CN105816702A - 一种甘蔗多酚提取物及其制备方法和应用 - Google Patents
一种甘蔗多酚提取物及其制备方法和应用 Download PDFInfo
- Publication number
- CN105816702A CN105816702A CN201610196161.1A CN201610196161A CN105816702A CN 105816702 A CN105816702 A CN 105816702A CN 201610196161 A CN201610196161 A CN 201610196161A CN 105816702 A CN105816702 A CN 105816702A
- Authority
- CN
- China
- Prior art keywords
- sacchari sinensis
- caulis sacchari
- extract
- preparation
- extraction
- 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
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 59
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 59
- 239000000284 extract Substances 0.000 title claims abstract description 56
- 238000000605 extraction Methods 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 240000000111 Saccharum officinarum Species 0.000 title abstract description 8
- 235000007201 Saccharum officinarum Nutrition 0.000 title abstract description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 54
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 53
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000003208 petroleum Substances 0.000 claims abstract description 16
- 238000010828 elution Methods 0.000 claims abstract description 12
- 239000000843 powder Substances 0.000 claims abstract description 12
- 239000004952 Polyamide Substances 0.000 claims abstract description 11
- 238000004440 column chromatography Methods 0.000 claims abstract description 11
- 229920002647 polyamide Polymers 0.000 claims abstract description 11
- 241000609240 Ambelania acida Species 0.000 claims abstract description 6
- 239000010905 bagasse Substances 0.000 claims abstract description 6
- 239000000706 filtrate Substances 0.000 claims abstract description 5
- 235000013376 functional food Nutrition 0.000 claims abstract description 5
- 238000001914 filtration Methods 0.000 claims abstract description 3
- 239000008346 aqueous phase Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000012071 phase Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 8
- 150000002989 phenols Chemical class 0.000 claims description 8
- 206010012601 diabetes mellitus Diseases 0.000 claims description 7
- 238000002481 ethanol extraction Methods 0.000 claims description 7
- HRGUSFBJBOKSML-UHFFFAOYSA-N 3',5'-di-O-methyltricetin Chemical compound COC1=C(O)C(OC)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 HRGUSFBJBOKSML-UHFFFAOYSA-N 0.000 claims description 6
- NGSWKAQJJWESNS-UHFFFAOYSA-N 4-coumaric acid Chemical compound OC(=O)C=CC1=CC=C(O)C=C1 NGSWKAQJJWESNS-UHFFFAOYSA-N 0.000 claims description 6
- IDDMFNIRSJVBHE-UHFFFAOYSA-N Piscigenin Natural products COC1=C(O)C(OC)=CC(C=2C(C3=C(O)C=C(O)C=C3OC=2)=O)=C1 IDDMFNIRSJVBHE-UHFFFAOYSA-N 0.000 claims description 6
- KCFYHBSOLOXZIF-UHFFFAOYSA-N dihydrochrysin Natural products COC1=C(O)C(OC)=CC(C2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 KCFYHBSOLOXZIF-UHFFFAOYSA-N 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- BMCJATLPEJCACU-UHFFFAOYSA-N tricin Natural products COc1cc(OC)c(O)c(c1)C2=CC(=O)c3c(O)cc(O)cc3O2 BMCJATLPEJCACU-UHFFFAOYSA-N 0.000 claims description 6
