CN104086664B - Gracilaria lemaneiformis polysaccharide extract and preparation method and application thereof - Google Patents
Gracilaria lemaneiformis polysaccharide extract and preparation method and application thereof Download PDFInfo
- Publication number
- CN104086664B CN104086664B CN201410252784.7A CN201410252784A CN104086664B CN 104086664 B CN104086664 B CN 104086664B CN 201410252784 A CN201410252784 A CN 201410252784A CN 104086664 B CN104086664 B CN 104086664B
- Authority
- CN
- China
- Prior art keywords
- thallus gracilariae
- extract
- solution
- extracting solution
- ethanol
- 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.)
- Expired - Fee Related
Links
- 239000000284 extract Substances 0.000 title claims abstract description 97
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 35
- 150000004676 glycans Chemical class 0.000 title claims abstract description 34
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 238000000605 extraction Methods 0.000 title claims description 11
- 241000206611 Gracilariopsis lemaneiformis Species 0.000 title 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 65
- 150000007524 organic acids Chemical class 0.000 claims abstract description 11
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000003456 ion exchange resin Substances 0.000 claims abstract description 10
- 229920003303 ion-exchange polymer Polymers 0.000 claims abstract description 10
- 239000003513 alkali Substances 0.000 claims abstract description 4
- 238000001035 drying Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 21
- 239000012466 permeate Substances 0.000 claims description 20
- 239000000843 powder Substances 0.000 claims description 16
- 238000001556 precipitation Methods 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 13
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 12
- 238000009835 boiling Methods 0.000 claims description 11
- 230000000694 effects Effects 0.000 claims description 11
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000000108 ultra-filtration Methods 0.000 claims description 9
- 238000002137 ultrasound extraction Methods 0.000 claims description 8
- 239000002893 slag Substances 0.000 claims description 7
- 238000005238 degreasing Methods 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 150000002772 monosaccharides Chemical group 0.000 claims description 6
- 229930182830 galactose Natural products 0.000 claims description 5
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims description 3
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 3
- 235000011090 malic acid Nutrition 0.000 claims description 3
- 239000001630 malic acid Substances 0.000 claims description 3
- 229940124213 Dipeptidyl peptidase 4 (DPP IV) inhibitor Drugs 0.000 claims description 2
- GZCGUPFRVQAUEE-KCDKBNATSA-N aldehydo-D-galactose Chemical compound OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-KCDKBNATSA-N 0.000 claims description 2
- 230000001186 cumulative effect Effects 0.000 claims description 2
- 239000003603 dipeptidyl peptidase IV inhibitor Substances 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 102000016622 Dipeptidyl Peptidase 4 Human genes 0.000 abstract description 11
- 101000930822 Giardia intestinalis Dipeptidyl-peptidase 4 Proteins 0.000 abstract description 11
- 235000005340 Asparagus officinalis Nutrition 0.000 abstract description 9
- 239000003472 antidiabetic agent Substances 0.000 abstract description 5
- 230000003078 antioxidant effect Effects 0.000 abstract description 4
- 230000036541 health Effects 0.000 abstract description 3
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 3
- 238000012545 processing Methods 0.000 abstract description 2
- 241000234427 Asparagus Species 0.000 abstract 7
- 238000009776 industrial production Methods 0.000 abstract 1
- 230000005764 inhibitory process Effects 0.000 abstract 1
- 230000003472 neutralizing effect Effects 0.000 abstract 1
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 66
- 229960004756 ethanol Drugs 0.000 description 16
- 239000000203 mixture Substances 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- 239000000047 product Substances 0.000 description 9
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 238000003809 water extraction Methods 0.000 description 8
- 230000001629 suppression Effects 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 150000003254 radicals Chemical class 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000009837 dry grinding Methods 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- BOVGTQGAOIONJV-BETUJISGSA-N 1-[(3ar,6as)-3,3a,4,5,6,6a-hexahydro-1h-cyclopenta[c]pyrrol-2-yl]-3-(4-methylphenyl)sulfonylurea Chemical compound C1=CC(C)=CC=C1S(=O)(=O)NC(=O)NN1C[C@H]2CCC[C@H]2C1 BOVGTQGAOIONJV-BETUJISGSA-N 0.000 description 3
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- 101100337060 Caenorhabditis elegans glp-1 gene Proteins 0.000 description 3
- 229920002307 Dextran Polymers 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000002292 Radical scavenging effect Effects 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 229960000346 gliclazide Drugs 0.000 description 3
- 230000031700 light absorption Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 230000002000 scavenging effect Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 244000003416 Asparagus officinalis Species 0.000 description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 2
- 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
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000003064 anti-oxidating 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
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 235000019504 cigarettes Nutrition 0.000 description 2
- 229960000935 dehydrated alcohol Drugs 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- -1 dpph free radical Chemical class 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- OBMZMSLWNNWEJA-XNCRXQDQSA-N C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 Chemical compound C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 OBMZMSLWNNWEJA-XNCRXQDQSA-N 0.000 description 1
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 1
- 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 description 1
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 1
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 1
- 241001134782 Gigartinales Species 0.000 description 1
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 description 1
- 101800000224 Glucagon-like peptide 1 Proteins 0.000 description 1
- 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 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical compound Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 1
- 208000013016 Hypoglycemia Diseases 0.000 description 1
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 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
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 101710176384 Peptide 1 Proteins 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 102100040918 Pro-glucagon Human genes 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 241000206572 Rhodophyta Species 0.000 description 1
- 241000195474 Sargassum Species 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 208000000260 Warts Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003570 air Substances 0.000 description 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 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 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
- 239000002585 base Substances 0.000 description 1
- 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 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000003653 coastal water Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 239000002027 dichloromethane extract Substances 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 206010013990 dysuria Diseases 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 201000001421 hyperglycemia Diseases 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000009318 large scale farming Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 201000010153 skin papilloma Diseases 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the field of deep processing of asparagus and discloses an asparagus polysaccharide extract and a preparation method and application thereof. The polysaccharide extract of the asparagus is prepared by drying and crushing the asparagus, and then extracting the dried and crushed asparagus by organic acid, neutralizing by alkali, extracting by ultrasonic, ultrafiltering, separating by ion exchange resin, concentrating and precipitating by alcohol. The preparation method of the asparagus polysaccharide extract is simple and efficient, and is suitable for industrial production, and the obtained asparagus polysaccharide extract has strong antioxidant activity and hypoglycemic activity, has high inhibition rate on dipeptidyl peptidase IV, and can be used for preparing hypoglycemic drugs or health care products.
