CN114836491A - Preparation method of anti-aging skin-brightening carnosine and application of carnosine in cosmetics - Google Patents
Preparation method of anti-aging skin-brightening carnosine and application of carnosine in cosmetics Download PDFInfo
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- CN114836491A CN114836491A CN202210604857.9A CN202210604857A CN114836491A CN 114836491 A CN114836491 A CN 114836491A CN 202210604857 A CN202210604857 A CN 202210604857A CN 114836491 A CN114836491 A CN 114836491A
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- Prior art keywords
- carnosine
- enzymolysis
- extraction method
- ultrasonic
- cosmetics
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- QRYRORQUOLYVBU-VBKZILBWSA-N Carnosic acid Natural products CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 title claims abstract description 83
- 108010087806 Carnosine Proteins 0.000 title claims abstract description 83
- CQOVPNPJLQNMDC-UHFFFAOYSA-N N-beta-alanyl-L-histidine Natural products NCCC(=O)NC(C(O)=O)CC1=CN=CN1 CQOVPNPJLQNMDC-UHFFFAOYSA-N 0.000 title claims abstract description 83
- CQOVPNPJLQNMDC-ZETCQYMHSA-N carnosine Chemical compound [NH3+]CCC(=O)N[C@H](C([O-])=O)CC1=CNC=N1 CQOVPNPJLQNMDC-ZETCQYMHSA-N 0.000 title claims abstract description 83
- 229940044199 carnosine Drugs 0.000 title claims abstract description 83
- 239000002537 cosmetic Substances 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims description 6
- 230000003712 anti-aging effect Effects 0.000 title abstract description 10
- 238000005282 brightening Methods 0.000 title abstract description 7
- 239000000843 powder Substances 0.000 claims abstract description 33
- 238000000034 method Methods 0.000 claims abstract description 19
- 210000003205 muscle Anatomy 0.000 claims abstract description 19
- 239000002994 raw material Substances 0.000 claims abstract description 14
- 238000001914 filtration Methods 0.000 claims abstract description 12
- 241001465754 Metazoa Species 0.000 claims abstract description 11
- 235000013305 food Nutrition 0.000 claims abstract description 6
- 230000036541 health Effects 0.000 claims abstract description 6
- 238000005119 centrifugation Methods 0.000 claims abstract description 4
- 102000004190 Enzymes Human genes 0.000 claims description 29
- 108090000790 Enzymes Proteins 0.000 claims description 29
- 229940088598 enzyme Drugs 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 238000000605 extraction Methods 0.000 claims description 19
- 239000000706 filtrate Substances 0.000 claims description 17
- 239000004365 Protease Substances 0.000 claims description 15
- 235000013372 meat Nutrition 0.000 claims description 15
- 239000000047 product Substances 0.000 claims description 14
- 102000004142 Trypsin Human genes 0.000 claims description 13
- 108090000631 Trypsin Proteins 0.000 claims description 13
- 239000012588 trypsin Substances 0.000 claims description 13
- 108090000145 Bacillolysin Proteins 0.000 claims description 12
- 102000035092 Neutral proteases Human genes 0.000 claims description 12
- 108091005507 Neutral proteases Proteins 0.000 claims description 12
- 108090000526 Papain Proteins 0.000 claims description 12
- 239000008367 deionised water Substances 0.000 claims description 12
- 229910021641 deionized water Inorganic materials 0.000 claims description 12
- 239000011259 mixed solution Substances 0.000 claims description 12
- 229940055729 papain Drugs 0.000 claims description 12
- 235000019834 papain Nutrition 0.000 claims description 12
- 239000012528 membrane Substances 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 9
- 230000002779 inactivation Effects 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 7
- 238000005520 cutting process Methods 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 150000001875 compounds Chemical class 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 3
- 235000020995 raw meat Nutrition 0.000 claims description 3
- 230000000415 inactivating effect Effects 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 14
- 230000003078 antioxidant effect Effects 0.000 abstract description 10
- 230000037303 wrinkles Effects 0.000 abstract description 6
- 230000037394 skin elasticity Effects 0.000 abstract description 5
- 230000000052 comparative effect Effects 0.000 description 13
- 239000000243 solution Substances 0.000 description 13
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 12
- 238000012360 testing method Methods 0.000 description 10
- 150000003254 radicals Chemical class 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 235000015277 pork Nutrition 0.000 description 7
- 238000002604 ultrasonography Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 229910000029 sodium carbonate Inorganic materials 0.000 description 6
- -1 superoxide anions Chemical class 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
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- 238000000108 ultra-filtration Methods 0.000 description 5
- 239000002585 base Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 4
- 230000007760 free radical scavenging Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000002000 scavenging effect Effects 0.000 description 4
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N squalane Chemical compound CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
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- 239000000126 substance Substances 0.000 description 3
- ASJSAQIRZKANQN-CRCLSJGQSA-N 2-deoxy-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)CC=O ASJSAQIRZKANQN-CRCLSJGQSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 206010015150 Erythema Diseases 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- 206010040829 Skin discolouration Diseases 0.000 description 2
- 229920002385 Sodium hyaluronate Polymers 0.000 description 2
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- TZCXTZWJZNENPQ-UHFFFAOYSA-L barium sulfate Chemical compound [Ba+2].[O-]S([O-])(=O)=O TZCXTZWJZNENPQ-UHFFFAOYSA-L 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 2
- 229940059958 centella asiatica extract Drugs 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000001947 glycyrrhiza glabra rhizome/root Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
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- 238000004020 luminiscence type Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- JXTPJDDICSTXJX-UHFFFAOYSA-N n-Triacontane Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC JXTPJDDICSTXJX-UHFFFAOYSA-N 0.000 description 2
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 229940010747 sodium hyaluronate Drugs 0.000 description 2
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 229940032094 squalane Drugs 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- WSGYTJNNHPZFKR-UHFFFAOYSA-N 3-hydroxypropanenitrile Chemical group OCCC#N WSGYTJNNHPZFKR-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 241000143060 Americamysis bahia Species 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- 241000272814 Anser sp. Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 208000007342 Diabetic Nephropathies Diseases 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- 241000272496 Galliformes Species 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940000635 beta-alanine Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000009920 chelation Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 208000033679 diabetic kidney disease Diseases 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
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- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 231100000321 erythema Toxicity 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229960002885 histidine Drugs 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 235000021190 leftovers Nutrition 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000011076 safety test Methods 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 230000009759 skin aging Effects 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 230000009967 tasteless effect Effects 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/10—Nitrogen as only ring hetero atom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
- A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides a method for extracting carnosine, which takes animal muscle tissues as raw materials, and obtains carnosine powder with high antioxidant activity through procedures of homogenate, ultrasonic enzymolysis, centrifugation, filtration and the like. The carnosine composition prepared by the invention can be used for preparing cosmetics, foods or health care products, especially for preparing the cosmetics, has obvious effects of resisting wrinkles, improving skin elasticity and brightening skin, can make the skin smooth and elastic after being used, and can be widely applied to anti-aging and brightening cosmetics.
Description
Technical Field
The invention relates to a preparation method of carnosine, in particular to a preparation method of cosmetic raw material grade carnosine, and belongs to the technical field of biological production.
Background
Carnosine was first discovered by russian scientists in 1900, and is a dipeptide composed of beta-alanine and L-histidine, which is very soluble in water, not easily soluble in organic solvents such as ethanol and acetone, is white powder, odorless and tasteless, and mainly exists in skeletal muscle and brain of vertebrates.
Carnosine has a wide range of effects, and has the effects of oxidation resistance, metal ion chelation, acid-base buffering, aging resistance and the like. Carnosine also has effects on many age-related diseases, such as promotion of wound healing, benefits for alzheimer's disease, parkinson's disease, stroke, and diabetic nephropathy.
Numerous research reports have demonstrated that carnosine can effectively scavenge active oxygen and free radicals, and that the scavenging capacity increases with increasing concentration. "Antioxidant effects of diabetes on lipids and beef homogenes", Beom Jun Lee et al, disclose a formulation consisting of Fe 3+ A system for degrading deoxyribose by catalyzing and generating hydroxyl free radicals, and carnosine is found to be capable of effectively inhibiting the degradation of deoxyribose, which indicates that the carnosine has the capacity of capturing the hydroxyl free radicals. "comparison of carnosine and SOD on free radical scavenging action", Wang' ei et al, disclose that barium sulfate is used as a stimulant, a stable whole blood polymorphous leukocyte (PMN) luminol-dependent chemiluminescence system is established, and the scavenging action of carnosine on free radicals is studied, when carnosine with different concentrations is added into the system, obvious inhibition on chemiluminescence can be seen, if the standard control luminescence value is set as 100, the inhibition luminescence action of carnosine under different concentrations of 2.5, 5, 10 and 15mmol/L is respectively 47.2, 71.4, 85.4 and 97. "The mechanism of interaction of camosine with superoxide raditions in water solutions", Andrev R et al, discloses The use of pulsed radiation to decompose aqueous solutions to produce superoxide anions, and monitoring shows that when carnosine is added, it is capable of concentration-dependent scavenging superoxide anions, which indicates that carnosine has The properties of superoxide dismutase, which is also responsible for The reduction of copper and zinc-initiated superoxide anion radicals by carnosine.
Free radicals can eventually destroy skin structure by activating mitogen-activated protein kinase pathways in the body, leading to aging of the skin, and thus, it is important to counteract aging by scavenging free radicals or reducing the production of free radicals. Carnosine can scavenge free radicals and some oxidation products, thereby playing a role in resisting skin aging, and thus, carnosine is widely applied to the field of cosmetics.
Carnosine is an endogenous active peptide mainly existing in animal muscle, the inoxidizability of a pure product of the carnosine is widely accepted, but artificially synthesized carnosine is expensive, a large amount of toxic or flammable and explosive organic solvent is used in the synthesis process, the operation is troublesome, the environment is polluted, the carnosine is harmful to human bodies, the carnosine is difficult to popularize in production, the development of the carnosine is limited, and the carnosine is extracted from animal muscle as a rich source of the carnosine and becomes an effective method for obtaining the carnosine.
Patent CN100465285C discloses a method for extracting carnosine, which takes animal meat as a raw material, and comprises the steps of grinding the meat, homogenizing, carrying out enzymolysis under an acidic condition, carrying out enzymolysis under an alkaline condition, inactivating, standing, adsorbing and eluting by a hydroxyapatite chromatographic column, carrying out ultrafiltration by a filter membrane, carrying out spray drying and the like. The above method has complicated steps and long operation period, and is not suitable for large-scale industrial production, and the above patent does not disclose the antioxidant activity of the obtained extract.
In view of the above, there is a need in the art for a carnosine extraction method suitable for industrial production, particularly for preparing cosmetic grade raw materials for preparing anti-aging and skin-lightening cosmetics.
Disclosure of Invention
The invention provides a method for extracting carnosine from animal meat, aiming at the problems in the prior art, the method has simple process, less steps and short time, and the obtained extract has good antioxidation when being applied to cosmetics.
In order to realize the purpose, the technical scheme adopted by the invention is as follows:
as a first aspect, the present invention provides a method for extracting carnosine, comprising the steps of:
(1) raw material treatment: taking muscle tissue of fresh animal meat, adding into deionized water for homogenizing treatment to obtain homogenate;
(2) ultrasonic enzymolysis: adding a compound enzyme into the homogenate obtained in the step (1) for enzymolysis under an ultrasonic condition to obtain an enzymolysis mixed solution, wherein the compound enzyme is trypsin, papain and neutral protease;
(3) inactivation: heating the enzymolysis mixed liquor obtained in the step (2) in a water bath condition at the temperature of 95-100 ℃ for 10-15min for inactivation;
(4) centrifugal filtration: carrying out centrifugal separation on the inactivated enzymolysis mixed liquor obtained in the step (3) to obtain supernatant; filtering the supernatant with a filter membrane, and collecting filtrate;
(5) and (3) drying: and (4) drying the filtrate obtained in the step (4) to obtain the carnosine powder.
According to one embodiment of the present invention, the animal meat of step (1) may be livestock, poultry or aquatic products. Domestic animals such as pork, beef or mutton, domestic fowls such as chicken, duck or goose, and aquatic animals such as various fishes, shrimps or shellfishes. Preferred animal meat is livestock, more preferably pork.
The invention can also adopt low-value meat such as meat on the processed waste meat bones, or eliminated egg chicken, or meat leftovers and the like to further reduce the production cost.
Removing non-muscle tissues such as fat meat, muscle and the like from raw meat, cleaning, cutting into blocks, adding deionized water according to the feed liquid ratio of 1g to 2-5ml, and crushing and homogenizing.
According to one embodiment of the invention, the temperature of the enzymolysis in the step (2) is 30-50 ℃; the pH value is 7.5-8.5; the enzymolysis time is 10-30 min; the adding mass of the complex enzyme is 0.5-2% of that of the raw meat; the mass ratio of trypsin, papain and neutral protease in the complex enzyme is 0.5-2: 11-13: 3-6; and carrying out ultrasonic treatment while carrying out enzymolysis, wherein the ultrasonic power is 100-400W, and the ultrasonic frequency is 15-40 kHz.
According to a preferred embodiment of the invention, the temperature of the enzymolysis in the step (2) is 30-40 ℃; the pH value is 7.5-8.0; the enzymolysis time is 15-20 min; the mass of the complex enzyme is 1-1.5% of that of the raw material meat; the mass ratio of trypsin, papain and neutral protease in the complex enzyme is 1-2: 12-13: 5-6; the ultrasonic power is 200-280W, and the ultrasonic frequency is 18-25 kHz.
According to one embodiment of the invention, the pH value of the solution is adjusted by using alkali in the step (2), wherein the alkali is sodium carbonate, sodium bicarbonate, sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonia water or the like.
According to one embodiment of the invention, the rotation speed of the centrifugation in the step (4) is 2000-8000rpm, and the centrifugation time is 10-15 min; the filter membrane is filtered by adopting a 0.22 mu m microporous filter membrane;
according to an embodiment of the present invention, the drying in step (5) may be spray drying or freeze drying.
The invention simultaneously carries out ultrasonic treatment in the enzymolysis process, and the ultrasonic wave improves the enzyme activity by changing the tertiary structure of enzyme molecules and simultaneously generates homogenization action on reaction substrates, thereby obviously improving the speed of the enzymolysis reaction. However, in the present invention, the conditions of the ultrasonic treatment and the enzymatic hydrolysis are strictly controlled in step (2), otherwise, the enzyme is inactivated, and the extraction of the target substance is affected. For example, for the ultrasonic time, when the ultrasonic time is too short, the enzymolysis is incomplete, and when the ultrasonic time is too long, the enzyme activity is inhibited, and other byproducts are generated; for ultrasonic frequency, when the frequency is too low, the collision frequency of enzyme and substrate is not enough, the enzymolysis reaction speed is slow, and when the ultrasonic frequency is too high, the enzyme molecular conformation is further changed, so that the enzyme activity is reduced, and the extraction of a target product is inhibited; for ultrasonic power, when the power is too small, the activity of the enzyme is not sufficiently improved, and when the power is too large, the activity of the enzyme is reduced.
The OPA method detection of the carnosine powder provided by the invention shows that the content of the carnosine is 50-60%. The invention discovers that when trypsin, papain and neutral protease are adopted and the proportion of the trypsin, the papain and the neutral protease is strictly controlled, the obtained extract has obviously higher inoxidizability and free radical scavenging capability than pure carnosine although the purity of the carnosine is not high, which probably leads the inoxidizability and the free radical scavenging capability of the extract to be greatly improved under the coordination of other active substances contained in the extract and the carnosine. The enzymolysis mixed liquid is centrifuged and filtered without further purification, and the product without purification has stronger inoxidizability and free radical scavenging capability, can be directly used for preparing cosmetics, and greatly improves the utilization rate of raw materials.
As a second aspect, the invention provides application of the muscle peptide powder prepared by the method in cosmetics, foods and health products, wherein the cosmetics, foods and health products have an anti-aging effect after being used.
When the carnosine powder is used for preparing cosmetics, the carnosine powder does not need to be further purified and can be directly used as a cosmetic raw material;
when the carnosine powder is used for preparing food or health care products, the carnosine powder needs to be further purified, the purification method can adopt a conventional ultrafiltration method in the field, specifically, the purification can be that after the filtration membrane is adopted in the step (4), an ultrafiltration membrane with the cut-off molecular weight of 3kD is used for ultrafiltration, and the obtained filtrate is dried to obtain the carnosine powder which can be directly used as a raw material of the food or health care products.
As a third aspect, the present invention provides an anti-aging skin-lightening cosmetic composition comprising carnosine powder obtained by the above extraction method.
According to one embodiment of the invention, the invention provides an anti-aging skin-brightening cosmetic composition which comprises the following raw materials in parts by weight: the composition comprises, by weight, 6-10 parts of carnosine powder extracted by the invention, 0.5-1 part of sodium hyaluronate, 2-5 parts of squalane, 7-12 parts of nicotinamide, 1-4 parts of centella asiatica extract and 1-4 parts of glycyrrhiza glabra root extract.
According to an embodiment of the present invention, a cosmetic base may be further added to the cosmetic composition, wherein the cosmetic base includes at least one of oil, emulsifier, humectant, solvent, skin conditioner, thickener, solubilizer, essence, and preservative, and the specific type of the cosmetic base is not particularly limited, and may be selected by one skilled in the art according to the specific dosage form of the product.
The anti-aging skin-brightening cosmetic composition provided by the invention can be used for preparing products such as astringent, emulsion, essence, mask, cream and the like.
Compared with the prior art, the invention has the following beneficial effects:
the carnosine extraction method provided by the invention has the advantages of simple process, short time, high carnosine extraction rate, high antioxidant activity of the obtained extract, no organic solvent adopted in the extraction process, safety, environmental protection and suitability for industrial production. The invention does not excessively purify the extract, retains other substances in the extract, simplifies the complicated purification process and simultaneously improves the antioxidant activity of the extract, and the obtained extract is used for preparing cosmetics, has obvious effects of resisting wrinkles and improving the elasticity and brightness of skin, can make the skin moist, smooth and elastic after being used, and can be widely applied to anti-aging and skin-brightening cosmetics.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. In addition, the starting materials of the present invention are all common commercial products unless otherwise specified.
Example 1
Removing fat and muscle of fresh pork to obtain 2kg of muscle tissue, cleaning, cutting into pieces, adding 6L of deionized water, and homogenizing for 5min with high-speed disperser; adding 0.1mol/L sodium carbonate solution into the homogenate to adjust the pH value to be about 7.5-7.8, controlling the temperature to be 30-35 ℃, uniformly stirring, adding 20g of complex enzyme, simultaneously starting ultrasound, controlling the ultrasonic power to be 230W, the frequency to be 20kHz and the temperature to be 30-35 ℃ for enzymolysis for 15min, wherein the complex enzyme used in the embodiment comprises 1g of trypsin, 13g of papain and 6g of neutral protease; after the enzymolysis is finished, heating the obtained enzymolysis mixed solution for 10-15min under the water bath condition of 95-100 ℃ for inactivation, and then naturally cooling to room temperature; centrifuging the inactivated enzymolysis mixed solution at 6000rpm for 10min, collecting supernatant, filtering with 0.22 μm microporous membrane, and collecting filtrate 6.2 kg; the filtrate was spray dried to give 11.8g of pale yellow carnosine powder.
Example 2
Removing fat and muscle of fresh pork to obtain 2kg of muscle tissue, cleaning, cutting into pieces, adding 10L of deionized water, and homogenizing for 5min with high-speed disperser; adding 0.1mol/L sodium carbonate solution into the homogenate to adjust the pH to be about 7.5-7.8, controlling the temperature to be 30-35 ℃, uniformly stirring, adding 30g of complex enzyme, simultaneously starting ultrasound, controlling the ultrasonic power to be 280W, the frequency to be 25kHz, and controlling the temperature to be 30-35 ℃ for enzymolysis for 18min, wherein the complex enzyme used in the embodiment comprises 3g of trypsin, 18g of papain and 9g of neutral protease; after enzymolysis, putting the obtained enzymolysis mixed solution into a water bath condition at the temperature of 95-100 ℃ for heating for 10-15min for inactivation, and then naturally cooling to room temperature; centrifuging the inactivated enzymolysis mixed solution at 4000rpm for 15min, filtering the supernatant with a 0.22 μm microporous filter membrane, and collecting filtrate to obtain 6.2 kg; the filtrate was spray dried to obtain 12.3g of pale yellow carnosine powder.
Comparative example 1
Removing fat and muscle of fresh pork to obtain 2kg of muscle tissue, cleaning, cutting into pieces, adding 6L of deionized water, and homogenizing for 5min with high-speed disperser; adding 0.1mol/L sodium carbonate solution into the homogenate to adjust the pH to 7.5-7.8, controlling the temperature to 30-35 ℃, stirring uniformly, adding 20g trypsin, simultaneously starting ultrasound, controlling the ultrasonic power to 230W, the frequency to 20kHz, and controlling the temperature to 30-35 ℃ for enzymolysis for 15 min; after enzymolysis, putting the obtained enzymolysis mixed solution into a water bath condition at the temperature of 95-100 ℃ for heating for 10-15min for inactivation, and then naturally cooling to room temperature; centrifuging the inactivated enzymolysis mixed solution at 6000rpm for 10min, collecting supernatant, filtering with 0.22 μm microporous membrane, and collecting filtrate 6.6 kg; the filtrate was spray dried to give a pale yellow carnosine powder (10.7 g).
Comparative example 2
Removing fat and muscle of fresh pork to obtain 2kg of muscle tissue, cleaning, cutting into pieces, adding 6L of deionized water, and homogenizing for 5min with high-speed disperser; adding 0.1mol/L sodium carbonate solution into the homogenate to adjust the pH to be about 7.5-7.8, controlling the temperature to be 30-35 ℃, stirring uniformly, adding 20g of complex enzyme, simultaneously starting ultrasound, controlling the ultrasonic power to be 230W, the frequency to be 20kHz, and controlling the temperature to be 30-35 ℃ for enzymolysis for 15min, wherein the complex enzyme used in the embodiment comprises 6g of trypsin, 8g of papain and 6g of neutral protease; after enzymolysis, putting the obtained enzymolysis mixed solution into a water bath condition at the temperature of 95-100 ℃ for heating for 10-15min for inactivation, and then naturally cooling to room temperature; centrifuging the inactivated enzymolysis mixed solution at 6000rpm for 10min, collecting supernatant, filtering with 0.22 μm microporous membrane, and collecting filtrate 6.2 kg; the filtrate was spray dried to give 12.8g of pale yellow carnosine powder.
Comparative example 3
Removing fat and muscle of fresh pork to obtain 2kg of muscle tissue, cleaning, cutting into pieces, adding 6L of deionized water, and homogenizing for 5min with high-speed disperser; adding 0.1mol/L sodium carbonate solution into the homogenate to adjust the pH to be about 7.5-7.8, controlling the temperature to be 30-35 ℃, stirring uniformly, adding 20g of complex enzyme, simultaneously starting ultrasound, controlling the ultrasonic power to be 400W, the frequency to be 30kHz, and controlling the temperature to be 30-35 ℃ for enzymolysis for 15min, wherein the complex enzyme used in the embodiment comprises 1g of trypsin, 13g of papain and 6g of neutral protease; after enzymolysis, putting the obtained enzymolysis mixed solution into a water bath condition at the temperature of 95-100 ℃ for heating for 10-15min for inactivation, and then naturally cooling to room temperature; centrifuging the inactivated enzymolysis mixed solution at 6000rpm for 10min, collecting supernatant, filtering with 0.22 μm microporous membrane, and collecting filtrate 6.0 kg; the filtrate was spray dried to give 11.6g of pale yellow carnosine powder.
Comparative example 4
The extraction is carried out by the method of example 1, after the filtration by a 0.22 μm microporous membrane, the filtrate is ultrafiltered by an ultrafiltration membrane with the cut-off molecular weight of 3kD, and the filtrate is collected and then is spray-dried to obtain 7.3g of light yellow carnosine powder.
Carnosine content detection
Measuring carnosine content by OPA-HPLC method, wherein Agilent 1200 high performance liquid chromatography is adopted as chromatographic equipment, and Shimadzu ODS-C is adopted as chromatographic column 18 A chromatographic column is arranged on the top of the chromatographic column,the mobile phase A is 12.5mmol/L acetic acid-sodium acetate buffer solution, and the mobile phase B is methanol-acetonitrile solution (7: 3, v/v); the gradient elution conditions were: 0min, 60% A; 15min, 40% A; 20-22min, 100% B; then running for 5 min; the sample feeding amount is 10 mu L, the sample flow rate is 1.0mL/min, the column temperature is 28 ℃, the detector is an ultraviolet detector, and the measurement is carried out at 228 nm; the preparation method of the OPA derivative reagent adopts 20mg phthalic acid, 1mL methanol is added for dissolution, 4mL boric acid buffer solution (pH 9.5) and 100mL beta-mercaptoethanol are added for even mixing, and the mixture is protected from light and is used as the reagent.
The carnosine contents of examples 1-2 and comparative examples 1-4 are as follows:
| example 1 | Example 2 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 | |
| Carnosine content | 54％ | 51％ | 36％ | 33％ | 49％ | 87％ |
Detection of antioxidant Activity
The antioxidant activity of the carnosine powders of the examples and comparative examples was examined by the DPPH method. Taking carnosine powder of examples 1-2 and comparative examples 1-4 respectively, preparing 1mg/mL sample solution by using deionized water, and preparing 0.1mmol/L DPPH reagent by using 85% ethanol; sample group: 100uL of sample solution is taken, 100uL of DPPH reagent is added, and the mixture is fully and evenly mixed and then is kept stand for 30min at room temperature. After the standing is finished, oscillating for 2min by using an oscillator, and detecting the absorbance A1 by using an enzyme-labeling instrument, wherein the detection wavelength is 519 nm; blank group: replacing the DPPH reagent with an 85% ethanol reagent, basically enabling the rest operations to be consistent with the sample group, and detecting the absorbance A2 by using an enzyme-labeling instrument; control group: replacing the sample solution with deionized water, basically conforming the rest operations to the sample group, and detecting the absorbance A0 by using an enzyme-labeling instrument;
DPPH radical clearance (%) (a0-a1+ a2)/a0 × 100%.
DPPH radical scavenging rates for examples 1-2 and comparative examples 1-4 are as follows:
the above results show that the extraction method of example 1-2 of the present application, by strictly controlling the mixture ratio of the three proteases, the conditions of enzymolysis and ultrasound, the carnosine content of the obtained carnosine powder is not high, but the carnosine powder has high antioxidant activity. When single protease is adopted for enzymolysis, or the proportion of three proteases is changed, or the parameter setting of ultrasound is changed, the content of carnosine in the obtained carnosine powder is reduced to different degrees, the antioxidant activity is also obviously reduced, the change of the parameters not only reduces the extraction rate of the carnosine in a sample, but also can reduce the antioxidant activity greatly due to the fact that the content of other active substances is different. From comparative example 4, after the obtained extract is further ultrafiltered, the carnosine content of the obtained carnosine powder is further improved, namely the content of other substances is reduced, but the oxidation resistance is obviously reduced, which shows that the other substances in the carnosine powder obtained in the examples 1-2 of the application play a role in enhancing the oxidation resistance activity, the oxidation resistance activity of the carnosine powder is far higher than that of the carnosine with higher purity, and the carnosine powder can be applied to the field of cosmetics.
The carnosine powder of example 1 was further used to prepare a cosmetic composition:
example 3
A cosmetic composition comprises the following raw materials: 8g of carnosine powder, 1g of sodium hyaluronate, 3g of squalane, 10g of nicotinamide, 2g of centella asiatica extract, 2g of glycyrrhiza glabra root extract, 30g of deionized water and 15g of glycerol in example 1.
To demonstrate the safety and efficacy of the cosmetic compositions described above, the following tests were carried out:
100 female volunteers between 30-35 years old were selected, and a control test was performed with 3cm × 3em of skin on the inner side of the left and right arms, and 0.5g of the cosmetic composition of example 3 was applied to the inner side of the left arm and 0.5g of glycerin was applied to the inner side of the right arm twice a day for 6 weeks.
Test example 1: safety test
The skin conditions of the arms of the volunteers after 24 hours and 72 hours were recorded, and the results showed that no allergic phenomena such as redness or erythema were observed after applying the cosmetic composition of example 3 of the present invention, indicating that the cosmetic composition of the present invention is safe and has no adverse phenomena.
Test example 2: skin wrinkle, elasticity test and lightness test
Skin wrinkles and skin elasticity were measured before and after use at weekly intervals on the left hand test area and the right hand control area, respectively. Wherein, the Skin wrinkles are measured by a Skin image analysis system (Skin Visiometer SV 600); the skin elasticity was measured using a skin elasticity tester MPA580 from CK, Germany, using a Revisometer RV600 probe;
respectively measuring the skin brightness conditions of a left-hand test area and a right-hand control area every other week before and after use, wherein the skin brightness is measured by adopting a skin chromaticity test probe and an MPA 9 tester;
the measurements were counted and the mean values calculated are shown in the following table:
the results of the tests in the table show that the cosmetic composition prepared from the carnosine powder has obvious effects of resisting wrinkles, improving skin elasticity and improving skin brightness, and the product can show obvious effects after being used for 6 weeks, thereby proving that the carnosine powder prepared by the invention can be used in anti-aging cosmetics.
Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, and that the simple modifications or equivalent substitutions of the technical solutions of the present invention by those of ordinary skill in the art can be made without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A method for extracting carnosine, comprising the steps of:
(1) raw material treatment: taking muscle tissue of fresh animal meat, adding into deionized water for homogenizing treatment to obtain homogenate;
(2) ultrasonic enzymolysis: adding a compound enzyme into the homogenate obtained in the step (1) for enzymolysis under an ultrasonic condition to obtain an enzymolysis mixed solution, wherein the compound enzyme is trypsin, papain and neutral protease;
(3) inactivation: heating and inactivating the enzymolysis mixed liquor obtained in the step (2);
(4) centrifugal filtration: carrying out centrifugal separation on the inactivated enzymolysis mixed liquor obtained in the step (3) to obtain supernatant; filtering the supernatant with a filter membrane, and collecting filtrate;
(5) and (3) drying: and (4) drying the filtrate obtained in the step (4) to obtain the carnosine powder.
2. The extraction method according to claim 1, characterized in that: the step (1) comprises removing non-muscle tissues from raw meat, cleaning, cutting into blocks, adding deionized water according to the feed-liquid ratio of 1 g: 2-5ml, and crushing and homogenizing.
3. The extraction method according to claim 1, characterized in that: the mass ratio of the trypsin, the papain and the neutral protease in the compound enzyme in the step (2) is 0.5-2: 11-13: 3-6.
4. The extraction method according to claim 3, characterized in that: the mass ratio of the trypsin, the papain and the neutral protease in the compound enzyme in the step (2) is 1-2: 12-13: 5-6.
5. The extraction method according to claim 1, characterized in that: and carrying out ultrasonic treatment while carrying out enzymolysis, wherein the ultrasonic power is 100-400W, and the ultrasonic frequency is 15-40 kHz.
6. The extraction method according to claim 5, characterized in that: the ultrasonic power is 200-280W, and the ultrasonic frequency is 18-25 kHz.
7. The extraction method according to any one of claims 3 to 6, characterized in that: the temperature of enzymolysis is 30-50 ℃; the pH value is 7.5-8.5; the enzymolysis time is 10-30 min; the mass of the complex enzyme is 0.5-2% of the raw material meat.
8. The extraction method according to claim 7, characterized in that: the temperature of the enzymolysis is 30-40 ℃; the pH value is 7.5-8.0; the enzymolysis time is 15-20 min; the mass of the complex enzyme added is 1-1.5% of the raw material meat.
9. The extraction method according to claim 1, characterized in that: the rotating speed of the centrifugation in the step (4) is 2000-; the filter membrane is filtered by adopting a 0.22 mu m microporous filter membrane.
10. Use of carnosine powder obtained by the extraction process according to any of claims 1 to 9 for the preparation of a cosmetic or food or health product.
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| K.M.CHAN 等: ""Extraction and Activity of Carnosine, a Naturally Occurring Antioxidant in Beef Muscle"", 《FOOD SCIENCE》 * |
| 赵赣 等: ""肌肽的提取及其抗氧化特性"", 《国外畜牧科技》 * |
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| Publication number | Publication date |
|---|---|
| CN114836491B (en) | 2023-03-14 |
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