Application of tomato callus extract in preparation of anti-aging cosmetics and cosmetics

Abstract

The invention relates to the field of cosmetics, in particular to application of tomato callus extract in preparation of anti-aging cosmetics and the cosmetics. According to the application of the tomato callus extract in preparation of the anti-aging cosmetics, the tomato callus extract is an aqueous extract of tomato calluses. The aqueous extract of the tomato callus can effectively reduce the concentration of inflammatory factors, can effectively promote healing of cell scratches, and can effectively improve the free radical scavenging rate of ABTS and DPPH (1, 1-diphenyl-2-picrylhydrazyl); the aqueous extract of the tomato calluses can effectively improve the tyrosinase inhibition effect, so that the aqueous extract of the tomato calluses can be applied to anti-aging cosmetics to achieve the anti-inflammatory, repairing, anti-oxidation and whitening effects, and the anti-aging effect can be achieved from the aspects of anti-inflammatory, repairing, anti-oxidation and whitening.

Description

Technical Field

[0001] The present application relates to the field of cosmetics, and in particular, to an application of a tomato callus extract in the preparation of anti-aging cosmetics and cosmetics. Background Art

[0002] At present, there is no application of tomato callus in the field of cosmetics. Summary of the invention

[0003] The purpose of the embodiments of the present application is to provide an application of a tomato callus extract in the preparation of anti-aging cosmetics and cosmetics.

[0004] In a first aspect, the present application provides a use of a tomato callus extract in the preparation of anti-aging cosmetics.

[0005] Tomato callus extract is a water extract of tomato callus.

[0006] The present application adopts the aqueous extract of tomato callus tissue for the preparation of anti-aging cosmetics. The aqueous extract of tomato callus tissue can effectively reduce the concentration of inflammatory factors, effectively promote the healing of cell scratches, and effectively improve the scavenging rate of ABTS and DPPH free radicals; and the aqueous extract of tomato callus tissue can effectively improve the tyrosinase inhibition effect. Therefore, the application of the aqueous extract of tomato callus tissue in anti-aging cosmetics can achieve the effects of anti-inflammatory, repair, antioxidant and whitening, thereby achieving anti-aging effects from multiple aspects of anti-inflammatory, repair, antioxidant and whitening.

[0007] Furthermore, the development of water extracts of tomato callus in the prior art has not yet been seen in the field of cosmetics. The present application successfully extracts water extracts of tomato callus, and finds that they can effectively reduce the concentration of inflammatory factors, promote cell scratch healing, and increase the scavenging rate of ABTS and DPPH free radicals; improve the tyrosinase inhibition effect; and thus can be effectively applied to cosmetics, greatly improving the development and utilization of tomato callus extracts in the field of cosmetics.

[0008] In other embodiments of the present application, the water extract of tomato callus is added to the anti-aging cosmetic in the form of an aqueous solution of tomato callus extract.

[0009] In other embodiments of the present application, the preparation of the aqueous solution of tomato callus extract includes: dissolving the tomato callus extract in water to obtain an aqueous solution; the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml.

[0010] In other embodiments of the present application, the concentration of the tomato callus extract in the aqueous solution is 5 mg / ml to 20 mg / ml.

[0011] In other embodiments of the present application, the preparation of tomato callus extract comprises:

[0012] The hypocotyls of sterile tomato seedlings are cultured to induce callus tissue; the callus tissue is taken and cut into small pieces, and then subcultured on the original culture medium to obtain grown tomato callus tissue; the grown tomato callus tissue is subjected to water extraction under the action of a high-voltage pulse electric field. Optionally, the supernatant after extraction is fully dried using a freeze dryer to obtain a tomato callus tissue extract.

[0013] In other embodiments of the present application, the culture medium includes: 0.2 mg / L-1.0 mg / L indoleacetic acid; 1.0 mg / L-2.0 mg / L zeatin.

[0014] In other embodiments of the present application, the induction culture medium includes: 0.5 mg / L-1.0 mg / L indoleacetic acid; 1.5 mg / L-2.0 mg / L zeatin.

[0015] In other embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulse electric field, including:

[0016] The grown tomato callus tissue is mixed with water at a ratio of 1:(5-20) by mass, and then water extraction is performed under the action of a high-voltage pulse electric field.

[0017] In other embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulse electric field, including:

[0018] The mixture of grown tomato callus and water is extracted in a high voltage pulse electric field of 30 kV / m-50 kV / m.

[0019] In other embodiments of the present application, the number of extractions is 60-120 times.

[0020] In a second aspect, the present application provides a cosmetic comprising a tomato callus extract; optionally, the tomato callus extract is an aqueous extract of tomato callus.

[0021] In other embodiments of the present application, when the tomato callus extract is added to the cosmetic, it is added in the form of an aqueous solution of the tomato callus extract; optionally, the preparation of the aqueous solution of the tomato callus extract includes: dissolving the tomato callus extract in water to obtain an aqueous solution; the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml.

[0022] In other embodiments of the present application, the preparation of tomato callus extract comprises:

[0023] The hypocotyls of sterile tomato seedlings are cultured to induce callus tissue; the callus tissue is taken and cut into small pieces, and then subcultured on the original culture medium to obtain grown tomato callus tissue; the grown tomato callus tissue is subjected to water extraction under the action of a high-voltage pulse electric field; optionally, the supernatant after the water extraction is fully dried in a freeze dryer to obtain a tomato callus tissue extract. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0025] Figure 1 In vitro efficacy evaluation of tomato callus extract (biochemical method); (The concentration shown in the figure is in cosmetics);

[0026] Figure 2 In vitro efficacy evaluation of tomato callus extract (cell method); (the concentration shown in the figure is that in cosmetics);

[0027] Figure 3 The effect of tomato callus extract on cell scratch healing. (The figure shows the concentration in cosmetics). DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments.

[0029] Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present application.

[0030] The present application provides an application of a tomato callus extract in the preparation of anti-aging cosmetics.

[0031] Tomato callus extract is a water extract of tomato callus.

[0032] The present application adopts the aqueous extract of tomato callus tissue for the preparation of anti-aging cosmetics. The aqueous extract of tomato callus tissue can effectively reduce the concentration of inflammatory factors, effectively promote the healing of cell scratches, and effectively improve the scavenging rate of ABTS and DPPH free radicals; and the aqueous extract of tomato callus tissue can effectively improve the tyrosinase inhibition effect. Therefore, the application of the aqueous extract of tomato callus tissue in anti-aging cosmetics can achieve the effects of anti-inflammatory, repair, antioxidant and whitening, thereby achieving anti-aging effects from multiple aspects of anti-inflammatory, repair, antioxidant and whitening.

[0033] Furthermore, the development of water extracts of tomato callus in the prior art has not yet been seen in the field of cosmetics. The present application successfully extracts water extracts of tomato callus, and finds that they can effectively reduce the concentration of inflammatory factors, promote cell scratch healing, and increase the scavenging rate of ABTS and DPPH free radicals; improve the tyrosinase inhibition effect; and thus can be effectively applied to cosmetics, greatly improving the development and utilization of tomato callus extracts in the field of cosmetics.

[0034] Further, in some embodiments of the present application, the water extract of tomato callus is added to the anti-aging cosmetic in the form of an aqueous solution of tomato callus extract.

[0035] Further, in some embodiments of the present application, the preparation of the aqueous solution of the tomato callus extract comprises:

[0036] The tomato callus extract is dissolved in water to obtain an aqueous solution; the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml.

[0037] In the above technical scheme, a tomato callus extract is dissolved in water to obtain an aqueous solution; the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml; the concentration of inflammatory factors can be effectively reduced, cell scratch healing can be promoted, the ABTS and DPPH free radical scavenging rates can be increased, and the tyrosinase inhibition effect can be improved.

[0038] Illustratively, in some embodiments of the present application, the concentration of the tomato callus extract in the aqueous solution is 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 11 mg / ml, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml or a range between any two of the foregoing values.

[0039] Further optionally, in some embodiments of the present application, in the aqueous solution of tomato callus extract, the concentration of the aqueous solution of tomato callus extract is 5 mg / ml to 20 mg / ml.

[0040] Further, in some embodiments of the present application, the preparation of tomato callus extract comprises:

[0041] The hypocotyls of sterile tomato seedlings are cultured to induce callus tissue; the callus tissue is taken and cut into small pieces, and then subcultured on the original culture medium to obtain grown tomato callus tissue; the grown tomato callus tissue is subjected to water extraction under the action of a high-voltage pulse electric field.

[0042] In the above technical scheme, the hypocotyls of sterile tomato seedlings are cultured to induce the production of callus tissue; the production of callus tissue can be induced quickly with high induction efficiency; and through the above technical scheme, tomato callus tissue with high dedifferentiation degree, good looseness and fast growth rate can be induced.

[0043] Furthermore, in some embodiments of the present application, the hypocotyls of sterile tomato seedlings are cultured to induce the production of callus tissue using a culture medium.

[0044] Further optionally, in some embodiments of the present application, the main components of the above-mentioned culture medium include: indoleacetic acid 0.2 mg / L-1.0 mg / L; zeatin 1.0 mg / L-2.0 mg / L.

[0045] The inventors have found through research that when the indoleacetic acid concentration is 0.2 mg / L-1.0 mg / L and the zeatin concentration is 1.0 mg / L-2.0 mg / L within the above range, callus tissue can be quickly induced with high induction efficiency; and through the above technical scheme, tomato callus tissue with a high degree of dedifferentiation, good looseness and fast growth rate can be induced.

[0046] Illustratively, in some embodiments of the present application, the main components of the above-mentioned culture medium include: indoleacetic acid is 0.2 mg / L, 0.3 mg / L, 0.4 mg / L, 0.5 mg / L, 0.6 mg / L, 0.7 mg / L, 0.8 mg / L, 0.9 mg / L, 1.0 mg / L or the range between any two of the aforementioned values; zeatin is 1.0 mg / L, 1.1 mg / L, 1.2 mg / L, 1.3 mg / L, 1.4 mg / L, 1.5 mg / L, 1.6 mg / L, 1.7 mg / L, 1.8 mg / L, 1.9 mg / L, 2.0 mg / L or the range between any two of the aforementioned values.

[0047] Further optionally, in some embodiments of the present application, the induction medium comprises: 0.5 mg / L-1.0 mg / L indoleacetic acid; 1.5 mg / L-2.0 mg / L zeatin.

[0048] Furthermore, in some embodiments of the present application, the pH value of the above-mentioned culture medium is 5.8-5.9.

[0049] Further, in some embodiments of the present application, the preparation of the sterile tomato seedlings includes:

[0050] After the tomato seeds are disinfected with ethanol, the ethanol is removed, and then they are disinfected with sodium hypochlorite, rinsed with sterile water, and the rinsed tomato seeds are soaked in sterile water overnight. The next day, they are sown on a 1 / 2MS solid culture medium without any growth regulator, cultured in the dark, and switched to long-day culture after germination.

[0051] For example, in some embodiments of the present application, the preparation of sterile tomato seedlings includes: selecting full tomato seeds, surface sterilizing with 75% to 85% ethanol for 20s to 60s in a clean bench, washing away the ethanol with sterile water, then sterilizing with 1% to 10% sodium hypochlorite solution for 6min to 12min, and washing with sterile water 4-5 times. The rinsed tomato seeds are soaked in sterile water overnight, sown on a 1 / 2MS solid culture medium without any growth regulator the next day, cultured in the dark, and switched to long-day culture after germination.

[0052] Exemplarily, in some embodiments of the present application, the preparation of sterile tomato seedlings includes: selecting full tomato seeds, surface disinfecting with 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84% or 85% ethanol in a clean bench for 20s, 25s, 30s, 35s, 40s, 45s, 50s, 55s or 60s, washing away the ethanol with sterile water, and then disinfecting with 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% sodium hypochlorite solution for 6min, 7min, 8min, 9min, 10min, 11min or 12min, and rinsing with sterile water 4-5 times. The rinsed tomato seeds are soaked in sterile water overnight, sown on 1 / 2MS solid culture medium without any growth regulator the next day, cultured in the dark, and switched to long-day culture after germination.

[0053] Further optionally, in some embodiments of the present application, the above-mentioned long-day culture conditions are: temperature 20°C ~ 30°C, relative humidity 50% ~ 80%, light intensity 1000lux ~ 3000lux, lighting time 12h / d ~ 20h / d.

[0054] Illustratively, in some embodiments of the present application, the long-day culture conditions are: temperature 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C or 30°C, relative humidity 50%, 55%, 60%, 65%, 70%, 75% or 80%, light intensity 1000lux, 1500lux, 2000lux, 2500lux, 2800lux or 3000lux, light time 12h / d, 13h / d, 14h / d, 15h / d, 16h / d, 17h / d, 18h / d, 19h / d, or 20h / d.

[0055] Further, in some embodiments of the present application, the above-mentioned culturing of the hypocotyls of sterile tomato seedlings comprises:

[0056] The tomato sterile seedlings cultured for 7 to 14 days under long-day light are used as materials, and the hypocotyls thereof are cut into small segments of 0.5 cm to 1.0 cm as explants and inoculated on an induction medium, and then cultured in the dark for 3 days before switching to long-day light culture. The induction medium contains the following components: 4.5 g / L to 5 g / L MS powder, 25 g / L to 35 g / L sucrose, 1.0 mg / L to 2.0 mg / L zeatin (ZT), 0.2 mg / L to 1.0 mg / L indoleacetic acid (IAA), 80 mg / L to 150 mg / L inositol, and 7.0 g / L to 8.0 g / L agar; the pH value of the culture medium is 5.8 to 5.9.

[0057] Illustratively, in some embodiments of the present application, the above-mentioned culturing of the hypocotyls of sterile tomato seedlings comprises:

[0058] Tomato sterile seedlings cultured under long-day conditions for 7, 8, 9, 10, 11, 12, 13 or 14 days are used as materials, and their hypocotyls are cut into small sections of 0.5 cm, 0.6 cm, 0.7 cm, 0.8 cm, 0.9 cm or 1.0 cm as explants, which are inoculated on an induction medium and cultured in the dark for 3 days before being switched to long-day culture. The induction medium comprises the following components: 4.5 g / L, 4.6 g / L, 4.7 g / L, 4.8 g / L, 4.9 g / L or 5 g / L of MS powder; 25 g / L, 26 g / L, 27 g / L, 28 g / L, 29 g / L, 30 g / L, 31 g / L, 32 g / L, 33 g / L, 34 g / L or 35 g / L of sucrose; 1.0 mg / L, 1.1 mg / L, 1.2 mg / L, 1.3 mg / L, 1.4 mg / L, 1.5 mg / L, 1.6 mg / L, 1.7 mg / L, 1.8 mg / L, 1.9 mg / L or 2.0 mg / L of zeatin (ZT); 0.2 mg / L of indoleacetic acid (IAA) , 0.3mg / L, 0.4mg / L, 0.5mg / L, 0.6mg / L, 0.7mg / L, 0.8mg / L, 0.9mg / L or 1.0mg / L; inositol 80mg / L, 85mg / L, 90mg / L, 95mg / L, 100mg / L, 110mg / L, 120mg / L, 130mg / L, 140mg / L or 150mg / L and agar 7.0g / L, 7.1g / L, 7.2g / L, 7.3g / L, 7.4g / L, 7.5g / L, 7.6g / L, 7.7g / L, 7.8g / L, 7.9g / L or 8.0g / L; the pH value of the culture medium is 5.8-5.9.

[0059] Furthermore, in some embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulse electric field, comprising:

[0060] The grown tomato callus tissue is mixed with water at a ratio of 1:(5-20) by mass, and then water extraction is performed under the action of a high-voltage pulse electric field.

[0061] In the above technical scheme, the grown tomato callus tissue is mixed with water in a ratio of 1:(5-20) by mass ratio, and then water extraction is performed under the action of a high-voltage pulse electric field; the tomato callus tissue extract can be effectively extracted; and the extraction efficiency is higher, the time consumption is short, and the cost is low; at the same time, water extraction is used, there is no organic solvent residue, and the safety is high and the extraction efficiency is high.

[0062] For example, in some embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulsed electric field, comprising:

[0063] The grown tomato callus is mixed with water in a mass ratio of 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20 or a range between any two of the foregoing values, and then subjected to water extraction under the action of a high-voltage pulse electric field.

[0064] Further optionally, in some embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulse electric field, comprising:

[0065] The grown tomato callus tissue is mixed with water at a ratio of 1:(5-10) by mass, and then water extraction is performed under the action of a high-voltage pulse electric field.

[0066] Furthermore, in some embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulse electric field, comprising:

[0067] The mixture of grown tomato callus and water is extracted in a high voltage pulse electric field of 30 kV / m-50 kV / m.

[0068] In the above technical solution, more tomato callus extract can be obtained by extracting the mixture of grown tomato callus and water in a high-voltage pulse electric field of 30kV / m-50kV / m, and the extraction efficiency is higher.

[0069] For example, in some embodiments of the present application, the above-mentioned water extraction of the grown tomato callus under the action of a high-voltage pulse electric field comprises:

[0070] The mixture of grown tomato callus and water is extracted in a high voltage pulse electric field of 30 kV / m, 32 kV / m, 35 kV / m, 38 kV / m, 40 kV / m, 42 kV / m, 45 kV / m, 48 kV / m, 50 kV / m or a range between any two of the aforementioned values.

[0071] Furthermore, in some embodiments of the present application, the above-mentioned extraction times is 60-120 times.

[0072] Illustratively, in some embodiments of the present application, the above-mentioned extraction times are 60 times, 65 times, 70 times, 75 times, 80 times, 85 times, 90 times, 95 times, 100 times, 105 times, 110 times, 115 times or 120 times.

[0073] For example, in some embodiments of the present application, the grown tomato callus is subjected to water extraction under the action of a high-voltage pulse electric field, comprising:

[0074] Take the vigorously growing tomato callus and homogenize it. Use pure water as solvent, and the solid-liquid ratio is 1:(5-20). Use high-voltage pulse electric field (PEF) extraction technology to extract the tomato callus, the field strength of the high-voltage pulse electric field is 30-50kV / m, and the number of pulses of the high-voltage pulse electric field is 60-120 times. After the extraction is completed, high-speed centrifugation is performed at 6000-12000rpm (preferably 10000rpm) for 5-15min, and the supernatant is taken and fully dried in a freeze dryer to obtain the tomato callus extract.

[0075] In a second aspect, some embodiments of the present application provide a cosmetic comprising a tomato callus extract.

[0076] Further optionally, in some embodiments of the present application, the tomato callus extract is a water extract of tomato callus.

[0077] Further optionally, in some embodiments of the present application, when the tomato callus extract is added to the cosmetic, it is added in the form of an aqueous solution of the tomato callus extract; optionally, the preparation of the aqueous solution of the tomato callus extract includes: dissolving the tomato callus extract in water to obtain an aqueous solution; the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml.

[0078] Illustratively, in some embodiments of the present application, the concentration of the tomato callus extract in the aqueous solution is 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 11 mg / ml, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, 20 mg / ml or a range between any two of the foregoing values.

[0079] Further optionally, in some embodiments of the present application, the preparation of tomato callus extract comprises:

[0080] The hypocotyls of sterile tomato seedlings are cultured to induce callus tissue; the callus tissue is taken and cut into small pieces, and then subcultured on the original culture medium to obtain grown tomato callus tissue; the grown tomato callus tissue is subjected to water extraction under the action of a high-voltage pulse electric field.

[0081] Further optionally, in some embodiments of the present application, a freeze dryer is used to fully dry the supernatant after extraction to obtain a tomato callus extract.

[0082] Further optionally, in some embodiments of the present application, when the tomato callus extract is added to the cosmetics in the form of an aqueous solution of the tomato callus extract, the added amount is 1% to 20% by mass. Exemplarily, the added amount is 1%, 2%, 3%, 4%, 5%, 6%, 8%, 10%, 12%, 15%, 18%, 20% or a range between any two of the foregoing values ​​by mass.

[0083] The features and performance of the present application are further described in detail below in conjunction with the embodiments:

[0084] Example 1

[0085] Induction of tomato callus:

[0086] (1) Seed disinfection and sowing:

[0087] Select plump tomato seeds, disinfect the surface with 75% ethanol for 30 seconds in a clean bench, wash off the ethanol with sterile water, and then disinfect with 5% sodium hypochlorite solution for 8 minutes, and rinse with sterile water 5 times. Soak the rinsed tomato seeds in sterile water overnight, sow them on 1 / 2MS solid culture medium without any growth regulator the next day, culture in the dark, and switch to long-day culture after germination. Long-day culture conditions: temperature 25℃, relative humidity 60%, light intensity 2000lux, light time 16h / d.

[0088] (2) Callus induction:

[0089] The tomato sterile seedlings cultured for 14 days under long-day light were used as materials, and the hypocotyls were cut into 0.8 cm small segments as explants and inoculated on the induction medium. After 3 days of dark culture, they were switched to long-day culture. The induction medium contained the following components: MS powder 4.74 g / L, sucrose 30 g / L, zeatin (ZT) 1.0 mg / L, indoleacetic acid (IAA) 0.2 mg / L, inositol 100 mg / L and agar 7.5 g / L; the pH value of the culture medium was 5.8-5.9. See Table 1 for details.

[0090] Embodiment 2-8

[0091] The difference from Example 1 is that the composition of the induction medium is different, see Table 1 for details.

[0092] Table 1 Effects of different culture media on tomato callus formation

[0093]

[0094] The above table tests the formation of tomato callus under different hormone concentrations and ratios, indicating that the formation of tomato callus requires a higher concentration of plant growth regulators. From the growth of callus, increasing the concentration of IAA is conducive to inducing callus with loose texture and yellowish color. Considering the callus rate and callus growth, the culture medium of Example 8, i.e., 2.0 mg / L ZT + 1.0 mg / L IAA, is selected as the best culture medium for tomato callus induction.

[0095] The tomato callus tissue with good growth status induced was cut into small pieces (0.5 cm × 0.5 cm × 0.5 cm), inoculated on the culture medium provided in Example 8 and continued to be subcultured (10 / 50 cm 2 ), and the results showed that its proliferation status was good.

[0096] Example 9

[0097] The tomato callus with good growth status induced in Example 8 was inoculated on the culture medium of Example 8 and continued to be subcultured:

[0098] The tomato callus tissue with good growth status obtained by induction in the above Example 8 was cut into small pieces (0.5 cm×0.5 cm×0.5 cm), inoculated on the subculture medium for subculture, and the inoculation amount was 10 / 50 cm 2 The subculture cycle is 14 days.

[0099] Take the vigorously growing tomato callus and homogenize it. Use pure water as the solvent, and the solid-liquid ratio is 1:(5-20) (preferably 1:10). Use high-voltage pulsed electric field (PEF) extraction technology to extract the tomato callus. The field strength of the high-voltage pulsed electric field is 30-50kV / m (preferably 40kV / m), and the number of pulses of the high-voltage pulsed electric field is 60-120 times (preferably 100 times). After the extraction is completed, high-speed centrifugation is performed at 6000-12000rpm (preferably 10000rpm) for 5-15min (preferably 10min), and the supernatant is taken. See Table 2 for details.

[0100] Examples 10-16

[0101] The difference from Example 9 is that the extraction conditions are different, see Table 2 for details.

[0102] The DPPH removal rate of the tomato callus extract supernatant obtained in Examples 10-16 was tested, and the test method was as follows:

[0103] Referring to the content of standard T / GDCA021-2023 "Chemical Evaluation Method for Evaluation of Antioxidant Activity of Cosmetics", the DPPH radical scavenging rate of each embodiment and comparative example was calculated.

[0104] The test results are shown in Table 2.

[0105] Table 2 Effects of different extraction conditions on DPPH scavenging rate of tomato callus extract supernatant

[0106]

[0107]

[0108] It can be seen from the above table that: taking into account the DPPH removal rate, extraction time and extraction cost, Example 15 is selected as the optimal extraction condition, that is, pure water is used as the solvent, the solid-liquid ratio is 1:10, the field strength of the high-voltage pulse electric field is 40 kV / m, and the number of pulses is 100 times.

[0109] Application Example 1

[0110] Provided is an anti-aging cosmetic, which is prepared according to the following steps:

[0111] The tomato callus extract supernatant prepared in the above Example 15 was fully dried in a freeze dryer to obtain a tomato callus extract, and the tomato callus extract was used to prepare the following anti-aging cosmetics.

[0112] The tomato callus extract was dissolved in water to prepare an aqueous solution of the tomato callus extract, wherein the concentration of the prepared aqueous solution of the tomato callus extract was 5 mg / mL.

[0113] Then compound it into anti-aging cosmetics according to the following formula:

[0114] Provided is an anti-aging cosmetic, comprising:

[0115] 3% glycerol; 2% butylene glycol; 0.1% xanthan gum; 0.5% 1,2-hexanediol; 0.5% p-hydroxyacetophenone; 10% aqueous solution of tomato callus extract; and the balance water. The concentration of the aqueous solution of tomato callus extract is 5 mg / mL; see Table 3 for details. At this time, the concentration of tomato callus extract in the anti-aging cosmetic is 0.5 mg / mL.

[0116] Application Examples 2-3

[0117] The difference from Application Example 1 is that the concentration of the aqueous solution of tomato callus extract is different, see Table 3 for details.

[0118] Negative control group: water.

[0119] Table 3

[0120]

[0121]

Performance test

[0122] 1. The in vitro efficacy of each of the above application examples was evaluated to evaluate the in vitro efficacy of the tomato callus extract.

[0123] (1) The biochemical test method is as follows:

[0124] ① ABTS free radical scavenging test: Referring to the content of standard T / GDCA 021-2023 "Chemical Evaluation Method for Evaluation of Antioxidant Activity of Cosmetics", calculate the ABTS free radical scavenging rate of each application example and negative control.

[0125] ②DPPH free radical scavenging test: Referring to the content of standard T / GDCA 021-2023 "Chemical Evaluation Method for Evaluation of Antioxidant Activity of Cosmetics", calculate the DPPH free radical scavenging rate of each application example and negative control.

[0126] ③Tyrosinase inhibition test: Referring to the content of standard T / GDCA006-2021 "Test method for inhibition of tyrosinase activity by cosmetic raw materials (in vitro method)", calculate the tyrosinase inhibition rate of each application example and negative control.

[0127] ④Collagenase inhibition test

[0128] The substance produced by collagenase acting on FALGPA absorbs at 335nm. The activity of collagenase is investigated by monitoring the absorbance value at 335nm. Using FALGPA as a substrate, the collagenase inhibition effect of a single sample is determined by spectrophotometry. The experimental design is shown in Table 4 below:

[0129] Table 4

[0130]

[0131] Calculation of inhibition rate:

[0132]

[0133] Where:

[0134] A0: Absorbance value of negative control group (mean)

[0135] A1: Negative blank group absorbance value (mean)

[0136] B0: Absorbance value of sample reaction group

[0137] B1: Absorbance value of sample blank group

[0138] The test results are shown in the attached manual. Figure 1 Among them, Figure 1A shows the scavenging effect of tomato callus extract on ABTS free radicals; B shows the scavenging effect of tomato callus extract on DPPH free radicals; C shows the inhibitory effect of tomato callus extract on tyrosinase; D shows the inhibitory effect of tomato callus extract on collagenase. Multiple comparison analysis was performed using Tukey's method, and lowercase letters indicate significant differences (n=3; P<0.05).

[0139] From the instruction manual Figure 1 It can be seen that: under the experimental conditions, when the concentration of tomato callus extract in anti-aging cosmetics is 0.5 mg / mL, 1.0 mg / mL, and 2.0 mg / mL, its scavenging rate for ABTS and DPPH free radicals increases linearly, and the sample solutions at each concentration point have significant scavenging effects on ABTS and DPPH free radicals ( Figure 1 A, B). When the concentration of tomato callus extract in anti-aging cosmetics was 1.0 mg / mL and 2.0 mg / mL, it had a significant inhibitory effect on tyrosinase, and when the concentration of tomato callus extract in anti-aging cosmetics was 2.0 mg / mL, it had a significant inhibitory effect on collagenase ( Figure 1 C,D).

[0140] (2) The cell method test method is as follows:

[0141] BJ cells in the logarithmic growth phase were inoculated into a 96-well plate at a density of 2.5*10^5 cells / well and cultured in an incubator for 24 hours. The sample was diluted to the corresponding concentration with serum-free medium and added to a 96-well plate. Three replicate wells were set up, and a positive control group (10% FBS) and a blank control group (2% FBS) were also set up. The culture was continued for 24 hours, and the cell morphology and quantity were observed regularly under a microscope. After 48 hours, CCK-8 was diluted 10 times with serum-free medium, 100 μL of CCK-8 diluent was added to each well, and then incubated in a 37°C incubator for 0.5 hours. The absorbance value was detected with an enzyme marker at a wavelength of 450nm.

[0142]

[0143] Where:

[0144] OD 试验 : Absorbance value of test well or solvent control well

[0145] OD 空白 : Absorbance value of blank well

[0146] RAW 264.7 (mouse macrophages) cells in the logarithmic growth phase were inoculated into 24-well plates with an inoculation density of 4*10^5 cells / well and cultured in an incubator for 24 hours. After the cells adhered to the wall, the culture medium was aspirated and 0.5 mL of the following samples were added: ① Blank control group: 2% serum cultured cells for 24 hours; ② Positive control group: 0.2 mg / mL dexamethasone + 2 μg / mL LPS acted on cells for 24 hours; ③ LPS inflammation model group: 1 μg / mL LPS continuously stimulated RAW 264.7 cells for 24 hours; ④ Sample group: Dilute the sample to the corresponding concentration solution with 2 μg / mL LPS to act on cells for 24 hours. Then use a pipette to aspirate the supernatant in the well plate into a centrifuge tube, centrifuge at 1000r / min for 5 minutes, discard the precipitate and aspirate the supernatant, dilute the supernatant 50 times, and use the TNF-ɑ inflammatory factor detection kit to detect the change in the amount of inflammatory factors in the supernatant. The detection method is strictly in accordance with the instructions on the detection kit.

[0147] The test results are shown in the attached manual. Figure 2 Among them, Figure 2 Figure A shows the effect of tomato callus extract on the survival rate of human skin fibroblasts (BJ cells); Figure 2 B in the figure shows the effect of tomato callus extract on inflammatory factor TNF-ɑ. Multiple comparison analysis was performed using Tukey's method, and lowercase letters indicate significant differences (n=3; P<0.05).

[0148] From the instruction manual Figure 2 It can be seen that: Cytotoxicity test ( Figure 2 A) showed that when the concentration of tomato callus extract in anti-aging cosmetics was 0.5mg / mL and 1.0mg / mL, it was safe for human skin fibroblasts (BJ cells) (cell survival rate was >100%), and when the concentration of tomato callus extract in anti-aging cosmetics was 1.0mg / mL, it significantly promoted the survival rate of BJ cells. Cell anti-inflammatory test showed that compared with the inflammatory model group, when the concentration of tomato callus extract in anti-aging cosmetics was 1.0mg / mL, the concentration of inflammatory factor TNF-ɑ was significantly reduced after treatment, indicating that it has anti-inflammatory effect ( Figure 2 B).

[0149] (3) Effects of tomato callus extract on cell scratch healing

[0150] The cell scratch healing test method is as follows:

[0151] BJ cells in the logarithmic growth phase were inoculated into a 24-well plate at a density of 2.5*10^5 cells / well. They were placed in a 37°C carbon dioxide incubator for 24 hours. When the cells were almost fully grown, a cross was made in the 24-well plate with a pipette tip, and then the plate was washed once with PBS. The sample was diluted with culture medium to the corresponding concentration and added to the wells, 500 μL was added to each well, and the plate was placed in a 37°C carbon dioxide incubator for the observation time set in the experiment. At the set time, the width of the scratch was observed and measured under a microscope and photographed.

[0152]

[0153] The test results are shown in the attached manual. Figure 3 Among them, Figure 3 Figure A shows the effect of tomato callus extract on the healing rate of BJ cells; Figure 3 B in the figure shows the comparison of the scratch of negative control cells at 0h (upper) and 24h (lower); Figure 3 C shows a comparison of cells scratched with 10% tomato callus extract at 0 h (upper) and 24 h (lower). Multiple comparison analysis was performed using Tukey's method, and lowercase letters indicate significant differences (n=3; P<0.05).

[0154] From the instruction manual Figure 3 It can be seen that when the concentration of tomato callus extract in anti-aging cosmetics is 5 mg / mL and 10 mg / mL, after 24 hours of action on cells, the cell scratch healing rates are 56.05% and 65.61%, respectively, which are significantly higher than the blank control group (48.11%), indicating that the extract at this concentration has a significant ability to promote the healing of BJ cells ( Figure 3 ).

[0155] In summary, the present application adopts the aqueous extract of tomato callus tissue for the preparation of anti-aging cosmetics. The aqueous extract of tomato callus tissue can effectively reduce the concentration of inflammatory factors, effectively promote the healing of cell scratches, and effectively improve the scavenging rate of ABTS and DPPH free radicals; and the aqueous extract of tomato callus tissue can effectively improve the tyrosinase inhibition effect. Therefore, the application of the aqueous extract of tomato callus tissue in anti-aging cosmetics can achieve the anti-inflammatory, repairing, antioxidant and whitening effects, thereby achieving anti-aging effects from multiple aspects of anti-inflammatory, repairing, antioxidant and whitening.

[0156] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A use of a tomato callus extract in the preparation of anti-aging cosmetics, characterized in that: The tomato callus extract is a water extract of tomato callus.

2. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 1, characterized in that: The tomato callus extract is added to the anti-aging cosmetic in the form of an aqueous solution of the tomato callus extract.

3. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 2, characterized in that: The preparation of the aqueous solution of the tomato callus extract comprises: The tomato callus extract is dissolved in water to obtain an aqueous solution; the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml.

4. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 3, characterized in that: The concentration of the tomato callus extract in the aqueous solution is 5 mg / ml to 20 mg / ml.

5. Use of the tomato callus extract according to any one of claims 1 to 4 in the preparation of anti-aging cosmetics, characterized in that: The preparation of the tomato callus extract comprises: The hypocotyls of sterile tomato seedlings are cultured to induce callus tissue; the callus tissue is taken and cut into small pieces, and then subcultured on the original culture medium to obtain grown tomato callus tissue; the grown tomato callus tissue is subjected to water extraction under the action of a high-voltage pulse electric field; Optionally, the supernatant after extraction is fully dried using a freeze dryer to obtain a tomato callus extract.

6. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 5, characterized in that: The culture medium comprises: 0.2 mg / L-1.0 mg / L indoleacetic acid; 1.0 mg / L-2.0 mg / L zeatin.

7. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 5, characterized in that: The culture medium comprises: 0.5 mg / L-1.0 mg / L indoleacetic acid; 1.5 mg / L-2.0 mg / L zeatin.

8. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 5, characterized in that: The method of extracting the grown tomato callus tissue by water under the action of a high-voltage pulse electric field comprises: The grown tomato callus tissue is mixed with water at a ratio of 1:(5-20) by mass, and then water extraction is performed under the action of a high-voltage pulse electric field.

9. The use of the tomato callus extract in the preparation of anti-aging cosmetics according to claim 5, characterized in that: The method of extracting the grown tomato callus tissue by water under the action of a high-voltage pulse electric field comprises: Extracting the mixture of the grown tomato callus and water in a high voltage pulse electric field of 30 kV / m-50 kV / m; Optionally, the number of extractions is 60-120 times.

10. A cosmetic, characterized in that: The cosmetic comprises a tomato callus extract; optionally, the tomato callus extract is a water extract of tomato callus; Optionally, when the tomato callus extract is added to the cosmetic, it is added in the form of an aqueous solution of the tomato callus extract; Optionally, the preparation of the aqueous solution of the tomato callus extract comprises: Dissolving the tomato callus extract in water to obtain an aqueous solution; wherein the concentration of the tomato callus extract in the aqueous solution is greater than 5 mg / ml; Optionally, the preparation of the tomato callus extract comprises: The hypocotyls of sterile tomato seedlings are cultured to induce callus tissue; the callus tissue is taken and cut into small pieces, and then subcultured on the original culture medium to obtain grown tomato callus tissue; the grown tomato callus tissue is subjected to water extraction under the action of a high-voltage pulse electric field; optionally, the supernatant after the water extraction is fully dried in a freeze dryer to obtain a tomato callus tissue extract.

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