Description
Technical field
The invention belongs to Thallus Gracilariae field of deep and in particular to a kind of Thallus Gracilariae polyoses extract and preparation method thereof with
Application.
Background technology
Thallus Gracilariae (gracialrialemaneofimrsi) belongs to Rhodophyta, Gigartinales, and river hedge belongs to (gracaialri), former
Originate in China's Shandong coastal waters and okinawa of japan.Because conventional Thallus Gracilariae cultivates small scale, yield is not high and palatability is poor, so
Seldom directly eat, be mainly used in extracting agar, separately have small part to be used as abalone culture feedstuff, among the people be used for being used as medicine." book on Chinese herbal medicine
Detailed outline " described in this flavour of a drug sweet cold in nature, have the effect of softening and eliminating sputum, clearing away heat and promoting diuresis, be used for controlling wart stagnation of pathogenic heat gas, dysuria, the moon
Empty interior-heat.At the beginning of 2000, Guangdong Province of Institute of Oceanology of the Chinese Academy of Sciences marine resources research centre of development and Nanao County science and technology are emerging
Extra large office cooperation development Thallus Gracilariae transplants the research of In The Eastern Guangdong Sea Area, and project succeeds, and has started south China sea area dragon
The successful precedent of dish nursery.After the popularization of 3 years, the cultivation of Nan'ao Thallus Gracilariae has reached large-scale production level, and average product reaches
To 525t/1000m2, and driven the rise of Guangdong Fujian coastal area large-scale farming.At present, Thallus Gracilariae has become as in China
The fourth-largest cultivation Sargassum after Thallus Laminariae (Thallus Eckloniae), Thallus Porphyrae, Thallus Laminariae.
Because early stage Thallus Gracilariae yield rareness is not taken seriously always, the research to Thallus Gracilariae is all little both at home and abroad.With near
The continuous expansion of artificial cultivation scale and increasing sharply of yield over year, the research about Thallus Gracilariae function and application is also gradually drawn
Play the interest of people, but research report relevant at present is concentrated mainly on plantation and processing aspect, to asparagus active material
Extract and the research report of physiological function is very few, only a small amount of patent is open.As State Oceanic Administration Bureau The Third Oceanography Institute is public
Open a kind of Thallus Gracilariae agaropectin oligose and preparation method thereof, can be applicable in antioxidation, uvioresistant health product and cosmetics (specially
Sharp application number 201210347105.5)；Li Xuejiao discloses a kind of method (number of patent application extracting cancer-resisting substance in Thallus Gracilariae
201210089080.3)；Zhengzhou Tobacco Research Institute of CNTC discloses the preparation method of asparagus rough polysaccharide and incites somebody to action
It is applied in Medicated cigarette, can improve humid keeping performance and the sucking quality (number of patent application 201010204566.8) of Medicated cigarette；Zhu Yi
Life discloses a kind of method (number of patent application 201010188414.3) of the antioxidation composition improving Thallus Gracilariae extract；Shanghai
Ocean university discloses a kind of extraction of Thallus Gracilariae polysaccharide and isolation and purification method, and gained polysaccharide has the immune work(of preferable raising
Effect (number of patent application 200810042987.8)；Shanghai Aquatic Products Univ. 9CN) discloses a kind of preparation method of Thallus Gracilariae polysaccharide, gained
Polysaccharides on Mice lymphocyte has higher cultivation effect (number of patent application 200710044610.1)；Guangdong Pharmaceutical University is open
A kind of Thallus Gracilariae and processing method, can be applicable to leisure food (number of patent application 200710027284.3).
