CN116694721B - Ganoderma lucidum glycopeptide and preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream - Google Patents
Ganoderma lucidum glycopeptide and preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream Download PDFInfo
- Publication number
- CN116694721B CN116694721B CN202310993467.XA CN202310993467A CN116694721B CN 116694721 B CN116694721 B CN 116694721B CN 202310993467 A CN202310993467 A CN 202310993467A CN 116694721 B CN116694721 B CN 116694721B
- Authority
- CN
- China
- Prior art keywords
- ganoderma lucidum
- glycopeptide
- ganoderma
- microemulsion
- skin
- 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.)
- Active
Links
- 102000002068 Glycopeptides Human genes 0.000 title claims abstract description 131
- 108010015899 Glycopeptides Proteins 0.000 title claims abstract description 129
- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 119
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 119
- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical compound NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 title claims abstract description 104
- 239000004530 micro-emulsion Substances 0.000 title claims abstract description 72
- 239000006071 cream Substances 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 49
- 241000222336 Ganoderma Species 0.000 claims abstract description 30
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 21
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 238000002156 mixing Methods 0.000 claims abstract description 16
- 206010051246 Photodermatosis Diseases 0.000 claims abstract description 12
- 230000008845 photoaging Effects 0.000 claims abstract description 12
- 238000000502 dialysis Methods 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 10
- 239000004064 cosurfactant Substances 0.000 claims abstract description 9
- 238000004108 freeze drying Methods 0.000 claims abstract description 8
- 239000002888 zwitterionic surfactant Substances 0.000 claims abstract description 8
- 239000006228 supernatant Substances 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims abstract description 5
- 239000000047 product Substances 0.000 claims abstract description 5
- 108010046516 Wheat Germ Agglutinins Proteins 0.000 claims abstract description 4
- 239000011347 resin Substances 0.000 claims abstract description 4
- 229920005989 resin Polymers 0.000 claims abstract description 4
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 30
- 239000003814 drug Substances 0.000 claims description 16
- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical group O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical group CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims description 6
- 239000003963 antioxidant agent Substances 0.000 claims description 5
- 230000003078 antioxidant effect Effects 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 5
- 239000000287 crude extract Substances 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 102000004142 Trypsin Human genes 0.000 claims description 3
- 108090000631 Trypsin Proteins 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 239000012588 trypsin Substances 0.000 claims description 3
- 229940083466 soybean lecithin Drugs 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract description 9
- 239000000284 extract Substances 0.000 abstract description 4
- 238000000605 extraction Methods 0.000 abstract description 3
- 230000001815 facial effect Effects 0.000 abstract description 2
- 238000007254 oxidation reaction Methods 0.000 abstract description 2
- 238000000751 protein extraction Methods 0.000 abstract description 2
- 238000010298 pulverizing process Methods 0.000 abstract description 2
- 239000008176 lyophilized powder Substances 0.000 abstract 1
- 238000003760 magnetic stirring Methods 0.000 abstract 1
- 230000003647 oxidation Effects 0.000 abstract 1
- 210000003491 skin Anatomy 0.000 description 74
- 241000700159 Rattus Species 0.000 description 48
- 239000012071 phase Substances 0.000 description 41
- 210000001519 tissue Anatomy 0.000 description 36
- 230000000694 effects Effects 0.000 description 28
- 239000003921 oil Substances 0.000 description 21
- 239000000243 solution Substances 0.000 description 18
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol group Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 17
- 238000012360 testing method Methods 0.000 description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- 238000002474 experimental method Methods 0.000 description 14
- 239000004094 surface-active agent Substances 0.000 description 14
- 229940079593 drug Drugs 0.000 description 13
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 12
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 10
- 229940118019 malondialdehyde Drugs 0.000 description 10
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 9
- 102000000380 Matrix Metalloproteinase 1 Human genes 0.000 description 9
- 108010016113 Matrix Metalloproteinase 1 Proteins 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- 210000004207 dermis Anatomy 0.000 description 9
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 9
- 229960002591 hydroxyproline Drugs 0.000 description 9
- 238000010587 phase diagram Methods 0.000 description 9
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 9
- VKUYLANQOAKALN-UHFFFAOYSA-N 2-[benzyl-(4-methoxyphenyl)sulfonylamino]-n-hydroxy-4-methylpentanamide Chemical compound C1=CC(OC)=CC=C1S(=O)(=O)N(C(CC(C)C)C(=O)NO)CC1=CC=CC=C1 VKUYLANQOAKALN-UHFFFAOYSA-N 0.000 description 8
- 102000004889 Interleukin-6 Human genes 0.000 description 8
- 108090001005 Interleukin-6 Proteins 0.000 description 8
- 102100030416 Stromelysin-1 Human genes 0.000 description 8
- 101710108790 Stromelysin-1 Proteins 0.000 description 8
- 210000002615 epidermis Anatomy 0.000 description 8
- 235000019441 ethanol Nutrition 0.000 description 8
- 239000008347 soybean phospholipid Substances 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
- 102000008186 Collagen Human genes 0.000 description 5
- 108010035532 Collagen Proteins 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 229920001436 collagen Polymers 0.000 description 5
- 239000002537 cosmetic Substances 0.000 description 5
- 210000004177 elastic tissue Anatomy 0.000 description 5
- 102000003777 Interleukin-1 beta Human genes 0.000 description 4
- 108090000193 Interleukin-1 beta Proteins 0.000 description 4
- 229920002565 Polyethylene Glycol 400 Polymers 0.000 description 4
- NWGKJDSIEKMTRX-AAZCQSIUSA-N Sorbitan monooleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O NWGKJDSIEKMTRX-AAZCQSIUSA-N 0.000 description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 4
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 4
- 230000001186 cumulative effect Effects 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 239000000835 fiber Substances 0.000 description 4
- 150000004676 glycans Chemical class 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 230000001953 sensory effect Effects 0.000 description 4
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 3
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 3
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 3
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 3
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 3
- 239000005642 Oleic acid Substances 0.000 description 3
- HVUMOYIDDBPOLL-XWVZOOPGSA-N Sorbitan monostearate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O HVUMOYIDDBPOLL-XWVZOOPGSA-N 0.000 description 3
- 230000001133 acceleration Effects 0.000 description 3
- 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 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 239000004359 castor oil Substances 0.000 description 3
- 235000019438 castor oil Nutrition 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 230000002500 effect on skin Effects 0.000 description 3
- 229960001031 glucose Drugs 0.000 description 3
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 3
- 229910001385 heavy metal Inorganic materials 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 3
- XUGNVMKQXJXZCD-UHFFFAOYSA-N isopropyl palmitate Chemical compound CCCCCCCCCCCCCCCC(=O)OC(C)C XUGNVMKQXJXZCD-UHFFFAOYSA-N 0.000 description 3
- 229940119170 jojoba wax Drugs 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 3
- JLFNLZLINWHATN-UHFFFAOYSA-N pentaethylene glycol Chemical compound OCCOCCOCCOCCOCCO JLFNLZLINWHATN-UHFFFAOYSA-N 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000009759 skin aging Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 3
- 230000037303 wrinkles Effects 0.000 description 3
- AXDJCCTWPBKUKL-UHFFFAOYSA-N 4-[(4-aminophenyl)-(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methyl]aniline;hydron;chloride Chemical compound Cl.C1=CC(=N)C(C)=CC1=C(C=1C=CC(N)=CC=1)C1=CC=C(N)C=C1 AXDJCCTWPBKUKL-UHFFFAOYSA-N 0.000 description 2
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 102000000589 Interleukin-1 Human genes 0.000 description 2
- 108010002352 Interleukin-1 Proteins 0.000 description 2
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 2
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 102000016611 Proteoglycans Human genes 0.000 description 2
- 108010067787 Proteoglycans Proteins 0.000 description 2
- BAECOWNUKCLBPZ-HIUWNOOHSA-N Triolein Natural products O([C@H](OCC(=O)CCCCCCC/C=C\CCCCCCCC)COC(=O)CCCCCCC/C=C\CCCCCCCC)C(=O)CCCCCCC/C=C\CCCCCCCC BAECOWNUKCLBPZ-HIUWNOOHSA-N 0.000 description 2
- PHYFQTYBJUILEZ-UHFFFAOYSA-N Trioleoylglycerol Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC(OC(=O)CCCCCCCC=CCCCCCCCC)COC(=O)CCCCCCCC=CCCCCCCCC PHYFQTYBJUILEZ-UHFFFAOYSA-N 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 238000002845 discoloration Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 2
- 229940093471 ethyl oleate Drugs 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 125000005456 glyceride group Chemical group 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 238000000265 homogenisation Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 230000003859 lipid peroxidation Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 230000003020 moisturizing effect Effects 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 210000004927 skin cell Anatomy 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid group Chemical group S(O)(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000002834 transmittance Methods 0.000 description 2
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 2
- 229940117972 triolein Drugs 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 208000032544 Cicatrix Diseases 0.000 description 1
- AEMOLEFTQBMNLQ-DTEWXJGMSA-N D-Galacturonic acid Natural products O[C@@H]1O[C@H](C(O)=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-DTEWXJGMSA-N 0.000 description 1
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 208000002197 Ehlers-Danlos syndrome Diseases 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241001480597 Ganoderma tsugae Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 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
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 241000222341 Polyporaceae Species 0.000 description 1
- 206010063493 Premature ageing Diseases 0.000 description 1
- 208000032038 Premature aging Diseases 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000219053 Rumex Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- FUQHZPGSFAWKGW-UHFFFAOYSA-N [Na+].[Na+].[Na+].[O-]B([O-])[O-].OS(O)(=O)=O Chemical compound [Na+].[Na+].[Na+].[O-]B([O-])[O-].OS(O)(=O)=O FUQHZPGSFAWKGW-UHFFFAOYSA-N 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- IAJILQKETJEXLJ-RSJOWCBRSA-N aldehydo-D-galacturonic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-RSJOWCBRSA-N 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229910052785 arsenic Inorganic materials 0.000 description 1
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 1
- 238000003287 bathing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- AEMOLEFTQBMNLQ-UHFFFAOYSA-N beta-D-galactopyranuronic acid Natural products OC1OC(C(O)=O)C(O)C(O)C1O AEMOLEFTQBMNLQ-UHFFFAOYSA-N 0.000 description 1
- 238000007068 beta-elimination reaction Methods 0.000 description 1
- 102000023732 binding proteins Human genes 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- UBXYXCRCOKCZIT-UHFFFAOYSA-N biphenyl-3-ol Chemical group OC1=CC=CC(C=2C=CC=CC=2)=C1 UBXYXCRCOKCZIT-UHFFFAOYSA-N 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 239000001045 blue dye Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 230000023402 cell communication Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000011382 collagen catabolic process Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- TWNIBLMWSKIRAT-VFUOTHLCSA-N levoglucosan Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@H]2CO[C@@H]1O2 TWNIBLMWSKIRAT-VFUOTHLCSA-N 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 210000002445 nipple Anatomy 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 230000037311 normal skin Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 150000002482 oligosaccharides Polymers 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 238000001139 pH measurement Methods 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 150000003214 pyranose derivatives Chemical class 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 230000037387 scars Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000037075 skin appearance Effects 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 210000000434 stratum corneum Anatomy 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- -1 trioleate Natural products 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000036269 ulceration Effects 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/04—Dispersions; Emulsions
- A61K8/06—Emulsions
- A61K8/068—Microemulsions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Birds (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Epidemiology (AREA)
- General Engineering & Computer Science (AREA)
- Water Supply & Treatment (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Dispersion Chemistry (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
Abstract
Ganoderma glycopeptide, its preparation method and application, and Ganoderma glycopeptide microemulsion and facial cream, and relates to the technical field of protein extraction, which improves Ganoderma application value and solves skin photoaging problem. Pulverizing dried Ganoderma fruiting body into coarse powder, mixing with pure water, heating for extraction, performing ultrasonic extraction, and mixing the supernatants to obtain coarse extract of Ganoderma glycopeptide; removing free protein, and dialyzing with dialysis bag; enzymolysis, eluting with wheat germ lectin resin until the molecular weight cut-off is less than 10000, and freeze drying to obtain Ganoderma glycopeptide lyophilized powder. The ganoderma lucidum glycopeptide and water are prepared into a water phase, a zwitterionic surfactant and a cosurfactant are mixed according to the proportion, then the mixture is mixed with an oil phase, and the water phase is added dropwise under magnetic stirring to obtain the ganoderma lucidum glycopeptide microemulsion. The invention can be applied to the preparation of products for resisting oxidation or skin photoaging.
Description
Technical Field
The invention relates to the technical field of protein extraction, in particular to a ganoderma lucidum glycopeptide and a preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream.
Background
Skin is the first line of defense of human body to the external environment, and in daily life, normal skin tissue structure and function are extremely important for body health. Ultraviolet rays, blue light, ozone, etc., which are exposed to the external environment, cause a series of premature aging phenomena of the skin, and ultraviolet rays are the factors which are most easily contacted by people. Ultraviolet rays in sunlight are mainly classified into three types, namely ultraviolet rays A (UVA), ultraviolet rays B (UVB) and ultraviolet rays C (UVC), wherein UVA (320-400 nm) and UVB (275-320 nm) have strong penetrating power, can directly reach the epidermis layer and dermis layer of skin, and are main causes of photoaging of skin.
Glycopeptides are binding proteins formed by connecting an oligosaccharide chain and a polypeptide chain through a covalent bond, and can be divided into two main types, namely N-glycosidic bonds and O-glycosidic bonds according to different glycosidic bonds formed by dehydration of hydroxyl groups on the carbon atoms of the hetero heads of the glycosyl groups and amide groups or hydroxyl groups on amino acid residues of the peptide chains. Glycopeptides are widely present in animals, plants and microorganisms, and are indispensable biological macromolecules for vital phenomena such as cell communication, immune regulation and the like in organisms. Because of the properties of both protein and sugar, the protein has various biological activities, such as antioxidation, anti-tumor, blood sugar and blood lipid reduction, organism immunity improvement and the like.
Ganoderma, also called Ganoderma, herba Mesonae chinensis, rumex, herba Hedyotidis Diffusae, herba Linderae Caesalpiniae, herba Cannabifoliae, and herba Cannabifoliae, is a fungus of Polyporaceae, and is widely distributed in nature, and its cultivation area is mainly distributed in southeast coast, middle and downstream of Yangtze river, and great Khingan area. The fresh ganoderma lucidum is rich in six nutritional elements of protein, carbohydrate, fat, vitamins, inorganic matters and water, also contains components such as ganoderma lucidum adenosine, ganoderma lucidum total alkali and the like, and is a fungus medicament which contains rich nutritional components and various development potentials. The ganoderma lucidum has wide application, can resist radiation, aging and oxidization, and has obvious effects of enhancing human immunity, regulating blood sugar, controlling blood pressure, protecting liver, promoting sleep and the like.
Unlike ganoderan (GL-PS), less research is carried out on ganoderan peptide at present, and patent CN1629186A (ganoderan protein, preparation method and application thereof) discloses ganoderan protein Gl-PP obtained by extracting from ganoderma, which is obtained by taking ganoderma as a raw material matrix to extract liquid, concentrating the extract liquid, and then, carrying out ultrafiltration or dialysis to intercept high molecular weight polysaccharide protein; also relates to a preparation method of the ganoderan protein capable of being industrially produced and application of the ganoderan protein Gl-PP in preparing anti-tumor drugs. The ultrafiltration method provided by the patent can be used for extracting ganoderan protein, so that the angiogenesis of tumor and the invasion of cancer cells can be inhibited, and the growth of tumor can be inhibited. Patent CN1715298 discloses that the crude product of ganoderma tsugae proteoglycan is a mixture containing three single proteoglycans P1, P2 and P3, the polysaccharide parts of the crude product are uniform glucosan, and the glucosidic bonds are beta- (1-3) configuration, which has the physiological effects of improving the immune function of the organism, resisting cell mutation, reducing blood fat and blood sugar, protecting liver, soothing the nerves, calming the nerves, resisting tumors and the like; through molecular biology and cell pharmacology research, it can regulate cell division and differentiation, and regulate cell growth and death. Is used for preparing antitumor drugs. In summary, the currently disclosed ganoderma lucidum glycopeptides are mostly used for anti-tumor aspects, and do not relate to the application in anti-aging and skin photoaging directions.
Disclosure of Invention
In order to further improve the use value of ganoderma lucidum and solve the problems of skin photoaging and the like, the invention provides ganoderma lucidum glycopeptide, a preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream.
