CN117771141A - Cactus fermentation filtrate with free radical scavenging effect - Google Patents
Cactus fermentation filtrate with free radical scavenging effect Download PDFInfo
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Abstract
The invention relates to a cactus fermentation filtrate with a free radical removal effect, which belongs to the technical field of application biology, and comprises the following raw materials: compound microorganism, auxiliary materials and cactus; the compound microorganism comprises saccharomycetes, aspergillus oryzae and mucor circinelloides; the auxiliary materials comprise rice and deionized water. The invention combines the cactus, the rice and the compound microorganism, and the finished product of the cactus fermentation filtrate has excellent DPPH free radical elimination effect. The saccharomycete, aspergillus oryzae and mucor circinelloides are combined, so that the fermentation of the cactus can be fully and completely achieved in a synergistic way, and the elimination rate of the finished product of the fermentation filtrate of the cactus on DPPH free radicals is further improved. The finished product of the fermented cactus filtrate prepared by the invention has no skin irritation, allergy and acute oral toxicity, and can be safely used for skin care products.
Description
Technical Field
The invention belongs to the technical field of applied biology, and particularly relates to a cactus fermentation filtrate with a free radical removal effect.
Background
Cactus stems (opuntia) are a generic term for a class of Cactaceae plants of the order Cactales, the varieties of which are widely distributed throughout the world. According to statistics, nearly hundred different cactus stem varieties are shared worldwide, and a considerable part of varieties have unique edible and medicinal values. The Chinese traditional medicine has been focused on the medicinal value of the cactus stems for a long time, and the medical value of the cactus stems is mentioned in the Chao gang mu Shi Yi of a Qing Dynasty doctor Zhao Xuemin, the cactus stems are light in taste and cold in nature, enter heart, lung and stomach meridians, and can promote qi and activate blood, clear heat and detoxify, reduce swelling and relieve pain, invigorate spleen and stop diarrhea, tranquillize mind and induce diuresis. The cactus stem is widely applied to the cosmetic industry besides being applied to the food and medicine industries, and the natural juice, the dry powder and the extract thereof have various effects of relieving, scavenging free radicals, preserving moisture, resisting bacteria, protecting hair and the like in cosmetics, and are good natural substances.
Although the cactus and its extract are applied in cosmetics industry, most of the active substances in the stems of cactus are extracted by physical or chemical methods such as solvent extraction, ultrasonic or microwave-assisted extraction, physical squeezing, etc., the application of biotechnology as an extraction process is rarely seen, i.e. the application of biotechnology as an extraction process for cactus is blank in the field of extracting the active substances of cactus, and even if the technology exists in the market, the effect of eliminating DPPH free radicals in the fermentation filtrate of the cactus extracted by the technology is extremely poor and is difficult to meet the application requirement.
Disclosure of Invention
The invention aims to provide the cactus fermentation filtrate with the function of removing free radicals, which combines the cactus, the rice and the compound microorganism, and the finished product of the cactus fermentation filtrate has excellent DPPH free radical removing effect. The compound combination of the saccharomycetes, the aspergillus oryzae and the mucor circinelloides can synergistically lead the fermentation of the cactus to be more complete, thereby further improving the elimination rate of the finished product of the fermentation filtrate of the cactus on DPPH free radicals and solving the problem of poor elimination rate of the DPPH free radicals in the existing technology of the fermentation filtrate of the cactus.
The aim of the invention can be achieved by the following technical scheme:
a cactus fermentation filtrate with free radical scavenging effect, the cactus fermentation filtrate comprises the following raw materials: compound microorganism, auxiliary materials and cactus.
The compound microorganism comprises saccharomycetes, aspergillus oryzae and mucor circinelloides; the auxiliary materials comprise rice and deionized water.
As a preferable technical scheme of the invention, the cactus fermentation filtrate comprises the following raw materials in percentage by mass:
yeast, which is Saccharomyces cerevisiae or other yeasts traditionally used in food production. Aspergillus oryzae and Mucor circinelloides are non-toxic and harmless microorganisms which are conventionally used in food production and can degrade starch into glucose for fermentation growth of yeast.
As a preferable technical scheme of the invention, the mass percentages of the saccharomycetes, the aspergillus oryzae and the mucor circinelloides are all 4-6%.
More preferably, the mass percentage of the saccharomycetes is 5%, the mass percentage of the aspergillus oryzae is 4%, and the mass percentage of the mucor circinelloides is 6%.
A method for preparing a cactus fermentation filtrate with a free radical scavenging effect, the method comprising the following steps:
s10, respectively carrying out pretreatment on the compound microorganism and the cactus to obtain a pretreated compound microorganism and a pretreated cactus;
s20, uniformly mixing rice and deionized water with the mass percentage of 10%, controlling temperature and pressure, cooling to below 30 ℃ after the treatment is completed, obtaining treated rice materials, uniformly inoculating the treated rice materials into pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides according to mass, performing first temperature control fermentation, uniformly mixing the fermented pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides, inoculating the mixture into pretreatment cactus, adding the balance deionized water, and performing second temperature control fermentation to obtain intermediate cactus fermentation liquid;
s30, filtering the middle cactus fermentation liquid, removing large-particle impurities in the tubular centrifugal core, and performing sterile filtration to obtain a finished product of the cactus fermentation filtrate with the effect of removing free radicals.
As a preferred technical solution of the present invention, the step S10 specifically includes the following steps:
s11, pretreatment of compound microorganisms:
respectively picking saccharomycetes, aspergillus oryzae and mucor circinelloides from the composite microorganism flat plate in a sterile mode, respectively inoculating the saccharomycetes, aspergillus oryzae and mucor circinelloides into a sterile YEPD culture medium, and culturing in a temperature-controlled shake flask to obtain pretreated saccharomycetes, pretreated aspergillus oryzae and pretreated mucor circinelloides;
s12, pretreatment of cactus: cleaning radix et caulis Opuntiae Dillenii, pulverizing, heating at controlled temperature, and cooling below 30deg.C to obtain pretreated radix et caulis Opuntiae Dillenii.
The heating treatment of the cactus is equivalent to the heating disinfection of the cactus, eliminates the possible microorganism carried by the cactus, and is convenient for fermenting the saccharomycete, the aspergillus oryzae and the mucor circinelloides.
As a preferred embodiment of the present invention, the components of the YEPD medium in step S11 include: yeast powder, peptone, glucose and distilled water.
As a preferable technical scheme of the invention, the mass ratio of the yeast powder to the peptone to the glucose to the distilled water is 1:2:2:95.
As a preferable technical scheme of the invention, the temperature of the temperature-controlled shake flask culture in the step S11 is 28-30 ℃ and the time is 16-30 h.
As a preferable technical scheme of the invention, the temperature of the temperature-controlled heating treatment in the step S12 is 70-80 ℃ and the time is 2-3 h.
As a preferable technical scheme of the invention, the temperature and pressure control treatment in the step S20 is carried out at 120-122 ℃, the pressure is 0.11Mpa, and the treatment time is 20min; the temperature of the first temperature-controlled fermentation is 28-30 ℃ and the time is 3-4 days; the temperature of the second temperature-controlled fermentation is 28-30 ℃ and the time is 5-6 days.
As a preferable technical scheme of the invention, the filtering in the step S30 is 40-200 mesh coarse filtering; the rotation speed of the tube type centrifugation is 8000-12000 rpm; the sterile-filtered microporous membrane was 0.22 μm.
The invention has the beneficial effects that:
(1) The microbial fermentation is used as a biological extraction technology, the decomposition of enzymes in the microorganism is utilized under proper conditions to convert the cactus into a product which is easier to be absorbed and utilized by human skin through a specific metabolic pathway, and the separation of active components of the cactus is promoted in the fermentation process so as to generate new efficacy or enhance the original efficacy. On one hand, the extraction of the cactus is enhanced by a fermentation technology, on the other hand, more active substances are generated, the utilization rate of raw materials is greatly improved, and a new way is added for the development and utilization of cactus resources.
(2) The invention combines the cactus, the rice and the compound microorganism to improve the elimination rate of the finished product of the cactus fermentation filtrate on DPPH free radicals; providing carbon source nutrient substances for rice to assist the growth of compound microorganisms so as to effectively ferment the cactus; the compound microorganism plays a main role in fermentation, a feedback system is further used for generating a large amount of nutrient substances, and meanwhile, the elimination rate of the finished product of the cactus fermentation filtrate on DPPH free radicals is further assisted; the three are combined, so that the finished product of the fermented filter liquor of the cactus has excellent DPPH free radical elimination effect.
(3) The invention combines the saccharomycetes, the aspergillus oryzae and the mucor circinelloides can effectively act on rice, and carbon sources such as sugar and the like for the growth of the saccharomycetes are supplied, so that the saccharomycetes are greatly propagated; on the other hand, aspergillus oryzae and mucor circinelloides have a large amount of enzymes capable of hydrolyzing plant cell walls such as cactus, so that nutrients in cactus cells can be released sufficiently, and yeast can ferment the cactus sufficiently; therefore, the combination of the saccharomycetes, the aspergillus oryzae and the mucor circinelloides can synergistically lead the fermentation of the cactus to be more complete, thereby further improving the elimination rate of the finished product of the fermentation filtrate of the cactus on DPPH free radicals.
(4) The finished product of the cactus fermentation filtrate has pH acidity after fermentation, is not easy to pollute microorganisms, has physical and chemical indexes within the quality specification requirements, and can be stored for 1 year at normal temperature.
(5) The finished product of the fermented cactus filtrate prepared by the invention has no skin irritation, allergy and acute oral toxicity, and can be safely used for skin care products.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The saccharomycetes, aspergillus oryzae and mucor circinelloides adopted by the invention are purchased from China institute of microbiology; the rice and the cactus are both commercially available, the cactus is 11 months and 12 days in 2012, and the 19 th file issued by the Ministry of health is approved as an ordinary food, namely an Opuntia ficus-indica (Linn.) Mill, mibangta variety; the details are not described in detail later.
Example 1
A cactus fermentation filtrate with free radical scavenging effect, the cactus fermentation filtrate comprises the following raw materials: compound microorganism, auxiliary materials and cactus;
the compound microorganism comprises saccharomycetes, aspergillus oryzae and mucor circinelloides; the auxiliary materials comprise rice and deionized water;
the cactus fermentation filtrate comprises the following raw materials in percentage by mass:
the preparation method of the cactus fermentation filtrate with the free radical scavenging effect comprises the following steps:
s10, respectively preprocessing the compound microorganism and the cactus:
s11, pretreatment of compound microorganisms:
respectively picking yeast, aspergillus oryzae and mucor circinelloides from the composite microorganism flat plate in a sterile way, respectively inoculating the yeast, aspergillus oryzae and mucor circinelloides into a sterile YEPD culture medium, and carrying out shake flask culture at a temperature of 30 ℃ for 30 hours to obtain pretreated yeast, pretreated aspergillus oryzae and pretreated mucor circinelloides;
the components of the YEPD medium include: yeast powder, peptone, glucose and distilled water;
the mass ratio of the yeast powder to the peptone to the glucose to the distilled water is 1:2:2:95;
s12, pretreatment of cactus: cleaning and pulverizing radix et caulis Opuntiae Dillenii, heating at 80deg.C for 2.5 hr, cooling to below 30deg.C, and collecting pretreated radix et caulis Opuntiae Dillenii;
s20, uniformly mixing rice and deionized water with the mass percentage of 10%, controlling the temperature to 121 ℃, controlling the pressure to 0.11Mpa, treating for 20min, cooling to below 30 ℃ after treatment to obtain treated rice materials, uniformly inoculating the treated rice materials into pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides according to mass, controlling the temperature to 29 ℃ for fermentation for 4 days, uniformly mixing the fermented pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides, inoculating the mixture into pretreatment cactus, adding the rest deionized water, controlling the temperature to 30 ℃ for fermentation for 6 days, and obtaining intermediate cactus fermentation liquid;
s30, performing 80-mesh coarse filtration on the middle cactus fermentation liquid, performing tubular centrifugation at 10000rpm to remove large-particle impurities, and performing sterile filtration at 0.22 mu m after removing the impurities to obtain a finished product of the cactus fermentation filtrate with the effect of removing free radicals.
Example 2
A cactus fermentation filtrate with free radical scavenging effect, the cactus fermentation filtrate comprises the following raw materials: compound microorganism, auxiliary materials and cactus;
the compound microorganism comprises saccharomycetes, aspergillus oryzae and mucor circinelloides; the auxiliary materials comprise rice and deionized water;
the cactus fermentation filtrate comprises the following raw materials in percentage by mass:
the preparation method of the cactus fermentation filtrate with the free radical scavenging effect comprises the following steps:
s10, respectively preprocessing the compound microorganism and the cactus:
s11, pretreatment of compound microorganisms:
respectively picking yeast, aspergillus oryzae and mucor circinelloides from the composite microorganism flat plate in a sterile way, respectively inoculating the yeast, aspergillus oryzae and mucor circinelloides into a sterile YEPD culture medium, and carrying out shake flask culture at the temperature of 28 ℃ for 23 hours to obtain pretreated yeast, pretreated aspergillus oryzae and pretreated mucor circinelloides;
the components of the YEPD medium include: yeast powder, peptone, glucose and distilled water;
the mass ratio of the yeast powder to the peptone to the glucose to the distilled water is 1:2:2:95;
s12, pretreatment of cactus: cleaning and pulverizing radix et caulis Opuntiae Dillenii, heating at 75deg.C for 2 hr, cooling to below 30deg.C, and collecting pretreated radix et caulis Opuntiae Dillenii;
s20, uniformly mixing rice and deionized water with the mass percentage of 10%, controlling the temperature to 120 ℃, controlling the pressure to 0.11Mpa, treating for 20min, cooling to below 30 ℃ after treatment to obtain treated rice materials, uniformly inoculating the treated rice materials into pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides according to mass, controlling the temperature to 28 ℃ for fermentation for 3 days, uniformly mixing the fermented pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides, inoculating the mixture into pretreatment cactus, adding the rest deionized water, and controlling the temperature to 28 ℃ for fermentation for 6 days to obtain intermediate cactus fermentation liquid;
s30, performing 40-mesh coarse filtration on the middle cactus fermentation liquid, performing tube centrifugation at 12000rpm to remove large-particle impurities, and performing sterile filtration at 0.22 mu m after removing the impurities to obtain a finished product of the cactus fermentation filtrate with the effect of removing free radicals.
Example 3
A cactus fermentation filtrate with free radical scavenging effect, the cactus fermentation filtrate comprises the following raw materials: compound microorganism, auxiliary materials and cactus;
the compound microorganism comprises saccharomycetes, aspergillus oryzae and mucor circinelloides; the auxiliary materials comprise rice and deionized water;
the cactus fermentation filtrate comprises the following raw materials in percentage by mass:
the preparation method of the cactus fermentation filtrate with the free radical scavenging effect comprises the following steps:
s10, respectively preprocessing the compound microorganism and the cactus:
s11, pretreatment of compound microorganisms:
respectively picking yeast, aspergillus oryzae and mucor circinelloides from the composite microorganism flat plate in a sterile way, respectively inoculating the yeast, aspergillus oryzae and mucor circinelloides into a sterile YEPD culture medium, and carrying out shake flask culture at the temperature of 29 ℃ for 16 hours to obtain pretreated yeast, pretreated aspergillus oryzae and pretreated mucor circinelloides;
the components of the YEPD medium include: yeast powder, peptone, glucose and distilled water;
the mass ratio of the yeast powder to the peptone to the glucose to the distilled water is 1:2:2:95;
s12, pretreatment of cactus: cleaning and pulverizing radix et caulis Opuntiae Dillenii, heating at 70deg.C for 3 hr, cooling to below 30deg.C, and collecting pretreated radix et caulis Opuntiae Dillenii;
s20, uniformly mixing rice and deionized water with the mass percentage of 10%, controlling the temperature to 122 ℃, controlling the pressure to 0.11Mpa, treating for 20min, cooling to below 30 ℃ after treatment to obtain treated rice materials, uniformly inoculating the treated rice materials into pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides according to mass, controlling the temperature to 30 ℃ for fermentation for 4 days, uniformly mixing the fermented pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides, inoculating the mixture into pretreatment cactus, adding the rest deionized water, and controlling the temperature to 29 ℃ for fermentation for 5 days to obtain intermediate cactus fermentation liquid;
s30, carrying out 200-mesh coarse filtration on the middle cactus fermentation liquid, then carrying out tubular centrifugation at 8000rpm to remove large-particle impurities, and carrying out aseptic filtration of 0.22 mu m after removing the impurities to obtain a finished product of the cactus fermentation filtrate with the effect of free radical removal.
Comparative example 1
Compared with the example 1, the difference is that the rice is not added for fermentation in the comparative example 1, namely, the raw materials of the cactus fermentation filtrate do not contain rice, and the mass percentages of the raw materials of the cactus fermentation filtrate are as follows:
7.5% of saccharomycetes, 6.5% of aspergillus oryzae, 8.5% of mucor circinelloides, 42.5% of cactus and 35% of deionized water;
the step S20 specifically includes:
s20, uniformly mixing pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides, inoculating the mixture into the pretreatment cactus, adding deionized water, and fermenting at the temperature of 30 ℃ for 6 days to obtain an intermediate cactus fermentation liquid;
the rest operation steps and parameters are unchanged.
Comparative examples 2 to 4
The mass percentages of yeasts, aspergillus oryzae and Mucor circinelloides in comparative examples 2-4 compared to example 1 are shown in Table 1, with the remaining operating steps and parameters unchanged.
TABLE 1
Test example 1
The finished cactus fermentation filtrate products prepared in examples 1-3 and comparative examples 1-4 were tested for their ability to scavenge free radicals according to standard T/SHRH 006-2018 cosmetic-free radical (DPPH) scavenging assay, the results of which are shown in Table 2.
TABLE 2
| DPPH radical scavenging Rate (%) | Standard Deviation (SD) | |
| Example 1 | 84.33 | 0.62 |
| Example 2 | 82.98 | 0.40 |
| Example 3 | 83.40 | 0.58 |
| Comparative example 1 | 13.44 | 1.25 |
| Comparative example 2 | 18.86 | 0.63 |
| Comparative example 3 | 16.31 | 0.88 |
| Comparative example 4 | 15.17 | 0.45 |
As can be seen from the above examples and comparative example 1, the fermentation of the individual cactus is performed, only the cactus is in a poor growth state of the strain due to the lack of the rice, which can provide a large amount of carbon source, the effective fermentation of the cactus is not possible, and the elimination rate of the filtrate to DPPH free radicals is difficult to improve, so that the three are combined, the combination of the cactus, the rice and the compound microorganism can be optimally cooperated, and the finished product of the fermentation filtrate of the cactus has an excellent DPPH free radical elimination effect.
The above examples and comparative examples 2-4 show that the invention combines saccharomycetes, aspergillus oryzae and mucor circinelloides, and can cooperate to make the fermentation of cactus more complete, thereby further improving the elimination rate of the finished product of the fermentation filtrate of the cactus on DPPH free radicals.
Test example 2
Acute skin irritation test
The detection method comprises the following steps: acute skin irritation/corrosiveness test 5.3 in chapter 4 of cosmetic safety Specification (2015 edition).
Detecting an object: the finished products of the fermented filtrate of cactus obtained in examples 1 to 3 and comparative examples 1 to 4.
Detection environment: ordinary environment animal house, temperature: 18-26 ℃, relative humidity: 40-70%.
Experimental animals and feed: each group of 4 normal New Zealand rabbits, male, weight 2.04-2.27 kg.
The test method comprises the following steps:
a) Animal treatment: about 24 hours before the test, the hairs on both sides of the back spine of the experimental animal are cut off, the epidermis can not be damaged, and the left and right hair removing ranges are about 3cm x3cm respectively.
b) Contamination: 0.5mL of the test substance is directly smeared on the skin of the dehairing area, then covered by two layers of medical gauze (2.5 cm x2.5 cm) and one layer of oilpaper, and then fixed by a non-irritating medical bandage, and the skin on the other side is used as a control. Sealing and applying for 4 hours. After the test, the residual test substance was removed with warm water.
c) Animal observation: and observing skin reaction of the smearing part 1h, 24h, 48h and 72h after the test object is removed, scoring skin reaction according to the skin irritation reaction scoring standard, comprehensively evaluating the average value of the integral of the test animal, and judging the skin irritation intensity according to the highest integral average value of each observation time point of 24h, 48h and 72h and the skin irritation intensity grading table. The time of observation should be determined to be sufficient to observe the whole process of reversible or irreversible stimulation, typically not exceeding 14d.
Wherein the skin irritation response score and skin irritation intensity scale are as follows:
skin irritation response score
Skin irritation intensity grading
| Integral mean (n) | Strength of |
| 0≤n≤0.5 | No irritation |
| 0.5≤n≤2.0 | Light irritation |
| 2.0≤n≤6.0 | Mid-irritation |
| 6.0≤n≤8.0 | Strong irritation |
As a result of the test, the highest integral mean value at each observation time point of each group of 24h, 48h and 72h is 0.
The results of the skin irritation response integral test for each of the examples and comparative examples are shown in tables 3-9 below:
test results of acute skin irritation test of the test object on rabbits:
TABLE 3 Table 3
TABLE 4 Table 4
TABLE 5
TABLE 6
TABLE 7
TABLE 8
TABLE 9
The test conclusion is that the highest integral average value of each observation time point of each group of test objects in 24h, 48h and 72h is 0, and each test object has no irritation to the acute skin of the rabbit.
Test example 3
Skin allergy Test (Buehler Test local seal coating)
1. Test method 5.1 local seal coat test in cosmetic safety Specification (2015 edition) chapter 6 skin allergy test.
2. Test object: the finished products of the fermented filtrate of cactus obtained in examples 1 to 3 and comparative examples 1 to 4.
3. The detection environment is an animal house in a common environment, and the temperature is 18-26 ℃ and the relative humidity is 40-70%.
4. Experimental animals and feeds, wherein each group of 30 normal grade FMMU guinea pigs, male, weight of 301.0-359.5 g.
5. Sample treatment-the sample is directly applied without treatment.
6. The positive substance is 2.4-dinitrochlorobenzene, the solvent is 95% ethanol and water, the induction concentration is 5g/L, the excitation concentration is 1g/L, and the dosage is 0.2 mL/L.
7. Test method
a) 30 guinea pigs were randomly divided into a test group and a negative control group, the test group was 20, and the negative control group was 10
b) About 24 hours before the experiment, the left side of the back of the guinea pig is dehaired, and the dehairing range is 4 cm-6 cm.
c) The contact is induced by coating about 0.2L of the test object on the skin of the dehairing area of the experimental animal, covering the test object with two layers of gauze and one layer of cellophane, and sealing and fixing the test object with a non-irritating adhesive tape for 6 hours. The 7 th and 14 th steps are repeated once in the same way. The negative control group was operated in the same manner as a 0.9% sodium chloride injection.
d) Approximately 0.2mL of the test substance was applied to the right 2cm x2cm dehairing area (dehairing 24 hours prior to contact) on the back of guinea pigs in the test group and the negative control group 14d after the last induction of the excitation contact, then covered with two layers of gauze and one layer of cellophane, and then fixed with a non-irritating adhesive tape for 6 hours.
e) Animals were observed for skin reactions 24h and 48h after challenge and scored according to the table allergy test skin response score.
8. Determination criteria:
skin response score for allergy test
When the integral of skin reaction of the animals in the test object group is more than 2, the animals are judged to be positive in skin allergy, and the sensitization intensity of the test object is judged according to the sensitization intensity judgment standard in the table.
Sensitization intensity criterion
| Sensitization rate% | Sensitization intensity |
| 0～8 | Weak and weak |
| 9～28 | Light weight |
| 29～64 | In (a) |
| 65～80 | Strong strength |
| 81～100 | Extremely strong |
When the sensitization rate is 0, it can be judged that skin allergy is not seen.
9. The test results are shown in table 10 below:
table 10
| Group of | Results |
| Example 1 | Skin allergy is not seen |
| Example 2 | Skin allergy is not seen |
| Example 3 | Skin allergy is not seen |
| Comparative example 1 | Skin allergy is not seen |
| Comparative example 2 | Skin allergy is not seen |
| Comparative example 3 | Skin allergy is not seen |
| Comparative example 4 | Skin allergy is not seen |
As can be seen from Table 10, no skin allergy was seen in each group, and the raw materials used in the present invention were safe.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing is merely illustrative and explanatory of the invention, as various modifications and additions may be made to the particular embodiments described, or in a similar manner, by those skilled in the art, without departing from the scope of the invention or exceeding the scope of the invention as defined in the claims.
Claims (10)
1. The cactus fermentation filtrate with the free radical scavenging effect is characterized by comprising the following raw materials: compound microorganism, auxiliary materials and cactus;
the compound microorganism comprises saccharomycetes, aspergillus oryzae and mucor circinelloides; the auxiliary materials comprise rice and deionized water.
2. The cactus fermentation filtrate with the free radical scavenging effect as claimed in claim 1, wherein the cactus fermentation filtrate comprises the following raw materials in percentage by mass:
3. the fermented cactus filtrate with free radical scavenging effect as claimed in claim 2, wherein the mass percentage of the yeast, aspergillus oryzae and mucor circinelloides is 4-6%.
4. The fermented cactus filtrate with free radical scavenging effect as claimed in claim 3, wherein the yeast is 5% by mass, the aspergillus oryzae is 4% by mass, and the mucor circinelloides is 6% by mass.
5. A process for preparing a fermented filtrate of cactus having radical scavenging effect as claimed in any one of claims 1 to 4, characterized in that the process comprises the steps of:
s10, respectively carrying out pretreatment on the compound microorganism and the cactus to obtain a pretreated compound microorganism and a pretreated cactus;
s20, uniformly mixing rice and deionized water with the mass percentage of 10%, controlling temperature and pressure, cooling to below 30 ℃ after the treatment is completed, obtaining treated rice materials, uniformly inoculating the treated rice materials into pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides according to mass, performing first temperature control fermentation, uniformly mixing the fermented pretreatment saccharomycetes, pretreatment aspergillus oryzae and pretreatment mucor circinelloides, inoculating the mixture into pretreatment cactus, adding the balance deionized water, and performing second temperature control fermentation to obtain intermediate cactus fermentation liquid;
s30, filtering the middle cactus fermentation liquid, removing large-particle impurities in the tubular centrifugal core, and performing sterile filtration to obtain a finished product of the cactus fermentation filtrate with the effect of removing free radicals.
6. The method for preparing a fermented filtrate of cactus with free radical scavenging effect according to claim 5, wherein the step S10 specifically comprises the steps of:
s11, pretreatment of compound microorganisms:
respectively picking saccharomycetes, aspergillus oryzae and mucor circinelloides from the composite microorganism flat plate in a sterile mode, respectively inoculating the saccharomycetes, aspergillus oryzae and mucor circinelloides into a sterile YEPD culture medium, and culturing in a temperature-controlled shake flask to obtain pretreated saccharomycetes, pretreated aspergillus oryzae and pretreated mucor circinelloides;
s12, pretreatment of cactus: cleaning radix et caulis Opuntiae Dillenii, pulverizing, heating at controlled temperature, and cooling below 30deg.C to obtain pretreated radix et caulis Opuntiae Dillenii.
7. The method for preparing a fermented filtrate of cactus with radical scavenging effect according to claim 6, wherein the components of the YEPD medium in step S11 comprise: yeast powder, peptone, glucose and distilled water.
8. The method for preparing a fermented filtrate of cactus with radical scavenging effect according to claim 7, wherein the mass ratio of yeast powder, peptone, glucose and distilled water is 1:2:2:95.
9. The method for preparing a fermented filtrate of Opuntia Dillenii having the effect of scavenging free radicals as claimed in claim 6, wherein the temperature of the flask culture is controlled to be 28-30deg.C for 16-30 h in step S11; and step S12, the temperature of the temperature-controlled heating treatment is 70-80 ℃ and the time is 2-3 h.
10. The method for preparing a fermented filtrate of Opuntia Dillenii having free radical scavenging effect according to claim 5, wherein the temperature and pressure controlled treatment in step S20 is 120-122 deg.C, 0.11Mpa, and treatment time is 20min; the temperature of the first temperature-controlled fermentation is 28-30 ℃ and the time is 3-4 days; the temperature of the second temperature-controlled fermentation is 28-30 ℃ and the time is 5-6 days.
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