Dipeptidyl peptidase iv (dipeptidyl peptidase iv, dpp-iv) can fast and effeciently degrade pancreas hyperglycemia
Plain sample peptide 1 (glp-1, glucagon-like peptide-1), glp-1 is that insulin generates and secretes maximally effective stimulant
One of, therefore suppression dpp-iv can strengthen the effect of endogenouss glp-1, thus improving the level of insulin in blood, and then drops
Blood sugar level that is low and maintaining diabetes patient.Medical science has confirmed that dpp-iv inhibitor is that a kind of new anti-diabetic is controlled at present
Treat medicine, clinical effectiveness shows that such medicine has good hypoglycemic effect, do not find that other diabetes medicaments are produced simultaneously
Common body weight increase and the untoward reaction such as hypoglycemia.
Content of the invention
In order to improve the deep process technology of Thallus Gracilariae, widen the range of application of Thallus Gracilariae, the primary and foremost purpose of the present invention is
A kind of Thallus Gracilariae polyoses extract is provided；
Another object of the present invention is to providing the preparation method of above-mentioned Thallus Gracilariae polyoses extract；
It is still another object of the present invention to provide the application of above-mentioned Thallus Gracilariae polyoses extract.
The purpose of the present invention is achieved through the following technical solutions:
A kind of Thallus Gracilariae polyoses extract, described Thallus Gracilariae polyoses extract is prepared from by Thallus Gracilariae is extracted, mainly
Monosaccharide component is d- galactose and 3,6- inner ether-l galactose；
Described Thallus Gracilariae polyoses extract is prepared from by the preparation method comprising the steps of:
(1) Thallus Gracilariae dried, pulverize and sieve, degreasing decoloring obtains Thallus Gracilariae powder；
(2) extract Thallus Gracilariae powder with organic acid soln, then neutralized with alkali, filter, obtain filtering residue and first time extracting solution；
(3) step (2) gained filtering residue is carried out ultrasonic extraction method in water, filter to obtain second extracting solution, will be described
First time extracting solution is merged with second extracting solution and obtains final extracting solution；
(4) described for step (3) final extracting solution is carried out ultrafiltration, collect permeate；
(5) spent ion exchange resin separates described permeate, takes high-activity component to carry out concentrated in vacuo, obtains concentrated solution；
(6) described concentrated solution is mixed with ethanol, standing, be filtrated to get polysaccharide precipitation；
(7) described polysaccharide precipitation is dried, obtains described Thallus Gracilariae polyoses extract.
The preparation method of above-mentioned Thallus Gracilariae polyoses extract, comprises the steps:
(1) Thallus Gracilariae dried, pulverize and sieve, degreasing decoloring obtains Thallus Gracilariae powder；
Preferably, described pulverizing and sieving crosses 20～60 mesh sieves for after pulverizing；
Wherein, the concretely comprising the following steps of described degreasing decoloring: using ethanol solution or acetone soln, micro-boiling is carried out to Thallus Gracilariae
Extract, filter to obtain slag, dry for standby；Preferably, the quality of the volume of described ethanol solution or acetone soln and described Thallus Gracilariae
Volume mass than be (2～6): 1；Described ethanol solution is the ethanol solution of volume fraction 95%；Described micro-boiling extract when
Between be 2～4 hours；
(2) extract Thallus Gracilariae powder with organic acid soln, then neutralized with alkali, filter, obtain filtering residue and first time extracting solution；
Preferably, the ph value of described organic acid soln is 1.5～3.5；
Preferably, described organic acid soln is citric acid solution or malic acid solution；
Preferably, the quality of described organic acid soln is 20～50 times of described Thallus Gracilariae powder quality；
(3) step (2) gained filtering residue is carried out ultrasonic extraction method in water, filter to obtain second extracting solution, will be described
First time extracting solution is merged with second extracting solution and obtains final extracting solution；
Preferably, the control condition of described ultrasonic extraction method is: ultrasonic power 320～640w, Extracting temperature be 30～
70 DEG C, extraction time is 20～60 minutes；
(4) described for step (3) final extracting solution is carried out ultrafiltration, collect permeate；
Preferably, described ultrafiltration adopts the ultrafilter membrane of molecular cut off 20kda；
(5) spent ion exchange resin separates described permeate, takes high-activity component to carry out concentrated in vacuo, obtains concentrated solution；
Preferably, the model deae-fast flow or 717 of described ion exchange resin；
Preferably, described gained concentrated solution concentrated in vacuo is the 1/5～1/3 of described permeate volume；
(6) described concentrated solution is mixed with ethanol, standing, be filtrated to get polysaccharide precipitation；
Preferably, the consumption of described ethanol accounts for 50%～80% of cumulative volume after described concentrated solution is mixed with ethanol；
Preferably, the concrete operations of described standing are: stand 8～14 hours at 0～4 DEG C；
(7) described polysaccharide precipitation is dried, obtains described Thallus Gracilariae polyoses extract；
Preferably, described drying is vacuum lyophilization or spray drying.
Application in preparing dipeptidyl peptidase iv inhibitor for the above-mentioned Thallus Gracilariae polyoses extract；
Application in preparing hypoglycemic drug or health product for the above-mentioned Thallus Gracilariae polyoses extract.
The present invention has such advantages as with respect to prior art and effect:
(1) preparation method of the present invention adopts organic acid to combine ultrasonic method and extracts polysaccharide, and its extraction ratio is than traditional water extraction
Method improves 2～4 times, can significantly shorten extraction time, reducing energy consumption, antioxidant effect is better than traditional water extraction simultaneously.
(2) Thallus Gracilariae polyoses extract of the present invention can significantly inhibit the activity of dipeptidyl peptidase iv, has blood sugar lowering
Effect, is the reported first of this area, and the research having filled up this respect both at home and abroad at present is blank.
Specific embodiment
With reference to embodiment, the present invention is described in further detail, but embodiments of the present invention not limited to this.
Embodiment 1
A kind of Thallus Gracilariae polyoses extract, its preparation method is as follows:
(1) Thallus Gracilariae crosses 20 mesh sieves after shining dry grinding, plus the volume fraction 95% with respect to 2 times of volume mass ratios of Thallus Gracilariae
Ethanol solution, micro-boiling extract 3 hours, filter to obtain slag, dry at 60 DEG C Thallus Gracilariae powder is standby；
(2) take 50g Thallus Gracilariae powder to be placed in container, mix homogeneously by the quality ratio of 1:30 with the citric acid of ph2.0, micro-boiling
Extract 3 hours, it is neutral for being neutralized to ph value with naoh, filters to obtain filtering residue and first time extracting solution；
(3) step (2) gained filtering residue is used ultrasonic extraction method in water, ultrasonic power 500w, 40 DEG C of Extracting temperature, carry
Take 30 minutes, filter to obtain second extracting solution, merge with step (2) gained first time extracting solution, obtain final extracting solution；
(4) the final extracting solution of step (3) gained is carried out ultrafiltration with the film of molecular cut off 20kda, collect permeate；
(5) spent ion exchange resin deae-fast flow model separation permeate, takes active highest component, and vacuum is dense
It is reduced to the 1/4 of permeate volume, obtain concentrated solution；
(6) add ethanol in concentrated solution, after mix homogeneously, at 2 DEG C, stand 12 hours, filter, remove the supernatant
Part, obtains polysaccharide precipitation；
(7) by gained polysaccharide precipitation lyophilization, obtain described Thallus Gracilariae polyoses extract, be designated as 1# extract.
Embodiment 2
A kind of Thallus Gracilariae polyoses extract, its preparation method is as follows:
(1) Thallus Gracilariae crosses 60 mesh sieves after shining dry grinding, plus the volume fraction 95% with respect to 6 times of volume mass ratios of Thallus Gracilariae
Ethanol solution, micro-boiling extract 2 hours, filter to obtain slag, dry at 40 DEG C Thallus Gracilariae powder is standby；
(2) take 50g Thallus Gracilariae powder to be placed in container, mix homogeneously by the quality ratio of 1:50 with the malic acid of ph1.5, micro-boiling
Extract 1 hour, it is neutral for being neutralized to ph value with naoh, filters to obtain filtering residue and first time extracting solution；
(3) step (2) gained filtering residue is used ultrasonic extraction method in water, ultrasonic power 320w, 60 DEG C of Extracting temperature, carry
Take 60 minutes, filter to obtain second extracting solution, merge with step (2) gained first time extracting solution, obtain final extracting solution；
(4) the final extracting solution of step (3) gained is carried out ultrafiltration with the film of molecular cut off 20kda, collect permeate；
(5) spent ion exchange resin 717 model separation permeate, takes active highest component, is concentrated in vacuo to permeate
The 1/3 of volume, obtains concentrated solution；
(6) add ethanol in concentrated solution, after mix homogeneously, at 4 DEG C, stand 14 hours, filter, remove the supernatant
Part, obtains polysaccharide precipitation；
(7) by gained polysaccharide precipitation lyophilization, obtain described Thallus Gracilariae polyoses extract, be designated as 2# extract.
Embodiment 3
A kind of Thallus Gracilariae polyoses extract, its preparation method is as follows:
(1) Thallus Gracilariae crosses 40 mesh sieves after shining dry grinding, plus the volume fraction 95% with respect to 5 times of volume mass ratios of Thallus Gracilariae
Ethanol solution, micro-boiling extract 4 hours, filter to obtain slag, dry at 60 DEG C Thallus Gracilariae powder is standby；
(2) take 50g Thallus Gracilariae powder to be placed in container, mix homogeneously by the quality ratio of 1:20 with the citric acid of ph3.5, micro-boiling
Extract 3 hours, it is neutral for being neutralized to ph value with naoh, filters to obtain filtering residue and first time extracting solution；
(3) step (2) gained filtering residue is used ultrasonic extraction method in water, ultrasonic power 640w, 30 DEG C of Extracting temperature, carry
Take 20 minutes, filter to obtain second extracting solution, merge with step (2) gained first time extracting solution, obtain final extracting solution；
(4) the final extracting solution of step (3) gained is carried out ultrafiltration with the film of molecular cut off 20kda, collect permeate；
(5) spent ion exchange resin deae-fast flow model separation permeate, takes active highest component, and vacuum is dense
It is reduced to the 1/5 of permeate volume, obtain concentrated solution；
(6) add ethanol in concentrated solution, after mix homogeneously, at 0 DEG C, stand 8 hours, filter, remove supernatant portion
Point, obtain polysaccharide precipitation；
(7) by gained polysaccharide precipitation lyophilization, obtain described Thallus Gracilariae polyoses extract, be designated as 3# extract.
The traditional water extraction preparation Thallus Gracilariae polyoses extract of comparative example 1
A kind of Thallus Gracilariae polyoses extract, is prepared using traditional water extraction, its preparation method is as follows:
(1) Thallus Gracilariae crosses 20 mesh sieves after shining dry grinding, plus the volume fraction 95% with respect to 2 times of volume mass ratios of Thallus Gracilariae
Ethanol solution, micro-boiling extract 3 hours, filter to obtain slag, dry at 60 DEG C Thallus Gracilariae powder is standby；
(2), after taking Thallus Gracilariae powder hot water in the container to boil 2 hours, filtering residue and first time extracting solution are filtered to obtain；Described filter
Slag boils 1 hour, filters to obtain second extracting solution；Then first time extracting solution is merged with second extracting solution, obtain and finally carry
Take liquid；
(3) the final extracting solution of step (2) gained is carried out ultrafiltration with the film of molecular cut off 20kda, collect permeate；
(4) spent ion exchange resin deae-fast flow model separation permeate, takes active highest component, and vacuum is dense
It is reduced to the 1/4 of permeate volume, obtain concentrated solution；
(5) add ethanol in concentrated solution, after mix homogeneously, at 0～4 DEG C, stand 12 hours, filter, remove upper strata clear
Liquid part, obtains polysaccharide precipitation；
(6) by gained polysaccharide precipitation lyophilization, obtain Thallus Gracilariae polyoses extract, be designated as 4# extract.
Experimental example 1 measures polysaccharide extract rate
Measure the polysaccharide extract rate of each Thallus Gracilariae polyoses extract in embodiment and comparative example.
Polyoses content in 1#～4# extract is measured using phend-sulphuric acid, calculates polysaccharide extract rate, formula is: many
Sugared extraction ratio (%)=(polyoses content (g)/raw material weight (g) in extracting solution) × 100.
Each Thallus Gracilariae polyoses extract measure and calculation result is as shown in table 1:
The polysaccharide extract rate of each extract of table 1.
| Extract | 1# extract | 2# extract | 3# extract | 4# extract |
| Polysaccharide extract rate (%) | 14.12±0.67 | 10.57±0.58 | 12.24±0.49 | 4.36±0.65 |
Note: in table, data is meansigma methodss ± standard deviation (n=3)
As can be known from the table data, the Thallus Gracilariae polysaccharide extract rate of the method for the invention is many apparently higher than traditional water extraction
Sugared extraction ratio.
Experimental example 2 measures dpph radical scavenging activity
Measure the Scavenging activity of the dpph free radical of each Thallus Gracilariae polyoses extract in embodiment and comparative example, dpph is freely
The Scavenging activity of base is a kind of method of more universal survey antioxidant activity.
The Scavenging activity that 1～4# extract carries out dpph free radical is taken to measure, assay method is as follows:
Take the 0.2mmol/l of the aqueous solution of Thallus Gracilariae polyoses extract that 2ml concentration is 0.1～2.0mg/ml and 2ml
Dpph solution mixes, vibration, avoid light place 30min under room temperature, surveys light absorption value (ai) under 517nm wavelength；Identical dense with 2ml
The sample of degree and 2ml dehydrated alcohol mix as a control group (aj), are in addition mixed with 2ml dpph solution and 2ml dehydrated alcohol
Survey light absorption value (ac) afterwards.
Computing formula is: dpph clearance rate (%)=[1- (ai-aj)/ac] × 100.Measure the analysis each dragon of gained
The dpph radical scavenging activity ic of dish polyoses extract50Value is as shown in table 2:
The dpph radical scavenging activity ic of each extract of table 2.50Value
From table, data analysiss understand, the dpph free radical scavenging activity of Thallus Gracilariae polyoses extract of the present invention is better than
The polyoses extract of traditional water extraction gained.
Experimental example 3 measures oxygen-derived free radicals absorbability (orac)
Measure the oxygen-derived free radicals absorbability of each Thallus Gracilariae polyoses extract in embodiment and comparative example.
Take 1～4# extract to carry out oxygen-derived free radicals absorbability (orac) respectively to measure, surveyed using fluorescence microplate reader
Fixed, concrete steps include the following:
It is separately added into the Thallus Gracilariae polyoses extract sample that 20 μ l concentration are 0.05mg/ml in each micropore of 96 orifice plates, then
Adding the phosphate buffered solution 20 μ l of ph7.4 and concentration is the luciferin solution 20 μ l of 7nmol/l, preset at 37 DEG C
After 15min, start reaction with the aaph140 μ l that multichannel pipettor adds rapidly 12mmol/l in each hole, and microwell plate is put
With excitation wavelength 485nm at 37 DEG C in microplate reader, launch wavelength 538nm carries out METHOD FOR CONTINUOUS DETERMINATION, and every 2min measures once each
Hole fluorescence intensity, the common 2h of minute.Each extract oxygen-derived free radicals absorbability (orac value) measurement result is as shown in table 3:
The oxygen-derived free radicals absorbability measurement result of each extract of table 3.
