Thick many candies；
(2) carboxy methylation of Fructus Schisandrae Polysaccharide：Fruit of Chinese magnoliavine Thick many candies are dissolved in aqueous isopropanol, stirring makes it fully molten
Solution, fruit of Chinese magnoliavine 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
15-20min, mixing speed is 60-240r/min, adds mass fraction to be 15- in the Fructus Schisandrae Polysaccharide solution after dissolving
20% NaOH solution is alkalized, and the addition of NaOH solution is the 20-30% of isopropanol volume, and alkalization temperature is 20-25
DEG C, alkalization time is 1-2h, then continues to add chloroethene in the fruit of Chinese magnoliavine Thick many candies reactant liquor after alkalization under stirring
Acid carries out carboxy methylation, and monoxone addition is 0.005-0.08g/mL, and bath temperature is 40-80 DEG C, during carboxymethylation reaction
Between be 1-8h, question response liquid cooling after adjust reactant liquor pH to neutrality, using ethanol alcohol precipitation, alcohol precipitation product is obtained after centrifugation, will
Using being dialysed after the redissolution of a small amount of distilled water, dialysis carries out vacuum drying and obtains the carboxy methylation fruit of Chinese magnoliavine alcohol precipitation product after terminating
Thick many candies；
(3) purify：The carboxy methylation fruit of Chinese magnoliavine Thick many candies that step (2) is obtained are dissolved in the NaCl of 0.05-0.1mol/L
In solution, then it is slowly added dropwise into DEAE-52 chromatographic columns, and the NaCl solution using 0.05-0.2mol/L carries out gradient and washes
It is de-, elution curve is drawn, merging eluent is collected respectively according to elution curve, obtain carboxy methylation Fructus Schisandrae Polysaccharide difference group
Point, the elution fraction for choosing 0.05mol/L NaCl merges, and Jing molecular cut offs are dialysed for the bag filter of 3000Da, vacuum drying
Afterwards, carboxy methylation Fructus Schisandrae Polysaccharide is obtained.
Further, the yield of fruit of Chinese magnoliavine Thick many candies is 15.9-19.8% in the step (1), and polyoses content is 54.4-
65.7%, solubility is 0.08-0.12mg/mL.
Further, in the step (2) carboxy methylation fruit of Chinese magnoliavine Thick many candies yield be 60.2-86.4%, carboxymethyl
Change substitution value is 0.41-0.71, and solubility is 0.46-0.54mg/mL.
Further, the temperature of backflow degreasing is 70-80 DEG C in the step (1), and degreasing time is 1-2h, water extraction temperature
For 90-100 DEG C, extraction time is 4-6h.
Further, 80% ethanol alcohol precipitation 24-48h is adopted in the step (2), precipitation solution is with 4000-5000rpm rotating speeds
Centrifugation 5-10min.
Further, adopt Jing molecular cut offs in the step (2) to dialyse initially with stream for the bag filter of 3000Da
Water is dialysed, and 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 for-
0.085--0.1Mpa。
Further, carboxy methylation fruit of Chinese magnoliavine Thick many candies Jing concussions heating for dissolving in NaCl solution in the step (3),
The temperature of shaking water bath is 50-60 DEG C, and concussion frequency is 120-160 beat/min, the time 10-12h of dissolving is shaken, then to molten
Solution after solution is centrifuged, and the rotating speed of solution centrifugal is：4000-5000r/min, centrifugation time is 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 is -0.085--0.1MPa.
Further, the carboxy methylation Fructus Schisandrae Polysaccharide can be used for functional food and food additive as immunopotentiator
Plus in agent.
Preparation method of the present invention is simple, and step is easily operated, and using the method for molecular modification the carboxy methylation five tastes are prepared
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 substitution value are higher, and dissolubility is good, and solubility is respectively 5 times of former Fructus Schisandrae Polysaccharide and 125 times, the carboxylic for preparing
The Fructus Schisandrae Polysaccharide that methylates can be effectively improved the immunosuppressive action of persistence organic pollutant PCB126, can be used as immunity
Reinforcing agent, is applied to functional food and food additives.
Description of the drawings
Fig. 1 is that reaction temperature, reaction time, the interphase interaction of monoxone final concentration are obtained to carboxy methylation Fructus Schisandrae Polysaccharide
Response surface figure (A, B, C) and contour map (D, E, F) that rate affects.
Fig. 2 is that reaction temperature, reaction time, the interphase interaction of monoxone final concentration take to carboxy methylation Fructus Schisandrae Polysaccharide
The response surface figure (A, B, C) affected for degree and contour map (D, E, F).
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), graphics (B) and the strand height distribution map of carboxy methylation Fructus Schisandrae Polysaccharide
(C)。
Fig. 6 is impact figure 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 groups.
Fig. 7 is impact figure of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice thymus indexs,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 groups.