The technical scheme of the invention is as follows:
a preparation method of ganoderma lucidum glycopeptides comprises the following steps:
s1, taking dried ganoderma lucidum fruiting bodies and crushing the dried ganoderma lucidum fruiting bodies into ganoderma lucidum coarse powder;
s2, mixing the ganoderma lucidum coarse powder with pure water according to 1g: uniformly mixing (50-70) mL of feed liquid ratio by shaking through a shaking table, heating and extracting for 10-12 h at 60-80 ℃, then performing ultrasonic extraction for at least 3 times for 15-30 min each time, and combining the supernatant to obtain ganoderma lucidum glycopeptide crude extract;
s3, removing free protein from the ganoderma lucidum glycopeptide crude extract, and then carrying out dialysis treatment by using a dialysis bag;
s4, carrying out enzymolysis on the dialyzed ganoderma lucidum glycopeptide, eluting with wheat germ lectin resin until the molecular weight cutoff is less than 10000, and freeze-drying to obtain ganoderma lucidum glycopeptide freeze-dried powder.
Preferably, the shaking table oscillates at a rate of 120 r/min-180 r/min.
Preferably, the removal of the free protein in step S3 is specifically performed by the Sevage method; the dialysis treatment specifically comprises the following steps: the solution was dialyzed in distilled water using a 0.1kDa dialysis bag for 48 hours.
Preferably, the enzymolysis is specifically: treating dialyzed ganoderma lucidum glycopeptides in a water bath at 50-60 ℃ for 30-35 min, adding trypsin with an enzyme-to-bottom ratio of 4%, magnetically stirring for 5-6 h, then placing in a constant-temperature water bath at 80-85 ℃ for 15-20 min, and finally cooling to room temperature.
Preferably, the temperature of the freeze-drying in step S4 is-86 ℃ and the time of the freeze-drying is at least 48 hours.
The invention also provides the ganoderma lucidum glycopeptide which is prepared by the preparation method, and the combination mode of the glycopeptide bond of the ganoderma lucidum glycopeptide is O-glycosidic bond.
The invention also provides application of the ganoderma lucidum glycopeptide, which is used for preparing an antioxidant or skin photo-aging resistant skin care product.
The invention also provides a ganoderma lucidum glycopeptide microemulsion which is prepared by using the ganoderma lucidum glycopeptide as the raw material.
The preparation method of the ganoderma lucidum glycopeptide microemulsion comprises the following steps:
the ganoderma lucidum glycopeptide and water are prepared into a water phase with the concentration of the medicine of 10 mg/mL-20 mg/mL, and a zwitterionic surfactant and a cosurfactant are mixed according to the proportion of 1: mixing (0.5-2) by mass ratio, mixing with oil phase, magnetically stirring at 23-27 ℃ and 500-1000 rpm, and dropwise adding water phase to obtain the ganoderma lucidum glycopeptide microemulsion.
Preferably, the zwitterionic surfactant is soybean phospholipid, the cosurfactant is ethanol, and the oil phase is isopropyl myristate.
The invention also provides a face cream which comprises the ganoderma lucidum glycopeptide microemulsion.
Compared with the prior art, the invention has the following specific beneficial effects:
1. the extraction method of the ganoderma lucidum glycopeptides is simple to operate, and the obtained ganoderma lucidum glycopeptides are high in extraction rate and stable in structure, so that the use value of ganoderma lucidum medicinal materials is further improved, and the development channel of the novel cosmetic field is expanded;
2. the ganoderma lucidum glycopeptide (GL-Gp) provided by the invention can slow down skin aging caused by ultraviolet rays, improve the activity and HYP content of SOD, GSH-Px in skin tissues, and reduce the content of MDA, MMP-1, MMP-3, IL-1 beta, IL-6 and TNF-alpha in skin tissues; pharmacological experiments show that the composition has remarkable antioxidant and skin photoaging repairing effects, and can be used for research and application of photoaging-resistant skin care products.
Drawings
FIG. 1 is a graph of ultraviolet spectra of ganoderma lucidum glycopeptides before and after alkali treatment;
FIG. 2 is an infrared spectrum of ganoderma lucidum glycopeptides;
FIG. 3 is an SDS-PAGE electrophoresis of ganoderma lucidum glycopeptides;
FIG. 4 is a diagram showing the amino acid composition of ganoderan;
fig. 5 is a pseudo ternary phase diagram of each oil phase with mixed surfactant when km=1:1;
FIG. 6 is a pseudo ternary phase diagram of isopropyl myristate and mixed surfactant formation at different Km;
FIG. 7 is a pseudo-ternary phase diagram at different drug concentrations;
FIG. 8 is a contour plot of factors A, B, C versus OD;
FIG. 9 is a three-dimensional effect plane graph of the factors A, B, C versus OD values;
FIG. 10 is a flow chart of a process for preparing the ganoderma lucidum glycopeptide microemulsion cream;
FIG. 11 is a graph showing the moisture retention of a ganoderma lucidum glycopeptide microemulsion face cream, wherein A is a glycerol group, B is a blank group, C is sample 1, D is sample 2, E is sample 3, F is sample 4, and G is sample 5;
FIG. 12 is an external view of a ganoderma lucidum glycopeptide microemulsion cream;
FIG. 13 is a comparison of the appearance of a stability test, wherein, A is 25 ℃ for 24 hours, B is 40 ℃ for 24 hours, C is-20 ℃ for 24 hours, and D is after cold and heat resistance acceleration test;
FIG. 14 is a graph showing the morphological effects of ganoderan on the photoaging appearance of rats;
FIG. 15 is a HE staining chart of a pathological section of the photoaged skin of a rat;
FIG. 16 is a graph of aldehyde fuchsin staining of pathological sections of photoaged skin of rats;
FIG. 17 is a graph showing the effect of ganoderan on MDA content in photoaged skin tissue of rats;
FIG. 18 is a graph showing the effect of ganoderan on SOD activity in rat skin tissue;
FIG. 19 is a graph showing the effect of ganoderan on GSH-Px content in rat skin tissue;
FIG. 20 is a graph showing the effect of ganoderan on HYP content in rat skin tissue;
FIG. 21 is a graph showing the effect of ganoderan on MMP-1 content in rat skin tissue;
FIG. 22 is a graph showing the effect of ganoderan on MMP-3 content in rat skin tissue;
FIG. 23 is a graph showing the effect of ganoderan on IL-6 content in rat skin tissue;
FIG. 24 is a graph showing the effect of ganoderan on IL-1β content in rat skin tissue;
FIG. 25 is a graph showing the effect of ganoderan on TNF- α levels in rat skin tissue.
Detailed Description
In order to make the technical solution of the present invention clearer, the technical solution of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings of the present invention, and it should be noted that the following embodiments are only used for better understanding of the technical solution of the present invention, and should not be construed as limiting the present invention.
Example 1.
In the embodiment, the ganoderma lucidum glycopeptide GL-Gp is extracted, and the specific operation is as follows:
pulverizing dried fruiting body of Ganoderma, mixing with 54mL pure water at shaking table vibration rate of 167r/min, heating at 71 deg.C for extracting for 12 hr, ultrasonic extracting for 30min for 3 times, and mixing the supernatants to obtain coarse extract of Ganoderma glycopeptide;
removing free protein from the obtained crude extract of ganoderma lucidum glycopeptide by adopting a Sevage method to obtain ganoderma lucidum glycopeptide solution, placing the obtained ganoderma lucidum glycopeptide solution in a dialysis bag of 0.1kDa, dialyzing for 48h, carrying out enzymolysis on the dialyzate, treating for 35min in a water bath at 60 ℃, adding trypsin with the enzyme bottom ratio of 4%, magnetically stirring for 6h, then placing in a constant-temperature water bath at 85 ℃ for 20min, cooling to room temperature, eluting with wheat germ lectin resin until the molecular weight cutoff is less than 10000, and freeze-drying to obtain ganoderma lucidum glycopeptide powder.
The ganoderma lucidum glycopeptides obtained in this example are characterized as follows:
(1) Beta-elimination experiment:
weighing 2mg of ganoderma lucidum glycopeptide, dissolving in 10ml of 0.4mol/L NaOH, carrying out water bath for 30min at 60 ℃, and scanning at 190-350 nm by using an ultraviolet spectrophotometer; the ganoderma lucidum glycopeptides which were not treated with 0.4mol/L NaOH were scanned under the same conditions. The results are shown in FIG. 1, wherein curve A represents the glossy ganoderma glycopeptides treated by NaOH, curve B represents the glossy ganoderma glycopeptides not treated by NaOH, and the light absorption value of the glossy ganoderma glycopeptides solution treated by NaOH is increased at the wavelength of 240nm, which indicates that polysaccharide and protein in the glossy ganoderma glycopeptides are connected in a covalent bond mode, and the connection mode is O-glycosidic bond.
(2) And (3) infrared spectrum analysis:
grinding small amount of Ganoderma glycopeptide and KBr, tabletting, and infrared scanning to obtain scanning result shown in figure 2, which can be seen in 3300 and 3300 cm -1 、2900 cm -1 、1640 cm -1 、1400 c cm -1 And 1000 cm -1 Absorption peaks are arranged on the left and right sides, which indicates that polysaccharide and protein molecules are contained; at 2930 cm -1 There is an absorption, characteristic absorption peaks indicative of methyl, methylene and other alkyl vibrations; amide at 1640 and 1640 cm -1 Has absorption peak, and the stretching vibration of sulfuric acid group is 1400 cm -1 At 1050 cm -1 The presence of an absorption peak indicates the presence of a pyranose configuration in the sugar chain.
The above results demonstrate that the present example successfully extracts ganoderma lucidum glycopeptides having O-glycosidic bonds as the binding mode of glycopeptides.
(3) Molecular weight determination:
the relative molecular weight of the obtained ganoderma lucidum glycopeptides was determined by gel electrophoresis in combination with coomassie brilliant blue dye pairs, and the result is shown in fig. 3. It is obvious that after SDS polyacrylamide gel electrophoresis separation, the ganoderma lucidum glycopeptides contain proteins with different molecular weights and are mainly distributed in 55 kD-70 kD, 35 kD-55 kD and 15 kD-25 kD.
(4) Analysis of amino acid composition results:
further determination of the amino acid composition of Gl-Gp using an amino acid analyzer, the results are shown in FIG. 4, and it can be seen that Gl-Gp consists of 18 amino acids: asp (5.86 mg.g) -1 ），Glu（4.08mg·g -1 ），Ser（2.65mg·g -1 ），His（0.23mg·g -1 ），Gly（1.95mg·g -1 ），Thr（1.36mg·g -1 ），Arg（0.54mg·g -1 ），Ala（2.35mg·g -1 ），Tyr（0.28mg·g -1 ），Cys（0.64mg·g -1 ），Val（0.85mg·g -1 ），Met（0.049mg·g -1 ），Phe（0.83mg·g -1 ），Ile（0.55mg·g -1 ），Lys（0.43mg·g -1 ），Leu（1.45mg·g -1 ），Pro（1.32mg·g -1 ），Trp（1.01mg·g -1 ）。
(5) Protein content in ganoderma lucidum glycopeptides was determined using BCA method:
preparing a protein standard solution of 0.5mg/mL, respectively sucking 0 mu L, 1 mu L, 2 mu L, 4 mu L, 8 mu L, 12 mu L, 16 mu L and 20 mu L, adding the protein standard solution into standard wells of a 96-well plate, adding a diluent to make up 20 mu L, adding 200 mu LBCA working solution into each well, standing for 20-30 min at 37 ℃, and measuring absorbance at 562nm wavelength. Drawing a standard curve by taking absorbance as an ordinate and the standard mass concentration of the protein as an abscissa, and obtaining a regression curve equation of y=0.8305x+0.2149, R 2 =0.9993。
(6) Acid sugar content determination:
accurately sucking 0.5mg/mL glucuronic acid quasi-product solution 0mL, 0.1mL, 0.25mL, 0.4mL, 0.5mL, 0.75mL, 1.0mL into 10mL test tubes, adding distilled water to 1.0mL, cooling in ice water bath, adding four materials2.5mL of sodium borate-sulfuric acid solution, uniformly mixing, heating in a boiling water bath for 10min, cooling to room temperature, adding 50 mu L of m-hydroxybiphenyl solution, carrying out vortex oscillation for 5min, standing at room temperature for 1h, taking distilled water as a blank group, and measuring absorbance at a wavelength of 525 nm. Drawing a standard curve by taking the concentration of D-galacturonic acid as an abscissa and the absorbance value as an ordinate to obtain a regression curve equation of y=0.001x+0.0557, R 2 =0.9952。
(7) Neutral sugar content determination:
accurately weighing 1mg of anhydrous glucose as a standard substance, fixing the volume by a volumetric flask with 25mL to prepare 40 mug/mL of standard substance solution, respectively sucking 0mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL, 1.4mL and 1.6mL of glucose standard substance solution into a test tube, adding distilled water to supplement 2mL, adding 1.0mL of 6% phenol solution, adding 5.0mL of concentrated sulfuric acid, standing for 10min, shaking uniformly, reacting at room temperature for 20min again, and measuring absorbance at 490nm wavelength. Drawing a standard curve by taking absorbance value A as an ordinate and glucose concentration as an abscissa to obtain a regression curve equation of y=0.0096 x-0.017, R 2 =0.9962。
According to calculation, the yield of the ganoderma lucidum glycopeptides (namely the amount of the obtained glycopeptides freeze-dried powder/3 g ganoderma lucidum coarse powder) in the embodiment is 9.05 percent, and the purity is 86.12 percent (sugar content+protein content).
Example 2.
In the embodiment, the area size of the formed pseudo ternary phase diagram is assumed to be used as an investigation index, and the following single-factor investigation is carried out on the oil phase, the zwitterionic surfactant, the cosurfactant, the Km value and the drug concentration before the microemulsion is prepared:
(1) Oil phase selection:
ethyl oleate, oleic acid, isopropyl myristate, isopropyl palmitate, trioleate, castor oil, jojoba oil and neodecanoate glyceride were initially selected as the oil phase. The mass ratio of the fixed soybean phospholipids to the absolute ethyl alcohol is 1:1 (Km=1:1) as a mixed surfactant, the fixed soybean phospholipids and the absolute ethyl alcohol are mixed with each oil phase according to the mass ratio of 1:1, the water added at the critical point is recorded, the mass percentage of the water phase is calculated, and the mass percentage of the water phase when each oil phase reaches the critical point is obtained as shown in table 1.
TABLE 1
From the results, the water phase mass percentages of the triolein, the castor oil, the jojoba oil, the caprylic/capric glyceride and the oleic acid are too small, so that the triolein, the castor oil, the jojoba oil, the caprylic/capric glyceride and the oleic acid are not considered to be used as oil phases; when ethyl oleate, isopropyl myristate and isopropyl palmitate are used as oil phases to form the microemulsion, the mass percentage of the water phase is equivalent, and screening is carried out continuously through a pseudo ternary phase diagram.
When km=1:1, the pseudo-ternary phase diagram formed by each oil phase and the mixed surfactant is shown in fig. 5, and the area of the microemulsion region is compared by AutoCAD2015 software, it was found that the area of the microemulsion region formed is the largest when the oil phase is isopropyl myristate.
(2) Zwitterionic surfactant and cosurfactant selection:
the mutual dissolution of different types of zwitterionic surfactants (soybean phospholipid, span 80 and span 60) and cosurfactants (ethanol, propylene glycol, glycerol and PEG 400) is examined, the composition of the mixed surfactants is primarily screened, km=1 and isopropyl palmitate are fixed and mixed according to a certain proportion, then the water carrying capacity residual microemulsion is examined to form difficulty, and the composition of the microemulsion surfactant is determined by taking the difficulty as an index.
Through testing, span 60 is insoluble in propylene glycol, glycerol and PEG400, and after span 60 is mixed with ethanol, the water content in isopropyl myristate is 0.49%, 0.33% and 0.39%; layering phenomenon can occur after span 80 is uniformly mixed with propylene glycol, glycerol and PEG400, water is added to the span to be turbid immediately after the span 80 is mixed with oil phase, and the water content in isopropyl myristate is 3.56% after the span 80 is mixed with ethanol; the soybean phospholipid is dissolved very slowly with glycerol and PEG400, is dissolved relatively slowly with propylene glycol, is viscous after being mixed with oil phase, is not easy to observe phase inversion phenomenon, is dissolved rapidly in ethanol, has a water content of 28.15% in isopropyl myristate after being mixed with ethanol, has a high water content, and has the characteristics of good biocompatibility, safety and no toxicity. Therefore, soybean phospholipids are selected as the surfactant and ethanol is selected as the cosurfactant.
(3) Selection of Km values:
the area of the microemulsion region formed by the mixed surfactant and the preferable oil phase when the Km value is 1:2, 1:1 and 2:1 is examined respectively, and the optimal Km value is determined.
When km=2:1, 1:1, 1:2, the pseudo-ternary phase diagram of isopropyl myristate and mixed surfactant is shown in fig. 6, the areas of the micro-emulsion areas are compared by AutoCAD2015 software, and the areas of the micro-emulsion areas formed when km=1:1 are larger than km=2:1 and km=1:2, and km=1 is selected as the best.
(4) Drug concentration selection:
the sizes of the microemulsion areas formed when the concentrations are 5mg/mL, 15mg/mL and 25mg/mL are respectively examined, and the optimal drug concentration is determined according to the state of the ganoderma lucidum glycopeptide aqueous solution after centrifugation. The test shows that the pseudo-ternary phase diagram formed under different drug concentrations is shown in figure 7, and the drug concentration has little influence on the formation area of the microemulsion pseudo-ternary phase diagram, so that 15mg/mL is selected as the optimal drug concentration according to the state of the drug solution after centrifugation shown in Table 2.
TABLE 2
| Drug concentration | Phenomenon after centrifugation |
| 5mg/mL | No insoluble matter |
| 15 mg/mL | No insoluble matter |
| 25 mg/mL | Presence of insolubilityArticle (B) |
(5) Optimally mixing and designing:
according to the single factor experimental investigation result, the preparation process is determined as follows: isopropyl myristate is used as an oil phase, the mass ratio of soybean phospholipid to absolute ethyl alcohol is 1:1 (Km=1), the drug concentration is 15mg/mL, and the sum of the three phases of water phase, oil phase and mixed surfactant is set to be 1 according to the microemulsion forming area, so that the optimal limit range of the mass fractions of each phase is obtained. The D-optimal mixing test in Design Expert 13 software is adopted, the water phase (A), the oil phase (B) and the mixed surfactant (C) are used as optimal components, and the experiment is designed to determine the optimal prescription of the ganoderma lucidum glycopeptide microemulsion, and experimental factors and levels are shown in Table 3.
TABLE 3 Table 3
| Component (A) | Water phase (A) | Oil phase (B) | Mixed surfactant (C) |
| Minimum value of | 0.13 | 0.23 | 0.26 |
| Maximum value of | 0.27 | 0.61 | 0.51 |
Taking OD as a response value of process optimization, and carrying out mathematical conversion on the encapsulation efficiency EE and the drug loading DL to obtain a normalized value (d), wherein the formula is as follows:
，/>，
wherein d 1 And d 2 Respectively represent glossy ganoderma glycopeptides EE and DL, Y 1 As measured value, Y max For the maximum value of EE and DL, Y mix And d values of all indexes are calculated according to a formula to obtain a total normalized value OD for the minimum value of the measured EE and DL. The experimental design and results are shown in Table 4.
TABLE 4 Table 4
Fitting and optimizing a model:
taking OD as a response value and A, B, C as a variable, the obtained fitting equation is OD=2.26A+0.0621B+1.15C-4.89 AB-1.63AC-2.24BC+21.25ABC-4.77AB (A-B) +3.19AC (A-C) +3.79BC (B-C). The model has significanceP<0.0001 No significance of the mismatching termP=0.7670>0.05），R 2 =0.9985，R adj 2 As can be seen from 0.9963, the model has a good fitting degree with the actual microemulsion prescription, the contour plot and the three-dimensional effect plot of the factors a, B, and C versus the OD values are shown in fig. 8 and 9, respectively, and the model fitting results are shown in table 5.
TABLE 5
(6) Optimal prescription and verification experiment:
according to the result, the optimal prescription of the ganoderma lucidum glycopeptide microemulsion is: the mass percent of the water phase is 0.260, the mass percent of the oil phase is 0.365, the mass percent of the mixed surfactant is 0.375, and the predicted OD value is 0.991.
The results of three verifiable experiments on the optimal prescription are shown in Table 6, and the stability of the prescription can be proved to be good.
TABLE 6
Thus, in this example, ganoderma glycopeptide was formulated with water to give an aqueous phase with a pharmaceutical concentration of 15mg/mL, and soybean phospholipid and absolute ethanol were mixed at a ratio of 1:1, mixing with isopropyl myristate, magnetically stirring at 25 ℃ at 800rpm, dropwise adding water phase to make the mass percent of the mixed water phase be 0.260, the mass percent of the oil phase be 0.365 and the mass percent of the mixed surfactant be 0.375, and stirring until clear emulsion is formed, thus obtaining the ganoderma lucidum glycopeptide microemulsion.
Effect example 1.
In vitro transdermal experiments were performed on the ganoderma lucidum glycopeptide microemulsion prepared in example 2:
removing epidermal hair and subcutaneous connective tissue from the isolated abdominal mouse skin, washing with physiological saline, and sucking to dryness for later use; in vitro transdermal experiments were performed using a Franz diffusion cell, contacting the stratum corneum of the isolated murine skin with a drug supply cell, the dermis layer facing downward with a receiving cell, and ensuring no air bubbles between the dermis layer and the receiving solution (normal saline); adding 1mL of Ganoderma glycopeptide water solution and Ganoderma glycopeptide microemulsion into the drug supply tank, respectively, and water-bathing at 37deg.C, 3000rmin -1 At the rotational speeds, 0.5mL was sampled at 2h, 4h, 8h, 12h, 18h, 24h, 36h in the EP tube, and the cumulative transdermal rates at the respective time points were measured and calculated according to the protein content test conditions in example 1.
The calculation formula is as follows:
，/>，
in which Q n Cumulative transmission per unit area (μg cm) -2 ) Cn is the concentration of ganoderan (mg.mL) at each sampling time point -1 ），V 0 For receiving pool volume (mL), ci is the concentration of ganoderan at a time point prior to each sampling time point, V 1 A is the effective permeation area (cm) of the diffusion cell for the sample volume (mL) 2 ) W is the dosage (mg.cm) -2 ）。
The experimental results are shown in Table 7, and the skin cumulative transmittance of the ganoderma lucidum glycopeptide microemulsion in 36h is (20.98 + -0.34)%. Experimental results show that the microemulsion can remarkably improve the percutaneous permeability of the ganoderma lucidum glycopeptides within 2-36 hours.
TABLE 7
| Time | Cumulative transmittance (%) |
| 0 | 0 |
| 2 | 3.99±0.52* |
| 4 | 5.39±0.28** |
| 8 | 6.57±0.19*** |
| 12 | 9.31±0.48** |
| 18 | 13.90±0.90** |
| 24 | 17.34±0.38*** |
| 36 | 20.98±0.34*** |
Example 3.
The ganoderma lucidum glycopeptide microemulsion cream is prepared in the embodiment, the formula is shown in Table 8, the preparation process is shown in FIG. 10, A, B phases are respectively weighed in beakers, heated in water bath at 80-85 ℃ and stirred for dissolution, B phase is poured into A phase for homogenization, C phase is added for homogenization again after the temperature is reduced to 60 ℃, and D phase is added finally, and stirring is continued until the temperature is 38 ℃ to obtain the ganoderma lucidum glycopeptide microemulsion cream.
TABLE 8
In the embodiment, the ganoderma lucidum glycopeptide microemulsion is taken as a variable of the experiment, 1mL, 2mL, 3mL, 4mL and 5mL of ganoderma lucidum glycopeptide microemulsion are added into 100g of cosmetics, and the ganoderma lucidum glycopeptide microemulsion face cream is prepared respectively according to the preparation process.
Effect example 2.
The ganoderma lucidum glycopeptide microemulsion creams of different addition amounts of ganoderma lucidum glycopeptide microemulsion prepared in example 3 were evaluated by using sensory evaluation and skin moisturization as indexes.
(1) The sensory evaluation index is shown in Table 9.
TABLE 9
Taking the ganoderma lucidum glycopeptide microemulsion as a variable, adding a sample No. 1mL, adding a sample No. 2mL and the like, and summarizing sensory scores of 30 testers on ganoderma lucidum glycopeptide microemulsion face cream, wherein the sample No. 1 and the sample No. 2 are adopted, and the sensory scores are summarized in Table 10. The score of sample No. 3 was optimal in terms of appearance, smoothness, spreadability, wetness, thickness, and absorbency, and the score of sample No. 4 was optimal in terms of spreadability, but there was no significant difference (P > 0.05) from sample No. 3.
Table 10
(2) Skin moisturization evaluation:
test conditions: the temperature is (21+/-1) DEG C, and the humidity is 40% -60%.
Two hours before testing, the test area was kept dry and cleaned with dry facial tissues; the subject needs to sit still for 30min before applying the sample; the whole test period is monitored in real time, and the moisture index of the test area is kept at 15% -45% as effective.
The test area is set to be 3cm multiplied by 3cm, the interval between the areas is 1cm, the same experimenter smears different samples in equal quantity, and after smearing for 1h, 2h and 3h, the skin moisture indexes are measured respectively, and the moisture retention rate is calculated. 2% aqueous glycerol was used as a control group and distilled water was used as a blank control group.
The calculation formula is as follows:
。/>
the evaluation results are shown in fig. 11, wherein a is a glycerol group, B is a blank group, C is sample 1, d is sample 2, e is sample 3, f is sample 4, g is sample 5, and the moisture retention after 1 hour of application is 3 # 5 # 4 >2 # 1 > Gan Youzu > blank group; after smearing 2h and 3h, the moisture retention rate is 3 No. 4 >5 No. 2 >1 > Gan Youzu > blank. The 5 groups of creams added with the ganoderma lucidum glycopeptide microemulsion have obvious moisturizing capability (P is less than 0.001), and the moisturizing performance of the No. 3 sample added with the 3mL ganoderma lucidum glycopeptide microemulsion is the best.
Effect example 3.
Quality evaluation was performed on the ganoderma lucidum glycopeptide microemulsion cream prepared in example 3 to which 3mL of ganoderma lucidum glycopeptide microemulsion was added:
(1) Appearance characteristics:
the glossy ganoderma glycopeptide microemulsion cream is described in terms of appearance, texture, smell and use feeling. The texture of the cream is shown in figure 12, and the glossy ganoderma glycopeptide microemulsion cream is in a milky cream shape, and has uniform and glossy texture; the face cream has no special smell, and the face is moistened and moisturized.
(2) And (3) pH value measurement:
the pH value of the cream was measured using a pH meter, and the glossy ganoderma glycopeptide microemulsion cream was mixed with distilled water at a ratio of 1:9, and carrying out pH measurement on the ganoderma lucidum glycopeptide microemulsion face cream diluted solution, wherein the pH value of the ganoderma lucidum glycopeptide microemulsion face cream is (7.44+/-0.08) according with the specified range of the pH value of the cosmetic between 4.5 and 8.5.
(3) Heavy metal and microbiological examination:
heavy metals and microorganisms in ganoderma lucidum glycopeptide microemulsion face cream are detected according to cosmetic safety technical Specification (2015 edition). The results are shown in Table 11, and the heavy metal and microorganism detection of the ganoderma lucidum glycopeptide micro-emulsion cream meets the cosmetic standard.
TABLE 11
| Test item | Test results | Standard value |
| Lead (mg kg-1) | Not detected<2） | ≤10 |
| Mercury (mg kg-1) | Not detected<0.01） | ≤1 |
| Arsenic (mg kg-1) | Not detected<0.01） | ≤2 |
| Cadmium (mg kg-1) | Not detected<0.5） | ≤5 |
| Colony count (CFU g-1) | <10 | ≤1000 |
| Total number of mold and yeast (CFU g-1) | <10 | ≤100 |
| Staphylococcus aureus (g) | Not detected | Cannot be detected |
| Heat resistant E.coli group (g) | Not detected | Cannot be detected |
| Pseudomonas aeruginosa (g) | Not detected | Cannot be detected |
Effect example 4.
Stability experiments were performed on the ganoderma lucidum glycopeptide microemulsion cream prepared in example 3 to which 3mL ganoderma lucidum glycopeptide microemulsion was added:
(1) Heat resistance experiment:
placing the sealed Ganoderma glycopeptide microemulsion face cream in a 40 deg.C oven, taking out respectively at 1, 7, 15 days, comparing with Ganoderma glycopeptide microemulsion face cream placed at room temperature, and observing layering, dilution, and discoloration.
(2) Cold resistance experiment:
placing the sealed Ganoderma glycopeptide microemulsion face cream in a refrigerator at-20deg.C, taking out respectively at 1, 7, and 15 days, comparing with Ganoderma glycopeptide microemulsion face cream placed at room temperature, and observing layering, dilution, and discoloration.
(3) Cold and heat resistant acceleration experiment:
placing the sealed ganoderma lucidum glycopeptide microemulsion face cream in a baking oven at 40 ℃ for 24h, placing at room temperature for 12h, placing at a refrigerator at-20 ℃ for 24h and placing at room temperature for 12h, completing cold-resistant and heat-resistant acceleration experiments, comparing with the ganoderma lucidum glycopeptide microemulsion face cream placed at room temperature, and observing the phenomena of layering, dilution, color change and the like.
(4) Centrifugal experiment
At room temperature, the ganoderma lucidum glycopeptide microemulsion face cream is processed by the steps of 3000r min -1 Centrifuging for 30min, comparing with non-centrifuged Ganoderma glycopeptide microemulsion cream, and observing layering.
The stability test results are shown in table 12 and fig. 13, and it can be seen that the ganoderma lucidum glycopeptide microemulsion cream has no layering, dilution, color change and other phenomena, which indicates that the ganoderma lucidum glycopeptide microemulsion cream has a feasible preparation process and good stability.
Table 12
Effect example 5.
Skin photoaging resistance effect test is carried out on ganoderma lucidum glycopeptide microemulsion face cream:
1. building an animal photo-aging model:
removing back hair from the rat to expose back skin, wherein the area is about 4cm×4cm; radiating the shaved rats under an ultraviolet lamp (UVA+UVB), radiating for 0.5h on the 1 st day, radiating for 1h on the 2 nd to 4 th days, radiating for 2h on the 5 th to 10 th days, radiating for 3h on the 11 th to 21 th days, and finishing molding; after the molding is finished, the back of the positive control group is smeared with 0.5mL vitamin E10 mg/mL, and the back of the ganoderma lucidum glycopeptide microemulsion high/medium/low dose group (Gl-Gp H/M/L) is smeared with 25mg/mL, 10mg/mL and 1mg/mL respectively with 0.5mL for 7 days.
The skin morphology of the rats after ultraviolet injury is observed in fig. 14, the back skin state of the rats in the blank group is smooth, no wrinkles and no visible scars are formed; the rough and thickened epidermis of the model group can be seen as obvious red swelling or slight ulceration, and transverse rough wrinkles can be seen to exist continuously and cannot disappear along with the movement of rats; the skin of the rat treated by VE and ganoderan is improved to a certain extent, the skin wrinkles and skin lifting are reduced compared with those of the model group, and the skin appearance of the photo-aged rat is improved more obviously along with the increase of the concentration of the ganoderan.
2. Skin treatment:
after administration, the rat spine is dislocated, skin tissues are sheared, subcutaneous connective tissues are removed, and the rat spine is weighed; adding 9 times of physiological saline, grinding into homogenate, centrifuging at 4000r/min for 20min, collecting tissue supernatant, and preserving at-80deg.C.
3. Tissue section:
taking 1cm multiplied by 1cm of skin on the back of a rat, placing the rat in 4% paraformaldehyde fixing solution for fixing for more than 48 hours, then dehydrating skin tissues with a gradient of 50% -100% ethanol, degreasing in dimethylbenzene, embedding in paraffin, and cutting the embedded skin tissue wax blocks into slices with the thickness of 5 mu m for HE dyeing; HE staining was performed according to the kit instructions and morphological changes of skin cells were observed as shown in fig. 15. It can be seen that the blank group skin has compact and thin epidermis structure and obvious dermal papilla, so that the dermis and epidermis are tightly connected, and the dermis reticulation layer is tightly and uniformly arranged; the cuticle of the skin of the model group is obviously thickened, the structure is loose, the connection between the cuticle layer and the dermis layer is lost, the dermis nipple disappears, and the reticular layer is loosely arranged; the thickness of the epidermis of the VE group and the ganoderma lucidum glycopeptide low-dose group is slightly improved, but the structure is still loose, and the dermal papilla is not obvious yet; the epidermis is thinned in the ganoderma lucidum glycopeptide and is visible in the high-dose group, the papilla of the dermis is recovered, and the reticular layers are arranged tightly, wherein the dermis of the ganoderma lucidum glycopeptide high-dose group is connected with the epidermis more tightly, and the ganoderma lucidum glycopeptide high-dose group is close to the normal group.
The paraffin sections were then subjected to aldehyde fuchsin staining to observe the changes in the number and morphology of dermal spandex, as shown in fig. 16. The reticular structure of the skin elastic fiber of the blank group rat is relatively clear, the fiber shape is slender, and the fibers are orderly arranged; the arrangement rule of the elastic fibers of the skin of the rat in the model group disappears, which is the result of massive breakage and accumulation of the elastic fibers; the VE group and the ganoderma lucidum glycopeptide group can see wavy elastic fibers, the fibers are rarely broken, crushed, curled and twisted and aggregated into clusters, and the repairing effect of the ganoderma lucidum glycopeptide group is positively related to the dosage.
4. Determination of SOD Activity, MDA, HYP, GSH-Px content in rat skin tissue:
skin tissue supernatants of a blank group (CON), a model group (MOD), a positive control group (VE), a high-dose group (Gl-Gp H) of ganoderan, a medium-dose group (Gl-Gp M) of ganoderan, and a low-dose group (Gl-Gp L) of ganoderan were taken respectively, and the total protein content was measured by using a BCA kit, and the SOD activity and MDA, HYP, GSH-Px content were measured according to the kit instructions.
As shown in fig. 17, malondialdehyde (MDA) is the final product of lipid peroxidation, and to some extent, the extent of damage to cells by lipid peroxidation can be assessed. The MDA content of the model group is obviously higher than that of the blank group, and the model group has obvious difference (P is less than 0.05), which indicates that the MDA content of the skin tissue of the rat can be obviously increased by ultraviolet irradiation; the MDA content of the Gl-Gp L group is reduced compared with the model group, but no significant difference (P is more than 0.05), the MDA content of the skin tissue of the rat is obviously reduced compared with the model group in the Gl-Gp M group and the Gl-Gp H group, the significant difference exists, and the reduction of the Gl-Gp H group is most significant (P is less than 0.01), so that the dose dependency exists.
The antioxidant enzyme system is used as an important component of an organism for resisting oxidative stress, and is one of important indexes for evaluating the antioxidant capacity of a human body. As shown in fig. 18, the SOD activity of the model group is significantly lower than that of the normal control group, and the difference (P < 0.001) is significant, which indicates that the SOD activity of the skin tissue of the rat can be significantly inhibited by the ultraviolet irradiation; the groups Gl-Gp L, gl-Gp M and Gl-Gp H showed significantly improved SOD activity compared with the model group, and showed significant difference (P < 0.05) and dose dependence.
As shown in fig. 19, the GSH-Px content of the model group was significantly lower than that of the blank group, and there was a significant difference (P < 0.01), indicating that the GSH-Px content of the rat skin tissue was significantly reduced by the ultraviolet irradiation. The Gl-Gp L group and the model group have increased GSH-Px content and significant difference (P < 0.05), and the Gl-Gp M and Gl-Gp H groups and the model group have significantly increased GSH-Px content and significant difference (P < 0.01) and have dose dependence.
Hydroxyproline (HYP) is a precursor amino acid for synthesizing skin collagen, can promote skin collagen regeneration and repair damaged skin metabolism, and meanwhile, the content of HYP is an important measure for reflecting the skin aging degree. As shown in fig. 20, the skin HYP content of the rats in the model group was significantly reduced (P < 0.05) compared with the blank group, which indicates that the skin of the rats in the model group had reduced collagen synthesis ability, the network structure of collagen and elastin was broken, the collagen degradation was serious, and the typical characteristics of aging were shown. The content of HYP in the skin of the Gl-Gp group rat is increased, the concentration dependence trend is presented, the content of HPY in the skin of the Gl-Gp group rat is obviously improved (P is less than 0.05) compared with the model group, and the content of Gl-Gp M and Gl-Gp H groups is obviously different (P is less than 0.01) compared with the model group.
5. Determination of MMP-1, MMP-3, IL-1. Beta., IL-6, TNF-. Alpha.content in rat skin tissue:
skin tissue supernatants of a blank group (CON), a model group (MOD), a positive control group (VE), a high-dose group (Gl-Gp H) of ganoderan, a medium-dose group (Gl-Gp M) of ganoderan, and a low-dose group (Gl-Gp L) of ganoderan were taken, and the contents of MMP-1, MMP-3, IL-1. Beta., IL-6, and TNF-alpha in rat skin were measured by referring to ELISA kit instructions.
During skin aging, a very distinct feature is the degradation of collagen fibers and the breakdown of elastic fibers, which has been shown to be the result of matrix metalloproteinase action. As shown in fig. 21, the skin tissue of rats in the model group had significantly increased MMP-1 content (P < 0.01) compared to the blank group, indicating that uv irradiation was able to stimulate the expression of MMPs in cells. Compared with the model group, the content of MMP-1 in the skin tissues of rats in the Gl-Gp L, gl-Gp M and Gl-Gp H groups is reduced, but the content of MMP-1 in the Gl-Gp H group is reduced more obviously (P < 0.01).
As shown in fig. 22, MMP-3 levels were significantly elevated in skin tissues of rats in the model group compared to the blank group (P < 0.0001); compared with the model group, the content of MMP-3 in the skin tissues of rats in the Gl-Gp L, gl-Gp M and Gl-Gp H groups is reduced, and the content of MMP-3 in the ginseng total saponin liposome is reduced more (P < 0.0001). Taken together, the ganoderma lucidum glycopeptides have a certain inhibition effect on the increase of MMP-1 and MMP-3 content induced by ultraviolet rays.
When the skin is stimulated by ultraviolet rays, the skin epidermis and the dermal cells can be promoted to secrete a large amount of pro-inflammatory factors IL-6, IL-1 beta, tumor necrosis factor TNF-alpha and the like. As shown in fig. 23, the skin tissue of the model rats had significantly increased IL-6 content (P < 0.001) compared to the blank rats, which suggests that the uv irradiation could significantly increase IL-6 content in the skin tissue. The amount of IL-6 in the skin of rats in the Gl-Gp L, gl-Gp M and Gl-Gp H groups was reduced compared to the model group; however, MMP-1 content was reduced more significantly in the Gl-Gp M and Gl-Gp H groups (P < 0.001).
As shown in fig. 24, the skin tissue of the model group rats had significantly increased IL-1 β content (P < 0.0001) compared to the blank group, which also suggests that the ultraviolet irradiation could significantly increase the IL-1 β content in the skin tissue. The content of IL-1β in the skin of rats in the Gl-Gp L, gl-Gp M and Gl-Gp H groups was reduced compared to the model group; however, the decrease in IL-1β content was more pronounced in the Gl-Gp H group (P < 0.001).
As shown in fig. 25, the TNF- α content in the skin tissue of rats in the model group was significantly increased (P < 0.01) compared to the blank group, suggesting that the uv irradiation resulted in an increase in TNF- α content in the skin tissue. Compared with the model group, the TNF-alpha content in the skin of rats in the groups of Gl-Gp L, gl-Gp M and Gl-Gp H is reduced, and the rats have significance (P < 0.05).