- 238000002137 ultrasound extraction Methods 0.000 claims description 6
- ZCOLJUOHXJRHDI-FZHKGVQDSA-N Genistein 7-O-glucoside Natural products O([C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1)c1cc(O)c2C(=O)C(c3ccc(O)cc3)=COc2c1 ZCOLJUOHXJRHDI-FZHKGVQDSA-N 0.000 claims description 5
- CJPNHKPXZYYCME-UHFFFAOYSA-N Genistin Natural products OCC1OC(Oc2ccc(O)c3OC(=CC(=O)c23)c4ccc(O)cc4)C(O)C(O)C1O CJPNHKPXZYYCME-UHFFFAOYSA-N 0.000 claims description 5
- YCUNGEJJOMKCGZ-UHFFFAOYSA-N Pallidiflorin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC(O)=C2C1=O YCUNGEJJOMKCGZ-UHFFFAOYSA-N 0.000 claims description 5
- 239000012141 concentrate Substances 0.000 claims description 5
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- WEPBGSIAWZTEJR-UHFFFAOYSA-N 3',4',5,7-tetrahydroxy-3-methoxyflavone Chemical compound O1C2=CC(O)=CC(O)=C2C(=O)C(OC)=C1C1=CC=C(O)C(O)=C1 WEPBGSIAWZTEJR-UHFFFAOYSA-N 0.000 claims description 3
- NGSWKAQJJWESNS-ZZXKWVIFSA-M 4-Hydroxycinnamate Natural products OC1=CC=C(\C=C\C([O-])=O)C=C1 NGSWKAQJJWESNS-ZZXKWVIFSA-M 0.000 claims description 3
- DFYRUELUNQRZTB-UHFFFAOYSA-N Acetovanillone Natural products COC1=CC(C(C)=O)=CC=C1O DFYRUELUNQRZTB-UHFFFAOYSA-N 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- RXHLADDBOLOIJO-UHFFFAOYSA-N quercetin 3-methyl ether Natural products COC1=C(Oc2c(O)cc(O)cc2C1=O)c3ccc(O)c(O)c3 RXHLADDBOLOIJO-UHFFFAOYSA-N 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 125000003976 glyceryl group Chemical group [H]C([*])([H])C(O[H])([H])C(O[H])([H])[H] 0.000 claims description 2
- 238000000227 grinding Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 229920002554 vinyl polymer Polymers 0.000 claims description 2
- 239000007795 chemical reaction product Substances 0.000 claims 1
- 239000000047 product Substances 0.000 abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 9
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 5
- 238000000746 purification Methods 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 2
- 235000013373 food additive Nutrition 0.000 abstract description 2
- 239000002778 food additive Substances 0.000 abstract description 2
- 235000019441 ethanol Nutrition 0.000 abstract 3
- 230000006837 decompression Effects 0.000 abstract 2
- 238000013375 chromatographic separation Methods 0.000 abstract 1
- 239000006185 dispersion Substances 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 26
- 239000000523 sample Substances 0.000 description 16
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 10
- 238000002835 absorbance Methods 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 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 8
- 239000008103 glucose Substances 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 7
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 description 7
- 235000005487 catechin Nutrition 0.000 description 7
- 229950001002 cianidanol Drugs 0.000 description 7
- 229930003944 flavone Natural products 0.000 description 7
- 235000011949 flavones Nutrition 0.000 description 7
- 229940074391 gallic acid Drugs 0.000 description 7
- 235000004515 gallic acid Nutrition 0.000 description 7
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 150000002213 flavones Chemical class 0.000 description 6
- 239000008055 phosphate buffer solution Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 230000001629 suppression Effects 0.000 description 5
- 229910021642 ultra pure water Inorganic materials 0.000 description 5
- 239000012498 ultrapure water Substances 0.000 description 5
- XUFXOAAUWZOOIT-SXARVLRPSA-N (2R,3R,4R,5S,6R)-5-[[(2R,3R,4R,5S,6R)-5-[[(2R,3R,4S,5S,6R)-3,4-dihydroxy-6-methyl-5-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)-1-cyclohex-2-enyl]amino]-2-oxanyl]oxy]-3,4-dihydroxy-6-(hydroxymethyl)-2-oxanyl]oxy]-6-(hydroxymethyl)oxane-2,3,4-triol Chemical compound O([C@H]1O[C@H](CO)[C@H]([C@@H]([C@H]1O)O)O[C@H]1O[C@@H]([C@H]([C@H](O)[C@H]1O)N[C@@H]1[C@@H]([C@@H](O)[C@H](O)C(CO)=C1)O)C)[C@@H]1[C@@H](CO)O[C@@H](O)[C@H](O)[C@H]1O XUFXOAAUWZOOIT-SXARVLRPSA-N 0.000 description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 4
- 229960002632 acarbose Drugs 0.000 description 4
- XUFXOAAUWZOOIT-UHFFFAOYSA-N acarviostatin I01 Natural products OC1C(O)C(NC2C(C(O)C(O)C(CO)=C2)O)C(C)OC1OC(C(C1O)O)C(CO)OC1OC1C(CO)OC(O)C(O)C1O XUFXOAAUWZOOIT-UHFFFAOYSA-N 0.000 description 4
- 102000016679 alpha-Glucosidases Human genes 0.000 description 4
- 108010028144 alpha-Glucosidases Proteins 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 108010051210 beta-Fructofuranosidase Proteins 0.000 description 4
- 238000000589 high-performance liquid chromatography-mass spectrometry 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
- 235000011073 invertase Nutrition 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 3
- GLEVLJDDWXEYCO-UHFFFAOYSA-N Trolox Chemical compound O1C(C)(C(O)=O)CCC2=C1C(C)=C(C)C(O)=C2C GLEVLJDDWXEYCO-UHFFFAOYSA-N 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 3
- 229940020947 fluorescein sodium Drugs 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000009533 lab test Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 238000004885 tandem mass spectrometry Methods 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 description 2
- YASAKCUCGLMORW-UHFFFAOYSA-N Rosiglitazone Chemical compound C=1C=CC=NC=1N(C)CCOC(C=C1)=CC=C1CC1SC(=O)NC1=O YASAKCUCGLMORW-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- QARRSDFIUQJIHX-QPZIMESWSA-O malvidin 3-glucoside-4-vinylguaiacol Chemical compound C1=C(O)C(OC)=CC(C=2OC3=CC(O)=CC=4[O+]=C(C(O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)=C(C3=4)C=2)C=2C=C(OC)C(O)=C(OC)C=2)=C1 QARRSDFIUQJIHX-QPZIMESWSA-O 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- UGNWTBMOAKPKBL-UHFFFAOYSA-N tetrachloro-1,4-benzoquinone Chemical compound ClC1=C(Cl)C(=O)C(Cl)=C(Cl)C1=O UGNWTBMOAKPKBL-UHFFFAOYSA-N 0.000 description 2
- 238000003828 vacuum filtration Methods 0.000 description 2
- QUTFFEUUGHUPQC-ILWYWAAHSA-N (2r,3r,4s,5r)-3,4,5,6-tetrahydroxy-2-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]hexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](C=O)NC1=CC=C([N+]([O-])=O)C2=NON=C12 QUTFFEUUGHUPQC-ILWYWAAHSA-N 0.000 description 1
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 description 1
- 208000010392 Bone Fractures Diseases 0.000 description 1
- 230000028937 DNA protection Effects 0.000 description 1
- 206010017076 Fracture Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 1
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003472 antidiabetic agent Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- KZNIFHPLKGYRTM-UHFFFAOYSA-N apigenin Chemical compound C1=CC(O)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 KZNIFHPLKGYRTM-UHFFFAOYSA-N 0.000 description 1
- XADJWCRESPGUTB-UHFFFAOYSA-N apigenin Natural products C1=CC(O)=CC=C1C1=CC(=O)C2=CC(O)=C(O)C=C2O1 XADJWCRESPGUTB-UHFFFAOYSA-N 0.000 description 1
- 235000008714 apigenin Nutrition 0.000 description 1
- 229940117893 apigenin Drugs 0.000 description 1
- 238000000889 atomisation Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 125000004403 catechin group Chemical group 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 description 1
- 229940114124 ferulic acid Drugs 0.000 description 1
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 description 1
- 235000001785 ferulic acid Nutrition 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 239000006199 nebulizer Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 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
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 229960001285 quercetin Drugs 0.000 description 1
- 235000005875 quercetin Nutrition 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 229960004586 rosiglitazone Drugs 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 description 1
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 description 1
- 235000012141 vanillin Nutrition 0.000 description 1
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
本发明属于保健食品技术领域，公开了一种甘蔗多酚提取物及其制备方法和应用。所述制备方法为：取甘蔗或蔗渣，粉碎后过筛得粗粉，然后将粗粉用乙醇溶液提取，过滤除去滤渣，滤液减压浓缩至浸膏；所得的浸膏加水分散，采用石油醚进行萃取，得到石油醚相和水相，用乙酸乙酯对水相进行萃取，收集乙酸乙酯相，减压浓缩至浸膏；然后通过聚酰胺柱层析分离，乙醇梯度洗脱，收集30％乙醇体积浓度的洗脱物，浓缩干燥得到甘蔗多酚提取物。本发明来源于大宗农作物甘蔗，通过多级萃取及柱层析梯度洗脱分离纯化，产品具有更强的降血糖活性，可用于食品添加剂和功能食品。
Description
技术领域
本发明属于保健食品技术领域，具体涉及一种甘蔗多酚提取物及其制备方法和应用。
背景技术
糖尿病是一种多因素引起的内分泌失调综合征，包括糖类、蛋白质以及脂肪的代谢紊乱。糖尿病患者分布在全世界各国家，患病人数也在逐年上升。糖尿病以及并发症不仅严重影响了患者的生活质量，也给个人和社会带来了沉重的经济负担。在所有的糖尿病确诊病例中，Ⅱ型糖尿病约占其中的90-95％。Ⅱ型糖尿病的发生，在很大程度上是由于生活习惯以及饮食方式的不合理，摄入的糖类过量，从而导致胰岛素及其相关受体的功能丧失。当前的降血糖药物在Ⅱ型糖尿病的治疗中起着重要的作用，但是，长期服用这些药物会对人体健康产生潜在的威胁，如诱发癌症、心肌梗塞以及骨折等。临床研究发现，二糖水解酶在糖尿病患者的小肠绒毛中异常活跃。蔗糖酶和麦芽糖酶是将淀粉水解物分解成游离的葡萄糖的两个关键酶。因此抑制蔗糖酶和麦芽糖酶的活性是治疗Ⅱ型糖尿病的关键所在；在降低多糖分解水平的同时，促进机体相关组织和器官对游离葡萄糖的摄取和转化，对于提高糖尿病治疗的效果具有重要的作用。因此，探寻对人体损伤小，抑制二糖水解酶的同时，能提高人体对葡萄糖的吸收的药物是一项具有意义的研究。
甘蔗广泛种植于全球热带地区和亚热带地区，是世界生产蔗糖的重要原料。甘蔗的年产量超过1亿吨。自古以来，甘蔗广泛用于中医治疗中，《本草纲目》有“蔗，脾之果也，其浆甘寒，能泻火热”的记载。经研究发现，甘蔗中含有丰富的天然多酚类物质，如槲皮素、阿魏酸、芹菜素等。甘蔗提取物在抗氧化、抗肿瘤、抑菌、DNA保护以及抗突变等方面有着潜在的生物活性。在甘蔗加工和食用的过程中，甘蔗往往会被当做废弃物，其生物价值被低估，因此探寻甘蔗的生物活性，尤其是甘蔗的降血糖生物活性具有重要的意义。
发明内容
为了解决现有技术的缺点和不足之处，本发明的首要目的在于提供一种甘蔗多酚提取物及其制备方法。
本发明的另一目的在于提供一种通过上述方法制备得到的甘蔗多酚提取物。
本发明的再一目的在于提供上述甘蔗多酚提取物在制备预防和治疗糖尿病的功能食品和药物中的应用。
本发明目的通过以下技术方案实现：
一种甘蔗多酚提取物的制备方法，包括以下步骤：
(1)取甘蔗或蔗渣，粉碎后过筛得粗粉，然后将粗粉用乙醇溶液提取，过滤除去滤渣，滤液减压浓缩至浸膏；
(2)将步骤(1)所得的浸膏加水分散，采用石油醚进行萃取，得到石油醚相和水相，用乙酸乙酯对水相进行萃取，收集乙酸乙酯相，减压浓缩至浸膏；
(3)将步骤(2)中获得的浸膏通过聚酰胺柱层析分离，乙醇梯度洗脱，收集30％乙醇浓度(体积比)的洗脱物，浓缩干燥得到甘蔗多酚提取物。
需要说明的是，上述提取物的制备方法中，还可直接对甘蔗或蔗渣用乙酸乙酯进行萃取后，采用聚酰胺柱层析分离，同样可以得到含量较低的甘蔗多酚提取物。
优选地，步骤(1)中所述粗粉与乙醇溶液的固液质量比为1:(5～15)，所述的乙醇溶液是指体积分数为60％～95％的乙醇水溶液。
优选地，步骤(1)中所述的提取是指在静态或动态下，进行浸渍提取或加热提取或回流提取或超声辅助提取。更优选超声辅助提取，超声辅助提取温度为50～70℃，提取时间为60～90min，超声功率为350W。
优选地，步骤(2)中所述的石油醚进行萃取是指用1～2倍体积的石油醚萃取，所述的用乙酸乙酯对水相进行萃取是指用1～2倍体积的乙酸乙酯萃取。
优选地，步骤(3)中所述聚酰胺柱层析分离是指采用30～60目聚酰胺柱层析分离；所述的乙醇梯度洗脱是指依次用体积比为30％、50％、70％和95％乙醇水溶液进行梯度洗脱，得到4个不同的极性部位，分别进行减压蒸发浓缩，得到30％、50％、70％和95％(30％E、50％E、70％E和95％E)共四个乙醇提取物，其中30％乙醇提取物即为具有高降血糖活性的甘蔗多酚提取物。
一种甘蔗多酚提取物，通过上述方法制备得到，所述甘蔗多酚提取物中总酚的质量百分含量为20％～98％，所述总酚包括二甲花翠素-4-乙烯基愈创木酚-3-葡萄糖苷(malvidin3-glucoside-4-vinylguaiacol，MGV)、槲皮素-3-甲基醚(quercetin3-methylether，Q3M)、苜蓿素(tricin)、苜蓿素-4-O-愈创木基丙三酯-7-O-吡喃葡萄糖苷(tricin4-O-(threoorerythro)guaiacylglycerylether-7-O-glucopyranoside，TGG)、p-香豆酸(p-coumaricacid,PCA)和染料木苷(genistin)这6种多酚的混合物，其中上述6种多酚占总酚质量的15％～85％。
上述6种多酚占总酚的质量百分含量为：1％～12％的MGV、5％～7％的Q3M、5％～20％的tricin、2％～35％的TGG、1％～5％的PCA、1％～6％的genistin。上述多酚的结构式分别如下所示：
TGG:R1＝R3＝R5＝H,R2＝OH,R4＝OGlc,R6＝R8＝OCH3,R7＝guaiacylglyceryl(threoorerythro)；
tricin:R1＝R3＝R5＝H,R2＝R4＝R7＝OH,R6＝R8＝OCH3；
Q3M:R1＝OCH3,R2＝R4＝R6＝R7＝OH,R3＝R5＝R8＝H。
上述甘蔗多酚提取物在制备预防和治疗糖尿病的功能食品和药物中的应用。
相对于现有技术，本发明具有如下优点及有益效果：
本发明来源于大宗农作物甘蔗，通过不同极性的有机溶剂对甘蔗多酚进行萃取，以纯化降血糖活性组分，减少或去除与降血糖活性无关的成分，纯化后的产品具有更强的降血糖活性，可用于食品添加剂和功能食品，具有安全、高效、生产工艺简单、成本低、原料易得的优点，具有广阔的市场开发前景。
附图说明
图1为本发明实施例1和2的甘蔗多酚提取物的提取技术路线图；
图2为本发明实施例所得产物甘蔗多酚提取物的葡萄糖吸收实验结果图；
图3为本发明实施例所得产物甘蔗多酚提取物的MS总粒子流图；
图4为本发明实施例所得产物甘蔗多酚提取物的MS/MS总离子流图。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述，但本发明的实施方式不限于此。
实施例1
本实施例的一种甘蔗多酚提取物的制备方法，其提取技术路线图如图1所示，具体提取步骤如下：
(1)选购新鲜的原料蔗，剥皮后将甘蔗粉碎，40℃烘干。过40目筛后得到粗粉，于-20℃保存。取300g上述甘蔗粗粉，用3000mL体积比为60％的乙醇水溶液，水浴回流温度60℃，提取90min，过滤，提取3次，真空抽滤后，合并滤液，减压浓缩至浸膏A1；
(2)将步骤(1)所得浸膏A1加水100mL分散，用石油醚萃取3次，每次加入石油醚100mL，分别收集石油醚萃取液和水相，将水相用100mL的乙酸乙酯萃取3次，收集乙酸乙酯相，减压浓缩至浸膏，制得浸膏A2；
(3)将浓缩浸膏A2通过30目聚酰胺柱层析，依次用体积比为30％、50％、70％和95％乙醇水溶液进行梯度洗脱，得到4个不同的极性部位，收集30％乙醇提取物组分，进行减压蒸发浓缩，冷冻干燥，得到产物甘蔗多酚提取物A3。
实施例2
本实施例的一种甘蔗多酚提取物的制备方法，其提取技术路线图如图1所示，具体提取步骤如下：
(1)取干燥蔗渣300g，粉碎后过40目筛得到粗粉，用1500mL体积比为95％的乙醇水溶液，在50℃超声辅助提取90min，超声功率为350w，提取3次。真空抽滤后，合并滤液，减压浓缩至浸膏B1；
(2)将步骤(1)所得浸膏B1加水100mL分散，用石油醚萃取3次，每次加入石油醚100mL，分别收集石油醚萃取液和水相，将水相用100mL的乙酸乙酯萃取3次，收集乙酸乙酯相，减压浓缩至浸膏，制得浸膏B2；
(3)将浓缩浸膏B2通过60目聚酰胺柱层析，依次用体积比为30％、50％、70％和95％乙醇水溶液进行梯度洗脱，得到4个不同的极性部位，收集30％乙醇提取物组分，进行减压蒸发浓缩，冷冻干燥，得到产物甘蔗多酚提取物B3。
实施例所得甘蔗多酚提取物理化性质测定
(1)总多酚含量的测定：
采用福林酚法。没食子酸标准曲线的制作：准确称量没食子酸标准品25.00mg，用超纯水溶解配制成1mg/mL的溶液，避光4℃保存备用。用超纯水稀释上述溶液得到0.1mg/mL的没食子酸标准溶液。将0.1mg/mL的没食子酸标准品配制成浓度依次为：0、20、60、100、150、200、300、400、500、600(μg/mL)的标准液。在试管中添加100μL的标准品，400μL的超纯水和100μLFolin-Ciocalteu试剂混合后静置6min。静置后加入7％Na2CO3溶液1mL和纯水0.8mL混合均匀。静置1.4h后在760nm波长下测定样品的吸光度值。至少重复三组平行实验。以没食子酸浓度(X)为横坐标，以吸光度(Y)为纵坐标绘制出没食子酸的标准曲线。
得到回归方程：Y＝0.0036X+0.0249，R2＝0.9995(式中Y为吸光度值，X为没食子酸浓度，0＜X＜600μg/mL)。
准确吸取化合物溶液100μL加入400μL的超纯水于试管中，采用福林酚法在760nm波长下测定样品的吸光度值，重复三组平行实验。根据上述回归方程计算出样品中多酚含量。多酚含量如下式计算。
多酚含量％＝(Y－0.0249)×100％/(0.0036×C×1000)
Y-吸光值；C-样品浓度(mg/mL)
(2)黄酮含量测定：
黄酮含量的测定采用硼氢化钠/四氯代苯对醌法(SBC)。称量29.0mg儿茶素用四氢呋喃：乙醇(1:1)溶解配制成10.0mmol/L的溶液。以四氢呋喃：乙醇(1:1)为溶剂将上述儿茶素溶液配置成浓度为0.3，0.5，1.0，2.0，4.0，5.0，6.0，8.0，10.0mmol/L的儿茶素标准品。在试管中加入1mL的儿茶素标准品，并做三个平行。加0.5mLNaBH4溶液和0.5mLAlCl3·6H2O溶液，混合均匀，在振荡器上震荡30min。加入0.5mLNaBH4溶液，再震荡30min。每个试管中加入2mL冰乙酸，再震荡15min(避光)。向每个试管加入1mL四氯代苯对醌溶液，将试管置于95℃的油浴锅中，进行1h的油浴。自然冷却，用甲醇定容至4mL。向每个试管加入1mL的16％香草醛甲醇溶液，再加入2mL12M的HCl，在室温下避光放置15min。后将试管中溶液转移至离心管，在2500rpm转速下，离心3min。之后在490nm波长下测其吸光度。以儿茶素浓度(X)为横坐标，以吸光度(Y)为纵坐标绘制出儿茶素的标准曲线。得到回归方程：Y＝0.1429X+0.0179，R2＝0.9923(式中X为儿茶素浓度，Y为吸光度值，0＜X＜10mmol/L)。
准确吸取化合物溶液100μL到试管中用氮吹仪干燥，用四氢呋喃：乙醇(1：1)定容到1mL。在490nm波长下测其吸光度，重复三组平行实验。根据上述回归方程计算出黄酮含量。黄酮含量按下式计算。
黄酮含量％＝(Y－0.0179)×290.27×100％/(0.1429×C×1000)；
Y-吸光值；C-样品浓度(mg/mL)。
经检测各提取阶段提取物中多酚和黄酮的含量如表1所示：
表1甘蔗提取物多酚和黄酮含量
(3)抗氧化活性(ORAC)测定：
用pH为7.4的75mMNaH2P04-Na2HPO4缓冲液配制浓度为500μM的荧光素钠储备液，配制完成后在－80℃下避光保存。用磷酸缓冲溶液稀释上述荧光素钠储备液，即得到使用液(50μM)。取4℃下保存的ABAP，用磷酸缓冲溶液配制成119.4μM，每次使用的ABAP均为新鲜配制。精确称量Trolox，溶于磷酸缓冲溶液，配成50μM溶液，并用磷酸缓冲溶液稀释成不同浓度，以磷酸缓冲溶液稀释样品溶液。
将酶标仪预温至37℃，设定激发波长为485nm，发射波长为538nm。在96孔板中，每孔加入20μL样品溶液或Trolox溶液，或磷酸缓冲溶液(空白对照)，然后加入200μL荧光素钠使用液，将孔板放入酶标仪中，37℃孵育20分钟。加入20μLABAP后，开始计时反应并读数，共读35次(共计反应155分钟)，将每次读数连成曲线。每个样品设置3个复孔。AUC表示曲线下的面积。Trolox浓度与其NetAUC是成正比的，将样品NetAUC代入所得标准曲线，换算得到ORAC值，即得每毫克粗提物(或浸膏)相当于Torlox的量(μmolTE/g)。实施例1所得A3，抗氧化活性结果如表2所示。
表2实施例1所得甘蔗多酚提取物A3抗氧化活性
(4)体外降血糖实验
将实施例2中所取得的样品用乙醇溶解，用磷酸盐缓冲液稀释成不同浓度的待测液，储存于-20℃冰箱备用，阿卡波糖(AB)为阳性对照。取1g鼠小肠丙酮粉用10mL0.9％NaCl溶解，依次进行超声(1min×3)，离心(3000rpm×30min)处理，取上清液用水稀释4倍作为酶溶液备用。蔗糖、麦芽糖和样品均用100mM磷酸盐缓冲溶液(pH6.9)配成所需浓度。取底物(0.2mL的3.5mM麦芽糖溶液或0.4mL的56mM蔗糖溶液)于10mL比色管中，加入50μL待测液，于37℃的水浴中孵育5min。然后，添加0.2mL酶溶液，37℃继续反应20min。最后，将比色管放入沸水浴，5min进行灭酶。冷却后用葡萄糖检测试剂盒检测样品吸光度。以阿卡波糖为阳性对照，同时设定空白组(缓冲液+酶液+底物)，空白对照组(缓冲液+缓冲液+底物)，样品测定组(样品+酶液+底物)，样品对照组(样品+缓冲液+底物)。
蔗糖酶/麦芽糖酶抑制率％＝(1–(A1–A0)/(A3–A2))×100％
式中A0，A1，A2，A3分别为505nm处空白对照组、空白组、样品对照组和样品测定组的吸光值。其结果如表3，表4所示。
表3各组分不同浓度提取物蔗糖酶抑制率
表4各组分不同浓度提取物麦芽糖酶抑制率
(5)葡萄糖吸收实验
细胞培养：人肝癌HepG2细胞用高糖Dulbeccominimumessentialmedium(DMEM)培养基添加10％胎牛血清培养于含有5％CO2，37℃的培养箱中。
葡萄糖吸收：将用高糖DMEM培养基(含10％胎牛血清)培养的人肝癌HepG2细胞以1.5×104个/孔的密度种植于黑色96孔板上，24h之后将实施例3中所制备的30％乙醇洗脱相(30％E)溶解于DMSO中，用高糖培养基(无血清，含有50μM葡萄糖类似物2-NBDG，1μM胰岛素)稀释至一定的浓度，阿卡波糖(10μM)和罗格列酮(10μM)为阳性对照。药物作用24h之后，去除培养基，用冷的磷酸盐缓冲液冲洗细胞3次，将96孔板至于荧光酶标仪下读数。激发波长设定于485nm，发射波长设定于535nm。结果如图2所示。
(6)HPLC-MS/MS测定甘蔗多酚成分
用HPLC-MS/MS对实施例所得产物甘蔗多酚提取物进行成分分析。取10mg甘蔗多酚提取物A3或B3溶于10mL甲醇溶液中，离心(12,000g)，用0.45μm的滤膜过滤后，用HPLC-MS/MS系统分析。
色谱条件：进样体积为20μL，流动相A：含有0.05％乙酸的超纯水，流动相B：色谱甲醇；梯度洗脱条件：0-12min,70A/30B；12-13min,30A/70B；13-15min,100B。
质谱条件：离子源：ESI；离子化模式：正离子模式；毛细管电压：2000V；干燥气：N2；雾化气流量：4L/min；雾化器压力：1bar；干燥器温度：180℃。液质分析结果如表5所示。
表5实施例所得产物甘蔗多酚提取物的HPLC-MS/MS分析结果
a为MS或者MS/MS离子片段。其MS总粒子流图和MS/MS总离子流图分别如图3和图4所示。