Fig. 8 is impact figure of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice serum TNF-cc levels,ap<0.05,
Compared with blank control group；bp<0.05, compared with model group.
Fig. 9 is impact figure of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice serum I NF- γ levels,ap<
0.05, compared with blank control group；bp<0.05, compared with model group.
Figure 10 is impact figure of the carboxy methylation Fructus Schisandrae Polysaccharide to PCB126 exposed Mice serum IL-2 levels,ap<0.05,
Compared with blank control group；bp<0.05, compared with model group；cp<0.05, compared with CSP1 high dose groups.
Specific embodiment
The preparation of fruit of Chinese magnoliavine Thick many candies
Embodiment 1：
New fresh schisandra chinensis are dried after 12h in 55 DEG C of baking ovens and are crushed, using petroleum ether to the dried fruit of Chinese magnoliavine
Flow back degreasing 1h in 70 DEG C, 2 backflow degreasings is carried out, then by the fruit of Chinese magnoliavine dregs of a decoction after degreasing with solid-liquid ratio 1:10 (g/mL) add
Entering distilled water carries out water extraction, in 90 DEG C of water extraction 4h, extracts 2 times, and extract merges after reduced pressure concentration, 80% ethanol alcohol repeatedly
Heavy 3 times, after centrifugation, supernatant Jing molecular cut offs are bag filter dialysis 12h, the vacuum drying 2h of 3000Da, and the fruit of Chinese magnoliavine is obtained
Thick many candies, polysaccharide yield is 15.9 ± 0.3%, and polyoses content is 54.4 ± 2.7%.
Embodiment 2
New fresh schisandra chinensis are dried after 24h in 65 DEG C of baking ovens and are crushed, using petroleum ether to the dried fruit of Chinese magnoliavine
Flow back degreasing 2h in 80 DEG C, 3 backflow degreasings is carried out, then by the fruit of Chinese magnoliavine dregs of a decoction after degreasing with solid-liquid ratio 1:15 (g/mL) add
Entering distilled water carries out water extraction, in 100 DEG C of water extraction 6h, extracts 3 times, and extract merges after reduced pressure concentration, and 80% ethanol is repeatedly
Alcohol precipitation 4 times, after centrifugation, supernatant Jing molecular cut offs are bag filter dialysis 36h, the vacuum drying 3h of 3000Da, and the five tastes are obtained
Sub- Thick many candies, polysaccharide yield is 19.8 ± 0.9%, and polyoses content is 65.7 ± 2.7%, and solubility is 0.10 ± 0.02mg/mL.
Raw material with the fruit of Chinese magnoliavine Thick many candies of the preparation of embodiment 2 as next step.
Compared with Fructus Schisandrae Polysaccharide preparation technology in existing document (Zhao Ting. the isolating and purifying of Fructus Schisandrae Polysaccharide, structural characterization
And its activity research [D]. Jiangsu University, 2009.), the preparation technology of the Thick many candies adopts alcohol precipitation 3-4 time repeatedly of 80% ethanol,
The polyoses content of fruit of Chinese magnoliavine Thick many candies is significantly improved, polyoses content improves about 21% or so；And adopt the boulton process can
Effectively shorten drying time, drying time reduces about 93% or so, the drying means can energy saving, be conducive to industrial production
In application.
The preparation of carboxy methylation fruit of Chinese magnoliavine Thick many candies
Embodiment 3：
Fruit of Chinese magnoliavine Thick many candies sample is weighed with isopropanol with solid-liquid ratio 1:30 (g/mL) mix, and at 20 DEG C, are with rotating speed
60r/min mechanical agitations 15min, are fully dissolved；Addition mass fraction is in the Fructus Schisandrae Polysaccharide solution after dissolving
15% NaOH solution is alkalized, and the addition of NaOH solution is the 20% of isopropanol volume, and machinery is continued at 20 DEG C of temperature
Stirring 1h；Monoxone is added in above-mentioned reactant liquor, makes chloroethene acid concentration reach 0.005g/mL, be placed in 40 DEG C of waters bath with thermostatic control
In pot, continue mechanic whirl-nett reaction 1h, product is adjusted to into neutrality, the ethanol alcohol precipitation 24h of Jing 80%, alcohol precipitation with hydrochloric acid after cooling
After liquid is with 4000rpm rotating speeds centrifugation 5min, precipitation Jing redissolves, and Jing molecular cut offs are the bag filter flowing water dialysis 36h of 3000Da,
After distilled water dialysis 24h, 2h is vacuum dried under 20 DEG C, vacuum -0.085MPa, obtains final product carboxy methylation fruit of Chinese magnoliavine Thick many candies
(CSP), polysaccharide yield is 75.9 ± 1.1%, and carboxy methylation substitution value is 0.41 ± 0.02.
