KR20060070846A - Mathod of extracting lycopene from tomato and the anti-oxidation cosmetic composition containing the above - Google Patents

Abstract

The present invention relates to a method for extracting lycopene from tomatoes and to a cosmetic composition containing the lycopene. More specifically, the present invention is to extract lycopene from the tomato using supercritical extraction method by inhibiting the production of free radicals in the process of oxygen respiration metabolism, using water as a co-solvent, squalene (squalene) The present invention relates to an oil-soluble antioxidant component that gives oleophilicity to be applied to makeup cosmetics in which solvent such as ethanol is not blended.

Tomato * Lycopene * Suppresses free radical production * Supercritical extraction * Makeup cosmetics * Oleophilicity * Oil-soluble antioxidants * Squalene

Description

Method of extracting lycopene from tomato and the anti-oxidation cosmetic composition containing the above}

1 is a test result showing the antioxidant effect by the tomato extract.

2 is a test result showing the excellent sensitivity of squalene.

3 shows the results of HPLC analysis of lycopene standard.

Figure 4 is an HPLC analysis of lycopene extracted using a supercritical extraction method in tomato.

5 is a result of HPLC analysis of lycopene extracted using ethanol extraction method in tomato.

The present invention relates to a method for extracting lycopene from tomatoes and to a cosmetic composition containing the lycopene. More specifically, the present invention uses squalene as a co-solvent in extracting lycopene from the tomato using supercritical extraction method by inhibiting the production of free radicals in the process of oxygen respiration metabolism The present invention relates to a cosmetic composition containing lycopene into which oleophilicity is introduced.

Skin is damaged by environmental pollution, chemicals, ultraviolet rays, and stress. Among the causes of biological damage, active oxygen is now emerging as the biggest factor. Humans breathe oxygen in the air through the respiration process and maintain life using the energy obtained by oxidation. At this time, active oxygen is a major component that is constantly produced in various metabolic processes and attacks biological tissues and oxidizes and damages cells. . Active oxygen refers to oxygen radicals and various oxygen compounds derived therefrom, all of which have high reactivity. Free radicals are atoms or molecules that contain electrons that are not chemically paired to the outermost electron orbit. They are unstable because they have the property of losing these unpaired electrons or getting one more electron from their surroundings to make it more stable. Therefore, they have high reactivity because they try to lose or gain electrons by reacting with surrounding compounds. Oxygen has two electrons that are not originally paired (they are also called bi-radical), but unlike other free radicals, they are very reactive. This is because the two unpaired electrons have the same spin (↑↑), and the pair of electrons into the oxygen has the opposite spin (↑ ↓) according to Pauli's principle. But when oxygen overcomes this spin restriction and accepts electrons, the active oxygen in question is produced. All molecules can form radicals, but oxygen radicals are particularly problematic because the use of oxygen is indispensable in aerobic life and electrons released from the intracellular electron transfer system are transferred to oxygen to produce oxygen radicals. The oxygen thus produced oxidizes in the human body quite differently from the oxygen we breathe. This causes damage to all other cellular structures, depending on the extent of the damage, causes the cells to lose function or deteriorate and to damage cells or organelles that occur in the presence of oxygen. When antioxidants do not remove these free radicals, the free radicals that accumulate damage the DNA and cellular tissues of the human genome, destroying the DNA's genetic information, and disrupt cell membranes to form abnormal cellular proteins that promote cancer development and aging. Cause. The Free Radical Theory of Aging was proposed by Harman in 1956, and the oxidative damage of biological components is caused by various free radicals that are incidental to normal metabolism. These damages accumulate and lead to aging and death. Damages caused by free radicals are coped with the defense ability of living organisms, but are not 100% complete defenses, and thus some of the free radicals are adversely affected. Therefore, in order to solve the problems related to skin beauty and physiological function protection, research and development of a substance that inhibits the production of free radicals, that is, various antioxidant enzymes and antioxidant factors, and the study of complex antioxidant system in vivo, Many research efforts have been actively made in the field of materials.

A component to be noted for inhibiting the production of active oxygen is lycopene represented by the following formula (1).

Lycopene, a component found in ripe tomatoes, watermelons, etc., is a type of carotenoids, one of the plant pigments. Lycopene, a potent antioxidant, plays a role in bright colors on plant leaves and vegetables. In particular, tomato lycopene is a dark red needle crystal containing 20ppm in biological tomatoes. It is fat-soluble and can easily penetrate into fat-rich tissue to function and remove harmful oxygen generated in biological tissue (Free Radical Scavenger). Its antioxidant capacity is about twice that of beta-carotenes and about one hundred times that of alpha-tocopherol.

On the other hand, tomato (Tomato, Lycopersicon esculentum Mill. ) Is a perennial herb belonging to the branch family, and about 90% of the fruit is composed of moisture and Lycopene, Beta-Carotenes, Vitamin C, E, and dietary fiber. It is a green-yellow vegetable with a lot of efficacy. Of particular note is lycopene, the red pigment in tomatoes.

In order to extract lycopene having a strong antioxidant capacity from these tomatoes, ethanol extraction method has been mainly used in the past, but in recent years, researches on supercritical extraction methods have been conducted to secure the stability and maximize the extraction yield of tomato extracts.

As a specific example, US Pat. No. 5,897,866 discloses partially dehydrated tomato plant fruit in supercritical conditions using an aliphatic hydrocarbon, aromatic hydrocarbon or water immiscible solvent as surfactant and stabilizer in the presence of phospholipids and using a multistage column. A method of extracting a lipophilic extract by concentrating a solvent in an extract by a concentrated concentration method through carbon dioxide backflow has been proposed.

US Patent Publication No. 2003-0180435 also discloses a method for extracting lycopene from tomatoes using a supercritical extraction method. Specifically, a carotenoid containing lycopene from edible oil is mixed using a powder of at least one plant and vegetable, including tomatoes, with edible oil and then using supercritical carbon dioxide fluid extraction. A method of separating is presented.

Recently published U.S. Patent Application Publication No. 2004-0131733 also describes a method for extracting concentrated lycopene using supercritical extraction, and more specifically, a lycopene-containing substance such as tomato or watermelon can be solubilized. After mixing with a critical fluid, a method of separating concentrated lycopene free of organic solvent by reducing the pressure to 10 Mpa or less is shown.

Accordingly, the present inventors conducted a study to find an antioxidant effect among various natural product extracts that can ensure a smooth fragrance, stability, and skin safety so that cosmetics can be easily applied, lycopene contained in tomato (Tomato) Lycopene removes free radicals generated in biological tissues during oxygen respiration metabolism (Free Radical Scavenger), which has an antioxidant effect that prevents the oxidation of cells and inhibits carcinogenesis. It was found and completed the present invention.

It is therefore an object of the present invention to provide a method for extracting lycopene, a skin antioxidant active ingredient that prevents the oxidation of cells and inhibits carcinogenesis from tomatoes.

Another object of the present invention is to provide a cosmetic composition containing the lycopene.

In order to achieve the above object, the present invention is to provide a method for extracting lycopene from the plant through a supercritical extraction method using carbon dioxide as the extraction solvent and squalene as the co-solvent. The plants include tomatoes, grape seeds, thistle seeds and green tea.

The present invention provides a method of stably extracting lycopene, which is an active ingredient of tomato, and maximizing extraction yield by using a supercritical extraction method. In general supercritical extraction, the optimum point of extraction temperature and pressure is obtained through several experiments. If the extraction temperature and pressure are not optimized, a complete 100% reaction will not occur and either unreacted or overreacted will result in the desired extraction sample. The extraction temperature in the supercritical extraction method of the present invention is preferably about 30 ~ 50 ℃, the extraction pressure is preferably about 350 ~ 450 atm, the temperature of the extraction tank is about 40 ℃, the pressure is most preferably about 400 atm Do.

The supercritical extraction method used in the present invention is extracted in a super critical fluid state having both properties of gas and liquid at a specific pressure and a specific temperature condition, thereby effectively extracting natural freshness without denatured by heat of active ingredient and impurities. It can be called an environmentally friendly extraction method.

By using carbon dioxide as the extraction solvent in the supercritical extraction method of rod invention, it is easy to adjust the critical conditions for each extraction component, it is possible to minimize the denaturation of the extract by low-temperature process, non-oxidative process. In addition, as a co-solvent, squalene, a natural extract functionalizing agent having high stability as a co-solvent and adding lipophilic property, is used.

Supercritical extraction method of the present invention,

1) putting tomatoes in a supercritical extraction tank and injecting squalene using a high pressure liquid pump;

2) injecting carbon dioxide using a high pressure gas pump;

3) extracting carbon dioxide at 30 to 50 ° C. and 350 to 450 atmospheres to discharge squalene;

4) adding squalene to the extract to obtain an extract mother liquor; And                     

5) extracting the extract mother liquor using ethanol, concentrated and filtered through a filter;

It includes.

In more detail, the tomato is placed in the supercritical extraction tank and simultaneously squalene is added to the extraction tank as a co-solvent using a high pressure liquid pump. The amount of squalene to be added is 30 to 40% by weight of the extract, but generally the amount of co-solvent is not determined. Thereafter, only the carbon dioxide is extracted using a high pressure gas pump to discharge all the squalene remaining in the extraction tank. After adding squalene to the extract obtained here as an extractive mother liquor, the mixture was extracted four times with 95% ethanol, concentrated and dissolved, followed by filtration with a 0.45 μm membrane filter to obtain a tomato extract.

On the other hand, the tomato supercritical extract according to the present invention has imparted oleophilicity to the extract by containing squalene which is a moisturizing ingredient of cosmetics as a cosolvent, and thus a makeup cosmetic which cannot be mixed with a solvent such as water or ethanol. For example, it is easy to apply to a variety of formulations such as solid foundation, lipstick, powder foundation, powder.

Hereinafter, although an Example and a test example are given and this invention is demonstrated further more concretely, this invention is not necessarily limited only to these examples.

Test Example 1 Antioxidant Effect by Tomato Extracts                     

This experiment was to measure the antioxidant effect to measure the efficacy effect of tomato extract. Antioxidant effect was measured by the method according to Korean Patent Application No. 10-2002-0027050.

The measuring method is the DPPH method, wherein the DPPH (1,1-diphenyll-2-picryl-hydrazyl) reagent used was purchased from Sigma Corporation (St. Louis, MO, USA). In Vitro , it is primarily used in the process of identifying free radical scavenging action. The concentration of each plant extract measured by the DPPH free radical scavenging activity measuring method was 1% by volume. Extracts of the above concentrations were placed in 96 well plates, respectively, to which DPPH prepared in 100 M ethanol solution was added so that the total volume of the solution was 200 μl. After leaving it at 37 degrees for 30 minutes, absorbance was measured using a 520 nm ELISA reader. Free radical scavenging activity (%) was calculated by the following equation 1 and the results are shown in Table 1 and FIG.

% Free radical scavenging activity = 100- (B / A) * 100

A: absorbance of control wells not treated with the extract of the present invention

B: Absorbance of Experimental Wells Treated with Extracts of the Invention

Kinds tomato Grape seed Thistle Seeds (Silymarin) green tea % Free radical scavenging activity 94.5 80.75 87.75 92.0

Note: The data above are averages of four measurements.

All the samples used in this experiment were tested for extracts extracted under the same conditions, and the results of experiments with 1% aqueous solution of the sample extract known to be known to have high antioxidant effect were the greatest antioxidant effect in tomato extract.

TEST EXAMPLE 2 Evaluation of Sensitivity of Cosmetic Composition According to Type of Cosolvent

Squalene, hexyl laurate, di-C12-15 alkyl malate, cyclohexanan, isobutene as cosolvent in supercritical extraction 100 panelists evaluated the sensitivity of each cosmetic composition containing the extract extracted using ethanol, or ethanol. Sensibility is composed of the effect and the touch on the skin surface of the cosmetic composition, the average of the evaluation of 100 panelists after evaluating each intensity of 1 to 10 is shown in FIG. As can be seen from the results of Figure 2, when using a squalene as a co-solvent, it can be seen that the cosmetic composition has an excellent surface effect and feel compared to other hydrocarbon-based or ester-based oils have.

 Example 1 Preparation of Supercritical Tomato Extract

5.0 g of crushed tomatoes were placed in a 100 ml supercritical extraction tank, and the extraction tank was sealed. The temperature of the extraction tank was raised to 40 degreeC, the high pressure liquid pump was attached, 2.0 g of squalene was thrown in, while carbon dioxide was thrown in using the high pressure gas pump, and the pressure was raised to 400 atmospheres.

The pressure and temperature were kept constant at 400 atm and 40 ° C. while releasing the extract using a pressure regulating valve attached to the outlet of the extraction tank. After all 5.0 g of squalene was added over 17 minutes, the high pressure liquid pump was stopped. Extraction was performed only with carbon dioxide for 2 hours using a high pressure gas pump to discharge all squalene remaining in the extraction tank and the extraction was completed. Squalene was added to the obtained extract to make a total weight of 100 g, which was used as an extracting mother liquor. 10 g of 100 g was extracted, concentrated and extracted four times with 10 ml of 95% ethanol, dissolved in 10 g, and filtered through a 0.45 μm membrane filter. Got it.

Comparative Example 1 Preparation of Tomato Extract Extracted with Ethanol

1.0 g of crushed tomatoes were placed in a 1 L beaker, 5 ml of methanol was added thereto, and the mixture was extracted under reflux with ultrasonic treatment (Ultrasonication) for 1 hour in a water bath at 100 ° C. After the reflux extraction, the mixture was cooled to room temperature, centrifuged to obtain a filtrate, and the filtrate was filtered with a 0.45 μm membrane filter to obtain a tomato extract.

Test Example 3 Quantitative Analysis of Lycopene Extracted from Tomato

In order to compare the selective extraction yield of the active ingredient according to the extraction method, HPLC analysis for Lycopene in lycopene standard, the supercritical tomato extract obtained in Example 1, and the ethanol tomato extract obtained in Comparative Example 1, The results are shown in FIGS. 3, 4, 5 and Table 2. However, the measured value of Table 2 is a value when lycopene (Lycopene) content of the ethanol extract is set to 1.00 ppm.

FIG. 3 shows the results of HPLC analysis of lycopene standard (manufacturer: Sigma,> 95%), retention time of 9.34 min: Lycopene, injection volume of 1 μl, and FIG. 4 shows the results of HPLC analysis of supercritical tomato extract obtained in Example 1. The retention time is 9.47 min: Lycopene and the injection volume is 1 μl. 5 is a HPLC analysis result of the ethanol tomato extract obtained in Comparative Example 1, the retention time is 9.50 min: Lycopene, the injection volume is 10μl.

<Analytical Instruments and Conditions>

(1) Instrument: Agilent HP 1100, Ion Trap Mass

(2) Column: ZORBAX Eclipse XDB-C ₁₈ 250 x 2.1 mm ID, 5mm

(3) Guard column: ZORBAX Eclipse XDB-C ₈ 12.5 x 2.1 mm ID, 5mm

(4) Mobile Phase: MeOH: MTBE: 0.1% Acetic Acid = 50: 46: 4

(5) Detector: 470nm (Lycopene) (UV-DAD)

(6) Flow Rate: 0.30ml / min

(7) Injection Volume: 1, 10 μl

(8) Column Oven Temp .: 35 ° C

(9) Retention time: 9.5min

Comparison of Tomato Lycopene Contents by Supercritical Extraction and Ethanol Extraction Supercritical Extraction Ethanol Extraction Method 3.40 ppm 1.00 ppm

4, 5 and Table 2, it can be seen that the content of lycopene in the tomato extract by the supercritical extraction method is much higher than the cervical extract extracted by the ethanol extraction method. Comparing the results shown in FIGS. 4 and 5 shows that the peak intensity of lycopene is higher when extracted using a supercritical extraction method, and the quantitative analysis shows that the amount of lycopene is higher than that of tomato lycopene by ethanol extraction. 3.4 times higher. In addition, the results of Figure 4, it can be seen that during the supercritical extraction, the peak intensity of other components is lowered to increase the purity of lycopene, which is an indicator of the extract.

The difference in retention time of lycopene in FIGS. 3, 4 and 5 is due to the column difference used for HPLC analysis.

Test Example 4 Test of the Effect of Inhibiting Oxygen Generation on the Human Body

In order to test the antioxidant effect on the human body, free radicals were measured in 2-3 drops of blood (whole blood). Since the active oxygen disappears after about one hundredth of a second, it is possible to measure the amount of active oxygen in the human body by checking the amount of lipid peroxide remaining as the active oxygen disappears even though it cannot be measured directly. First, a small amount of blood was collected by a needle with a saliva, placed in a constant container, activated by adding a special solution, centrifuged, and its color was read by photo meta and the concentration of active oxygen was expressed as a numerical value (carr U). Fluorescence from chemicals that react with oxygen can also be measured using a luminometer. Then, the foundation formulation of Table 3 was applied twice a day (morning and evening) as a representative formulation example, and the concentration of active oxygen in the human body was measured again one month later.                     

The determination of the inhibitory effect on the generation of active oxygen was determined by calculating the numerical value "carr U" representing the amount of active oxygen in the human body.

When the test substance is applied, the carr U value gradually decreases, and the comparison between the test substances is expressed as Δ carr U (Equation 2), and the results are shown in Table 4 below.                     

△ carr U = ((carr U value after application-carr U value before application) / carr U value before application) * 100

division Treatment Concentration (%) Inhibitory effect on free radical formation (△ carr U,%) Formulation Example 1 1.0 75.6 Comparative Formulation Example 1 1.0 8.2

As shown in Table 3, when applying the formulation example 1 containing the lycopene (Lycopene), it can be seen that the active oxygen production inhibitory effect is superior to the comparative formulation example 1 does not contain the lycopene (Lycopene).

The cosmetic composition containing lycopene according to the present invention may be formulated as a flexible lotion, astringent lotion, nutrition lotion, nutrition cream, massage cream, essence, pack, foundation, lipstick, powder foundation, etc. It is not.

Formulation Example 2 Flexible Lotion

A flexible lotion containing lycopene was prepared in the composition shown in Table 5 below.                     

[Formulation Example 3] nutrition lotion

A flexible lotion containing lycopene was prepared in the composition shown in Table 6 below.                     

[Formulation Example 4] Foundation

A flexible lotion containing lycopene was prepared in the composition shown in Table 7 below.

As described above, the present invention provides a supercritical extraction method having a high extraction yield in extracting lycopene having an excellent antioxidant effect by inhibiting generation of active oxygen from oxygen respiration metabolism from tomatoes.                     

In addition, in the supercritical extraction method of the present invention, since squalene, which is a moisturizing ingredient of cosmetics, is used as a cosolvent, it is possible to apply lycopene, which is an extract, to a cosmetic composition of a dispersion formulation in which a solvent such as water or ethanol is difficult to be blended.

Claims (6)

A method of supercritical extraction of lycopene from a plant, wherein the supercritical extraction method of lycopene is characterized by using carbon dioxide as an extraction solvent and squalene as a cosolvent. The method of claim 1, wherein the plant is selected from the group consisting of tomato, grape seed, thistle seed and green tea supercritical extraction method of lycopene. 1) putting tomatoes in a supercritical extraction tank and injecting squalene using a high pressure liquid pump; 2) injecting carbon dioxide using a high pressure gas pump; 3) extracting carbon dioxide at 30 to 50 ° C. and 350 to 450 atmospheres to discharge squalene; 4) adding squalene to the extract to obtain an extract mother liquor; And 5) extracting the extract mother liquor using ethanol, concentrated and filtered through a filter; Supercritical extraction method of lycopene comprising a.        Cosmetic composition containing a lycopene extract extracted by the method according to any one of claims 1 to 3. The cosmetic composition according to claim 4, wherein the composition is for inhibiting generation of active oxygen. The cosmetic composition according to claim 4 or 5, wherein the cosmetic composition is a makeup formulation formulated with at least one selected from the group consisting of foundation, lipstick, and powder.

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