CN110638704A - Extraction method of rose extract, rose extract obtained by extraction, application and cosmetics - Google Patents
Extraction method of rose extract, rose extract obtained by extraction, application and cosmetics Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
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- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Extraction Or Liquid Replacement (AREA)
Abstract
The invention provides an extraction method of a rose extract, the rose extract obtained by extraction, application and cosmetics, and relates to the technical field of plant extraction. The extraction method provided by the invention is simple and convenient to operate, the effective extraction of the total polyphenol in the rose can be realized by avoiding the use of various complex extraction reagents and processes, and in addition, the extraction solvent, the macroporous resin and other purification materials used in the extraction method can be recycled, so that the energy consumption can be reduced, the cost can be saved, and the green production can be realized.
Description
Technical Field
The invention relates to the technical field of plant extraction, in particular to an extraction method of a rose extract, the rose extract obtained by extraction, application and cosmetics.
Background
The skin is gradually aged due to the continuous contact with the outside, including external pollution, radiation exposure and other factors, which continuously generate free radicals. Antioxidant is a substance that effectively inhibits the oxidation reaction of free radicals, either by acting directly on the free radicals or by indirectly consuming substances that readily form free radicals, preventing further reactions from occurring.
With the development of natural products, plant polyphenols have become the most potential source of natural antioxidants, and the research on antioxidant activity thereof is receiving much attention. The plant polyphenol as a natural antioxidant active factor has the characteristics of high safety, high antioxidant activity, high extraction rate and the like, has important research significance and development prospect when being applied to cosmetics with the effects of whitening, removing freckles, resisting aging, astringing and the like, and becomes a necessary trend for market development.
The rose contains micromolecular active substances such as polyphenol, soluble sugar, organic acid, flavonoid and the like, and has higher economic value. However, at present, the utilization of the polyphenol in the roses is limited, and an efficient and environment-friendly method for extracting the polyphenol in the roses is lacked.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
It is a first object of the present invention to provide a method for extracting rose extract to alleviate at least one of the technical problems of the prior art.
The second purpose of the invention is to provide the rose extract obtained by the extraction method, wherein the mass fraction of total polyphenols of the rose extract reaches 56.3%, and the rose extract has good antioxidant capacity.
The third purpose of the invention is to provide the extraction method or the application of the rose extract in preparing cosmetics.
The fourth purpose of the invention is to provide a cosmetic which comprises the rose extract provided by the invention and has strong oxidation resistance.
The invention provides an extraction method of a rose extract, which comprises the steps of adding rose into an organic solvent, carrying out ultrasonic extraction, carrying out solid-liquid separation treatment on the obtained ultrasonic extracting solution, and purifying filtrate by macroporous resin to obtain the rose extract;
wherein the power of ultrasonic is 200-250W, the time of ultrasonic extraction is 30-45min, and the temperature of ultrasonic extraction is 75-85 ℃;
preferably, the power of the ultrasonic is 220-240W, the time of the ultrasonic extraction is 35-40min, and the temperature of the ultrasonic extraction is 76-84 ℃;
preferably, the power of the ultrasonic is 228W, the time of the ultrasonic extraction is 40min, and the temperature of the ultrasonic extraction is 80 ℃.
Further, the organic solvent comprises a low carbon alcohol solvent, preferably comprises a C1-C4 alcohol solvent, preferably comprises an ethanol solvent;
preferably, the ethanol solvent is an aqueous ethanol solution, preferably an aqueous ethanol solution of 40% -60% v/v.
Furthermore, the material-liquid ratio of the rose to the organic solvent is 1:30-50g/ml, preferably 1:35-45g/ml, and more preferably 1:40 g/ml.
Further, the solid-liquid separation treatment comprises a centrifugal treatment;
preferably, the centrifugation speed is 4000-; centrifuging for 20-40min, preferably 30 min;
preferably, the solid-liquid separation treatment further comprises collecting supernatant after the centrifugation treatment, and performing suction filtration separation treatment to obtain filtrate.
Further, concentrating the filtrate, and purifying with macroporous resin;
preferably, the concentration is carried out by adopting a reduced pressure concentration mode;
preferably, the temperature of the reduced pressure concentration is 40-60 ℃, preferably 50 ℃; the pressure is-0.05 to-0.2 MPa, preferably-0.1 MPa;
preferably, the condensation temperature of the reduced pressure concentration is 1-5 ℃, preferably 2 ℃; the rotation speed is 120-150rpm, preferably 130 rpm.
Further, the macroporous resin comprises D101, HPD-100, HPD-400, AB-8 or ADS-17, preferably HPD-400;
preferably, the sample loading amount is 1.0-3.0BV, preferably 2.0BV, during the purification process of the macroporous resin; the sample loading speed is 1-3ml/min, preferably 2 ml/min;
preferably, the elution is carried out with 60% -80% v/v ethanol in water, preferably 70% v/v ethanol in water;
preferably, the ethanol is removed after elution, followed by freeze drying to obtain the rose extract.
Further, the method also comprises the step of pretreating the rose before adding the organic solvent;
the pre-treatment comprises drying and pulverizing the roses;
preferably, coarse fibers are removed after crushing to obtain dry rose powder;
preferably, the rose flower is crushed and sieved by a 75-85 mesh sieve to remove coarse fibers, so that dry rose flower powder is obtained.
The invention also provides a rose extract obtained by the extraction method;
preferably, the mass fraction of total polyphenols in the rose extract is 50% -60%.
The invention also provides the extraction method or the application of the rose extract in preparing cosmetics.
In addition, the invention also provides a cosmetic which comprises the rose extract.
The extraction method of the rose extract provided by the invention comprises the steps of adding rose into an organic solvent, carrying out ultrasonic extraction, carrying out solid-liquid separation treatment on the obtained ultrasonic extracting solution, and purifying the filtrate by macroporous resin to obtain the rose extract. Immersing rose in organic solvent, performing ultrasonic extraction at power of 200-250W and temperature of 75-85 ℃ for 30-45min, and performing ultrasonic extraction under specific conditions, so that total polyphenol in the rose can be extracted mildly and efficiently, and the mass fraction of the total polyphenol in the obtained rose extract is as high as 56.3%. The solid-liquid separation is carried out on the ultrasonic extracting solution, the extracted rose fiber waste can be effectively removed, and the purity of the extracted product is ensured. Meanwhile, the content of total polyphenol of effective components can be further improved by purifying the filtrate after solid-liquid separation by macroporous resin. The extraction method provided by the invention is simple and convenient to operate, the effective extraction of the total polyphenol in the rose can be realized by avoiding the use of various complex extraction reagents and processes, and in addition, the extraction solvent, the macroporous resin and other purification materials used in the extraction method can be recycled, so that the energy consumption can be reduced, the cost can be saved, and the green production can be realized.
The rose extract extracted by the extraction method provided by the invention has the mass fraction of total polyphenols as high as 56.3%, and the antioxidant determination of the rose extract, such as DPPH free radical and ABTS elimination, potassium ferricyanide reducing power and FRAP value determination, shows that the rose extract can show stronger antioxidant power.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a standard curve of the method for detecting polyphenol by the Folin-phenol method provided in Experimental example 1 of the present invention;
FIG. 2 is a graph showing DPPH scavenging ability of different concentrations of rose extract according to Experimental example 2 of the present invention;
FIG. 3 shows ABTS +. clearance of rose extract at different concentrations according to Experimental example 3;
FIG. 4 shows the potassium ferricyanide reducing power of different concentrations of rose extract according to Experimental example 4 of the present invention;
FIG. 5 shows FeSO provided in Experimental example 5 of the present invention4A standard curve;
FIG. 6 shows FRAP values of rose extracts of different concentrations provided in Experimental example 5 of the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
According to one aspect of the invention, the extraction method of the rose extract is provided, and comprises the steps of immersing rose in an organic solvent, carrying out ultrasonic extraction, carrying out solid-liquid separation treatment on the obtained ultrasonic extracting solution, and purifying the filtrate by macroporous resin to obtain the rose extract;
wherein, the power of the ultrasound is 200-250W, such as but not limited to 200W, 210W, 220W, 230W, 240W or 250W; the ultrasonic extraction time is 30-45min, such as but not limited to 30min, 32min, 35min, 38min, 40min, 42min or 45 min; the temperature of ultrasonic extraction is 75-85 deg.C, and can be, but is not limited to, 75 deg.C, 78 deg.C, 80 deg.C, 82 deg.C or 85 deg.C.
Immersing rose in organic solvent, and performing ultrasonic extraction at power of 200-250W and temperature of 75-85 deg.C for 30-45min under the optimum extraction condition determined by single factor and orthogonal test, wherein the extract obtained under the condition has the highest content of active polyphenol. By the extraction conditions, the total polyphenol in the rose can be extracted mildly and efficiently, and the mass fraction of the total polyphenol in the obtained rose extract is as high as 56.3%.
The solid-liquid separation is carried out on the ultrasonic extracting solution, the extracted rose fiber waste can be effectively removed, and the purity of the extracted product is ensured. The invention does not limit the solid-liquid separation mode, and all methods can play a role in solid-liquid separation so as to remove solid insoluble substances.
Macroporous resin is a kind of high molecular polymer which is characterized by adsorption and has the functions of concentrating and separating organic matters. The content of total polyphenol of effective components can be further improved by purifying the filtrate after solid-liquid separation by macroporous resin.
The extraction method provided by the invention is simple and convenient to operate, the effective extraction of the total polyphenol in the rose can be realized by avoiding the use of various complex extraction reagents and processes, and in addition, the extraction solvent, the macroporous resin and other purification materials used in the extraction method can be recycled, so that the energy consumption can be reduced, the cost can be saved, and the green production can be realized.
Preferably, the power of the ultrasonic is 220-240W, the time of the ultrasonic extraction is 35-40min, and the temperature of the ultrasonic extraction is 76-84 ℃;
preferably, the power of the ultrasonic is 228W, the time of the ultrasonic extraction is 40min, and the temperature of the ultrasonic extraction is 80 ℃.
By further adjusting and optimizing the ultrasonic extraction conditions, the extraction efficiency can be further improved, the content of the target product can be improved, and the cost can be effectively saved.
In some preferred embodiments, the organic solvent comprises a low carbon alcohol solvent, preferably a C1-C4 alcohol solvent, preferably an ethanol solvent.
The low-carbon alcohol solvent has good compatibility with polyphenol substances in roses, and when ethanol is selected as the solvent, the solvent is easier to remove on the basis of ensuring good compatibility, and is non-toxic and harmless.
Preferably, the ethanol solvent is an aqueous ethanol solution, preferably an aqueous ethanol solution of 40% -60% v/v.
The cost can be further saved on the basis of ensuring the extraction efficiency by taking the ethanol water solution as the organic solvent, wherein the concentration of the ethanol water solution can be, but is not limited to, 40%, 45%, 50%, 55% or 60%.
In some preferred embodiments, the rose flower and the organic solvent have a feed-to-liquid ratio of 1:30-50g/ml, such as but not limited to 1:30g/ml, 1:33g/ml, 1:38g/ml, 1:42g/ml, 1:48g/ml or 1:50g/ml, and extraction is performed at the feed-to-liquid ratio, so that the extraction efficiency can be effectively improved, and waste of resources can be avoided.
The material-liquid ratio of the rose to the organic solvent is preferably 1:35-45g/ml, and more preferably 1:40 g/ml. By further adjusting and optimizing the ratio of the rose to the organic solvent, the extraction efficiency can be further improved, the content of a target product can be improved, and the cost can be effectively saved.
In some preferred embodiments, the solid-liquid separation treatment comprises a centrifugation treatment.
The rose fiber waste which is extracted can be removed preliminarily by using centrifugal treatment, the purity of the extracted product is ensured, the centrifugal treatment is simple and convenient to operate, expensive and precise instruments are not needed, and the cost can be effectively saved.
Preferably, the centrifugation speed is 4000-; the centrifugation time is 20-40min, for example, but not limited to, 20min, 25min, 30min, 35min or 40min, preferably 30 min.
Preferably, the solid-liquid separation treatment further comprises collecting supernatant after the centrifugation treatment, and performing suction filtration separation treatment to obtain filtrate.
Insoluble substances which are not taken out after centrifugation can be further clarified through the suction filtration separation treatment, and the purity of the product is ensured.
In some preferred embodiments, the filtrate is concentrated and then purified by macroporous resin, and the filtrate is concentrated and then purified, so that the purification efficiency can be effectively improved.
Preferably, the concentration is carried out by adopting a reduced pressure concentration mode;
preferably, the temperature of the reduced pressure concentration is 40-60 ℃, for example, but not limited to, 40 ℃, 45 ℃, 50 ℃, 55 ℃ or 60 ℃, preferably 50 ℃; the pressure is-0.05 to-0.2 MPa, and may be, for example, but not limited to, -0.05MPa, -0.1MPa, -0.15MPa or-0.2 MPa, preferably-0.1 MPa.
Preferably, the reduced pressure concentration of condensation temperature is 1-5 ℃, for example can be, but is not limited to, 1 ℃, 2 ℃, 3 ℃, 4 ℃ or 5 ℃, preferably 2 ℃; the rotation speed is 120-150rpm, and can be, but is not limited to, 120rpm, 130rpm, 140rpm or 150rpm, preferably 130 rpm.
In some preferred embodiments, the macroporous resin comprises D101, HPD-100, HPD-400, AB-8, or ADS-17, preferably HPD-400. DPPH free radical scavenging experiments are respectively carried out on the purified extracting solutions, and the result shows that the DPPH free radical scavenging effect of the extracting solution obtained after the macroporous resin HPD-400 is more excellent than that of the extracting solution obtained after other resins are purified.
Preferably, the loading amount is 1.0-3.0BV during the macroporous resin purification, for example, but not limited to, 1.0BV, 2.0BV or 3.0BV, preferably 2.0 BV; the sample loading speed is 1-3ml/min, for example, but not limited to, 1ml/min, 2ml/min or 3ml/min, preferably 2 ml/min;
preferably, elution is with 60% -80% v/v aqueous ethanol, for example, but not limited to, 60%, 65%, 70%, 75% or 80%, preferably elution is with 70% v/v aqueous ethanol;
preferably, the ethanol is removed after elution, followed by freeze drying to obtain the rose extract.
The method for removing ethanol is not limited, and any method capable of effectively removing ethanol may be used, and examples thereof include, but are not limited to, a concentration method and an evaporation method.
In some preferred embodiments, the method further comprises the step of pretreating the rose before adding the organic solvent;
the pre-treatment comprises drying the roses and comminuting.
The dried rose can be better preserved, the preservation period is prolonged, and the crushed rose can be more effectively contacted with an organic solvent, so that the extraction rate is improved.
Preferably, coarse fibers are removed after crushing to obtain dry rose powder;
preferably, the rose flower is crushed and sieved by a 75-85 mesh sieve to remove coarse fibers, so that dry rose flower powder is obtained.
According to another aspect of the invention, the rose extract obtained by the extraction method provided by the invention is provided;
preferably, the mass fraction of total polyphenols in the rose extract is 50% -60%.
The rose extract extracted by the extraction method provided by the invention has the mass fraction of total polyphenols as high as 56.3%, and the antioxidant determination of the rose extract, such as DPPH free radical and ABTS elimination, potassium ferricyanide reducing power and FRAP value determination, shows that the rose extract can show stronger antioxidant power.
Based on the extraction method provided by the invention and the rose extract obtained by extraction, the total polyphenol content is high, and the oxidation resistance is strong, so that the rose extract can be applied to the preparation of cosmetics.
In addition, based on the inventive concept of the rose extract provided by the invention, the invention also provides a cosmetic containing the rose extract, so that the cosmetic provided by the invention has all the beneficial effects of the rose extract provided by the invention, and the description is omitted.
The invention is further illustrated by the following specific examples, which, however, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.
Example 1
The embodiment provides an extraction method of a rose extract, which comprises the following steps:
a) pulverizing flos Rosae Rugosae, and sieving with 80 mesh sieve to obtain powder;
b) adding 50% ethanol water solution into the powder, and performing ultrasonic extraction with power of 228W and material-liquid ratio of 1:40g/ml, the extraction time is 40min, and the extraction temperature is 80 ℃;
c) centrifuging the ultrasonic extract obtained in the step b), wherein the centrifugation speed is 4500rpm, the centrifugation time is 30min, and the centrifugation frequency is 1 time;
d) carrying out suction filtration separation on the centrifuged supernatant, carrying out primary purification, and removing most impurities;
e) concentrating the filtrate under reduced pressure until no ethanol smell; wherein the reduced pressure concentration temperature is 50 ℃, the pressure is-0.1 MPa, the temperature of condensed water is 2 ℃, and the rotating speed is 130 rpm;
f) selecting HPD-400 type macroporous resin for purification, wherein the sample loading amount is 2.0BV, and the sample loading speed is 2 mL/min; eluting with 70% ethanol water solution, concentrating the eluate to remove ethanol, and freeze drying to obtain flos Rosae Rugosae extract.
Example 2
The embodiment provides an extraction method of a rose extract, which comprises the following steps:
a) pulverizing flos Rosae Rugosae, and sieving with 75 mesh sieve to obtain powder;
b) adding 60% ethanol water solution into the powder, and performing ultrasonic extraction with the power of 200W and the material-liquid ratio of 1:50g/ml, the extraction time is 30min, and the extraction temperature is 85 ℃;
c) centrifuging the ultrasonic extracting solution obtained in the step b), wherein the centrifuging speed is 4000rpm, the centrifuging time is 40min, and the centrifuging times are 1 time;
d) carrying out suction filtration separation on the centrifuged supernatant, carrying out primary purification, and removing most impurities;
e) concentrating the filtrate under reduced pressure until no ethanol smell; wherein the reduced pressure concentration temperature is 40 ℃, the pressure is-0.2 MPa, the temperature of the condensed water is 1 ℃, and the rotating speed is 150 rpm;
f) selecting HPD-400 type macroporous resin for purification, wherein the sample loading amount is 1.0BV, and the sample loading speed is 3 mL/min; eluting with 60% ethanol water solution, concentrating the eluate to remove ethanol, and freeze drying to obtain flos Rosae Rugosae extract.
Example 3
The embodiment provides an extraction method of a rose extract, which comprises the following steps:
a) pulverizing flos Rosae Rugosae, and sieving with 85 mesh sieve to obtain powder;
b) adding 40% ethanol aqueous solution into the powder, and performing ultrasonic extraction with the power of 250W and the material-liquid ratio of 1:30g/ml, extraction time of 45min and extraction temperature of 75 ℃;
c) centrifuging the ultrasonic extracting solution obtained in the step b), wherein the centrifuging speed is 5000rpm, the centrifuging time is 20min, and the centrifuging times are 1 time;
d) carrying out suction filtration separation on the centrifuged supernatant, carrying out primary purification, and removing most impurities;
e) concentrating the filtrate under reduced pressure until no ethanol smell; wherein the reduced pressure concentration temperature is 60 ℃, the pressure is-0.05 MPa, the temperature of condensed water is 5 ℃, and the rotating speed is 120 rpm;
f) selecting HPD-400 type macroporous resin for purification, wherein the sample loading amount is 3.0BV, and the sample loading speed is 1 mL/min; eluting with 80% ethanol water solution, concentrating the eluate to remove ethanol, and freeze drying to obtain flos Rosae Rugosae extract.
Example 4
The embodiment provides an extraction method of a rose extract, which is different from the embodiment 1 in that the ultrasonic power is 220W, the ultrasonic extraction time is 40min, the ultrasonic extraction temperature is 76 ℃, and the material-liquid ratio of rose and an organic solvent is 1:45 g/ml.
Example 5
The embodiment provides an extraction method of a rose extract, which is different from the embodiment 1 in that the power of ultrasonic is 240W, the time of ultrasonic extraction is 35min, the temperature of ultrasonic extraction is 84 ℃, and the material-liquid ratio of rose and an organic solvent is 1:35 g/ml.
Example 6
This example provides a method for extracting rose extract, which differs from example 1 in that D101 type macroporous resin is selected for purification.
Example 7
The embodiment provides an extraction method of a rose extract, which is different from the embodiment 1 in that ADS-17 model macroporous resin is selected for purification.
Example 8
This example provides a method for extracting rose extract, which is different from example 1 in that rose is directly added to 50% ethanol aqueous solution.
Example 9
This example provides a method for extracting rose extract, which is different from example 1 in that 50% ethanol aqueous solution is replaced with 50% propanol aqueous solution.
Example 10
This example provides a method of extracting a rose extract, which differs from example 1 in that step c) is not included.
Example 11
This example provides a method of extracting a rose extract, which differs from example 1 in that step d) is not included.
Example 12
This example provides a method of extracting a rose extract, which differs from example 1 in that step e) is not included.
Comparative example 1
This comparative example provides a method for extracting a rose extract, which is different from example 1 in that the power of ultrasonic is 180W, the time of ultrasonic extraction is 50min, and the temperature of ultrasonic extraction is 70 ℃.
Comparative example 2
This comparative example provides a method for extracting a rose extract, which is different from example 1 in that the power of ultrasonic is 280W, the time of ultrasonic extraction is 25min, and the temperature of ultrasonic extraction is 90 ℃.
Comparative example 3
This comparative example provides a method of extracting rose extract, which differs from example 1 in that it does not include a step of macroporous resin purification.
Comparative example 4
The comparative example provides an extraction method of a rose extract, which is different from the extraction method of example 1 in that the ultrasonic step is not carried out, direct water bath is changed, the extraction time is 40min, the rose extract is placed in a water bath pot to be heated, the temperature is 80 ℃, and the rose extract is uniformly mixed by shaking every 10 min.
Experimental example 1
The mass content of polyphenol in the rose extracts prepared by the preparation methods provided in examples 1 to 12 and comparative examples 1 to 4 was measured by the forskolin-phenol method, and the results are shown in table 1 below. Wherein, the standard curve of the forskolin-phenol method for detecting polyphenol is shown in figure 1.
TABLE 1 quality content of polyphenols in the rose extracts of the groups
From the above results, it can be seen that the content of total polyphenols in the rose extract obtained by applying the extraction method of the rose extract provided by the present invention is higher than that of the comparative example, which indicates that the extraction efficiency can be effectively improved by the specific extraction method and extraction conditions of the present invention. Further, it can be seen from the above results that the content of polyphenol in the rose extract obtained by extraction under the preferred conditions of the present invention is higher, which indicates that the extraction efficiency can be further improved by adjusting and optimizing the extraction conditions of the present invention.
In order to save cost, the following experimental examples all select the example 1 with the highest polyphenol content for verification.
Experimental example 2
The specific operation method for testing the antioxidant capacity of the rose extract by adopting the DPPH scavenging capacity measurement in the experimental example is as follows:
(1) preparing 0.2mmol/L DPPH ethanol solution; preparing Vc and rose extract sample solutions to be detected with different concentrations.
(2) Putting 100 μ L of each sample solution and DPPH solution in a 96-well plate, mixing uniformly, reacting for 30min in a dark place, and measuring the light absorption value A1 at the wavelength of 517 nm;
(3) putting the sample solution and 100 μ L ethanol in a 96-well plate, mixing uniformly, reacting for 30min in a dark place, and measuring the light absorption value A2 at the wavelength of 517 nm;
(4) mixing DPPH solution and 100 μ L ethanol in 96-well plate, reacting in dark for 30min, and measuring the light absorption value A0 at wavelength of 517 nm;
the clearance of the liquid to be measured is calculated according to the following formula:
referring to fig. 2, it can be seen that the effect of removing DPPH radicals is dose-dependent for the rose extract of example 1, which also indicates that the rose extract of the present invention has the effect of removing DPPH radicals.
Experimental example 3
The total antioxidant capacity of the rose extract was tested in this experimental example using ABTS +. scavenging capacity assay. ABTS is oxidized into green ABTS +. under the action of a proper oxidant, the generation of ABTS + is inhibited in the presence of antioxidant, and the total antioxidant capacity of a sample can be measured and calculated by measuring the absorbance of ABTS at 734nm, wherein the specific operation method comprises the following steps:
(1) diluting the ABTS stock solution with 7mmol/L PBS (pH 7.4) by 40-50 times to obtain ABTS working solution with an absorbance value of 0.7 +/-0.02 (30 ℃) at 734 nm; preparing Vc and rose extract sample solutions to be detected with different concentrations;
(2) and (3) sample determination: sucking 6mL of ABTS working solution, adding 60 mu L of sample solution, oscillating for 10s, standing for 6min at 30 ℃, and measuring the light absorption value at 734 nm;
referring to fig. 3, it can be seen that the cleaning effect of the rose extract of example 1 is obviously dose-dependent, and the rose extract of the present invention has obvious antioxidant ability. In this experiment, the IC50 value of the rose extract is lower than Vc, and the ABTS +. removing ability of the rose extract is slightly higher than Vc.
Experimental example 4
The experimental example is a test for measuring the reduction capacity by a potassium ferricyanide method, and the oxidation resistance of the rose extract is tested. The antioxidant can give out electrons to scavenge free radicals through self-reduction, and the stronger the reducing ability is, the stronger the oxidation resistance is. The sample can reduce ferric iron of potassium ferricyanide into ferrous iron (potassium ferrocyanide), and the ferrous iron (potassium ferrocyanide) further reacts with ferric trichloride to generate Prussian blue (Fe) with maximum absorbance at 700nm4[Fe(CN)6]3) Therefore, the reduction capability of the antioxidant can be indirectly reflected by measuring the height at 700nm, and the higher the absorbance is, the stronger the reduction capability is. The specific operation method comprises the following steps:
preparing Vc and rose extract sample solutions to be detected with different concentrations. 2mL of the sample solution was taken,5mL of 0.2mol/L phosphate buffer pH 6.6 was added, 5mL of 1% potassium ferricyanide solution was added, and the mixture was incubated at 50 ℃ for 20 min. Rapidly cooling, adding 5mL of 10% trichloroacetic acid solution, mixing, centrifuging at 3000r/min for 10min, collecting supernatant 2.5mL, adding distilled water 2.5mL and FeCl 0.1% 0.5mL3Reacting for 10min, measuring the absorbance value under the condition of 700nm wavelength, and using distilled water instead of sample solution as blank control. The greater the absorbance, the greater the reduction ability.
Referring to fig. 4, it can be seen that the cleaning effect of the rose extract of example 1 is obviously dose-dependent, and the rose extract of the present invention has obvious antioxidant ability.
Experimental example 5
This example is the FRAP method for determining the antioxidant capacity of rose extract. The FRAP method (the Ferric reducing antioxidant potential Assay) can detect the total antioxidant capacity of various body fluids, cell or tissue lysates and plant extracts. The principle is that Fe is in acidic condition3+-pyridinetriazazine (Fe)3+TPTZ) can be reduced by reducing substances in the sample to Fe which produces a blue color2+TPTZ, which has the maximum absorption at 593nm, and the antioxidant activity of the sample can be calculated according to the magnitude of the light absorption value.
(1) Preparing Vc and rose extract sample solutions to be detected with different concentrations. And (3) putting 0.1mL of sample solution into a test tube, adding 3mL of TPTZ working solution, adding 0.3mL of distilled water, uniformly mixing, keeping the temperature at 37 ℃ for 5min, and measuring the light absorption value at 593 nm. The absorbance values of the sample solution, positive control (VC or Trolox) and blank control were examined separately by zeroing with distilled water, and each group was assayed three times.
(2)FeSO4Standard curve
1mmol/L of FeSO4The solution is diluted into standard gradient solutions of 0.2, 0.4, 0.6 and 0.8mmol/L, 0.1mL of the solution is respectively absorbed and measured according to the experimental steps, and a regression curve between the light absorption value and the concentration is drawn. And calculating the FRAP values of the samples with different concentrations according to the standard curve. Wherein, FeSO4The standard curve of (2) is shown in fig. 5.
Referring to fig. 6, it can be seen that the dose-dependent inhibitory effect of the rose extract of example 1 also indicates that the rose extract of the present invention has significant antioxidant activity.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. An extraction method of a rose extract is characterized by comprising the steps of immersing rose in an organic solvent, carrying out ultrasonic extraction, carrying out solid-liquid separation treatment on an obtained ultrasonic extracting solution, and purifying filtrate by macroporous resin to obtain the rose extract;
wherein the power of ultrasonic is 200-250W, the time of ultrasonic extraction is 30-45min, and the temperature of ultrasonic extraction is 75-85 ℃;
preferably, the power of the ultrasonic is 220-240W, the time of the ultrasonic extraction is 35-40min, and the temperature of the ultrasonic extraction is 76-84 ℃;
preferably, the power of the ultrasonic is 228W, the time of the ultrasonic extraction is 40min, and the temperature of the ultrasonic extraction is 80 ℃.
2. The extraction process according to claim 1, characterized in that the organic solvent comprises a low carbon alcoholic solvent, preferably a C1-C4 alcoholic solvent, preferably an ethanol solvent;
preferably, the ethanol solvent is an aqueous ethanol solution, preferably an aqueous ethanol solution of 40% -60% v/v.
3. The extraction method according to claim 1, wherein the material-to-liquid ratio of the rose flowers to the organic solvent is 1:30-50g/ml, preferably 1:35-45g/ml, and more preferably 1:40 g/ml.
4. The extraction method according to claim 1, wherein the solid-liquid separation treatment comprises a centrifugation treatment;
preferably, the centrifugation speed is 4000-; centrifuging for 20-40min, preferably 30 min;
preferably, the solid-liquid separation treatment further comprises collecting supernatant after the centrifugation treatment, and performing suction filtration separation treatment to obtain filtrate.
5. The extraction method according to claim 1, wherein the filtrate is concentrated and then purified by macroporous resin;
preferably, the concentration is carried out by adopting a reduced pressure concentration mode;
preferably, the temperature of the reduced pressure concentration is 40-60 ℃, preferably 50 ℃; the pressure is-0.05 to-0.2 MPa, preferably-0.1 MPa;
preferably, the condensation temperature of the reduced pressure concentration is 1-5 ℃, preferably 2 ℃; the rotation speed is 120-150rpm, preferably 130 rpm.
6. The extraction process according to claim 1, wherein the macroporous resin comprises D101, HPD-100, HPD-400, AB-8 or ADS-17, preferably HPD-400;
preferably, the sample loading amount is 1.0-3.0BV, preferably 2.0BV, during the purification process of the macroporous resin; the sample loading speed is 1-3ml/min, preferably 2 ml/min;
preferably, the elution is carried out with 60% -80% v/v ethanol in water, preferably 70% v/v ethanol in water;
preferably, the ethanol is removed after elution, followed by freeze drying to obtain the rose extract.
7. The extraction method according to any one of claims 1 to 6, further comprising a step of pre-treating the rose before adding the organic solvent;
the pre-treatment comprises drying and pulverizing the roses;
preferably, coarse fibers are removed after crushing to obtain dry rose powder;
preferably, the rose flower is crushed and sieved by a 75-85 mesh sieve to remove coarse fibers, so that dry rose flower powder is obtained.
8. Rose extract obtained by extraction according to the extraction method of any one of claims 1 to 7;
preferably, the mass fraction of total polyphenols in the rose extract is 50% -60%.
9. Use of the extraction method according to any one of claims 1 to 7 or the rose extract according to claim 8 for the preparation of a cosmetic.
10. A cosmetic comprising the rose extract according to claim 8.
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