Embodiment 4：
Fruit of Chinese magnoliavine Thick many candies sample is weighed with isopropanol with solid-liquid ratio 1:35 (g/mL) mix, and at 25 DEG C, are with rotating speed
240r/min mechanical agitations 20min, are fully dissolved；Addition mass fraction is in the Fructus Schisandrae Polysaccharide solution after dissolving
20% NaOH solution is alkalized, and the addition of NaOH solution is the 25% of isopropanol volume, and machinery is continued at 25 DEG C of temperature
Stirring 2h；Monoxone is added in above-mentioned reactant liquor, makes chloroethene acid concentration reach 0.02g/mL, be placed in 60 DEG C of thermostat water baths
In, continue mechanic whirl-nett reaction 4h, product is adjusted to into neutrality, the ethanol alcohol precipitation 36h of Jing 80%, precipitation solution with hydrochloric acid after cooling
After with 4000rpm rotating speeds centrifugation 10min, precipitation Jing redissolves, and Jing molecular cut offs are the bag filter flowing water dialysis 48h of 3000Da,
After distilled water dialysis 36h, 3h is vacuum dried under 25 DEG C, vacuum -0.1MPa, obtains final product carboxy methylation fruit of Chinese magnoliavine Thick many candies
(CSP), polysaccharide yield is that polysaccharide yield is 86.4 ± 1.5%, and carboxy methylation substitution value is 0.71 ± 0.03, and solubility is 0.50
±0.04mg/mL。
Embodiment 5：
Fruit of Chinese magnoliavine Thick many candies sample is weighed with isopropanol with solid-liquid ratio 1:40 (g/mL) mix, and at 25 DEG C, are with rotating speed
240r/min mechanical agitations 20min, are fully dissolved；Addition mass fraction is in the Fructus Schisandrae Polysaccharide solution after dissolving
20% NaOH solution is alkalized, and the addition of NaOH solution is the 30% of isopropanol volume, and machinery is continued at 25 DEG C of temperature
Stirring 2h；Monoxone is added in above-mentioned reactant liquor, makes chloroethene acid concentration reach 0.08g/mL, be placed in 80 DEG C of thermostat water baths
In, continue mechanic whirl-nett reaction 8h, product is adjusted to into neutrality, the ethanol alcohol precipitation 48h of Jing 80%, precipitation solution with hydrochloric acid after cooling
After with 5000rpm rotating speeds centrifugation 10min, precipitation Jing redissolves, and Jing molecular cut offs are the bag filter flowing water dialysis 48h of 3000Da,
After distilled water dialysis 36h, 3h is vacuum dried under 25 DEG C, vacuum -0.1MPa, obtains final product carboxy methylation fruit of Chinese magnoliavine Thick many candies
(CSP), polysaccharide yield is 60.2 ± 2.2%, and carboxy methylation substitution value is 0.68 ± 0.02.
Response phase method optimizes carboxy methylation fruit of Chinese magnoliavine Thick many candies synthesis technique
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 chooses tri- levels of 0.01g/mL, 0.02g/mL and 0.04g/mL, utilizes
The softwares of Design Expert 8.0 carry out experimental design according to Box-Benhnken center combination designs principle, with reaction temperature
(A), reaction time (B) and monoxone final concentration (C) are independent variable, with the yield (Y of CSP1) and substitution value (Y2) it is response,
Optimize fruit of Chinese magnoliavine 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 the yield of table 3
Note:P ＜ 0.0001 are for extremely significantly, P ＜ 0.05 are notable.
Regression analysis is carried out to experimental data using the softwares of Design-Expert 8, yield test is tied according to each factor
The impact 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 is respectively provided with more significant impact (P<0.05).Three factors are suitable for the impact of yield
Sequence is：Monoxone final concentration>Reaction temperature>Reaction time.Test model P values<0.0001, it was demonstrated that model is notable.Lose and intend error
Not notable (F value=2.14 of value；P values=0.2374), illustrate that assay is not significantly different from 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 substitution value
Note:P ＜ 0.0001 are for extremely significantly, P ＜ 0.05 are notable
Regression analysis is carried out to experimental data using the softwares of Design-Expert 8, substitution value is tested according to each factor
As a result impact 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 substitution value.
The substitution value of each factor pair CSP has more significant impact (P<0.05).Three factors are for the impact of substitution value
Sequentially it is：Monoxone final concentration>Reaction time>Reaction temperature.The P values of test model<0.0001, it was demonstrated that model is notable.Lose and intend
Not notable (F value=6.37 of error amount；P values=0.0528), illustrate that assay is not significantly different from the model calculation, table
Bright regression model is fine with actual conditions fitting, can be used for the theoretical prediction of Fructus Schisandrae Polysaccharide carboxy methylation compound experiment.
Plot analysis are carried out according to multiple regression equation using the softwares of Design Expert 8.0, regression equation is obtained
Yield and substitution value of three factors of response surface figure (see Fig. 1 and Fig. 2) for CSP is respectively provided with significant impact, wherein monoxone
The impact of final concentration is most notable.
(5) interpretation and optimization：
Regression analysis calculating is carried out to experimental data using the softwares of Design-Expert 8, obtains synthesizing the optimal work of CSP
Skill condition is：Reaction temperature is 63 DEG C, and the reaction time is 4.5h, the final concentration of 0.022g/mL of monoxone.Under this condition, put down
Row experiment 5 times, the yield for obtaining CSP is 88.3 ± 1.65%, and substitution value is 0.72 ± 0.022.Actual measured value and response surface
Model prediction result is closer to, and illustrates that the response phase method optimizes the model for obtaining accurately and reliably, it is adaptable to fruit of Chinese magnoliavine Thick many candies
Carboxymethylated modification.With 2011, face upward the applications such as pomegranate green grass or young crops patent " a kind of carboxy methylation Blackfungus polyhexose, Thick many candies and its
Preparation method and application " (application number：CN201110219047.3) compare, monoxone consumption is reduced in the process conditions
46%, the reduction of monoxone consumption can reduce the generation of polysaccharide degraded side reaction in course of reaction so that carboxymethylated polysaccharides are obtained
Rate is significantly improved, and improves about 30% or so；Secondly, magnetic agitation is replaced from mechanical agitation in the process conditions, stirring is strong
Degree is big, good mixing effect, it is easy to adjusts and operates, is more suitable for a large amount of preparations of carboxy methylation Fructus Schisandrae Polysaccharide；And adopt true
Empty seasoning can effectively shorten drying time, and energy saving is conducive to the application in industrial production.
Embodiment 6：
The optimal processing parameter reaction temperature that response surface is obtained is adopted for 63 DEG C, the reaction time is 4.5h, and monoxone is dense eventually
Spend for 0.022g/mL, prepare carboxy methylation fruit of Chinese magnoliavine Thick many candies.
The purifying and architectural feature of carboxy methylation fruit of Chinese magnoliavine Thick many candies
Preparation carboxy methylation fruit of Chinese magnoliavine Thick many candies (CSP) 0.5g in above-described embodiment 6 is weighed, the carboxy methylation fruit of Chinese magnoliavine is carried out
The purifying of Thick many candies.
Embodiment 7：
0.05mol/LNaCl solution is dissolved in, is 160 beats/min, is filled in the concussion water bath with thermostatic control of temperature 50 C in concussion frequency
Divide dissolving 10h, lysate Jing 4000r/min centrifugation 10min, after removing insoluble matter, supernatant passes through DEAE-52 ion exchanges
Post (2.0 × 50cm) is separated；Eluted with 0.05,0.1,0.15 and 2mol/L NaCl solution, with automatic collector by pipe
Eluent is collected, by the flow control of constant flow pump in 1mL/min, often pipe collection 5mL.Collection liquid sulfuric acid-phynol method is examined by pipe
Survey, merging eluent, the eluent Jing reduced pressure concentrations of wherein 0.05mol/L NaCl, retention point are collected respectively according to elution curve
Son amount for 3000Da bag filter flowing water dialysis 48h, distilled water dialysis 24h after, in 20 DEG C of temperature, vacuum be -0.085MPa
Vacuum drying chamber in be dried 2h, obtain carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1).
Embodiment 8：
0.08mol/LNaCl solution is dissolved in, is 150 beats/min, is filled in the concussion water bath with thermostatic control of 55 DEG C of temperature in concussion frequency
Divide dissolving 11h, lysate Jing 4500r/min centrifugation 9min, after removing insoluble matter, supernatant passes through DEAE-52 ion exchange columns
(2.0 × 50cm) is separated；Eluted with 0.05,0.1,0.15 and 2mol/L NaCl solution, received by pipe with automatic collector
Collection eluent, by the flow control of constant flow pump in 1mL/min, often pipe collection 5mL.Collection liquid sulfuric acid-phynol method by pipe detect,
Merging eluent is collected respectively according to elution curve, the eluent Jing reduced pressure concentrations of wherein 0.05mol/L NaCl retain molecule
Measure for 3000Da bag filter flowing water dialysis 40h, distilled water dialysis 30h after, in 21 DEG C of temperature, vacuum be -0.090MPa's
2.5h is dried in vacuum drying chamber, carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) is obtained.
Embodiment 9：
0.1mol/LNaCl solution is dissolved in, is 120 beats/min, is filled in the concussion water bath with thermostatic control of temperature 60 C in concussion frequency
Divide dissolving 12h, lysate Jing 5000r/min centrifugation 8min, after removing insoluble matter, supernatant passes through DEAE-52 ion exchange columns
(2.0 × 50cm) is separated；Eluted with 0.05,0.1,0.15 and 2mol/L NaCl solution, received by pipe with automatic collector
Collection eluent, by the flow control of constant flow pump in 1mL/min, often pipe collection 5mL.Collection liquid sulfuric acid-phynol method by pipe detect,
Merging eluent is collected respectively according to elution curve, the eluent Jing reduced pressure concentrations of wherein 0.05mol/L NaCl retain molecule
Measure for 3000Da bag filter flowing water dialysis 36h, distilled water dialysis 36h after, in 25 DEG C of temperature, vacuum be the true of -0.1MPa
3h is dried in empty drying box, carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) is obtained.
With existing document (Zhao Ting. the isolating and purifying of Fructus Schisandrae Polysaccharide, structural characterization and its activity research [D]. Jiangsu is big
Learn, 2009.) with patent (application number：CN201110219047.3 and CN201210132340.0) compare, it is many in the purge process
Sugared Jing isothermal vibrations are fully dissolved, and reduce the waste of carboxy methylation Fructus Schisandrae Polysaccharide raw material；And post purge process is adopted
The ion exchange column being relatively large in diameter so that applied sample amount increased one times, dramatically speeds up on the basis of purification effect is not changed
Purifying speed；And adopting boulton process effectively to shorten drying time, energy saving is conducive to the application in industrial production.
Example 8 prepares carboxy methylation Fructus Schisandrae Polysaccharide (CSP1), and solubility is 12.50mg/mL.
The measure of solubility is carried out according to 2005 editions pharmacopeia, the solubility of fruit of Chinese magnoliavine Thick many candies is 0.10 ± 0.03mg/mL, carboxylic
Methylate fruit of Chinese magnoliavine Thick many candies solubility be 0.50 ± 0.04mg/mL, the solubility of carboxy methylation Fructus Schisandrae Polysaccharide (CSP1)
For 12.50 ± 0.26mg/mL.
The measure of molecular weight：Using efficient gel size exclusion chromatography-multi-angle laser light scattering instrument-differential pulse polarograpll
Instrument combination analysis method (HPSEC-MALLS-RI System) is measured to polysaccharide relative molecular mass.Chromatographic condition is with TSK-
GEL series G6000PWXL and G4000PWXL chromatographic columns (300mm, 7.8mm, TOSOH, Japan) are analytical column, with 0.15mol/L
NaNO3 and 0.05mol/L NaH2PO4 (pH=7) are mobile phase, under conditions of column temperature is for 35 DEG C, with the stream of 0.5mL/min
Speed carries out elution analysis.By CSP1 sample preparations into the solution that mass concentration is 5mg/mL, the high speed centrifugation of Jing 12000r/min
Supernatant is taken after machine centrifugation 10min cross the μ L of sample introduction 100 after film.CSP1 molecular weight is 1.698 × 104Da.With existing document (Zhao
It is graceful. the isolating and purifying of Fructus Schisandrae Polysaccharide, structural characterization and its activity research [D]. Jiangsu University, 2009.) and patent (application number：
CN201110219047.3 and CN201210132340.0) compare, CSP1 molecular weights, and the polysaccharide of molecular weight dissolves
Spend performance that is high and being easier to 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 plus after pyrohydrolysis 8h, hydrolyzate adds brium carbonate to be neutralized to neutrality, and with 4000r/min BaSO is centrifuged off4
Precipitation, supernatant freeze-drying.Hydrolysate is analyzed Jing after acetylation with the gas chromatography system of Shimadzu GC 2010.The list of CSP1
Sugar is consisted of：Mannose, glucose and galactolipin, its mol ratio is 1:44.84:3.71.It can be seen that CSP1 is made up of three kinds of monose,
Respectively mannose, glucose and galactolipin, and existing document (Zhao Ting. the isolating and purifying of 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 monose structures
Into (rhamnose, arabinose, wood sugar, mannose, glucose and galactolipin).
Infrared analysis：With KBr (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 Jing carboxy methylations
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 absworption peak of-C-O- at left and right),
Show carboxy methylation success.
Atomic force Electronic Speculum：Polysaccharide sample is configured to into the solution of 1mg/mL, film is crossed and is collected filtrate, taking 20 μ L filtrates makes it
Uniformly it is attached on clean mica sheet, anti-gray is dried under room temperature.Sample is seen with scanning probe microscopy (MFP-3D)
Survey.In aqueous in aggregation shape, the thickness of strand is in 0.1-1nm.With existing document (Zhao Ting. the structure of Fructus Schisandrae Polysaccharide,
Biologically active and its immunologic mechanism research [D]. Jiangsu University, 2013.) to compare, carboxy methylation Fructus Schisandrae Polysaccharide is in aqueous
Aggregation reduce, and polysaccharide chain thickness significantly reduces compared with Fructus Schisandrae Polysaccharide component (1-5nm).
Carboxy methylation Fructus Schisandrae Polysaccharide causes the intervention effect of immunity of organism toxicity to PCB126
Animal used as test
ICR mouse (cleaning grade), male and female half and half, body weight is 20 ± 2g, is purchased from Jiangsu University's animal experimental center, tests it
Front quarantine 3 days, raising temperature scope is 21 ± 1 DEG C, and it is 60 ± 5% to raise humidity range.
Material and reagent
The carboxy methylation Fructus Schisandrae Polysaccharide that the purifying of Example 8 is obtained；
PCB126, physiological saline, picric acid；
TNF-α enzyme linked immunological kit, IL-2 enzyme linked immunological kits, IFN-γ enzyme linked immunological kit.
Experiment packet and administration
Adapt to feed 70 ICR mouse before experiment after 3 days, be randomly divided into 5 groups, 14 per group, male and female half and half simultaneously separate
Feed, blank control group, PCB126 model groups, the high, medium and low three kinds of concentration of carboxy methylation Fructus Schisandrae Polysaccharide component (CSP1) are set
Dosage group.It is administered once daily, continuous 14 days.And in administration the 7th day, PCB126 model groups, CSP1 were high, in CSP1, CSP1 it is low by three
Simultaneously gavage gives 50 μ g/kg PCB126 (gavage amount is every 0.1mL) to plant concentration dose group, persistently to malicious 5 days.
Packet, dosage and method of administration are as follows：
Blank control group：Daily the physiological saline (0.1mL/20g/d) of gavage same dose, takes food and drinks water freely；
PCB126 model groups：Daily the physiological saline (0.1mL/20g/d) of gavage same dose, in the 7th day gavage was started
PCB126 is given, is given 5 days, feed and drinking-water are freely；
CSP1 administration groups：Divide high, medium and low three kinds of concentration dose groups,
CSP1 high dose groups：Daily the CSP1 (400mg/kg/d) of difference gavage same dose, starts to fill on the 7th day in administration
Stomach gives PCB126, gives 5 days, and feed and drinking-water are freely；
CSP1 middle dose groups：Daily the CSP1 (200mg/kg/d) of difference gavage same dose, starts to fill on the 7th day in administration
Stomach gives PCB126, gives 5 days, and feed and drinking-water are freely；
CSP1 low dose groups：Daily the CSP1 (100mg/kg/d) of difference gavage same dose, starts to fill on the 7th day in administration
Stomach gives PCB126, gives 5 days, and feed and drinking-water are freely.
(1) Immune Organs Index
After last dose, in next day fasting 8h, weigh the weight of animals and record.Extract eyeball and take execution after blood, take out chest
Gland, spleen are blotted Jing after physiological saline cleaning with filter paper, are weighed and are recorded.It is calculated as follows each organ index：
(2) physiochemical indice
The content of IL-2, INF- γ, TNF-α in mice serum is determined with ELISA enzyme linked immunological kits.
Statistical procedures
Data are analyzed with SPSS16.0 statistical softwares.As a result it is expressed asGroup difference adopts single factor test variance
Analysis (One-way ANOVA), using Tukey methods of inspection the significance of difference between each group is compared, and P ﹤ 0.05 indicate that conspicuousness is poor
It is different.Experimental result
CSP1 is shown in respectively Fig. 6 and Fig. 7 to index and spleen index, the thymus index of exposed Mice.Compared with blank control group, model
The mouse spleen and thymus index of group is remarkably decreased (P ＜ 0.05), illustrates that PCB126 causes mouse immune inhibition to be successfully established；
And the high, medium and low dosage of CSP1 has recovered in varying degrees the Thymus and spleen index of mouse, wherein CSP1 high dose groups 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 levels
Fig. 8, Fig. 9 and Figure 10 are shown in respectively in impact.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 contents are equal in the high, medium and low dosage mice serums of CSP1
The content for having different degrees of rising, the wherein TNF-α in CSP1 high dose groups mice serum significantly rises (P ＜ compared with model group
, and the content of IL-2 significantly rises (P ＜ 0.05) compared with model group in CSP1 high dose groups and middle dosage mice serum 0.05).Mesh
Before, existing document report, Fructus Schisandrae Polysaccharide (Zhao Ting. the structure of Fructus Schisandrae Polysaccharide, biologically active and its immunologic mechanism research [D].
Jiangsu University, 2013.) with Blackfungus polyhexose (Song Guanglei. Blackfungus polyhexose separates preparation and bioactivity research [D]. Zhejiang
Industrial and commercial university, 2011.) with immunoregulatory activity, can significantly improve the immunosuppressive action of caused by cyclophosphamide, but for holding
Long the intervention effect of immunity of organism toxicity has no relevant report caused by property organic pollution PCB126.
Claims (10)
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 monose of the carboxy methylation Fructus Schisandrae Polysaccharide is consisted of：Mannose, glucose and galactolipin, mol ratio is 1:
44.84:3.71, the solubility of the carboxy methylation Fructus Schisandrae Polysaccharide is 12.24-12.76 mg/mL, the carboxy methylation five tastes
Sub- polysaccharide forms strand in aggregation shape in aqueous, and the thickness of the strand is 0.1-1 nm.
2. the preparation method of the carboxy methylation Fructus Schisandrae Polysaccharide described in claim 1, it is characterised in that carry out as steps described below：
（1）The preparation of fruit of Chinese magnoliavine Thick many candies：New fresh schisandra chinensis are dried after 12-24h in 55-65 DEG C of baking oven and are crushed, adopted
Degreasing 2-3 time that flows back is carried out to the dried fruit of Chinese magnoliavine with petroleum ether, then by the fruit of Chinese magnoliavine dregs of a decoction after degreasing with solid-liquid ratio 1:
10-1:15 addition distilled water carry out water extraction, take 2-3 time and obtain Fructus Schisandrae Polysaccharide extract, Fructus Schisandrae Polysaccharide extract Jing decompressions
Concentration, using 80% ethanol alcohol precipitation 3-4 time, the alcohol hypostasis for obtaining redissolves, waves alcohol Jing after centrifugation, and Jing molecular cut offs are 3000
The bag filter dialysis 12-36h of Da, vacuum drying 2-3h obtain fruit of Chinese magnoliavine Thick many candies；
（2）The carboxy methylation of Fructus Schisandrae Polysaccharide：Fruit of Chinese magnoliavine Thick many candies are dissolved in aqueous isopropanol, mechanical agitation makes it fully molten
Solution, fruit of Chinese magnoliavine 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 is 60-240 r/min, adds mass fraction to be 15- in the Fructus Schisandrae Polysaccharide solution after dissolving
20% NaOH solution is alkalized, and the addition 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 to add monoxone in the fruit of Chinese magnoliavine Thick many candies reactant liquor after alkalization under stirring
Carboxy methylation is carried out, monoxone addition is 0.005-0.08 g/mL, and bath temperature is 40-80 DEG C, the carboxymethylation reaction time
For 1-8 h, reactant liquor pH is adjusted after the cooling of question response liquid to neutrality, using ethanol alcohol precipitation, alcohol precipitation product is obtained after centrifugation, by alcohol
, using being dialysed after the redissolution of a small amount of distilled water, dialysis carries out vacuum drying after terminating, and to obtain the carboxy methylation fruit of Chinese magnoliavine thick for heavy product
Polysaccharide；
（3）Purifying：By step（2）The carboxy methylation fruit of Chinese magnoliavine Thick many candies for obtaining are dissolved in the NaCl solution of 0.05-0.1mol/L
In, then it is slowly added dropwise into DEAE-52 chromatographic columns, and the NaCl solution using 0.05-0.2 mol/L carries out gradient elution,
Elution curve is drawn, merging eluent is collected respectively according to elution curve, obtain carboxy methylation Fructus Schisandrae Polysaccharide different component, selected
The elution fraction for taking 0.05mol/L NaCl merges, Jing after bag filter dialysis, vacuum drying that molecular cut off is 3000 Da,
Obtain carboxy methylation Fructus Schisandrae Polysaccharide.
3. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（1）
The yield of middle fruit of Chinese magnoliavine Thick many candies is 15.9-19.8%, and polyoses content is 54.4-65.7%, and solubility is 0.08-0.12 mg/
mL。
4. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（2）
The yield of middle carboxy methylation fruit of Chinese magnoliavine Thick many candies is 60.2-86.4%, and carboxy methylation substitution value is 0.41-0.71, and solubility is
0.46-0.54 mg/mL。
5. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（1）
The temperature of middle backflow degreasing is 70-80 DEG C, and degreasing time is 1-2 h, and water extraction temperature is 90-100 DEG C, and extraction time is 4-6 h.
6. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（2）
In adopt 80% ethanol alcohol precipitation 24-48h, precipitation solution is centrifuged 5-10 min with 4000-5000rpm rotating speeds.
7. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（2）
It is middle adopt Jing molecular cut offs for 3000 Da bag filter dialysis initially with flowing water dialysis, finally carry out distilled water dialysis, flow
Water dialysis time is 36-48h, and distilled water dialysis time is 24-36h.
8. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（2）
Middle vacuum drying time is 2-3h, and temperature is 20-25 DEG C, and vacuum is-0.085-- 0.1Mpa.
9. the preparation method of carboxy methylation Fructus Schisandrae Polysaccharide according to claim 2, it is characterised in that：The step（3）
Middle carboxy methylation fruit of Chinese magnoliavine Thick many candies Jing concussions heating for dissolving in NaCl solution, the temperature of shaking water bath is 50-60 DEG C, concussion
Frequency is 120-160 beat/min, shakes the time 10-12h of dissolving, then the solution after dissolving is centrifuged, solution
The rotating speed of centrifugation is：4000-5000 r/min, centrifugation time is 8-10 min；
The step（3）Middle flowing water dialysis time is 36-48h, and distilled water dialysis time is 24-36h, and vacuum drying time is 2-
3h, vacuum drying temperature is 20-25 DEG C, and vacuum drying vacuum is-0.085-- 0.1MPa.
10. the application of carboxy methylation Fructus Schisandrae Polysaccharide described in claim 1, it is characterised in that：The carboxy methylation fruit of Chinese magnoliavine is more
Sugar can be used in functional food and food additives as immunopotentiator.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610313007.8A CN106632709B (en) | 2016-05-12 | 2016-05-12 | A kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610313007.8A CN106632709B (en) | 2016-05-12 | 2016-05-12 | A kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106632709A true CN106632709A (en) | 2017-05-10 |
| CN106632709B CN106632709B (en) | 2019-05-31 |
Family
ID=58848684
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610313007.8A Active CN106632709B (en) | 2016-05-12 | 2016-05-12 | A kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106632709B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112294691A (en) * | 2020-11-04 | 2021-02-02 | 赵玉龙 | Pumpkin-extracted firming anti-wrinkle molecular mask and preparation method thereof |
| CN115746159A (en) * | 2022-11-29 | 2023-03-07 | 广西大学 | Preparation process and application of carboxymethylated abrus cantoniensis hance polysaccharide |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
-
2016
- 2016-05-12 CN CN201610313007.8A patent/CN106632709B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
|---|
| 孟宪军等: "五味子多糖的分离纯化及清除自由基研究", 《食品科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112294691A (en) * | 2020-11-04 | 2021-02-02 | 赵玉龙 | Pumpkin-extracted firming anti-wrinkle molecular mask and preparation method thereof |
| CN115746159A (en) * | 2022-11-29 | 2023-03-07 | 广西大学 | Preparation process and application of carboxymethylated abrus cantoniensis hance polysaccharide |
| CN115746159B (en) * | 2022-11-29 | 2024-08-16 | 广西大学 | Preparation process and application of carboxymethylated abrus herb polysaccharide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106632709B (en) | 2019-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gao et al. | Structural characterization and immunomodulatory activity of a water-soluble polysaccharide from Ganoderma leucocontextum fruiting bodies | |
| Shen et al. | Structural characterization and immunomodulatory activity of novel polysaccharides from Citrus aurantium Linn. variant amara Engl | |
| Wang et al. | Structure and chain conformation of water-soluble heteropolysaccharides from Ganoderma lucidum | |
| CN111285939B (en) | Artemisia annua polysaccharide with effects of resisting oxidation and regulating intestinal flora and preparation method and application thereof | |
| CN105001352B (en) | Beta-1,3/1,6-glucan, preparation method therefor, and application thereof in preparing immune enhancement and anti-tumor medicine and functional food | |
| CN104672339B (en) | Cordyceps cicadae rhzomorph as well as preparation method and application thereof | |
| US10835552B2 (en) | Method for preparing linseed polysaccharide having antiviral activity and immunological activity, and use of the linseed polysaccharide | |
| Li et al. | Three kinds of Ganoderma lucidum polysaccharides attenuate DDC-induced chronic pancreatitis in mice | |
| CN107541533A (en) | A kind of preparation method of medicine food hypha polysaccharide polypeptide immunopotentiator | |
| CN108727509A (en) | A kind of moso bamboo bamboo shoot shell arabogalactan and its preparation and use | |
| CN113278091A (en) | Porphyridium polysaccharide and preparation method and application thereof | |
| CN114907494B (en) | Rosa roxburghii polysaccharide with significant lipid and cholesterol reducing effects and preparation method and application thereof | |
| CN106632709B (en) | A kind of carboxy methylation Fructus Schisandrae Polysaccharide and its preparation method and application | |
| Geng et al. | Effects of different extraction methods on the physico-chemical characteristics and biological activities of polysaccharides from Clitocybe squamulosa | |
| CN115160450B (en) | Rapid preparation method and application of Pholiota nameko polysaccharide | |
| CN108359021B (en) | Method for rapidly preparing flaxseed polysaccharide with antiviral and immunoregulatory activities | |
| CN114874346B (en) | Lepista nuda polysaccharide and preparation method and application thereof | |
| Pan et al. | Ultrasonic-assisted extraction of a low molecular weight polysaccharide from Nostoc commune Vaucher and its structural characterization and immunomodulatory activity | |
| CN112794925B (en) | Amomum villosum polysaccharide and preparation method and application thereof | |
| CN105753998B (en) | A kind of peach gum polysaccharide catabolite PGP 1 and its preparation method and application | |
| CN104287032B (en) | Ulva juice product containing ulva oligosaccharide and preparation method thereof | |
| CN112778430A (en) | Sulfated modified cyclocarya paliurus polysaccharide and preparation method and application thereof | |
| Teng et al. | Structural characterization, physicochemical properties and hypoglycemic activity of sulfated polysaccharides from Porphyra yezoensis | |
| CN113388046B (en) | Citron fruit polysaccharide and preparation method and application thereof | |
| CN111690073B (en) | Lactarius deliciosus polysaccharide and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |