A fermentation anti-hair loss composition and preparation method thereof

Abstract

The present invention proposes a fermentation anti-hair loss composition and a preparation method thereof, and belongs to the technical field of hair care products. Wheat protein / biotin-ornithine / nicotinamide condensation complex is prepared by dehydration condensation, Zn ions are chelated to obtain chelated zinc complex, tea polysaccharide is extracted, and licorice, astragalus, ginger, and ginseng are extracted by ethanol aqueous solution to obtain extract, tea residues and solid residues are fermented by lactic acid bacteria to obtain fermentation products, and fermentation products, tea polysaccharides, extracts, chelated zinc complexes, oleanolic acid, pantothenic acid, and hydroxyethylpiperazineethanesulfonic acid are mixed uniformly to obtain a fermentation anti-hair loss composition. The fermentation anti-hair loss composition prepared by the present invention has good antioxidant, antibacterial, mite removal, and anti-inflammatory effects, protects scalp health and maintains scalp homeostasis, promotes hair growth, reduces the effect of hair loss, improves the toughness and elasticity of hair, nourishes hair, has good anti-hair loss and solid hair growth effects, and has broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of hair care products, and in particular to a fermented anti-hair loss composition and a preparation method thereof. Background Art

[0002] Hair loss is a common dermatological disease, and androgenic alopecia is the most common. Hair loss affects the appearance and brings psychological pressure to patients. At present, the main solutions to prevent hair loss and hair growth are hair growth drugs and surgery. Surgical treatment is expensive, traumatic, and requires a certain recovery period; drug treatment mainly uses finasteride and minoxidil, which often have more serious side effects.

[0003] In recent years, there have been more and more reports on the use of plant extracts to treat hair loss. However, a single Chinese herbal medicine has limited effect on promoting hair growth in vitro, and according to the theory of complementarity in traditional Chinese medicine, the use of Chinese herbal medicine compounding is effective. Existing plant-based hair loss prevention and growth products often contain more than a dozen plant ingredients, which are costly, and the safety of multiple ingredients is unknown, and they cannot effectively prevent hair loss. Summary of the invention

[0004] The purpose of the present invention is to provide a fermented anti-hair loss composition and a preparation method thereof, which has excellent antioxidant, antibacterial, mite removal and anti-inflammatory effects, protects scalp health and maintains scalp homeostasis, promotes hair growth, reduces hair loss, improves the strength and elasticity of hair, nourishes hair, has excellent anti-hair loss and hair growth effects, and has broad application prospects.

[0005] The technical solution of the present invention is achieved in this way:

[0006] The invention provides a preparation method of a fermented hair loss prevention composition. The method comprises the following steps: preparing a wheat protein / biotin-ornithine / nicotinamide condensation complex by dehydration condensation, chelating Zn ions to obtain a chelated zinc complex, extracting tea polysaccharides, extracting liquorice, astragalus, ginger and ginseng by ethanol aqueous solution, fermenting tea residues and solid residues by lactic acid bacteria to obtain a fermentation product, and uniformly mixing the fermentation product, tea polysaccharides, the extract, the chelated zinc complex, oleanolic acid, pantothenic acid and hydroxyethylpiperazineethanesulfonic acid to prepare the fermented hair loss prevention composition.

[0007] As a further improvement of the present invention, the following steps are included:

[0008] S1. Add wheat protein and biotin to water, add EDC and NHS, stir to activate, add ornithine and nicotinamide, stir to react, centrifuge, wash, freeze-dry, and prepare a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0009] S2. adding a soluble zinc salt to water, adding the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stirring to react, dialyzing, and freeze-drying to obtain a chelated zinc complex;

[0010] S3. Add the crushed tea leaves to water, heat to boiling for extraction, filter, dry the tea residue and retain it, add ethanol to the filtrate for precipitation, centrifuge, wash, and dry to obtain tea polysaccharides;

[0011] S4. Grind and mix licorice, astragalus, ginger and ginseng, add to a mixed solvent of ethanol and water, heat to boiling for extraction, filter, dry the solid residue and retain it, dry the filtrate to obtain an extract;

[0012] S5. The tea residue in step S3 and the solid residue in step S4 are added to water, inoculated with lactic acid bacteria seed solution for fermentation, filtered, and the filtrate is dried to obtain a fermentation product;

[0013] S6. The oleanolic acid, pantothenic acid, hydroxyethylpiperazine ethanesulfonic acid were mixed to obtain a mixture;

[0014] S7. Evenly mix the chelated zinc complex prepared in step S2, the tea polysaccharide prepared in step S3, the extract prepared in step S4, the fermentation product prepared in step S5, and the mixture prepared in step S6 to prepare a fermented anti-hair loss composition.

[0015] As a further improvement of the present invention, the mass ratio of wheat protein, biotin, EDC, NHS, ornithine and nicotinamide in step S1 is 10-12:3-5:7-10:8-12:3-5:2-4, the activation time is 20-40 minutes, and the stirring reaction time is 18-24 hours.

[0016] As a further improvement of the present invention, the soluble zinc salt in step S2 is selected from at least one of zinc gluconate, zinc chloride, zinc sulfate, zinc citrate, and zinc nitrate, the mass ratio of the soluble zinc salt wheat protein / biotin-ornithine / nicotinamide condensation complex is 1-2:15-20, the stirring reaction time is 20-30 minutes, the dialysis bag pore size used in the dialysis is 1000-1500Da, and the time is 24-30 hours.

[0017] As a further improvement of the present invention, the time for heating to boiling extraction in step S3 is 2-4 hours, ethanol is added to the system ethanol content of 65-75wt%, and precipitation is carried out for 1-2 hours.

[0018] As a further improvement of the present invention, the mass ratio of licorice, astragalus, ginger, ginseng, and ethanol-water mixed solvent in step S4 is 1-2:2-4:10-12:2-3:200-300, the volume ratio of ethanol and water in the ethanol-water mixed solvent is 1:2-3, and the time for heating to boiling extraction is 2-4h.

[0019] As a further improvement of the present invention, the mass ratio of tea residue, solid residue and water in step S5 is 10-13:17-20:200, the lactic acid bacteria seed liquid is Lactobacillus helveticus seed liquid and Lactobacillus plantarum seed liquid, the inoculation amounts are 1-3v / v% and 2-4v / v%, respectively, and the bacterial content of the seed liquid is 10 8 -10 9 cfu / mL, and the fermentation conditions are 36-38°C, 100-150r / min, and anaerobic fermentation for 24-36h.

[0020] As a further improvement of the present invention, the mass ratio of oleanolic acid, pantothenic acid and hydroxyethylpiperazineethanesulfonic acid in step S6 is 3-5:0.5-1:1-2; the mass ratio of chelated zinc complex, tea polysaccharides, extract, fermentation product and mixture in step S7 is 10:2-3:1-2:4-6:0.5-1.

[0021] As a further improvement of the present invention, the present invention specifically comprises the following steps:

[0022] S1. Add 10-12 parts by weight of wheat protein and 3-5 parts by weight of biotin to 200 parts by weight of water, add 7-10 parts by weight of EDC and 8-12 parts by weight of NHS, stir and activate for 20-40 minutes, add 3-5 parts by weight of ornithine and 2-4 parts by weight of nicotinamide, stir and react for 18-24 hours, centrifuge, wash, and freeze-dry to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0023] S2. Add 1-2 parts by weight of a soluble zinc salt to 100 parts by weight of water, add 15-20 parts by weight of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir the reaction for 20-30 minutes, dialyze for 24-30 hours using a dialysis bag with a pore size of 1000-1500Da, and freeze-dry to obtain a chelated zinc complex;

[0024] S3. 10 parts by weight of crushed tea leaves were added to 100 parts by weight of water, heated to boiling and extracted for 2-4h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 65-75wt% of the system, precipitated for 1-2h, centrifuged, washed, and dried to obtain tea polysaccharides;

[0025] S4. 1-2 parts by weight of licorice, 2-4 parts by weight of astragalus, 10-12 parts by weight of ginger, and 2-3 parts by weight of ginseng are crushed and mixed, and added to 200-300 parts by weight of a mixed solvent of ethanol and water, wherein the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 1:2-3, heated to boiling and extracted for 2-4h, filtered, the solid residue is dried and retained, and the filtrate is dried to obtain an extract;

[0026] S5. 10-13 parts by weight of the tea residue in step S3 and 17-20 parts by weight of the solid residue in step S4 are added to 200 parts by weight of water, inoculated with Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution, the inoculation amounts are 1-3v / v% and 2-4v / v%, respectively, at 36-38°C, 100-150r / min, and fermented anaerobically for 24-36h, filtered, and the filtrate is dried to obtain a fermentation product;

[0027] The bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL;

[0028] S6 3-5 parts by weight of oleanolic acid, 0.5-1 parts by weight of pantothenic acid, 1-2 parts by weight of hydroxyethylpiperazineethanesulfonic acid were mixed to obtain a mixture;

[0029] S7. Evenly mix 10 parts by weight of the chelated zinc complex obtained in step S2, 2-3 parts by weight of tea polysaccharides obtained in step S3, 1-2 parts by weight of the extract obtained in step S4, 4-6 parts by weight of the fermentation product obtained in step S5, and 0.5-1 parts by weight of the mixture obtained in step S6 to obtain a fermented anti-hair loss composition.

[0030] The present invention further protects a fermented anti-hair loss composition prepared by the above preparation method.

[0031] The present invention has the following beneficial effects:

[0032] Wheat protein is rich in glutamic acid, lysine, cystine and other amino acids required for hair growth. It can be adsorbed on damaged parts of hair to enhance the tension and elasticity of hair fibers. Biotin has a significant effect on the prevention and treatment of hair loss. After the carboxyl groups of wheat protein and biotin are activated by EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide) and NHS (N-hydroxysuccinimide), they can undergo condensation reactions with nicotinamide and ornithine with amino groups to obtain wheat protein / biotin-ornithine / nicotinamide condensation complex. Ornithine can promote keratin synthesis and interact with disulfide bonds in hair to have a certain protective effect on hair. It can also accelerate cell growth and promote protein synthesis. Nicotinamide can also promote blood circulation in the hair bulb. Therefore, the prepared wheat protein / biotin-ornithine / nicotinamide condensation complex has the effects of significantly improving hair elasticity, protecting hair, enhancing hair application, and preventing and treating hair loss.

[0033] The mineral protein formed by the covalent bonding of trace element Zn and protein can greatly improve the effect of metal ions being absorbed by cells. At the same time, zinc can form a chemical bond similar to a disulfide bond on the helical structure between cysteine ​​molecules in hair, which has a good effect of nourishing hair and thus preventing hair loss.

[0034] Tea polysaccharides have excellent antioxidant, anti-inflammatory and antibacterial effects. The products obtained by extracting licorice, astragalus, ginger and ginseng with water and ethanol are rich in active ingredients such as astragaloside I, glycyrrhizic acid, ginsenosides, curcumin, gingerol, etc. On the one hand, they have antioxidant and anti-inflammatory effects. At the same time, they can also stimulate the local skin and blood supply, activate hair follicles and promote hair growth, and have a good effect of fixing hair and preventing hair loss.

[0035] Mixing tea residues and solid residues of traditional Chinese medicine for fermentation can, on the one hand, convert ordinary ginsenosides into rare ginsenosides, which are rich in active ingredients, greatly enhancing the antioxidant, antibacterial, anti-mite and anti-inflammatory effects of the fermentation products, thereby significantly protecting the health of the scalp, protecting hair follicles, promoting hair growth and reducing hair loss.

[0036] Hydroxyethylpiperazineethanesulfonic acid can keep the scalp in a relatively stable weak acid range for a long time, improve skin condition, and increase the permeability of active ingredients to the skin. It is non-toxic, mild, and non-irritating. Oleanolic acid can play an anti-hair loss effect by antagonizing the receptors of corticotropin-releasing hormone (CRH), significantly inhibiting the phenomenon of growth phase hair turning into catagen phase hair caused by CRH, thereby preventing hair loss. Pantothenic acid is a vitamin necessary for hair growth metabolism, which helps to improve the strength of hair and has a synergistic effect.

[0037] The fermented anti-hair loss composition prepared by the present invention has excellent antioxidant, antibacterial, mite removal and anti-inflammatory effects, protects scalp health and maintains scalp homeostasis, promotes hair growth, reduces hair loss, improves hair toughness and elasticity, nourishes hair, has excellent anti-hair loss and hair growth effects, and has broad application prospects. DETAILED DESCRIPTION

[0038] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] Lactobacillus helveticus, CICC6024, 10 billion cfu / g, was purchased from Ruichu Biotechnology (Jiangsu) Co., Ltd. Lactobacillus plantarum, 10 billion cfu / g, was purchased from Baoji Liupanyun Biotechnology Co., Ltd.

[0040] Preparation method of Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution: a single strain is inoculated into a slant culture medium, cultured under anoxic conditions at 37°C and 100r / min for 18-24h to obtain a strain seed solution containing 10 8 -10 9 cfu / mL.

[0041] Wheat protein is hydrolyzed wheat protein with a content of >99%, CAS number 70084-87-6, purchased from Sichuan Century Star Biotechnology Co., Ltd.

[0042] Example 1

[0043] This embodiment provides a method for preparing a fermented anti-hair loss composition, which specifically comprises the following steps:

[0044] S1. 10 g wheat protein and 3 g biotin were added to 200 g water, 7 g EDC and 8 g NHS were added, and the mixture was stirred and activated at 0°C for 20 min, 3 g ornithine and 2 g nicotinamide were added, and the mixture was stirred and reacted for 18 h, centrifuged, washed, and freeze-dried to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0045] S2. Add 1 g of zinc chloride to 100 g of water, add 15 g of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir and react for 20 min, dialyze for 24 h using a dialysis bag with a pore size of 1000 Da, and freeze-dry to obtain a chelated zinc complex;

[0046] S3. 10 g of crushed tea leaves were added to 100 g of water, heated to boiling and extracted for 2 h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 65 wt % of the system, precipitated for 1 h, centrifuged, washed, and dried to obtain tea polysaccharides;

[0047] S4. 1g licorice, 2g astragalus, 10g ginger, 2g ginseng were crushed and mixed, added to 200g ethanol-water mixed solvent, heated to boiling and extracted for 2h, filtered, the solid residue was dried and retained, and the filtrate was dried to obtain an extract;

[0048] The volume ratio of ethanol to water in the ethanol-water mixed solvent is 1:2;

[0049] S5. 10 g of the tea residue in step S3 and 17 g of the solid residue in step S4 were added to 200 g of water and inoculated with Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution, respectively, inoculated at 1 v / v% and 2 v / v%, 36 ° C, 100 r / min, anoxic fermentation for 24 h, filtered, and the filtrate was dried to obtain a fermentation product;

[0050] S6. 3 g of oleanolic acid, 0.5 g of pantothenic acid and 1 g of hydroxyethylpiperazineethanesulfonic acid were stirred and mixed for 10 min to obtain a mixture;

[0051] S7. Stir and mix 10 g of the chelated zinc complex prepared in step S2, 2 g of tea polysaccharides prepared in step S3, 1 g of the extract prepared in step S4, 4 g of the fermentation product prepared in step S5, and 0.5 g of the mixture prepared in step S6 for 15 minutes to prepare a fermented anti-hair loss composition.

[0052] Example 2

[0053] This embodiment provides a method for preparing a fermented anti-hair loss composition, which specifically comprises the following steps:

[0054] S1. 12 g wheat protein and 5 g biotin were added to 200 g water, 10 g EDC and 12 g NHS were added, and the mixture was stirred and activated at 0°C for 40 min, 5 g ornithine and 4 g nicotinamide were added, and the mixture was stirred and reacted for 24 h, centrifuged, washed, and freeze-dried to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0055] S2. Add 2 g of zinc citrate to 100 g of water, add 20 g of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir and react for 30 min, dialyze for 30 h using a dialysis bag with a pore size of 1500 Da, and freeze-dry to obtain a chelated zinc complex;

[0056] S3. 10 g of crushed tea leaves were added to 100 g of water, heated to boiling and extracted for 4 h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 75 wt % of the system, precipitated for 2 h, centrifuged, washed, and dried to obtain tea polysaccharides;

[0057] S4. 2g licorice, 4g astragalus, 12g ginger, 3g ginseng were crushed and mixed, added to 300g of ethanol and water mixed solvent, heated to boiling and extracted for 4h, filtered, the solid residue was dried and retained, and the filtrate was dried to obtain an extract;

[0058] The volume ratio of ethanol to water in the ethanol-water mixed solvent is 1:3;

[0059] S5. 13 g of the tea residue in step S3 and 20 g of the solid residue in step S4 were added to 200 g of water and inoculated with Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution, the inoculation amounts were 3 v / v% and 4 v / v%, respectively, at 38 ° C, 150 r / min, anoxic fermentation for 36 h, filtered, and the filtrate was dried to obtain a fermentation product;

[0060] S6. 5 g of oleanolic acid, 1 g of pantothenic acid and 2 g of hydroxyethylpiperazineethanesulfonic acid were stirred and mixed for 10 min to obtain a mixture;

[0061] S7. Stir and mix 10 g of the chelated zinc complex prepared in step S2, 3 g of the tea polysaccharide prepared in step S3, 2 g of the extract prepared in step S4, 6 g of the fermentation product prepared in step S5, and 1 g of the mixture prepared in step S6 for 15 minutes to prepare a fermented anti-hair loss composition.

[0062] Example 3

[0063] This embodiment provides a method for preparing a fermented anti-hair loss composition, which specifically comprises the following steps:

[0064] S1. 11 g wheat protein and 4 g biotin were added to 200 g water, 8.5 g EDC and 10 g NHS were added, and the mixture was stirred and activated at 0°C for 30 min, 4 g ornithine and 3 g nicotinamide were added, and the mixture was stirred and reacted for 21 h, centrifuged, washed, and freeze-dried to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0065] S2. Add 1.5 g of zinc nitrate to 100 g of water, add 17 g of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir and react for 25 min, dialyze for 26 h using a dialysis bag with a pore size of 1200 Da, and freeze-dry to obtain a chelated zinc complex;

[0066] S3. 10 g of crushed tea leaves were added to 100 g of water, heated to boiling and extracted for 3 h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 70 wt % of the system, precipitated for 1.5 h, centrifuged, washed, and dried to obtain tea polysaccharides;

[0067] S4. 1.5 g licorice, 3 g astragalus, 11 g ginger, 2.5 g ginseng were crushed and mixed, added to 250 g of ethanol and water mixed solvent, heated to boiling and extracted for 3 h, filtered, the solid residue was dried and retained, and the filtrate was dried to obtain an extract;

[0068] The volume ratio of ethanol to water in the ethanol-water mixed solvent is 1:2-3;

[0069] S5. 11 g of the tea residue in step S3 and 18 g of the solid residue in step S4 were added to 200 g of water and inoculated with Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution, the inoculation amounts were 2 v / v% and 3 v / v%, respectively, at 37 ° C, 125 r / min, anoxic fermentation for 30 h, filtered, and the filtrate was dried to obtain a fermentation product;

[0070] S6. 4 g of oleanolic acid, 0.7 g of pantothenic acid and 1.5 g of hydroxyethylpiperazineethanesulfonic acid were stirred and mixed for 10 min to obtain a mixture;

[0071] S7. Stir and mix 10 g of the chelated zinc complex prepared in step S2, 2.5 g of tea polysaccharides prepared in step S3, 1.5 g of the extract prepared in step S4, 5 g of the fermentation product prepared in step S5, and 0.7 g of the mixture prepared in step S6 for 15 minutes to prepare a fermented anti-hair loss composition.

[0072] Comparative Example 1

[0073] Compared with Example 3, the difference is that wheat protein is not added in step S1.

[0074] The details are as follows:

[0075] S1. Add 15g of biotin to 200g of water, add 8.5g of EDC and 10g of NHS, stir and activate at 0℃ for 30min, add 4g of ornithine and 3g of nicotinamide, stir and react for 21h, centrifuge, wash, and freeze-dry to obtain a biotin-ornithine / nicotinamide condensation complex.

[0076] Comparative Example 2

[0077] Compared with Example 3, the difference is that biotin is not added in step S1.

[0078] The details are as follows:

[0079] S1. Add 15g wheat protein to 200g water, add 8.5g EDC and 10g NHS, stir and activate at 0℃ for 30min, add 4g ornithine and 3g nicotinamide, stir and react for 21h, centrifuge, wash, freeze-dry, and obtain wheat protein-ornithine / nicotinamide condensation complex.

[0080] Comparative Example 3

[0081] Compared with Example 3, the difference is that ornithine is not added in step S1.

[0082] The details are as follows:

[0083] S1. Add 11g wheat protein and 4g biotin to 200g water, add 8.5g EDC and 10g NHS, activate with stirring at 0℃ for 30min, add 7g nicotinamide, react with stirring for 21h, centrifuge, wash, freeze-dry, and obtain wheat protein / biotin-nicotinamide condensation complex.

[0084] Comparative Example 4

[0085] Compared with Example 3, the difference is that nicotinamide is not added in step S1.

[0086] The details are as follows:

[0087] S1. Add 11g wheat protein and 4g biotin to 200g water, add 8.5g EDC and 10g NHS, activate with stirring at 0℃ for 30min, add 7g ornithine, react with stirring for 21h, centrifuge, wash, freeze-dry, and obtain wheat protein / biotin-ornithine condensation complex.

[0088] Comparative Example 5

[0089] Compared with Example 3, the difference is that step S2 is not performed.

[0090] The details are as follows:

[0091] S1. 11 g wheat protein and 4 g biotin were added to 200 g water, 8.5 g EDC and 10 g NHS were added, and the mixture was stirred and activated at 0°C for 30 min, 4 g ornithine and 3 g nicotinamide were added, and the mixture was stirred and reacted for 21 h, centrifuged, washed, and freeze-dried to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0092] S2. 10 g of crushed tea leaves were added to 100 g of water, heated to boiling and extracted for 3 h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 70 wt % of the system, precipitated for 1.5 h, centrifuged, washed, and dried to obtain tea polysaccharides;

[0093] S3. 1.5 g licorice, 3 g astragalus, 11 g ginger, 2.5 g ginseng were crushed and mixed, added to 250 g ethanol-water mixed solvent, heated to boiling and extracted for 3 h, filtered, the solid residue was dried and retained, and the filtrate was dried to obtain an extract;

[0094] The volume ratio of ethanol to water in the ethanol-water mixed solvent is 1:2-3;

[0095] S4. 11g of the tea residue in step S2 and 18g of the solid residue in step S3 were added to 200g of water, inoculated with Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution, the inoculation amounts were 2v / v% and 3v / v%, respectively, at 37°C, 125r / min, and fermented anaerobically for 30h, filtered, and the filtrate was dried to obtain a fermentation product;

[0096] S5. 4 g of oleanolic acid, 0.7 g of pantothenic acid and 1.5 g of hydroxyethylpiperazineethanesulfonic acid were stirred and mixed for 10 min to obtain a mixture;

[0097] S6. Stir and mix 10 g of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, 2.5 g of tea polysaccharides prepared in step S2, 1.5 g of the extract prepared in step S3, 5 g of the fermentation product prepared in step S4, and 0.7 g of the mixture prepared in step S5 for 15 minutes to prepare a fermented anti-hair loss composition.

[0098] Comparative Example 6

[0099] Compared with Embodiment 3, the difference is that step S5 is not performed.

[0100] The details are as follows:

[0101] S1. 11 g wheat protein and 4 g biotin were added to 200 g water, 8.5 g EDC and 10 g NHS were added, and the mixture was stirred and activated at 0°C for 30 min, 4 g ornithine and 3 g nicotinamide were added, and the mixture was stirred and reacted for 21 h, centrifuged, washed, and freeze-dried to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex;

[0102] S2. Add 1.5 g of zinc nitrate to 100 g of water, add 17 g of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir and react for 25 min, dialyze for 26 h using a dialysis bag with a pore size of 1200 Da, and freeze-dry to obtain a chelated zinc complex;

[0103] S3. 10 g of crushed tea leaves were added to 100 g of water, heated to boiling and extracted for 3 h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 70 wt % of the system, precipitated for 1.5 h, centrifuged, washed, and dried to obtain tea polysaccharides;

[0104] S4. 1.5 g licorice, 3 g astragalus, 11 g ginger, 2.5 g ginseng were crushed and mixed, added to 250 g of ethanol and water mixed solvent, heated to boiling and extracted for 3 h, filtered, the solid residue was dried and retained, and the filtrate was dried to obtain an extract;

[0105] The volume ratio of ethanol to water in the ethanol-water mixed solvent is 1:2-3;

[0106] S5. 4 g of oleanolic acid, 0.7 g of pantothenic acid and 1.5 g of hydroxyethylpiperazineethanesulfonic acid were stirred and mixed for 10 min to obtain a mixture;

[0107] S6. 10 g of the chelated zinc complex prepared in step S2, 2.5 g of the tea polysaccharide prepared in step S3, 1.5 g of the extract prepared in step S4, and 0.7 g of the mixture prepared in step S5 were stirred and mixed for 15 minutes to prepare a fermented anti-hair loss composition.

[0108] Comparative Example 7

[0109] Compared with Example 3, the difference is that hydroxyethylpiperazineethanesulfonic acid is not added in step S6.

[0110] The details are as follows:

[0111] S6. 4 g of oleanolic acid and 0.7 g of pantothenic acid were stirred and mixed for 10 min to obtain a mixture.

[0112] Comparative Example 8

[0113] Compared with Example 3, the difference is that no chelated zinc complex is added in step S7.

[0114] The details are as follows:

[0115] S7. 2.5 g of the tea polysaccharide prepared in step S3, 1.5 g of the extract prepared in step S4, 5 g of the fermentation product prepared in step S5, and 0.7 g of the mixture prepared in step S6 were stirred and mixed for 15 minutes to prepare a fermented anti-hair loss composition.

[0116] Comparative Example 9

[0117] Compared with Example 3, the difference is that no extract is added in step S7.

[0118] The details are as follows:

[0119] S7. 10 g of the chelated zinc complex prepared in step S2, 2.5 g of tea polysaccharides prepared in step S3, 5 g of the fermentation product prepared in step S5, and 0.7 g of the mixture prepared in step S6 were stirred and mixed for 15 minutes to prepare a fermented anti-hair loss composition.

[0120] Comparative Example 10

[0121] Compared with Example 3, the difference is that no mixture is added in step S7.

[0122] The details are as follows:

[0123] S7. Stir and mix 10 g of the chelated zinc complex prepared in step S2, 2.5 g of tea polysaccharides prepared in step S3, 1.5 g of the extract prepared in step S4, and 5 g of the fermentation product prepared in step S5 for 15 minutes to prepare a fermented anti-hair loss composition.

[0124] Test Example 1 In vitro 5α-reductase inhibition assay

[0125] The fermented anti-hair loss compositions prepared in Examples 1-3 and Comparative Examples 1-10 were formulated into 0.05 mg / mL test solutions.

[0126] Preparation of 5α-reductase: Male SD rats were fasted overnight but not water-deprived and then killed. The prostate was removed and cut into pieces on an ice table. A 1:5 homogenate was made in a glass homogenizer at 0°C using a buffer solution (containing 0.32 mol / L sucrose, 0.1 mmol / L dithiothreitol, 1 mmol / L EDTA and 0.2 mol / L phosphate buffer), and the mixture was centrifuged. After removing the upper white fat layer, the supernatant was taken and added with a buffer solution to a constant volume of 20 mL, which was then dispensed into EP tubes and stored in a -80°C refrigerator.

[0127] 5α-reductase activity determination: Take 0.5mL phosphate buffer, 0.1mL test solution, 200μL 300mg / L testosterone solution and 200μL 0.8g / L NADPH (reduced nicotinamide adenine dinucleotide phosphate) solution, add 0.5mL 5α-reductase extract, react at 37℃ for 30min, add 2.5mL dichloromethane to stop and extract testosterone, then add 0.25mL 100mg / L propyl hydroxybenzoate as internal standard, centrifuge, remove the upper aqueous phase, remove about 1mL organic phase, evaporate to dryness, dissolve the residue in 1.5mL methanol, and use high performance liquid chromatography to determine the residual testosterone. Set up 5α-reductase reaction tube and 5α-reductase blank tube. Finasteride (0.05mg / L) was used as the control group. The results are shown in Table 1.

[0128] 5α-reductase inhibition rate (%) = (R 样品 -R 反应 ) / (R 空白 -R 反应 )×100%

[0129] Where R is the ratio of the target peak area to the internal standard peak area.

[0130] Table 1

[0131]

[0132]

[0133] Most people with hair loss suffer from androgenic alopecia, which is caused by the conversion of the male hormone testosterone into more active dihydrotestosterone under the action of 5α-reductase, which inhibits the binding of testosterone to receptors and blocks the hair follicles in the resting phase from entering the growth phase. Therefore, the fermented anti-hair loss composition prepared in Examples 1-3 of the present invention has a good 5α-reductase inhibitory effect, thereby playing a good anti-hair loss role.

[0134] Test Example 2 DPPH clearance test:

[0135] The fermented anti-hair loss composition prepared in Examples 1-3 and Comparative Examples 1-10 was formulated into a 0.05 mg / mL test solution. A 0.2 mmol / L DPPH solution was prepared on the same day, 200 μL of the test solution was added to 800 μL of the DPPH solution, incubated at room temperature for 60 min in the dark, and the absorbance value (A value) was measured at a wavelength of 517 nm. The test was repeated 3 times. Vitamin C (0.05 mg / mL) was used as the control group. The results are shown in Table 2.

[0136] DPPH free radical scavenging rate (%) = 1-(A 样品 -A 空白 ) / A 对照 ×100%

[0137] Table 2

[0138]

[0139]

[0140] It can be seen from the above table that the fermented anti-hair loss compositions prepared in Examples 1-3 of the present invention have good antioxidant properties.

[0141] Test Example 3 Mouse hair growth experiment

[0142] SPF grade C57BL / 6 black mice were randomly divided into Example 1-3 groups, Comparative Example 1-10 groups, positive control group (2% minoxidil solution) and blank group (normal saline), 10 mice in each group, and the hair on the back of the mice was removed one day before the test, and 1% of the corresponding fermented anti-hair loss composition sample aqueous solution, 2% minoxidil solution and normal saline were applied respectively, and the application was continued for 33 days, 0.2mL / mouse / day. The back skin of the mice was observed every day, and the time from pink to black was recorded, which was recorded as T1; the time from black to hair growth was recorded as T2.

[0143] The results are shown in Table 3.

[0144] Table 3

[0145] Group T1(d) T2(d) Blank group 18.9±4.2 24.4±3.9 Positive control group 9.9±2.6* 12.3±1.8* Example 1 10.4±1.9* 13.8±2.8* Example 2 10.5±2.1* 13.6±2.4* Example 3 10.1±2.3* 13.0±2.2* Comparative Example 1 11.5±3.3 14.5±3.6 Comparative Example 2 12.2±3.9 15.7±3.2 Comparative Example 3 12.9±2.8 16.4±2.9 Comparative Example 4 13.1±3.4 15.8±3.5 Comparative Example 5 14.8±3.2 16.9±3.1 Comparative Example 6 14.1±3.8 16.2±3.7 Comparative Example 7 14.4±3.4 16.4±3.9 Comparative Example 8 15.3±4.2 17.6±4.0 Comparative Example 9 14.7±3.3 16.8±4.1 Comparative Example 10 14.9±4.1 17.1±3.8

[0146] Note: *Compared with the blank group, P < 0.05.

[0147] It can be seen from the above table that the fermented anti-hair loss compositions prepared in Examples 1-3 of the present invention have a good effect of shortening the time for hair to enter the growth phase and the time for hair to grow out, which helps to promote hair growth.

[0148] On the 14th day, mice were randomly killed 24 hours after application, and complete skin specimens were taken from the same part of the shaved area, fixed and made into paraffin sections, stained with HE, and the number of hair follicles and subcutaneous hair follicles was analyzed under a microscope. Five fields of view were randomly selected from each slide for analysis. The results are shown in Table 4.

[0149] Growth phase induction rate (%) = number of subcutaneous hair follicles / total number of hair follicles × 100%.

[0150] Table 4

[0151] Group Growth period induction rate (%) Blank group 55.2 Positive control group 67.8 Example 1 65.4 Example 2 65.9 Example 3 66.1 Comparative Example 1 63.5 Comparative Example 2 62.9 Comparative Example 3 63.1 Comparative Example 4 61.8 Comparative Example 5 58.2 Comparative Example 6 59.7 Comparative Example 7 60.2 Comparative Example 8 56.1 Comparative Example 9 62.3 Comparative Example 10 58.9

[0152] It can be seen from the above table that the fermented anti-hair loss compositions prepared in Examples 1-3 of the present invention have a higher growth phase induction rate.

[0153] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a fermented anti-hair loss composition, characterized in that: The following steps are involved: S1. Add wheat protein and biotin to water, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and N-hydroxysuccinimide, stir to activate, add ornithine and nicotinamide, stir to react, centrifuge, wash, freeze-dry, and prepare a wheat protein / biotin-ornithine / nicotinamide condensation complex; the mass ratio of the wheat protein, biotin, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, N-hydroxysuccinimide, ornithine and nicotinamide is 10-12:3-5:7-10:8-12:3-5:2-4; S2. Add a soluble zinc salt to water, add the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir to react, dialyze, and freeze-dry to obtain a chelated zinc complex; the mass ratio of the soluble zinc salt to the wheat protein / biotin-ornithine / nicotinamide condensation complex is 1-2:15-20; S3. Add the crushed tea leaves to water, heat to boiling for extraction, filter, dry the tea residue and retain it, add ethanol to the filtrate for precipitation, centrifuge, wash, and dry to obtain tea polysaccharides; S4. Grind and mix licorice, astragalus, ginger and ginseng, add them to a mixed solvent of ethanol and water, heat them to boiling for extraction, filter them, dry the solid residue and retain it, dry the filtrate to obtain an extract; the mass ratio of licorice, astragalus, ginger, ginseng and ethanol-water mixed solvent is 1-2:2-4:10-12:2-3:200-300; S5. The tea residue in step S3 and the solid residue in step S4 are added to water, inoculated with lactic acid bacteria seed solution for fermentation, filtered, and the filtrate is dried to obtain a fermentation product; the mass ratio of the tea residue, solid residue and water is 10-13:17-20:200; S6. The oleanolic acid, pantothenic acid, hydroxyethylpiperazine ethanesulfonic acid were mixed to obtain a mixture; the mass ratio of the oleanolic acid, pantothenic acid, hydroxyethylpiperazine ethanesulfonic acid was 3-5:0.5-1:1-2; S7. The chelated zinc complex prepared in step S2, the tea polysaccharide prepared in step S3, the extract prepared in step S4, the fermentation product prepared in step S5, and the mixture prepared in step S6 are uniformly mixed to prepare a fermented anti-hair loss composition; the mass ratio of the chelated zinc complex, tea polysaccharide, extract, fermentation product, and mixture is 10:2-3:1-2:4-6:0.5-1.

2. The preparation method according to claim 1, characterized in that: The activation time in step S1 is 20-40 minutes, and the stirring reaction time is 18-24 hours.

3. The preparation method according to claim 1, characterized in that: The soluble zinc salt in step S2 is selected from at least one of zinc gluconate, zinc chloride, zinc sulfate, zinc citrate, and zinc nitrate. The stirring reaction time is 20-30 minutes. The dialysis bag used in the dialysis has a pore size of 1000-1500Da and the time is 24-30 hours.

4. The preparation method according to claim 1, characterized in that: The time of heating to boiling extraction in step S3 is 2-4 hours, ethanol is added to the system ethanol content of 65-75wt%, and precipitation is carried out for 1-2 hours.

5. The preparation method according to claim 1, characterized in that: In step S4, the volume ratio of ethanol to water in the ethanol-water mixed solvent is 1:2-3, and the time for heating to boiling extraction is 2-4 hours.

6. The preparation method according to claim 1, characterized in that: The lactic acid bacteria seed solution in step S5 is a Lactobacillus helveticus seed solution and a Lactobacillus plantarum seed solution, and the inoculation amounts are 1-3 v / v% and 2-4 v / v%, respectively. The bacterial content of the seed solution is 10 8 -10 9 cfu / mL, and the fermentation conditions are 36-38°C, 100-150r / min, and anaerobic fermentation for 24-36h.

7. The preparation method according to claim 1, characterized in that: The specific steps include: S1. Add 10-12 parts by weight of wheat protein and 3-5 parts by weight of biotin to 200 parts by weight of water, add 7-10 parts by weight of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 8-12 parts by weight of N-hydroxysuccinimide, stir and activate for 20-40 minutes, add 3-5 parts by weight of ornithine and 2-4 parts by weight of nicotinamide, stir and react for 18-24 hours, centrifuge, wash, and freeze-dry to obtain a wheat protein / biotin-ornithine / nicotinamide condensation complex; S2. Add 1-2 parts by weight of a soluble zinc salt to 100 parts by weight of water, add 15-20 parts by weight of the wheat protein / biotin-ornithine / nicotinamide condensation complex prepared in step S1, stir the reaction for 20-30 minutes, dialyze for 24-30 hours using a dialysis bag with a pore size of 1000-1500Da, and freeze-dry to obtain a chelated zinc complex; S3. 10 parts by weight of crushed tea leaves were added to 100 parts by weight of water, heated to boiling and extracted for 2-4h, filtered, the tea residue was dried and retained, ethanol was added to the filtrate to an ethanol content of 65-75wt% of the system, precipitated for 1-2h, centrifuged, washed, and dried to obtain tea polysaccharides; S4. 1-2 parts by weight of licorice, 2-4 parts by weight of astragalus, 10-12 parts by weight of ginger, and 2-3 parts by weight of ginseng are crushed and mixed, and added to 200-300 parts by weight of a mixed solvent of ethanol and water, wherein the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 1:2-3, heated to boiling and extracted for 2-4h, filtered, the solid residue is dried and retained, and the filtrate is dried to obtain an extract; S5. 10-13 parts by weight of the tea residue in step S3 and 17-20 parts by weight of the solid residue in step S4 are added to 200 parts by weight of water, inoculated with Lactobacillus helveticus seed solution and Lactobacillus plantarum seed solution, the inoculation amounts are 1-3v / v% and 2-4v / v%, respectively, at 36-38°C, 100-150r / min, and fermented anaerobically for 24-36h, filtered, and the filtrate is dried to obtain a fermentation product; The bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL; S6 3-5 parts by weight of oleanolic acid, 0.5-1 parts by weight of pantothenic acid, 1-2 parts by weight of hydroxyethylpiperazineethanesulfonic acid were mixed to obtain a mixture; S7. Evenly mix 10 parts by weight of the chelated zinc complex obtained in step S2, 2-3 parts by weight of tea polysaccharides obtained in step S3, 1-2 parts by weight of the extract obtained in step S4, 4-6 parts by weight of the fermentation product obtained in step S5, and 0.5-1 parts by weight of the mixture obtained in step S6 to obtain a fermented anti-hair loss composition.

8. A fermented anti-hair loss composition obtained by the preparation method according to any one of claims 1 to 7.

Images