Claims (8)
1. The application of the ganoderma lucidum glycopeptide is characterized by being used for preparing an antioxidant or skin photoaging resisting skin care product;
the combination mode of the glycopeptide bond of the ganoderma lucidum glycopeptide is O-glycosidic bond;
the ganoderma lucidum glycopeptide is prepared by the following process:
s1, taking dried ganoderma lucidum fruiting bodies and crushing the dried ganoderma lucidum fruiting bodies into ganoderma lucidum coarse powder;
s2, mixing the ganoderma lucidum coarse powder with pure water according to 1g: uniformly mixing (50-70) mL of feed liquid ratio by shaking through a shaking table, heating and extracting for 10-12 h at 60-80 ℃, then performing ultrasonic extraction for at least 3 times for 15-30 min each time, and combining the supernatant to obtain ganoderma lucidum glycopeptide crude extract;
s3, removing free protein from the ganoderma lucidum glycopeptide crude extract, and then carrying out dialysis treatment by using a dialysis bag;
s4, carrying out enzymolysis on the dialyzed ganoderma lucidum glycopeptides, carrying out water bath treatment at 60 ℃ for 35min, adding trypsin with the enzyme-to-bottom ratio of 4%, magnetically stirring for 6h, then placing in a constant-temperature water bath at 85 ℃ for 20min, and cooling to room temperature; then eluting with wheat germ agglutinin resin until the molecular weight cut-off is less than 10000Da, and freeze-drying to obtain Ganoderma glycopeptide freeze-dried powder.
2. The use of a ganoderma lucidum glycopeptide according to claim 1, wherein the shaking speed of the shaking table is 120r/min to 180r/min.
3. The use of a ganoderma lucidum glycopeptide according to claim 1, wherein the removing of the free protein in step S3 is performed by a Sevage method; the dialysis treatment specifically comprises the following steps: the solution was dialyzed in distilled water using a 0.1kDa dialysis bag for 48 hours.
4. The use of a ganoderma lucidum glycopeptide according to claim 1 wherein the temperature of the freeze drying in step S4 is-86 ℃ and the time of the freeze drying is at least 48 hours.
5. A ganoderma lucidum glycopeptide microemulsion, characterized in that the ganoderma lucidum glycopeptide microemulsion is prepared by taking the ganoderma lucidum glycopeptide as the raw material in any one of claims 1-4.
6. A method for preparing the ganoderma lucidum glycopeptide microemulsion according to claim 5, comprising the following steps:
the ganoderma lucidum glycopeptide and water are prepared into a water phase with the concentration of the medicine of 10 mg/mL-20 mg/mL, and a zwitterionic surfactant and a cosurfactant are mixed according to the proportion of 1: mixing (0.5-2) by mass ratio, mixing with oil phase, magnetically stirring at 23-27 ℃ and 500-1000 rpm, and dropwise adding water phase to obtain the ganoderma lucidum glycopeptide microemulsion.
7. The method of claim 6, wherein the zwitterionic surfactant is soybean lecithin, the cosurfactant is ethanol, and the oil phase is isopropyl myristate.
8. A cream comprising the ganoderma lucidum glycopeptide microemulsion of claim 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310993467.XA CN116694721B (en) | 2023-08-09 | 2023-08-09 | Ganoderma lucidum glycopeptide and preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310993467.XA CN116694721B (en) | 2023-08-09 | 2023-08-09 | Ganoderma lucidum glycopeptide and preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116694721A CN116694721A (en) | 2023-09-05 |
| CN116694721B true CN116694721B (en) | 2023-10-20 |
Family
ID=87837947
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310993467.XA Active CN116694721B (en) | 2023-08-09 | 2023-08-09 | Ganoderma lucidum glycopeptide and preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116694721B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1429837A (en) * | 2001-12-29 | 2003-07-16 | 福州绿谷生物药业技术研究所 | Grass planted ganoderma polysaccharide peptide and its extraction method |
| CN1786023A (en) * | 2005-07-27 | 2006-06-14 | 西安绿谷制药有限公司 | Lucid ganoderma poly saccharide peptide standard product, sts preparation method and application |
| CN105418781A (en) * | 2015-11-23 | 2016-03-23 | 宁国千方中药发展有限公司 | Ultrasonic extraction method of ganoderan |
| CN110016086A (en) * | 2019-03-14 | 2019-07-16 | 合肥工业大学 | A kind of process for refining of ganoderma lucidum polysaccharide |
| CN113151389A (en) * | 2021-04-24 | 2021-07-23 | 长春中医药大学 | Ginseng glycopeptide, preparation method and medical application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2405923A1 (en) * | 2009-03-10 | 2012-01-18 | Palmed Teva Ltd | Novel coprinus comatus and tremella mesenterica mushroom strains, products and extracts thereof and compositions comprising them |
-
2023
- 2023-08-09 CN CN202310993467.XA patent/CN116694721B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1429837A (en) * | 2001-12-29 | 2003-07-16 | 福州绿谷生物药业技术研究所 | Grass planted ganoderma polysaccharide peptide and its extraction method |
| CN1786023A (en) * | 2005-07-27 | 2006-06-14 | 西安绿谷制药有限公司 | Lucid ganoderma poly saccharide peptide standard product, sts preparation method and application |
| CN105418781A (en) * | 2015-11-23 | 2016-03-23 | 宁国千方中药发展有限公司 | Ultrasonic extraction method of ganoderan |
| CN110016086A (en) * | 2019-03-14 | 2019-07-16 | 合肥工业大学 | A kind of process for refining of ganoderma lucidum polysaccharide |
| CN113151389A (en) * | 2021-04-24 | 2021-07-23 | 长春中医药大学 | Ginseng glycopeptide, preparation method and medical application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 灵芝多糖肽GL-PPS 的结构研究;王赛贞等;西南农业学报;第28卷(第2期);793-796 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116694721A (en) | 2023-09-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2492850C2 (en) | Fibroblast growth agent | |
| EP2175833B1 (en) | Dermocosmetic compositions based on tamarind seed polysaccharide and vegetable extracts | |
| CN111135117A (en) | Small-molecule skin softening essence and preparation method thereof | |
| CN115531272A (en) | Antioxidant composition derived from alpine plants and preparation method and application thereof | |
| KR101834795B1 (en) | Cosmetic composition containing nanoliposome encapsulating extracts of Tremella fuciformis and Dioscorea japonica Thunb | |
| CN1823724A (en) | Salamander face beautifying skin protection product and its preparation method | |
| CN116694721B (en) | Ganoderma lucidum glycopeptide and preparation method and application thereof, and ganoderma lucidum glycopeptide microemulsion and face cream | |
| KR102130149B1 (en) | Cosmetic composition containing peach gum extract | |
| KR101073284B1 (en) | A composition for skin external application | |
| CN116869898A (en) | Protective cream for sensitive skin with enhanced barrier and preparation method thereof | |
| KR101794419B1 (en) | Osmotic enzyme fermentation natural liposome containing mushroom, process for the preparation thereof, and cosmetic, food or pharmaceutical composition comprising the same | |
| KR101662272B1 (en) | Natural cosmetic composition for skin moisturizing comprising Mixed extract of barley and germ and rice bran of black rice and preparation method thereof | |
| CN107174536A (en) | A kind of salamander face beautifying skin protection product and preparation method thereof | |
| CN114317651B (en) | Polygonatum sibiricum polypeptide, preparation method and application thereof | |
| CN115040448A (en) | Skin care product with enhanced anti-aging effect and preparation method thereof | |
| KR102595837B1 (en) | Cosmetic composition comprising spore oil extracted from antler-shaped Ganoderma lucidum and the method for preparing the same | |
| KR20200001686A (en) | Anti-aging and anti-wrinkle of skin improving cosmetic composition comprising extracts of Moschus, Angelica gigas Nakai, Cornus officinalis, Cervi Parvum Cornu and Phellinus linteus, Ganoderma Lucidum, Inonotus obliquus, Grifola Frondosa, Sparassis crispa | |
| KR100532633B1 (en) | Cosmetic composition having anti-inflammatory, skin-protecting, skin-elastic effects which comprise mixed plants extract | |
| CN114652628A (en) | Anti-aging cream for beautifying chest and body and enhancing skin immune barrier and preparation method thereof | |
| KR102157210B1 (en) | The functional cosmetic composition comprising extract of natural micro organism-fermented black ginseng | |
| KR20060104807A (en) | A method for producing extract of germinated brown rice and cosmetic composition comprising the extract | |
| KR102073943B1 (en) | A skin-care agent or cosmetics containig Lyophyllum ulmarium fruit body extract or Lyophyllum ulmarium mycelium extract | |
| KR20210092587A (en) | Liposome composition comprising skin physiologically active substances and method of preparing the same | |
| CN111700818A (en) | Wrinkle-removing and anti-aging ginseng polypeptide toning lotion and preparation method thereof | |
| CN111419779A (en) | Pure natural stretch mark protective cream and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |