CN113288847A

CN113288847A - Preparation method and application of rhizoma typhonii fermentation extracting solution

Info

Publication number: CN113288847A
Application number: CN202110570354.XA
Authority: CN
Inventors: 蔡宝国; 荣绍丰; 管世敏; 李茜茜; 黄煜玲
Original assignee: Shanghai Institute of Technology
Current assignee: Shanghai Institute of Technology
Priority date: 2021-05-25
Filing date: 2021-05-25
Publication date: 2021-08-24

Abstract

The invention discloses a preparation method and application of a giant typhonium rhizome fermentation extracting solution. The preparation method comprises the following steps: inoculating the strain into a fermentation culture medium, culturing, filtering, and collecting filtrate to obtain fermentation extract; the fermentation medium comprises: 0.1-3 wt% of giant typhonium rhizome powder, 0.1-2 wt% of glucose and 0.1-1 wt% of yeast powder. The content of protein, amino acid, total sugar and polypeptide in the giant typhonium rhizome extracting solution prepared by the preparation method is higher than that of the giant typhonium rhizome extracting solution extracted by the water extraction method, and when the giant typhonium rhizome fermenting extracting solution prepared by the preparation method is used for preparing the skin care product, the hyaluronidase inhibition rate, the lipoxygenase inhibition rate and the tyrosinase inhibition rate of the obtained skin care product are obviously higher than those of the giant typhonium rhizome extracting solution prepared by the water extraction method.

Description

Preparation method and application of rhizoma typhonii fermentation extracting solution

Technical Field

The invention relates to a preparation method and application of a giant typhonium rhizome fermentation extracting solution, belonging to the technical field of cosmetics.

Background

Rhizoma Typhonii is underground tuber of Typhonium giganteum of genus Typhonium of family Araceae, and is collected in 2010 pharmacopoeia of China. In the traditional Chinese medicine beauty treatment prescription, the appearance frequency of rhizoma typhonii is higher and accounts for more than 20%. The main components of rhizoma typhonii are fatty acid, sterol compound, volatile oil component, polypeptide, amino acid and polysaccharide, etc., and have strong moisturizing, antioxidant and whitening effects.

Based on the increasing demand of natural cosmetic raw materials in the market, the current method for extracting the active ingredients from rhizoma typhonii is water extraction or organic solvent extraction. When the two methods are adopted to extract the rhizoma typhonii active substances, the raw materials are in a high-temperature state of more than 50 ℃ for a long time, the main active ingredients of the rhizoma typhonii are damaged, and meanwhile, a series of problems of high energy consumption, low extraction rate, large wastewater discharge amount, poor smell of an extracting solution, poor color stability and the like exist. The prior art urgently needs a method which has low energy consumption, high extraction rate, no pollution and stability and does not destroy the biological active ingredients of the rhizoma typhonii to meet the defects of the prior art.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: the existing method for extracting the active ingredients from the giant typhonium rhizome has the problems of high energy consumption, low extraction rate, large wastewater discharge, poor smell of extracting solution, poor color stability and the like.

In order to solve the technical problems, the invention provides a preparation method of a giant typhonium rhizome fermentation extracting solution, which comprises the following steps: inoculating lactobacillus buchneri in a fermentation culture medium for fermentation culture, filtering, and taking filtrate to obtain a fermentation extract; the fermentation medium comprises 0.1-3 wt% of giant typhonium rhizome powder, 0.1-2 wt% of glucose and 0.1-1 wt% of yeast powder.

Preferably, the lactobacillus buchneri is lactobacillus buchneri MF060, which is classified and named as: lactobacillus buchneri, named latin: lactobacillus buchneri, deposited in units of: the China general microbiological culture Collection center has the following preservation addresses: xilu No.1 Hospital No. 3, Beijing, Chaoyang, with a preservation date of: 12 months and 12 days in 2019, the preservation number is: CGMCC No. 19125.

Preferably, the method for activating lactobacillus buchneri comprises: inoculating the lactobacillus buchneri into a seed culture medium, and performing activated culture for 48h at room temperature, wherein the seed culture medium comprises the following components: 0.1-0.3 wt% of sodium chloride, 1-3 wt% of yeast powder, 3-5 wt% of glucose, 0.1-0.3 wt% of dipotassium hydrogen phosphate, 0.05-0.2 wt% of magnesium chloride and the balance of distilled water, wherein the pH value of the mixture is 6.2 +/-0.2.

Preferably, the process conditions of the fermentation culture are as follows: the inoculation amount is 0.5-3 wt%, the temperature is 28-32 ℃, and the time is 1-7 d.

Preferably, the filtration comprises a coarse filtration and a micro filtration performed in sequence.

More preferably, the rough filtration is performed by adopting a filter membrane with the aperture of 38-42 μm; the microfiltration adopts a filter membrane with the aperture of 0.2-0.24 mu m for filtration.

Preferably, the particle size of the giant typhonium rhizome powder is 60-100 meshes.

Preferably, the fermentation extract is preserved by adding a preservative.

More preferably, the preservative is hexanediol, and the added volume of the hexanediol is 50-200 times of the volume of the fermentation medium.

The invention also provides application of the giant typhonium rhizome fermented extract prepared by the preparation method of the giant typhonium rhizome fermented extract in preparing cosmetics.

Compared with the prior art, the invention has the beneficial effects that:

1. the protein, amino acid, sugar and polypeptide components contained in the giant typhonium rhizome fermentation extracting solution obtained by adopting lactobacillus buchneri fermentation are respectively 4.7-7.6 times, 3.2-5.8 times, 2.4-4.6 times and 8.7-15.18 times higher than those obtained by a water extraction method;

2. the invention adopts a filtering method for sterilization, and has the advantages of retaining the active ingredients in the giant typhonium rhizome and being closer to the skin micro-ecology compared with a high-temperature sterilization process;

3. compared with skin cream and skin lotion prepared from giant typhonium rhizome extract obtained by a water extraction method, the skin cream and the skin lotion prepared from giant typhonium rhizome extract are respectively improved in inhibition rate of hyaluronic acid, lipoxidase and tyrosinase by 1.9-2.4 times, 3.2-3.3 times and 3.0-3.3 times.

Detailed Description

In order to make the invention more comprehensible, preferred embodiments are described in detail below.

In the following examples, Lactobacillus buchneri is used as Lactobacillus buchneri MF060, which was classified and designated as: lactobacillus buchneri, named latin: lactobacillus buchneri, deposited in units of: the China general microbiological culture Collection center has the following preservation addresses: xilu No.1 Hospital No. 3, Beijing, Chaoyang, with a preservation date of: 12 months and 12 days in 2019, the preservation number is: CGMCC No. 19125.

Example 1

A method for preparing rhizoma Typhonii fermentation extract comprises the following steps:

step 1: preparing a fermentation medium from the following components in percentage by mass: 0.1 wt% of giant typhonium rhizome powder, 0.1 wt% of glucose and 0.1 wt% of yeast powder; wherein the rhizoma Typhonii powder is 60 mesh sieved powder.

Step 2: taking 0.2g of sodium chloride, 2g of yeast powder, 4g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium chloride and 100ml of distilled water, adjusting the pH value to 6.2 +/-0.2, sterilizing by high-pressure steam for 15min, cooling to room temperature, inoculating a preservation strain Lactobacillus buchneri CGMCC No.19125, and carrying out activated culture for 48 h.

And step 3: inoculating lactobacillus buchneri activated by the seed culture medium into the prepared fermentation culture medium according to 2% of the mass of the fermentation culture medium, and culturing at 30 ℃ for 1d to obtain a fermentation product.

And 4, step 4: filtering the fermentation product with a filter membrane with a pore size of 40 μm, collecting the filtrate, filtering with a filter membrane with a pore size of 0.22 μm, collecting the filtrate, and adding 200 times of hexanediol in the volume of the fermentation medium to obtain rhizoma Typhonii fermentation extract.

And detecting the polypeptide concentration, the amino acid concentration, the total sugar concentration and the protein concentration of the prepared rhizoma typhonii fermentation extracting solution. The results are shown in Table 1.

Wherein the method for measuring the concentration of the polypeptide is a Folin phenol method; the amino acid concentration is determined by QB/T2409-1998; the determination method of the total sugar concentration refers to SN/T4260-2015; the protein concentration was determined by reference to GB/T6432-2018.

Example 2

step 1: preparing a fermentation medium from the following components in percentage by mass: 1.5 wt% of giant typhonium rhizome powder, 1.2 wt% of glucose and 0.5 wt% of yeast powder; wherein the rhizoma Typhonii powder is sieved with 80 mesh sieve.

And step 3: inoculating lactobacillus buchneri activated by the seed culture medium into the prepared fermentation culture medium according to 0.5 percent of the mass of the fermentation culture medium, and culturing for 4d at 28 ℃ to obtain a fermentation product.

And 4, step 4: filtering the fermentation product with a filter membrane with the pore size of 38 μm, collecting the filtrate, filtering with a filter membrane with the pore size of 0.24 μm, collecting the filtrate, and adding 100 times of hexanediol in the volume of the fermentation medium to obtain rhizoma Typhonii fermentation extract.

Example 3

step 1: preparing a fermentation medium from the following components in percentage by mass: 3.0 wt% of giant typhonium rhizome powder, 2.0 wt% of glucose and 1.0 wt% of yeast powder; wherein the rhizoma Typhonii powder is powder sieved with 100 mesh sieve.

And step 3: inoculating lactobacillus buchneri activated by the seed culture medium into the prepared fermentation culture medium according to 3% of the mass of the fermentation culture medium, and culturing at 32 ℃ for 7d to obtain a fermentation product.

And 4, step 4: filtering the fermentation product with 42 μm filter membrane, collecting filtrate, filtering with 0.2 μm filter membrane, collecting filtrate, and adding 66.7 times of hexanediol to obtain rhizoma Typhonii fermentation extractive solution.

Comparative example 1

A method for preparing rhizoma Typhonii extractive solution comprises:

taking the giant typhonium rhizome powder which passes through a 100-mesh sieve, wherein the mass ratio of the giant typhonium rhizome powder to the water is 3 g: 100mL, extracting for 3h at 50 ℃ under reflux, and filtering with a 0.22 μm filter membrane after extraction to obtain rhizoma Typhonii extract.

And detecting the polypeptide concentration, the amino acid concentration, the total sugar concentration and the protein concentration in the prepared rhizoma typhonii extracting solution. The results are shown in Table 1.

TABLE 1 concentrations of active ingredients in extract solutions of rhizoma Typhonii of examples 1-3 and comparative example 1

Sample (I)	Protein (g/L)	Amino acid (g/L)	Total sugar (g/L)	Polypeptide (g/L)
					Example 1	0.43	2.43	2.74	0.96
Example 2	0.69	3.88	3.92	1.67
					Example 3	0.46	4.32	5.155	1.44
Comparative example 1	0.09	0.75	1.12	0.11

As is clear from Table 1, the extract solutions from the fermentation of rhizoma Typhonii of examples 1-3 all contained significantly higher concentrations of protein, amino acids, total sugars and polypeptides than the extract solution from rhizoma Typhonii of comparative example 1. Wherein the concentration of the protein in examples 1-3 is 4.7-7.6 times higher than that in comparative example 1; the concentration of the amino acid is 3.2 to 5.8 times higher than that of the amino acid in the comparative example 1; the concentration of total sugar is 2.4-4.6 times higher than that of the comparative example 1; the concentration of the polypeptide is 8.7-15.18 times higher than that of the polypeptide in comparative example 1.

Application example 1

The fermented extract of rhizoma typhonii obtained in example 1 was prepared into skin lotion according to the formulation shown in table 2.

TABLE 2 skin lotion formulation (content is mass fraction)

The preparation method comprises the following steps: adding water, glycerol, butanediol, water-soluble vitamin E, resveratrol, sodium citrate and hyaluronic acid into a water phase pot, heating to 80 ℃, preserving heat, stirring and dissolving uniformly; cooling to 40 ℃, adding the rhizoma typhonii fermented extract obtained in the example 1, and uniformly stirring to obtain the skin lotion.

The inhibition rates of the skin moisturizer prepared above on hyaluronidase, lipoxygenase and tyrosinase were determined, and the results are shown in table 4.

Application example 2

The extract of giant typhonium rhizome obtained in example 2 was prepared into skin cream according to the formulation shown in table 3.

TABLE 3 skin cream formulations

Composition (I)	Content (wt.)
		Monoglyceride	2wt％
Spermaceti stearic acid	2wt％
		Plant squalane	2.4wt％
Olive oil	3wt％
		Shea butter	2.8wt％
Plant sterol	1wt％
		Rhizoma typhonii fermented extract	16.8wt％
Phenoxyethanol	0.5wt％
		Water (W)	70wt％

The preparation method comprises the following steps: adding monoglyceride, cetostearyl alcohol, plant squalane, olive oil, shea butter and phytosterol into an oil phase pot, heating to 80 ℃, adding water, rhizoma typhonii fermentation extract and phenoxyethanol into a water phase pot, heating to 80 ℃, keeping the constant temperature for 30min, simultaneously pumping the raw materials in the oil phase pot and the water phase pot into a main emulsifying pot for emulsification, stirring at 25rpm, homogenizing at 3600 rpm for 6min, cooling to 40 ℃, uniformly stirring and mixing, discharging at the temperature lower than 30 ℃, and thus obtaining the skin cream.

And detecting the inhibition rate of the prepared skin cream on hyaluronidase, lipoxygenase and tyrosinase. The results are shown in Table 4.

Comparative example 2

The giant typhonium rhizome extract extracted in the comparative example 1 is prepared into the skin lotion according to the formula and the preparation method of the application example 1.

And detecting the inhibition rate of the prepared skin care lotion on hyaluronidase, lipoxygenase and tyrosinase. The results are shown in Table 4.

TABLE 4 inhibition of hyaluronidase, lipoxygenase and tyrosinase by the skin care products of examples 1-2 and comparative example 2

Sample (I)	Hyaluronidase inhibition rate	Lipoxygenase inhibition rate	Tyrosinase inhibition rate
				Application example 1	21.76％	27.61％	30.21％
Application example 2	26.53％	27.12％	33.32％
				Comparative example 2	11.12％	8.35％	10.28％

As can be seen from table 4, the skin care products obtained in application examples 1 to 2 all have higher inhibition rates for hyaluronidase, lipoxygenase and tyrosinase than the skin care product obtained in comparative example 2. Compared with the skin care lotion of comparative example 2, the skin care products obtained by applying the examples 1-2 have the advantages that the inhibition rates of hyaluronic acid, lipoxygenase and tyrosinase are respectively improved by 1.9-2.4 times, 3.2-3.3 times and 3.0-3.3 times. The different active ingredients of the giant typhonium rhizome extract obtained by different preparation methods are different, so that the corresponding effect of the giant typhonium rhizome skin care product can be influenced.

The above-described embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention in any way and substantially, it should be noted that those skilled in the art may make several modifications and additions without departing from the scope of the present invention, which should also be construed as a protection scope of the present invention.

Claims

1. The preparation method of the giant typhonium rhizome fermentation extracting solution is characterized by comprising the following steps of: inoculating the activated lactobacillus buchneri into a fermentation culture medium for fermentation culture, filtering, and taking filtrate to obtain a fermentation extract; the fermentation medium comprises 0.1-3 wt% of giant typhonium rhizome powder, 0.1-2 wt% of glucose and 0.1-1 wt% of yeast powder.

2. The method for preparing rhizoma typhonii fermented extract as claimed in claim 1, wherein said lactobacillus buchneri is lactobacillus buchneri MF060, which is classified and named as: lactobacillus buchneri, named latin: lactobacillus buchneri, deposited in units of: the China general microbiological culture Collection center has the following preservation addresses: xilu No.1 Hospital No. 3, Beijing, Chaoyang, with a preservation date of: 12 months and 12 days in 2019, the preservation number is: CGMCC No. 19125.

3. The method for preparing rhizoma typhonii fermented extract liquid according to claim 1, wherein the method for activating lactobacillus buchneri comprises: inoculating the lactobacillus buchneri into a seed culture medium, and performing activated culture for 48h at room temperature, wherein the seed culture medium comprises the following components: 0.1-0.3 wt% of sodium chloride, 1-3 wt% of yeast powder, 3-5 wt% of glucose, 0.1-0.3 wt% of dipotassium hydrogen phosphate, 0.05-0.2 wt% of magnesium chloride and the balance of distilled water, wherein the pH value of the mixture is 6.2 +/-0.2.

4. The method for preparing rhizoma typhonii fermentation extract according to claim 1, wherein the fermentation culture process conditions are as follows: the inoculation amount is 0.5-3 wt%, the temperature is 28-32 ℃, and the time is 1-7 d.

5. The method according to claim 1, wherein the filtering comprises straining and micro-filtering sequentially.

6. The method for preparing rhizoma typhonii fermentation extract according to claim 5, wherein the rough filtration is performed by a filter membrane with a pore size of 38-42 μm; the microfiltration adopts a filter membrane with the aperture of 0.2-0.24 mu m for filtration.

7. The method for preparing rhizoma typhonii fermented extract according to claim 1, wherein the particle size of the rhizoma typhonii powder is 60-100 mesh.

8. The method for preparing rhizoma typhonii fermented extract as claimed in claim 1, wherein the fermented extract is preserved by adding a preservative.

9. The method for preparing rhizoma typhonii fermentation extract as claimed in claim 8, wherein the preservative is hexylene glycol, and the volume of the hexylene glycol added is 50-200 times of the volume of the fermentation medium.

10. Use of the fermented extract of giant typhonium rhizome prepared by the method for preparing the fermented extract of giant typhonium rhizome according to any one of claims 1 to 9 in the preparation of cosmetics.