阅读说明：本技术 一种滋养头皮、防脱发的弗朗鼠李皮发酵液及其制备方法 (Francisella fragrans bark fermentation liquor capable of nourishing scalp and preventing hair loss and preparation method thereof ) 是由 郭智华 徐双华 史姣姣 于 2020-12-28 设计创作，主要内容包括：本发明涉及一种发酵液,具体涉及一种滋养头皮、防脱发的弗朗鼠李皮发酵液及其制备方法。发酵液的制备方法,包括以类植物乳杆菌GUFHSL-70(CCTCC M 2016002)对弗朗鼠李皮进行发酵。研究发现,经类植物乳杆菌GUFHSL-70(CCTCC M 2016002)发酵后,弗朗鼠李皮原材料中原有的活性成分、发酵产生的次级代谢产物以及类植物乳杆菌GUFHSL-70的溶胞物等协同作用,可以更好地滋养头皮、防脱发。(The invention relates to fermentation liquor, in particular to frangula rhamnus bark fermentation liquor for nourishing scalp and preventing alopecia and a preparation method thereof. The preparation method of the fermentation liquor comprises the step of fermenting frangula rhamnus with lactobacillus plantarum GUFHSL-70(CCTCC M2016002). Researches show that after fermentation of lactobacillus plantarum GUFHSL-70(CCTCC M2016002), the synergistic effects of the original active ingredients in the Franza rhamnus raw material, secondary metabolites generated by fermentation, lysate of the lactobacillus plantarum GUFHSL-70 and the like can better nourish the scalp and prevent alopecia.)

1. A preparation method of Franza rhamnoides fermentation liquor for nourishing scalp and preventing hair loss is characterized by comprising the step of fermenting Franza rhamnoides with Lactobacillus plantarum GUFHSL-70(CCTCC M2016002).

2. The method according to claim 1, wherein the culture medium used is MRS medium.

3. The preparation method according to claim 2, wherein the raw material for fermentation comprises frangula rhamnus bark and MRS medium at a weight ratio of 5-20:60-95, optionally 5-20: 85.

4. The preparation method as claimed in any one of claims 1 to 3, wherein Franza rhamnoides is used as pulverized to 80-100 mesh.

5. The process according to any one of claims 1 to 3, wherein the raw material for fermentation is directly added with freeze-dried powder of Lactobacillus plantarum GUFHSL-70 for fermentation; or activating Lactobacillus plantarum GUFHSL-70 to obtain seed solution, and inoculating into the raw materials for fermentation.

6. The method according to any one of claims 1 to 3, wherein the seed liquid of Lactobacillus plantarum GUFHSL-70 is inoculated into the raw material for fermentation; the weight ratio of the seed liquid to the Francisella Wilsonii to the MRS culture medium is 2-15:5-20: 60-95; wherein the effective viable count in the seed liquid is (1.0-5.0) x 10⁸CFU/mL, optionally 3.0X 10⁸CFU/mL。

7. The method of any one of claims 1 to 6, wherein the fermentation temperature is 34 to 39 ℃, optionally 37 ℃.

8. Franzenbuckthorn bark fermentation broth for nourishing scalp and preventing alopecia, which is prepared by the method of any one of claims 1-7; optionally, the frangula fermentation broth is a supernatant obtained after fermentation.

9. A fermented product which is a cell-free sterile liquid, or a sterile filtrate of the liquid, or a sterile supernatant, or a concentrate or a dried product thereof, further prepared from the scalp nourishing, hair loss preventing frangula rhamna cortex fermentation broth prepared by the method of any one of claims 1-7.

10. A hair product comprising the frangula rhamna fermentation broth of claim 8 for nourishing the scalp, preventing hair loss, or the fermentation product of claim 9; optionally, the hair product further comprises adjuvants useful in the cosmetic field.

Technical Field

The invention relates to fermentation liquor, in particular to frangula rhamnus bark fermentation liquor for nourishing scalp and preventing alopecia and a preparation method thereof.

Background

Hair is an important indicator of human health. It is desired to provide a hair product having excellent effects of preventing hair loss, nourishing hair, promoting hair growth, etc. At present, anti-dropping products added with chemical components, such as the traditional medicine minoxidil and the like, are available on the market, and if the products are improperly used, certain harm can be caused to the human body. Currently, alternative techniques have been developed for using Chinese herbal medicines as a therapeutic drug. Some traditional processing techniques and using methods may damage the effective components in the raw materials and fail to achieve the expected efficacy.

Disclosure of Invention

The invention provides a fermentation broth, in particular to a Franzenhamus skin fermentation broth for nourishing scalp and preventing alopecia, which can be used as a hair product.

The invention firstly provides a preparation method of frangula rhamnus fermentation broth for nourishing scalp and preventing alopecia, which comprises the step of fermenting frangula rhamnus with lactobacillus plantarum GUFHSL-70(CCTCC M2016002).

The Lactobacillus plantarum GUFHSL-70 is named as Lactobacillus parapralatarum GUFHSL-70, and the pure culture of the strain is preserved in China center for type culture Collection in 2016, 01 and 04 days, wherein the preservation unit address is as follows: china, Wuhan and Wuhan university, the preservation number is: CCTCC M2016002. The lactobacillus plantarum GUFHSL-70(CCTCC M2016002) mentioned in the invention is disclosed in Chinese patent publication No. CN 105695363A.

Franza Rhamnus is dried bark or branch bark of Rhamnus Frangula L (Frangula alnus Miller) belonging to Rhamnaceae. Francinna bark is rich in multiple active ingredients, mainly including gluco-rhamnetin and anthraquinone compounds. Most of the effective components in the frangula rhamnus are in a compound form, so that the components are very unstable and easy to decompose, the raw materials are expensive, and the waste of the raw materials is caused when the components exist in the compound form or are subjected to ineffective decomposition, so that the actual utilization rate of the frangula rhamnus is not high.

The inventor finds that the frangula rhamnus is fermented by lactobacillus plantarum GUFHSL-70(CCTCC M2016002), so that the main active ingredients of the frangula rhamnus raw material can be fully reserved, and a plurality of beneficial ingredients in the frangula rhamnus raw material can be dissolved in fermentation liquor after being hydrolyzed; further decomposing macromolecular active ingredients into small molecules which are more beneficial to the absorption of a human body through the fermentation effect of the lactobacillus plantarum GUFHSL-70; in addition, substances which are not contained or are contained in the frangula rhamna, such as a plurality of secondary metabolites, are also produced through fermentation. Researches show that after fermentation of lactobacillus plantarum GUFHSL-70(CCTCC M2016002), the synergistic effects of the original active ingredients in the Franza rhamnus raw material, secondary metabolites generated by fermentation, lysate of the lactobacillus plantarum GUFHSL-70 and the like can better nourish the scalp and prevent alopecia.

The inventor also finds that after the Franza rhamnoides is fermented by the Lactobacillus plantarum GUFHSL-70(CCTCC M2016002), various beneficial components are hydrolyzed and dissolved in fermentation liquor, and various components can be more stable and efficient after the action of microorganisms.

In some embodiments, the culture medium used in the preparation method of the frangula rhamnus fermentation broth for nourishing scalp and preventing alopecia is MRS culture medium.

Specifically, MRS medium: 10.0g/L of peptone, 10.0g/L of beef extract, 5.0g/L of yeast powder, 20.0g/L of glucose, 0.1g/L of magnesium sulfate, 5.0g/L of sodium acetate, 2.0g/L of ammonium citrate, 2.0g/L of dipotassium hydrogen phosphate, 0.05g/L of manganese sulfate and Tween-801.0 g/L; water make up to 1L.

In some embodiments, the raw material for fermentation comprises frangula rhamnus bark and MRS culture medium at a weight ratio of 5-20:60-95, optionally 5-20:85, such as 5: 85, 10: 85, 20: 85.

In some embodiments, frangula rhamnoides is pulverized and added to a culture medium for fermentation. Generally, it is pulverized to 80-100 mesh. The method has the advantages that the method is more favorable for releasing active ingredients of the frangula rhamnus, increases the contact area with microorganisms, and improves the bioavailability of raw materials.

In some embodiments, the freeze-dried powder of the lactobacillus plantarum GUFHSL-70 can be directly put into raw materials for fermentation to be fermented; or activating Lactobacillus plantarum GUFHSL-70 to obtain seed solution, and inoculating into raw materials for fermentation.

The freeze-dried powder of the lactobacillus plantarum GUFHSL-70 can be prepared by adopting a method commonly used in the field.

Seed liquid of the Lactobacillus plantarum GUFHSL-70 can be prepared by methods commonly used in the art. For example, the specific method comprises: preparing MRS liquid culture solution, sterilizing at high temperature under high pressure, inoculating the glycerol tube strain to MRS liquid culture medium, and sealing for culturing (37 deg.C, 48 hr). Thus obtaining the seed liquid.

In some embodiments, the seed liquid of the lactobacillus plantarum GUFHSL-70 is inoculated into a raw material used for fermentation to perform fermentation; the weight ratio of the seed liquid to the Francisella Wilsonii and the MRS culture medium is 2-15:5-20: 60-95; wherein the effective viable count in the seed liquid is (1.0-5.0) x 10⁸CFU/mL, e.g. 3.0X 10⁸CFU/mL。

In some embodiments, the frangula rhamnoides (for example, 80-100 meshes) is mixed with MRS liquid culture medium, sterilized, and inoculated with seed liquid of Lactobacillus plantarum GUFHSL-70 for fermentation.

The sterilization can be carried out by a method commonly used in the art, such as autoclaving at 126 ℃ for 15 min.

In some embodiments, the temperature of the fermentation is 34-39 ℃, e.g., 37 ℃.

In some embodiments, fermentation is carried out for 48-72 hours, followed by aeration, stirring (or shaking), sealing, and fermentation for 48-96 hours.

In some embodiments, the fermentation is conducted at a relative humidity of 33% to 60%, for example, a relative humidity of 50%.

In some embodiments, the supernatant obtained after fermentation is the frangula rhamnus bark fermentation broth for nourishing scalp and preventing alopecia according to the present invention. For convenience of storage and subsequent use, the supernatant obtained after fermentation may be further sterilized, for example, by autoclaving at 126 deg.C for 15 min.

In some embodiments, the fermentation is followed by filtration to obtain a clear filtrate, which is then sterilized (126 ℃ C., 15min) and filtered to obtain the desired fermentation broth.

The invention also discloses Francisella fragrans bark fermentation liquor which is prepared by the preparation method and is used for nourishing scalp and preventing alopecia.

The invention also provides a fermentation product, which is a cell-free sterile liquid, or a liquid sterile filtrate, or a sterile supernatant, or a concentrate or a dry product thereof, further prepared from the frangula buckthorn skin fermentation broth for nourishing the scalp and preventing alopecia, prepared by the method.

Typically, the fermentation broth may be concentrated or dried (e.g., lyophilized or spray dried) using methods conventional in the art.

The invention also provides application of the compound fermentation liquor or fermentation product for nourishing scalp and preventing alopecia, which is prepared by the method, in preparation of hair products, in particular application in preparation of hair products for nourishing scalp and preventing alopecia.

The invention also provides a hair product, which comprises the Francisella planckii fermentation liquor or the fermentation product prepared by the method for nourishing scalp and preventing alopecia. Further, the hair product also includes raw and auxiliary materials usable in the art (cosmetic field).

The experimental result shows that the fermentation liquor can remarkably promote the hair to enter the growth period, reduce the generation of the hair in the resting period, improve the hair density, and has the beneficial effects of nourishing the scalp and preventing alopecia.

Detailed Description

The present invention will be described in detail with reference to examples.

The seed solution of Lactobacillus plantarum GUFHSL-70(CCTCC M2016002) was prepared according to the method disclosed in CN105695363A, and the viable count in the seed solution was 3.0X 10⁸CFU/mL or so.

Francinna buckthorn bark used below was ground to 80 mesh.

The parts indicated below are parts by weight.

MRS medium used was as follows: 10.0g/L of peptone, 10.0g/L of beef extract, 5.0g/L of yeast powder, 20.0g/L of glucose, 0.1g/L of magnesium sulfate, 5.0g/L of sodium acetate, 2.0g/L of ammonium citrate, 2.0g/L of dipotassium hydrogen phosphate, 0.05g/L of manganese sulfate, 0.8978 g/L of tween-801.0 and the balance of water to 1L.

Example 1

A preparation method of Francisella planch skin fermentation liquor for nourishing scalp and preventing alopecia comprises the following raw materials:

frangula rhamna 10 parts

10 parts of lactobacillus plantarum GUFHSL-70 seed liquid

85 portions of MRS culture medium

The preparation method comprises the following steps:

crushing frangula rhamnoides and adding the crushed frangula rhamnoides into an MRS culture medium to be uniformly stirred; sterilizing at 126 deg.C for 15min under high temperature and high pressure; inoculating lactobacillus plantarum GUFHSL-70 seed liquid, and fermenting for 72h at 37 ℃ and 50% humidity; then deflating and stirring for 10min, sealing and continuing fermentation for 48 h. Filtering after fermentation to obtain clear filtrate, sterilizing (126 deg.C, 15min), and filtering to obtain desired fermentation liquid.

Example 2

A preparation method of Francisella planch skin fermentation liquor for nourishing scalp and preventing alopecia comprises the following raw materials:

20 portions of frangula rhamna

10 parts of lactobacillus plantarum GUFHSL-70 seed liquid

85 portions of MRS culture medium

The specific preparation method is as in example 1.

Example 3

A preparation method of Francisella planch skin fermentation liquor for nourishing scalp and preventing alopecia comprises the following raw materials:

frangula rhamna bark 5 parts

10 parts of lactobacillus plantarum GUFHSL-70 seed liquid

85 portions of MRS culture medium

The specific preparation method is as in example 1.

Example 4

A preparation method of Francisella planch skin fermentation liquor for nourishing scalp and preventing alopecia comprises the following raw materials:

frangula rhamna 10 parts

15 parts of lactobacillus plantarum GUFHSL-70 seed liquid

85 portions of MRS culture medium

The specific preparation method is as in example 1.

Comparative example 1

A preparation method of fermentation liquor comprises the following raw materials:

frangula rhamna 10 parts

10 portions of yeast seed liquid

85 portions of MRS culture medium

The preparation method comprises the following steps:

the preparation method of the yeast seed liquid comprises the following steps: preparing a PDA liquid culture medium (6 g/L of potato powder, 20g/L of glucose, 0.1g/L of chloramphenicol, and constant volume of sterile water to 1L). Sterilizing PDA liquid culture medium at 126 deg.C for 15min under high temperature and high pressure, cooling to room temperature, adding yeast powder 0.1%, and culturing at 28 deg.C for 5 days to obtain yeast seed solution.

Crushing frangula rhamnus peel, adding into an MRs liquid culture medium, and uniformly stirring; sterilizing at 126 deg.C for 15min under high temperature and high pressure; inoculating yeast seed liquid, and fermenting at 28 deg.C and 50% humidity for 72 hr; then deflating and stirring for 10min, sealing and continuing fermentation for 48 h. Filtering after fermentation to obtain clear filtrate, sterilizing (126 deg.C, 15min), and filtering to obtain desired fermentation liquid.

Comparative example 2

A preparation method of fermentation liquor comprises the following raw materials:

frangula rhamna 10 parts

Lactic acid bacteria seed liquid 10 parts

85 portions of MRS culture medium

The preparation method comprises the following steps: the preparation method of the lactobacillus seed liquid comprises the following steps: preparing MRS liquid culture medium, sterilizing at 126 deg.C for 15min under high temperature and high pressure, cooling to room temperature, adding 0.01% of commercially available lactobacillus powder, and culturing at 37 deg.C for 48 hr to obtain lactobacillus seed solution.

Crushing frangula rhamnus, adding into a newly prepared MRS liquid culture medium, and uniformly stirring; sterilizing at 126 deg.C for 15min under high temperature and high pressure; inoculating lactobacillus seed solution, and fermenting at 37 deg.C and 50% humidity for 72 hr; then deflating and stirring for 10min, sealing and continuing fermentation for 48 h. Filtering after fermentation to obtain clear filtrate, sterilizing (126 deg.C, 15min), and filtering to obtain desired fermentation liquid.

Comparative example 3

A method for preparing a fermentation broth is different from that of example 1 only in that Franza Rhamnus bark is replaced with an equal amount of Yunnan Rhamnus (Latin's name: Rhamnus aureus Heppl.) bark (pulverized to 80 mesh).

Comparative example 4

A mixed solution is prepared from 10 parts of frangula rhamnus and 85 parts of MRS culture medium; the preparation method comprises the following steps: adding frangula rhamnoides into an MRS culture medium and uniformly stirring; sterilizing at 126 deg.C for 15min under high temperature and high pressure, and standing at 37 deg.C and 50% humidity for 72 h; then stirring for 10min, sealing and then continuously standing for 48 h. Filtering to obtain clear filtrate, sterilizing (126 deg.C, 15min), and filtering to obtain the desired mixture.

Comparative example 5

A fermentation liquor is prepared by fermenting 10 parts of Lactobacillus plantarum GUFHSL-70 seed liquid and 85 parts of MRS culture medium; the preparation method comprises the following steps: adding the seed liquid of the Lactobacillus plantarum GUFHSL-70 into MRS liquid culture medium sterilized at 126 ℃ for 15min, and fermenting for 72h at 37 ℃ and 50% humidity; then deflating and stirring for 10mi, sealing and continuing to ferment for 48 h. Fermenting, filtering to obtain clear filtrate, sterilizing (126 deg.C, 15min), and filtering to obtain desired fermentation liquid.

Examples of the experiments

Experimental samples: the fermentation liquids prepared in examples 1 to 4, the fermentation liquids prepared in comparative examples 1 to 3 and 5, and the mixed liquid prepared in comparative example 4.

The experimental method comprises the following steps: referring to the laboratory procedure (12 week hair growth efficacy test protocol for hair product), 34 subjects with hair loss levels of grade 2 to 4 according to the AlviArmani type were selected, aged 24-50 years, selected on the head, shaved off the hair, and applied daily to the test samples. Subjects tested the growth phase hair ratio, resting phase hair ratio, hair density, respectively, before, after 4 weeks of continuous use, 8 weeks of continuous use, and 12 weeks of continuous use of the test product. The hair density is the result of examination of a round scalp test area having a diameter of 1.7 cm.

The results of the experiments are shown in tables 1-3 below:

note: in tables 1 to 3, P is 0.01, and the difference was very significant compared to the initial value. P ≦ 0.05, with a significant difference compared to the initial value. The unlabeled representation is not significantly different.

From the above experimental data: the effect of the fermentation liquid in the examples 1 to 4 is obviously better than that of the fermentation liquid in the comparative examples 1 to 5, so that the anti-hair loss effect of the obtained fermentation liquid is most obvious after the frangula and lactobacillus plantarum are mixed and fermented.

Example 1 the best effect of example 1 compared to examples 2 and 3 and the little difference in effect compared to example 4 demonstrate that the best effect of the mixed fermentation product of the present invention can be ensured and the unnecessary waste of raw materials can be effectively controlled by fermenting the mixed fermentation product according to a certain proportion.

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.