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CN-121987515-A - Preparation method of theaflavin self-assembled nano-particles and application of theaflavin self-assembled nano-particles in hair loss prevention

CN121987515ACN 121987515 ACN121987515 ACN 121987515ACN-121987515-A

Abstract

The invention discloses a preparation method of theaflavin self-assembled nano particles and application thereof in preventing alopecia. The preparation method of the theaflavin self-assembled nano-particles comprises the following steps of (1) soaking tea tree branches and leaves in water, adding laccase to perform enzymatic oxidation reaction, filtering and concentrating after the reaction, extracting chloroform and ethyl acetate to obtain theaflavin, and (2) dissolving the theaflavin and L-cysteine in the water, and dropwise adding formaldehyde solution to perform reaction to obtain the theaflavin self-assembled nano-particles. The self-assembled nano particles prepared by the invention are rich in sulfhydryl groups, can target hair follicles, stimulate hair growth, are rich in amino groups, carboxyl groups and phenolic hydroxyl groups, can be combined with hyaluronidase and 5 alpha-reductase, and have the effects of preventing androgenetic alopecia and improving scalp sensitivity.

Inventors

  • YE YONG
  • LIU XUELUAN
  • YUAN HANG

Assignees

  • 华南理工大学

Dates

Publication Date
20260508
Application Date
20260202

Claims (10)

  1. 1. The preparation method of the theaflavin self-assembled nano-particles is characterized by comprising the following steps of: (1) Preparing theaflavin, namely soaking tea tree branches and leaves in water, adding laccase, carrying out enzymatic oxidation reaction under the conditions of 30-50 ℃ and stirring, inactivating enzyme after the reaction is finished, cooling to room temperature, filtering and collecting filtrate, concentrating to obtain concentrated solution, extracting the concentrated solution with chloroform, collecting water phase, extracting with ethyl acetate, collecting ethyl acetate phase, finally removing ethyl acetate, and drying to obtain theaflavin; (2) And (3) preparing theaflavin self-assembled nano particles, namely dissolving the theaflavin and the L-cysteine obtained in the step (1) into water, then dropwise adding formaldehyde solution, continuously stirring at room temperature for reaction, centrifuging after the reaction is finished, collecting precipitate, washing and drying to obtain the theaflavin self-assembled nano particles.
  2. 2. The method for preparing the theaflavin self-assembled nanoparticle according to claim 1, wherein: the dosage ratio of tea tree branches and leaves to laccase in the step (1) is 1 g:250-750U; the mass ratio of the theaflavin to the L-cysteine in the step (2) is 1:0.2-1; The dosage ratio of theaflavin to water in the step (2) is 1 g:200-500 mL.
  3. 3. The method for preparing the theaflavin self-assembled nanoparticle according to claim 1, wherein: The dosage ratio of tea tree branches to leaves to water in the step (1) is 1 g:30-50 mL; the concentration of the formaldehyde solution in the step (2) is 37 percent by volume; the dosage ratio of the theaflavin to the formaldehyde solution in the step (2) is 1 g:2-5 mL.
  4. 4. The method for preparing the theaflavin self-assembled nanoparticle according to claim 1, wherein: the time of the enzymatic oxidation reaction in the step (1) is 6-10 hours; The stirring reaction condition in the step (2) is that the stirring speed is 300-600 r/min, and the reaction time is 2-6 h.
  5. 5. The method for preparing the theaflavin self-assembled nanoparticle according to claim 1, wherein: The tea tree branches and leaves in the step (1) are tea tree pruning branches and leaves; the stirring speed in the step (1) is 300-500 r/min; The enzyme deactivation in the step (1) is medium-high temperature enzyme deactivation, and the conditions are that the temperature is 90-100 ℃ and the time is 5-10 min; The drying condition in the step (1) is that the vacuum drying is carried out for 12-24 hours at 50-70 ℃; The centrifugation condition in the step (2) is that the centrifugation rotating speed is 8000-12000 r/min, and the centrifugation time is 5-10 min; And (3) the drying condition in the step (2) is freeze drying for 36-48 h.
  6. 6. The method for preparing the theaflavin self-assembled nanoparticle according to claim 1, wherein: The concentration condition in the step (1) is that the concentration is carried out in vacuum at 60-80 ℃ until the volume is 1/5-1/10 of the volume of the original reaction liquid; The dosage of the chloroform in the step (1) is 0.5-1.5 times of the volume of the concentrated solution; The ethyl acetate in the step (1) is used in an amount which is 0.5-1.5 times the volume of the water phase.
  7. 7. The method for preparing the theaflavin self-assembled nanoparticle according to claim 1, wherein: the times of extraction with chloroform in the step (1) are 2-3 times; The extraction times of ethyl acetate in the step (1) are 2-3 times; the washing in the step (2) is carried out by adopting deionized water.
  8. 8. A theaflavin self-assembled nanoparticle characterized by being prepared by the method of any one of claims 1 to 7.
  9. 9. Use of the theaflavin self-assembled nanoparticle of claim 8 in the preparation of an anti-hair loss, anti-oxidant, anti-allergic and/or anti-inflammatory product.
  10. 10. The use according to claim 9, characterized in that: the alopecia is androgenetic alopecia; the product comprises medicines or daily chemicals.

Description

Preparation method of theaflavin self-assembled nano-particles and application of theaflavin self-assembled nano-particles in hair loss prevention Technical Field The invention relates to the field of cosmetics, in particular to a preparation method of theaflavin self-assembled nano particles and application of the theaflavin self-assembled nano particles in hair loss prevention. Background Androgenic alopecia (AGA) is the most common type of hair loss, the core mechanism of which is that the hair follicle is affected by androgens (e.g. dihydrotestosterone, DHT), leading to miniaturization, shortened anagen phase and eventually atrophy. The existing treatment means (such as minoxidil and finasteride) have the problems of local irritation, systemic side effects or unstable curative effect and the like, and have limited applicability to scalp sensitive people (such as people accompanied with itching, erythema and seborrheic dermatitis), and are easy to cause scalp oxidative stress damage, thereby further exacerbating alopecia. Natural active ingredients become research and development hot spots for AGA treatment and scalp health management due to multi-target regulation and low toxicity advantages, but are limited by the problems of poor stability, low permeation efficiency and the like, and have limited practical application effects. Theaflavin is a polyphenol compound generated in the reaction process of black tea, and has remarkable anti-inflammatory, antioxidant and antibacterial biological functions. However, theaflavin molecules are susceptible to degradation by light, temperature and pH, and their hydrophilicity results in difficulty penetrating the scalp stratum corneum, limiting their immediate use in hair care products. In the prior art, as in chinese patent application CN119326128a, a preparation method of pH-responsive composite hydrogel carrying theaflavin is disclosed, CN116003367a discloses a preparation method of theaflavin derivative, and stability of the theaflavin derivative is improved by embedding technology or chemical modification respectively, but there are problems of complex process, potential toxicity or insufficient loading of carrier material, etc., so that effects of preventing alopecia and improving scalp sensitivity are difficult to achieve. Disclosure of Invention The primary aim of the invention is to overcome the defects and shortcomings of the prior art and provide a preparation method of theaflavin self-assembled nano particles. The invention also aims to provide the theaflavin self-assembled nano-particles prepared by the method. It is a further object of the present invention to provide the use of said theaflavin self-assembled nanoparticle. The aim of the invention is achieved by the following technical scheme: A preparation method of theaflavin self-assembled nano-particles comprises the following steps: (1) Preparing theaflavin, namely soaking tea tree branches and leaves in water, adding laccase, carrying out enzymatic oxidation reaction under the conditions of 30-50 ℃ and stirring, inactivating enzyme after the reaction is finished, cooling to room temperature, filtering and collecting filtrate, concentrating to obtain concentrated solution, extracting the concentrated solution with chloroform, collecting water phase, extracting with ethyl acetate, collecting ethyl acetate phase, finally removing ethyl acetate, and drying to obtain theaflavin; (2) And (3) preparing theaflavin self-assembled nano particles, namely dissolving the theaflavin and the L-cysteine obtained in the step (1) into water, then dropwise adding formaldehyde solution, continuously stirring at room temperature for reaction, centrifuging after the reaction is finished, collecting precipitate, washing and drying to obtain the theaflavin self-assembled nano particles. Preferably, the tea tree branches and leaves in the step (1) are tea tree pruning branches and leaves. Preferably, the water in steps (1) and (2) is deionized water. Preferably, the dosage ratio of tea tree branches and leaves to water in the step (1) is 1g (30-50) mL. Preferably, the dosage ratio of tea tree branches and leaves to laccase in the step (1) is 1g (250-750) U. Preferably, the stirring speed in the step (1) is 300-500 r/min. Preferably, the time of the enzymatic oxidation reaction in the step (1) is 6-10 hours. Preferably, the enzyme deactivation in the step (1) is medium-high temperature enzyme deactivation, and the condition is that the temperature is 90-100 ℃ and the time is 5-10 min. Preferably, the concentration in the step (1) is vacuum concentration, and more preferably, the concentration is vacuum concentration to the volume of 1/5-1/10 of the volume of the original reaction liquid at the temperature of 60-80 ℃. Preferably, the chloroform in the step (1) is used in an amount of 0.5 to 1.5 times the volume of the concentrated solution. Preferably, the number of times of extraction with chloroform in the step (1) is 2-