由上述结果可以看出，本发明的甘蔗多酚提取物(30％乙醇提取物)具有很好的体外降血糖效果。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其它的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。
Claims (10)
1.一种甘蔗多酚提取物的制备方法，其特征在于包括以下步骤：
(1)取甘蔗或蔗渣，粉碎后过筛得粗粉，然后将粗粉用乙醇溶液提取，过滤除去滤渣，滤液减压浓缩至浸膏；
(2)将步骤(1)所得的浸膏加水分散，采用石油醚进行萃取，得到石油醚相和水相，用乙酸乙酯对水相进行萃取，收集乙酸乙酯相，减压浓缩至浸膏；
(3)将步骤(2)中获得的浸膏通过聚酰胺柱层析分离，乙醇梯度洗脱，收集30％乙醇体积浓度的洗脱物，浓缩干燥得到甘蔗多酚提取物。
2.根据权利要求1所述的一种甘蔗多酚提取物的制备方法，其特征在于：步骤(1)中所述粗粉与乙醇溶液的固液质量比为1:(5～15)g/mL，所述的乙醇溶液是指体积分数为60％～95％的乙醇水溶液。
3.根据权利要求1所述的一种甘蔗多酚提取物的制备方法，其特征在于：步骤(1)中所述的提取是指在静态或动态下，进行浸渍提取或加热提取或回流提取或超声辅助提取。
4.根据权利要求3所述的一种甘蔗多酚提取物的制备方法，其特征在于：所述超声辅助提取温度为50～70℃，提取时间为60～90min，超声功率为350W。
5.根据权利要求1所述的一种甘蔗多酚提取物的制备方法，其特征在于：步骤(2)中所述的石油醚进行萃取是指用1～2倍体积的石油醚萃取，所述的用乙酸乙酯对水相进行萃取是指用1～2倍体积的乙酸乙酯萃取。
6.根据权利要求1所述的一种甘蔗多酚提取物的制备方法，其特征在于：步骤(3)中所述聚酰胺柱层析分离是指采用30～60目聚酰胺柱层析分离；所述的乙醇梯度洗脱是指依次用体积比为30％、50％、70％和95％乙醇水溶液进行梯度洗脱，分别进行减压蒸发浓缩，得到30％、50％、70％和95％共四个乙醇提取物，其中30％乙醇提取物即为最终产物。
7.根据权利要求1所述的一种甘蔗多酚提取物的制备方法，其特征在于：直接对甘蔗或蔗渣用乙酸乙酯进行萃取后，收集乙酸乙酯相，减压浓缩至浸膏；然后将获得的浸膏通过聚酰胺柱层析分离，乙醇梯度洗脱，收集30％乙醇体积浓度的洗脱物，浓缩干燥得到甘蔗多酚提取物。
8.一种甘蔗多酚提取物，其特征在于：通过权利要求1～6任一项所述的方法制备得到，所述甘蔗多酚提取物中总酚的质量百分含量为20％～98％，所述总酚包括二甲花翠素-4-乙烯基愈创木酚-3-葡萄糖苷、槲皮素-3-甲基醚、苜蓿素、苜蓿素-4-O-愈创木基丙三酯-7-O-吡喃葡萄糖苷、p-香豆酸和染料木苷这6种多酚的混合物，其中上述6种多酚占总酚质量的15％～85％。
9.根据权利要求8所述的一种甘蔗多酚提取物，其特征在于：上述6种多酚占总酚的质量百分含量为：1％～12％的MGV、5％～7％的Q3M、5％～20％的tricin、2％～35％的TGG、1％～5％的PCA、1％～6％的genistin。
10.权利要求8或9所述的甘蔗多酚提取物在制备预防和治疗糖尿病的功能食品和药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610196161.1A CN105816702B (zh) | 2016-03-31 | 2016-03-31 | 一种甘蔗多酚提取物及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610196161.1A CN105816702B (zh) | 2016-03-31 | 2016-03-31 | 一种甘蔗多酚提取物及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105816702A true CN105816702A (zh) | 2016-08-03 |
| CN105816702B CN105816702B (zh) | 2020-01-14 |
Family
ID=56525464
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610196161.1A Active CN105816702B (zh) | 2016-03-31 | 2016-03-31 | 一种甘蔗多酚提取物及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105816702B (zh) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106177555A (zh) * | 2016-08-18 | 2016-12-07 | 广东省生物工程研究所（广州甘蔗糖业研究所） | 一种从甘蔗浮渣中提取甘蔗多酚的方法 |
| CN107522754A (zh) * | 2017-08-31 | 2017-12-29 | 浙江大学 | 一种提取甘蔗皮多酚的方法 |
| CN108041599A (zh) * | 2017-12-15 | 2018-05-18 | 华南理工大学 | 凤眼果果皮提取物及其制备方法和应用 |
| CN113248382A (zh) * | 2021-05-26 | 2021-08-13 | 云南农业大学 | 一种抗氧化化合物及其制备方法和应用 |
| CN113367287A (zh) * | 2021-06-24 | 2021-09-10 | 福建达利食品科技有限公司 | 低gi豆奶配方及生产工艺 |
| CN116114876A (zh) * | 2023-02-28 | 2023-05-16 | 东莞市东南部中心医院 | 一种含甘蔗多酚与莱茵衣藻的降血糖组合物及其制备方法 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102516319A (zh) * | 2011-12-08 | 2012-06-27 | 广西大学 | 从甘蔗梢中提取酚类物质的方法 |
-
2016
- 2016-03-31 CN CN201610196161.1A patent/CN105816702B/zh active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102516319A (zh) * | 2011-12-08 | 2012-06-27 | 广西大学 | 从甘蔗梢中提取酚类物质的方法 |
Non-Patent Citations (1)
| Title |
|---|
| 陈纯等: "响应面法优化超声波提取甘蔗皮多酚的工艺条件", 《中国食品学报》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106177555A (zh) * | 2016-08-18 | 2016-12-07 | 广东省生物工程研究所（广州甘蔗糖业研究所） | 一种从甘蔗浮渣中提取甘蔗多酚的方法 |
| CN106177555B (zh) * | 2016-08-18 | 2019-07-12 | 广东省生物工程研究所（广州甘蔗糖业研究所） | 一种从甘蔗浮渣中提取甘蔗多酚的方法 |
| CN107522754A (zh) * | 2017-08-31 | 2017-12-29 | 浙江大学 | 一种提取甘蔗皮多酚的方法 |
| CN108041599A (zh) * | 2017-12-15 | 2018-05-18 | 华南理工大学 | 凤眼果果皮提取物及其制备方法和应用 |
| CN108041599B (zh) * | 2017-12-15 | 2021-08-10 | 华南理工大学 | 凤眼果果皮提取物及其制备方法和应用 |
| CN113248382A (zh) * | 2021-05-26 | 2021-08-13 | 云南农业大学 | 一种抗氧化化合物及其制备方法和应用 |
| CN113248382B (zh) * | 2021-05-26 | 2023-08-18 | 云南农业大学 | 一种抗氧化化合物及其制备方法和应用 |
| CN113367287A (zh) * | 2021-06-24 | 2021-09-10 | 福建达利食品科技有限公司 | 低gi豆奶配方及生产工艺 |
| CN116114876A (zh) * | 2023-02-28 | 2023-05-16 | 东莞市东南部中心医院 | 一种含甘蔗多酚与莱茵衣藻的降血糖组合物及其制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105816702B (zh) | 2020-01-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105816702A (zh) | 一种甘蔗多酚提取物及其制备方法和应用 | |
| Saggu et al. | Adaptogenic and safety evaluation of seabuckthorn (Hippophae rhamnoides) leaf extract: a dose dependent study | |
| CN101863871B (zh) | 蔷薇红景天总苷及其医药用途和制备方法 | |
| CN104661668A (zh) | 从山蚂蝗属制备植物提取物的方法及其提取物 | |
| Naik et al. | Physicochemical and Phytochemical Analysis of Different Plant Parts of Annona muricata L.(Annonaceae). | |
| Rao et al. | Quercetin 3-galactoside from Azadirachta indica | |
| CN109232491A (zh) | 一种华泽兰中苯并呋喃类化合物的制备方法及用途 | |
| Wang et al. | Pistafolin B is the major aldose reductase inhibitor of the pods of tara [Caesalpinia spinose (Molina) Kuntze] | |
| CN1989984B (zh) | 川芎有效组分、制备方法及其制剂与用途 | |
| Chaika et al. | Research in phytochemical composition and hypoglycemic activity screening of the dry extracts from bearberry leaves | |
| CN105030914A (zh) | 粗壮女贞苦丁茶提取物在α-葡萄糖苷酶抑制剂的应用 | |
| CN102151306B (zh) | 一种从荔枝果肉中提取的活性组合物及其制备方法和应用 | |
| Fecka et al. | In Vitro Antiglycation and Methylglyoxal Trapping Effect of Peppermint Leaf (Mentha× piperita L.) and Its Polyphenols | |
| CN102727593A (zh) | 金荞麦及其提取物在制备降血糖药物及保健食品中的新用途 | |
| Hsieh et al. | A rapid quantitative 1 H NMR analysis of kinsenoside and other bioactive principles from Anoectochilus formosanus | |
| CN102670698B (zh) | 千斤拔提取物在制备防治糖尿病药物中的应用 | |
| Sun et al. | Antioxidant activity and immunomodulatory of extracts from roots of Actinidia kolomikta | |
| Qamar | Antidiabetic activity, polyphenols-based characterization and molecular interaction of extract of un-ripe pods of Vinca rosea cv. Pink | |
| Li et al. | The chemical and pharmacological research progress on a kind of Chinese herbal medicine, Fructus Malvae | |
| Kutovaya et al. | Flavonoid-profile determination for a hypoglycemic collection by high-performance liquid chromatography | |
| Abd El-Aziz et al. | Biological and chemical evaluation of the use of Acacia nilotica (L.) in the Egyptian traditional medicine | |
| CN107174605A (zh) | 一种黄毛草莓正丁醇提取物及其制备方法和应用 | |
| CN113786423B (zh) | 一种金丝皇菊挥发油及其组合物在制备治疗糖脂代谢综合紊乱的产品方面的应用 | |
| CN108896500A (zh) | 一种白藤梨根抗肿瘤活性物质的筛选及提取方法 | |
| CN103739657B (zh) | 一种油茶皂苷化合物、其制备方法、应用及其制备的抗肿瘤药物 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |