CN104940049A - Method for extracting smallanthus sonchifolius plant extract, extract and application - Google Patents

Abstract

The invention discloses a method for extracting yacon plant extracts, comprising: adding ethanol or ethanol aqueous solution as an extractant to pretreated leaves or tubers of yacon, ultrasonicating, filtering, and concentrating the filtrate; passing the concentrate through The macroporous resin column chromatography is used for separation, and the column chromatography adopts gradient elution, which is first eluted with deionized water, and then sequentially eluted with ethanol aqueous solution with gradually increasing mass percentage concentration, and the extract is obtained after post-processing. The invention also discloses the yacon plant extract obtained by the above method and the application method of the extract. The present invention uses the leaves or tubers of yacon as raw materials, and through a simple extraction process, the extracted components with whitening and anti-aging effects of different activities can be obtained, which can be classified and collected according to actual needs, and further prepared into required The product is highly practical and has broad market prospects.

Description

Extraction method, extract and application of yacon plant extract

technical field

The invention belongs to the technical field of plant extraction, and in particular relates to a method for extracting yacon plant extract, the extract and its application as whitening and anti-aging actives and the whitening and anti-aging actives. the

Background technique

Among the determinants of human skin color, the activity of melanocytes, which produce melanin in the human body, occupies the most important position. Studies have shown that human melanocytes produce melanin through the activities of various enzymes (for example, tyrosinase), and the formation of melanin is also affected by genetic factors, hormone secretion, physiological factors related to stress and environmental factors (for example, effects of ultraviolet radiation). Melanin has the functions of blocking the sun's ultraviolet rays, protecting the skin organs under the dermis, removing free radicals produced in skin tissues, and protecting the proteins and genes in the skin. However, since melanin is a stable substance, it is not easy to be removed when the stress disappears, and it cannot be removed from the skin until it falls off with the aging of keratinocytes. Therefore, when an unnecessary amount of melanin is produced due to environmental factors, hyperpigmentation that detracts from beauty occurs, such as darkening, freckles, and blemishes. As the number of people who enjoy outdoor activities increases, so does the need to prevent hyperpigmentation caused by UV radiation. In order to meet this demand, substances having tyrosinase inhibitory activity such as ascorbic acid, kojic acid, arbutin and the like have been used in external skin preparations such as cosmetics. However, the use of these ingredients is limited due to unsatisfactory whitening effects and other problems (for example, skin irritation caused by adding the above-mentioned whitening actives to cosmetics, formulation stability problems, etc.). As a potential whitening active substance, tyrosinase inhibitors derived from plant extracts have safety and low skin irritation expectations. There have been various development reports (patent document CN201310411195: Baishui extract The detection method of whitening effect; CN201310382406.6: the application of the extract from the peel of Akiras sinensis in the preparation of whitening cosmetics). the

The causes of skin aging can be divided into photoaging and intrinsic aging. Photoaging caused by UV exposure can cause an increase in the activity of skin elastase. Elastase is distributed in a variety of tissues and cells. Increased elastase activity can lead to hydrolysis of skin elastic fibers, thereby accelerating the decomposition of collagen and elastin in the dermis, causing skin loss of elasticity and skin aging such as wrinkles. Currently known anti-aging active substances derived from plants have elastase inhibitory effect, such as Polygonum indigo (patent document CN200680023570.6: skin external preparation), sweet pea (patent document CN200680009020.9: cosmetics containing sweet pea extract) wait. the

Yacon (Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson) belongs to the genus Sunflower of the Compositae family. It is native to the Andes in South America and was introduced from Japan to Yunnan and other places in my country in the early 21st century. Yacon, formerly known as yacon, is also known as Yacon, chrysanthemum yam, and yacon. It is a traditional food of South American Indians. It is a South American medicinal and edible plant with multiple functions such as relieving diabetes. the

Yacon root contains a large amount of fructo-oligosaccharides, and has the title of "king of fructo-oligosaccharides". Yacon root also contains a variety of phenolic acids, terpenoids and flavonoids. It has been reported that yacon leaf extract has the effect of anti-oxidation and scavenging free radicals. the

Contents of the invention

The invention provides a method for extracting yacon plant extract, which is simple to operate and suitable for industrial production, and the extract obtained by the method has good skin care and whitening effects. the

The present invention also provides a yacon plant extract extracted by the above method, and the extract has good skin care and whitening effects. the

The present invention also provides an application method of the above-mentioned yacon plant extract, which can be used as an active ingredient of an external preparation for whitening and anti-aging skin. the

A method for extracting yacon plant extracts, comprising:

(1) In the leaves or tubers of the pretreated yacon, add ethanol or an aqueous ethanol solution as an extractant, ultrasonically, filter, and concentrate the filtrate;

(2) The concentrated solution obtained in step (1) is further separated and purified through a macroporous resin, preferably using macroporous resin column chromatography and gradient elution, first eluted with deionized water, and then successively with mass Gradient elution was carried out with ethanol aqueous solution with increasing percentage concentration, and the extract was obtained after post-processing. the

In step (1), in order to improve the extraction effect, the pretreatment mainly includes steps such as washing and crushing. Preferably, the extractant is an aqueous ethanol solution, and the concentration of the aqueous ethanol solution is 50-90% by mass, more preferably 60-80%, and even more preferably 70%. To ensure that as much active ingredient as possible is extracted. the

In the step (1), the amount of the extractant added once is generally 5-15 times of the volume of the leaves or roots of Yacon. The ultrasonic treatment time can be determined according to actual needs, generally 0.5-2 hours, further 1-2 hours. The number of times of extraction can generally also be determined according to actual needs. As a preference, extraction is performed two to three times, and ultrasonic treatment is performed for each extraction. The extraction temperature can be carried out at room temperature, or it can be properly heated. the

In step (2), during macroporous resin column chromatography, preferably, the macroporous resin is AB-8 macroporous adsorption resin. First, deionized water is used to elute the AB-8 macroporous adsorption resin, mainly to remove some water-soluble impurities. In step (2), preferably, the eluent of the gradient elution is an ethanol aqueous solution with a mass percent concentration of 20-40%, an ethanol aqueous solution with a mass percent concentration of 60-80%, and a mass percent concentration of 90-80%. 100% ethanol aqueous solution, and the ethanol eluate was collected respectively, concentrated and prepared respectively to obtain three kinds of extracts. As a further preference, the eluents are 30% ethanol aqueous solution, 70% ethanol aqueous solution and 95% ethanol aqueous solution respectively in sequence. The use of eluents with stepwise concentration changes can ensure that various active ingredients in the extract are separated, and the activities of the separated active ingredients are different. In practical applications, the amounts of the three ingredients are adjusted and selected according to the different activities. , to meet the needs of different occasions. the

The present invention also provides a yacon plant extract extracted by the above method. The yacon plant extract can be used in different occasions according to actual needs. the

The present invention also provides application methods of several above-mentioned yacon plant extracts as skin external preparations. the

An application of the yacon plant extract in the preparation of an external skin preparation with anti-oxidation and free radical scavenging effects, wherein the yacon plant extract is prepared by the above-mentioned various methods. the

When applying the above occasion, as a preference, when the yacon leaf is used as the raw material of the yacon plant extract, the eluted product with a mass percentage concentration of 20-100% ethanol aqueous solution is collected during gradient elution. When the yacon plant extract uses the tuber root of yacon as a raw material, the eluted product with a mass percent concentration of 20-80% ethanol aqueous solution is collected during gradient elution. As a further preference, the yacon plant extract uses yacon leaves as a raw material, and the eluted product of ethanol aqueous solution with a mass percentage concentration of 70-95% is collected during gradient elution. the

An application of the yacon plant extract in the preparation of an anti-aging skin external preparation, wherein the yacon plant extract is prepared by the above-mentioned various methods. the

When the above occasions are applied, preferably, the eluted product of ethanol aqueous solution with a mass percent concentration of 60-80% is collected during gradient elution. As a further preference, the root tuber of yacon is used as raw material. As a further preference, during gradient elution, the eluted product of ethanol aqueous solution with a mass percent concentration of 70% is collected; meanwhile, the root tuber of yacon is used as a raw material. the

An application of a yacon plant extract in the preparation of an external skin preparation with whitening effect, wherein the yacon plant extract is prepared by the above-mentioned various methods. the

When the above occasions are applied, preferably, the eluted product of ethanol aqueous solution with a mass percent concentration of 60-100% is collected during gradient elution. As a further preference, the leaves of yacon are used as raw materials, and the eluted products with a mass percentage concentration of 70-95% ethanol aqueous solution are collected during gradient elution. Even more preferably, the root or leaf of yacon is used as the raw material, and the eluted product with a concentration of 70% ethanol in water is collected during gradient elution. the

As a preference for the above several solutions, the external skin preparation is a cosmetic, especially an essence cosmetic, and the weight percentage of yacon extract in the essence is 1-10%, more preferably 1-7%. the

What used in the present invention is Compositae sunflower plant, preferably yacon (Smallanthus sonchifolius (Poepp. & Endl.) H.Robinson). the

Compared with the prior art, the beneficial effects of the present invention are reflected in:

The present invention uses the leaves or tubers of yacon as raw materials, and through a simple extraction process, the extracted components with whitening and anti-aging effects of different activities can be obtained, which can be classified and collected according to actual needs, and further prepared into required The product is highly practical and has broad market prospects. the

Detailed ways

Embodiment 1: the preparation of yacon extract

Yacon leaves or tubers produced in Yunnan are used in this embodiment, but not limited thereto. In addition, the percentages mentioned in the examples all represent mass percentage concentrations unless otherwise specified. the

(1) Add 1000 mL of 70% ethanol to 100 g of yacon leaves, ultrasonicate at room temperature for 1 hour, extract twice, and then filter. Concentrate the solvent of the filtrate to 50ml under reduced pressure, put it on the AB-8 resin column, and use deionized water, 30% ethanol, 70% ethanol and 95% ethanol to elute (each 500ml) successively, collect and remove the solvent under reduced pressure after collecting respectively, The dried solid was obtained and weighed to obtain 2.95 g of the 30% ethanol part of the yacon leaf, 9.89 g of the 70% ethanol part, and 2.45 g of the 95% ethanol part. the

(2) Add 1000 mL of 70% ethanol to 100 g of yacon root, ultrasonicate at room temperature for 1 hour, extract twice, and then filter. Concentrate the solvent of the filtrate to 50ml under reduced pressure, put it on the AB-8 resin column, and use deionized water, 30% ethanol, 70% ethanol and 95% ethanol to elute (each 500ml) successively, collect and remove the solvent under reduced pressure after collecting respectively, The dry solid was obtained and weighed to obtain 3.0 g of the 30% ethanol part of the yacon root, 3.6 g of the 70% ethanol part, and 1.0 g of the 95% ethanol part. the

Embodiment 2: the detection of antioxidant activity

The yacon extract was prepared according to Example 1, and its free radical scavenging activity was measured by the DPPH method. The determination steps are: the yacon extract is prepared into an aqueous solution (mass volume concentration: 0.05 mg/mL) with deionized water, pipette 2 mL into a 10 mL stoppered test tube, and add 2 mL of DPPH (1,1-diphenyl -2-trinitrophenylhydrazine) ethanol solution (molar volume concentration is 2*10 ^-4 mol/L), mix well, let stand at room temperature, measure the absorbance T of 517nm wavelength with a spectrophotometer after 30min; measure 2mL Absorbance C of DPPH solution mixed with 2mL ethanol, absorbance C ₀ of 2mL deionized water mixed with 2L ethanol, and absorbance T ₀ of 2mL extract mixed with 2mL ethanol solution. Three parallel measurements were taken, the average value was taken, and the free radical scavenging rate was calculated according to the following formula.

Free radical scavenging rate=[1-(TT ₀ )/(CC ₀ )]×100%.

The assay result of table 1 DPPH method free radical scavenging activity

As can be seen from Table 1, among the yacon extracts used in the present invention, any solvent extract has an excellent free radical scavenging effect and is useful as an anti-aging agent. Especially when yacon leaves are used as the extraction material, the free radical scavenging rate is the best when the eluent is 70%-95% ethanol aqueous solution. Therefore, yacon extract can be blended into external preparations and used as an anti-aging active substance that prevents skin aging and maintains a young and healthy skin state. the

Example 3: Detection of Elastase Inhibition

The yacon extracts prepared according to Example 1 were dissolved in deionized water to prepare an aqueous solution (mass volume concentration of 0.5 mg/ml) for the determination of elastase inhibitory activity. The assay steps are: add 10 μL sample solution and 130 μL containing 1.015 mM reaction substrate N-succinyl-propionamide-propionamide-propionamide-p-nitroaniline (English name Succ-Ala-Ala) to a 96-well plate -Ala-pNA, CAS number: 52299-14-6) in 0.1M Tris-Cl buffer solution (pH8.0), incubate at 25°C for 5 minutes, add 15μL elastase solution (0.5U/ml), continue Incubate at 25°C for 30 min, and then measure the absorbance A ₄₁₀ at a wavelength of 410 nm with a microplate reader. The sample aqueous solution was replaced with deionized water, and the absorbance was also measured as a reference solution, and the results are shown in Table 2.

The formula for calculating the inhibition rate of elastase activity is: inhibition rate (%)=(1-(A ₄₁₀ -C ₀ )/A ₀ )×100%, where A ₀ is the absorbance of the reference solution, and A ₄₁₀ is the absorbance of the sample solution , C ₀ is the absorbance of the sample solution background.

The assay result of table 2 elastase inhibition

As can be seen from Table 2, in the yacon extract used in the present invention, the 70% ethanol part of the root of the yacon and the 70% ethanol part of the leaf of the yacon all have the effect of inhibiting elastase, and the root of the yacon and the leaf of the yacon can be extracted Compounded in external preparations, it is used as an anti-aging agent to prevent skin aging and maintain young and healthy skin. the

Example 4: Detection of Tyrosinase Activity Inhibition

The yacon extract prepared according to Example 1 was dissolved in deionized water and formulated into an aqueous solution (mass volume concentration: 0.5 mg/ml) for the determination of tyrosinase inhibitory activity, with arbutin as a reference. The measurement steps are: phosphate buffer solution (pH6.8, 30mmol/L) 70μL, 1.66mmol/L tyrosine solution 80μL, sample solution 80μL, mix evenly, incubate in a constant temperature water tank at 37°C for 5 minutes, add tyrosinase 10 μL of the solution (1000 U/ml) was kept at 37° C. for 10 minutes, and the absorbance A ₄₇₅ at a wavelength of 475 nm was measured with a microplate reader. The sample aqueous solution was replaced with deionized water, and the absorbance was also measured as a reference solution, and the results are shown in Table 3.

The formula for calculating the inhibition rate of tyrosinase activity is: inhibition rate (%)=(1-(A ₄₇₅ -C ₀ )/A ₀ )×100%, where A ₀ is the absorbance of the reference solution, and A ₄₇₅ is the sample solution The absorbance of C ₀ is the absorbance of the sample solution background.

The assay result of table 3 tyrosinase inhibitory activity

As can be seen from Table 3, each yacon extract used in the present invention has tyrosinase activity inhibitory effect. Among them, the 70% ethanol part of the yacon root and the 70% ethanol part of the yacon leaf have a stronger inhibitory effect on tyrosinase, and both are better than the positive control substance arbutin at the same concentration, and the yacon root and yacon leaf can be extracted Compounded in external preparations, it is used as a whitening active to prevent skin pigmentation. the

Example 5: External skin preparation containing yacon extract

Add 1000 mL of 70% ethanol to 100 g of yacon root, ultrasonicate at room temperature for 1 hour, extract twice, and then filter. Concentrate the solvent of filtrate to 50ml under reduced pressure, go up AB-8 resin post, use deionized water, 30% ethanol, 70% ethanol and 95% ethanol elution (each 500ml) successively, collect 70% ethanol elution position and reduce Pressure distillation removes solvent, obtains the 70% ethanol part of yacon root tuber, is used for the preparation of skin external preparation. The weight percent of the yacon extract in the skin external preparation is 1-10%, preferably 1-7%. The following are examples of specific applications of yacon root in skin external preparations. "-" in Table 4 below means no addition. the

Embodiment 6: Contain the essence of yacon root

Get yacon root tuber extract, be used to prepare the essence with whitening effect, the component that described essence contains is as shown in table 4:

Table 4

The above are preferred embodiments of the invention and are not intended to limit the invention. the

Claims (10)

1. an extracting method for yacon plant extract, comprising:

(1) in the leaf or tuber of pretreated yacon, add ethanol or ethanol water as extractant, ultrasonic, filter, filtrate concentrates;

(2) concentrated solution that step (1) obtains is carried out the further separating-purifying of post layer through macroporous resin, and adopt gradient elution, first carry out eluting with deionized water, then carry out gradient elution with the ethanol water that mass percent concentration increases gradually respectively successively, post processing obtains extract.

2. the extracting method of yacon plant extract according to claim 1, is characterized in that, in step (2), described macroporous resin is AB-8 macroporous adsorbent resin.

3. the extracting method of yacon plant extract according to claim 2, it is characterized in that, the eluant of described gradient elution is followed successively by ethanol water that mass percent concentration is 20-40%, ethanol water that mass percent concentration is 60-80%, mass percent concentration be the ethanol water of 90-100%.

4. a yacon plant extract, is characterized in that, described yacon plant extract is extracted by the method described in the arbitrary claim of claim 1-3 and obtains.

5. yacon plant extract is preparing an application for skin preparations for extenal use, it is characterized in that, described yacon plant extract is prepared by the method described in the arbitrary claim of claim 1-3.

6. yacon plant extract according to claim 5 is preparing the application of skin preparations for extenal use, it is characterized in that, described skin preparations for extenal use is cosmetics.

7. yacon plant extract according to claim 6 is preparing the application of skin preparations for extenal use, it is characterized in that, described skin preparations for extenal use is essence cosmetics, and the percentage by weight that Smallanthus sonchifolius fruit extract accounts in essence is 1-10%.

8. yacon plant extract according to claim 5 is preparing the application of skin preparations for extenal use, it is characterized in that, when the leaf that described yacon plant extract is employing yacon is raw material, during gradient elution, collect the eluted products that mass percent concentration is the ethanol water of 20-100%; When the tuber that described yacon plant extract is employing yacon is raw material, during gradient elution, collect the eluted products that mass percent concentration is the ethanol water of 20-80%.

9. yacon plant extract according to claim 5 is preparing the application of skin preparations for extenal use, it is characterized in that, the tuber adopting yacon is raw material; The eluted products that mass percent concentration is the ethanol water of 60-80% is collected during gradient elution.

10. yacon plant extract according to claim 5 is preparing the application of skin preparations for extenal use, it is characterized in that, adopts the tuber of yacon or leaf to be raw material, collects the eluted products that mass percent concentration is the ethanol water of 70-95% during gradient elution.