Note: in table, data is meansigma methodss ± standard deviation (n=3)
From table, data analysiss understand, the antioxidant activity of Thallus Gracilariae polyoses extract of the present invention is better than traditional water extraction
The polyoses extract of method gained.
Experimental example 4 measures the molecular weight of gained Thallus Gracilariae polyoses extract product
Measure the molecular weight of each Thallus Gracilariae polyoses extract in embodiment and comparative example.
1～4# extract is taken to carry out molecular weight determination respectively, by the dextran standard of known molecular amount respectively with flowing
Mutually it is configured to the solution of 2.0mg/ml, molecular weight is measured using gel permeation chromatography (gpc), location parameter includes the following:
Chromatographic condition: tsk gel guard column (pw × l6.0 × 40), tsk g-4000k pw × l7.8 × 300 gel column,
Tskg-2500k pw × l7.8 × 300 gel column；Mobile phase is 0.2mol/lkh2po4 solution, ph6.0；Flow velocity 0.6ml/
min；35 DEG C of column temperature；Sample introduction 30 μ l；Standard specimen be known molecular amount dextran standard (dextran, molecular weight be 4400da,
9900da, 21400da, 43500da, 124000da, 196000da, 277000da, 401000da, 1285000da, sigma are public
Department)；Detector is waters2414 differential refraction detector.The average molecular weight results measuring are as shown in table 4:
The molecular weight of each extract of table 4.
From table 4, data analysiss understand, the molecular weight of Thallus Gracilariae polyoses extract of the present invention is significantly less than comparative example 1
The molecular weight of traditional water extraction gained polyoses extract.
Experimental example 5 measures the monosaccharide composition of gained Thallus Gracilariae polyoses extract product
Example 1～3 products therefrom Thallus Gracilariae polyoses extract carries out monosaccharide composition measuring, assay method and ginseng respectively
Number includes the following:
Weigh Thallus Gracilariae polyoses extract sample 20mg, add 4m trifluoroacetic acid 5ml, sealing, 110 DEG C of hydrolysis 2h；Hydrolysis
Liquid to dry, add methanol 3ml, then is spin-dried for, be repeated 3 times to obtain Thallus Gracilariae polysaccharide hydrolysis thing in 50 DEG C of rotary evaporation in vacuo.
Add oxammonium hydrochloride. 10mg, internal standard inositol six acetass 1mg and pyridine 2ml in Thallus Gracilariae polysaccharide hydrolysis thing, seal,
90 DEG C of water-bath 30min, add after 2ml acetic anhydride 90 DEG C of water-bath 30min again, add 2ml water terminating reaction, add 2ml dichloro
Methane extracts, and is repeated twice, discards water layer, is settled to 5ml, adds anhydrous sodium sulfate drying to cross film standby.
Gas chromatographic detection program: hp-5 quartz capillary column (30m × 0.32mm × 0.25 μm)；Constant voltage mode,
20psi；Temperature programming: 100 DEG C of initial column temperature, keep 0.5min；Then heated up with 20 DEG C/min, keep 5min；With 3 DEG C/min
Rise to 160 DEG C；It is raised to 250 DEG C with 10 DEG C/min again, keep 5min；Injection port is using not shunt mode, 250 DEG C of temperature, carrier gas
For nitrogen；250 DEG C of the temperature of fid detector, hydrogen, air and nitrogen flow rate are respectively 30,400 and 25ml/min；Sample introduction body
Long-pending 1 μ l.
Various standard monosaccharide (rhamnose, arabinose, fucose, xylose, mannose, glucose and galactose) are identical
Under the conditions of carry out saccharin acetass derivatization treatment, the same terms gas chromatographic analysiss.
Measure known to analysis result, the principal monosaccharides of 1#～3# extract consist of: galactose and 3,6- inner ether galactose.
Experimental example 6 measures the rejection ability to dipeptidyl peptidase iv for the Thallus Gracilariae polysaccharide
1～4# extract sample and control sample hypoglycemic drug Gliclazide is taken to carry out the suppression of dipeptidyl peptidase iv respectively
The mensure of ability processed, assay method and parameter include the following:
The sample of variable concentrations is added in the dipeptidyl peptidase iv solution that enzyme activity unit is 1u l/l, makes final concentration
It is followed successively by 1nmol/l, 10nmol/l, 20nmol/l, 100nmol/l and 500nmol/l, with water as negative control, ice bath is incubated
30min, is added in 96 orifice plates, every hole 100 μ l；Add 100 μ l substrate solutions, with 50mmol/l tris-hcl, 1mol/
It is blank that ledta, ph8.3 replace substrate solution, and every group sets 4 multiple holes, after 37 DEG C of incubation 1h, surveys light absorption value at 405nm,
And suppression ratio is calculated according to formula.
Computing formula: dipeptidyl peptidase iv suppression ratio=(aNegative control- aPolysaccharide)/aNegative control× 100%.
Each sample is as shown in table 5 to the rejection ability of dipeptidyl peptidase iv:
The dipeptidyl peptidase iv suppression ratio result of table 5. each sample
| Sample | 1# extract | 2# extract | 3# extract | 4# extract | Gliclazide |
| Suppression ratio (%) | 75.49±1.44 | 70.57±1.89 | 64.23±1.11 | 50.24±2.43 | 77.68±1.94 |
Note: in table, data is meansigma methodss ± standard deviation (n=3)
As can be known from the table data, products therefrom Thallus Gracilariae polyoses extract of the present invention is high to the suppression ratio of dipeptidyl peptidase iv
In traditional extraction process products therefrom, can be suitable with hypoglycemic drug Gliclazide.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not subject to above-described embodiment
Limit, other any spirit without departing from the present invention and the change made under principle, modification, replacement, combine, simplify,
All should be equivalent substitute mode, be included within protection scope of the present invention.
Claims (3)
1. application in preparing dipeptidyl peptidase iv inhibitor for the Thallus Gracilariae polyoses extract it is characterised in that: described Thallus Gracilariae
Polyoses extract is prepared from by Thallus Gracilariae is extracted, and principal monosaccharides group is divided into d- galactose and 3,6- inner ether-l galactose；
Described Thallus Gracilariae polyoses extract is prepared from by the preparation method comprising the steps of:
(1) Thallus Gracilariae dried, pulverize and sieve, degreasing decoloring obtains Thallus Gracilariae powder；
(2) extract Thallus Gracilariae powder with organic acid soln, then neutralized with alkali, filter, obtain filtering residue and first time extracting solution；
(3) step (2) gained filtering residue is carried out ultrasonic extraction method in water, filter to obtain second extracting solution, by described first
Secondary extracting solution is merged with second extracting solution and obtains final extracting solution；
(4) described for step (3) final extracting solution is carried out ultrafiltration, collect permeate；
(5) spent ion exchange resin separates described permeate, takes high-activity component to carry out concentrated in vacuo, obtains concentrated solution；
(6) described concentrated solution is mixed with ethanol, standing, be filtrated to get polysaccharide precipitation；
(7) described polysaccharide precipitation is dried, obtains described Thallus Gracilariae polyoses extract；
The described ultrafiltration of step (4) adopts the ultrafilter membrane of molecular cut off 20kda.
2. according to claim 1 application it is characterised in that: pulverize and sieve described in step (1) for pulverize after cross 20～60
Mesh sieve；The concretely comprising the following steps of step (1) described degreasing decoloring: using ethanol solution or acetone soln, micro-boiling is carried out to Thallus Gracilariae
Extract, filter to obtain slag, dry for standby；
The ph value of the described organic acid soln of step (2) is 1.5～3.5；The described organic acid soln of step (2) be citric acid solution or
Malic acid solution；The quality of the described organic acid soln of step (2) is 20～50 times of described Thallus Gracilariae powder quality；
The control condition of the described ultrasonic extraction method of step (3) is: ultrasonic power 320～640w, and Extracting temperature is 30～70 DEG C,
Extraction time is 20～60 minutes；
The model deae-fast flow or 717 of the described ion exchange resin of step (5)；The described institute concentrated in vacuo of step (5)
Obtaining concentrated solution is the 1/5～1/3 of described permeate volume；
The consumption of the described ethanol of step (6) accounts for 50%～80% of cumulative volume after described concentrated solution is mixed with ethanol；Step (6) institute
The concrete operations stating standing are: stand 8～14 hours at 0～4 DEG C；
The described drying of step (7) is vacuum lyophilization or spray drying.
3. according to claim 2 application it is characterised in that: in the concrete steps of the described degreasing decoloring of step (1): described
The volume of ethanol solution or acetone soln is (2～6) with the volume mass ratio of the quality of described Thallus Gracilariae: 1；Described ethanol solution
Ethanol solution for volume fraction 95%；The time that described micro-boiling is extracted is 2～4 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410252784.7A CN104086664B (en) | 2014-06-09 | 2014-06-09 | Gracilaria lemaneiformis polysaccharide extract and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410252784.7A CN104086664B (en) | 2014-06-09 | 2014-06-09 | Gracilaria lemaneiformis polysaccharide extract and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104086664A CN104086664A (en) | 2014-10-08 |
| CN104086664B true CN104086664B (en) | 2017-01-25 |
Family
ID=51634446
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410252784.7A Expired - Fee Related CN104086664B (en) | 2014-06-09 | 2014-06-09 | Gracilaria lemaneiformis polysaccharide extract and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104086664B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105287648B (en) * | 2015-11-05 | 2019-02-12 | 广东医学院 | Sea is delivered vegetables the extraction preparation method and applications of active constituent |
| CN105535031A (en) * | 2015-12-30 | 2016-05-04 | 大连海洋大学 | Preparation method and application of asparagus extract |
| CN108586632B (en) * | 2018-06-28 | 2021-08-06 | 华南理工大学 | Gracilaria lemaneiformis polysaccharide with significant blood fat reducing activity and preparation method and application thereof |
| CN112159827B (en) * | 2020-10-16 | 2022-03-08 | 福建环海生物科技股份有限公司 | Production process of agar oligosaccharide |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101019666B (en) * | 2007-03-23 | 2010-09-08 | 广东药学院 | Asparagus and its processing method and application |
| CN101397346B (en) * | 2008-09-17 | 2010-08-11 | 上海海洋大学 | Method for preparing asparagus pure polysaccharide having immunoregulation role |
| CN101845102B (en) * | 2010-06-22 | 2011-12-21 | 中国烟草总公司郑州烟草研究院 | Preparation of asparagus rough polysaccharide and application thereof in cigarette |
-
2014
- 2014-06-09 CN CN201410252784.7A patent/CN104086664B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN104086664A (en) | 2014-10-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102964461B (en) | Auxiliary extraction method of biological enzyme for improving dissolution rate of dendrobe bioactive polysaccharide | |
| CN101412703B (en) | Composite extracting technique for coproduction of mulberry tea flavone, polysaccharide and alkaloid | |
| CN101747446B (en) | Method for extracting anti-fatigue acidic ginseng polysaccharide | |
| CN104086664B (en) | Gracilaria lemaneiformis polysaccharide extract and preparation method and application thereof | |
| CN101961371B (en) | Method for extracting and separating ginsenoside, flavone and polysaccharide from sweet gynostemma pentaphylla | |
| CN106905440A (en) | The new method that a kind of tremella polysaccharides is extracted | |
| CN103059162B (en) | A kind of novel method of high efficiency extraction lentinan | |
| CN106046192A (en) | Process for extracting pachyman from poria coccus wolf | |
| CN105483183A (en) | Preparation method of sargassum oligosaccharide and application of sargassum oligosaccharide in hypoglycemic drugs | |
| CN107557421A (en) | A kind of method of the effective component extracting from ganoderma lucidum | |
| CN103638089B (en) | A kind of method utilizing microwave-compound enzyme coupled method to extract alpha-amylase inhibitor from Semen Phaseoli Vulgaris | |
| CN106720752A (en) | Camellia chrysantha beverage and preparation method thereof | |
| CN101544998A (en) | Separation and purification preparation method and antineoplastic activity of tea polysaccharide | |
| CN1858068A (en) | Method for preparing pollen pini polyose, pollen pipe polyose and its use in medicine | |
| CN101225421A (en) | Novel process for extracting golden fungus polysaccharides by step enzyme method | |
| CN106748923B (en) | A kind of method that alliin is extracted from black garlic | |
| CN104262500B (en) | It is a kind of that there is immunocompetent novel red support dictyophora fungus polysaccharide and its preparation method and application | |
| CN103739731A (en) | Method for removing pesticide residue and heavy metal residue out of lentinan by using resin adsorption method | |
| CN105777921A (en) | Astragalus polysaccharide extraction process and application thereof | |
| CN104774827A (en) | Method for preparing alginate lyase from abalone internal organs | |
| CN101899120A (en) | Method for refining jujube polysaccharide | |
| CN102885847B (en) | Uses of camellia oleifera cake polysaccharide | |
| CN110013508A (en) | A kind of Traditional Chinese medicinal liquor improving antitumor immunity | |
| CN104945533B (en) | A kind of preparation method of active corn stigma holosaccharide | |
| CN107163163A (en) | A kind of processing method of chondroitin sulfate product |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190718 Address after: 516600 Intersection of Industrial Avenue and Wenhua Road, Shanwei City, Guangdong Province (behind Laodetou Trading Company) Patentee after: SHANWEI WEIMING BIOTECHNOLOGY CO.,LTD. Address before: Nansha District Avenue South Ring of 511458 cities in Guangdong province Guangzhou City, No. 25 Hua Da Guangzhou production and Research Institute Patentee before: South China University of Technology |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A Gracilaria lemaneiformis polysaccharide extract and its preparation method and Application Effective date of registration: 20201225 Granted publication date: 20170125 Pledgee: Yuecai Inclusive Finance (Shanwei) Financing Guarantee Co.,Ltd. Pledgor: SHANWEI WEIMING BIOTECHNOLOGY Co.,Ltd. Registration number: Y2020440000421 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20220113 Granted publication date: 20170125 Pledgee: Yuecai Inclusive Finance (Shanwei) Financing Guarantee Co.,Ltd. Pledgor: SHANWEI WEIMING BIOTECHNOLOGY CO.,LTD. Registration number: Y2020440000421 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| CP03 | Change of name, title or address |
Address after: 516600 Intersection of Industrial Avenue and Wenhua Road in Shanwei City, Guangdong Province (behind Laodetou Trading Company) Patentee after: SHANWEI WEIMING BIOTECHNOLOGY CO.,LTD. Address before: 516600 intersection of TONGHANG road and Wenhua Road, Industrial Avenue, Shanwei City, Guangdong Province (behind laodetou trading company) Patentee before: SHANWEI WEIMING BIOTECHNOLOGY CO.,LTD. |
|
| CP03 | Change of name, title or address | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170125 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |