Emulsified system capable of stably carrying ergothioneine and preparation and application thereof

Abstract

The present application provides an emulsified system capable of stably carrying ergothioneine, which comprises an aqueous phase composed of a polyacrylic polymer and an aqueous carrier, an oil phase composed of cetylstearyl alcohol, cetearyl glucoside and isopropyl palmitate, and triethanolamine, wherein the weight ratio of the aqueous phase, the oil phase and triethanolamine is 80-95:8-10:0.01-0.5, and the content of the triethanolamine in the emulsified system is at least 0.1wt%. The present application also discloses the preparation of the emulsified system and the application of the emulsified system in skin external agents.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of fine chemicals and cosmetics, in particular to an emulsified system capable of stably carrying ergothioneine and its preparation and application. BACKGROUND

[0002] Ergothioneine (EGT) is a natural amino acid that exists in many plants and animals and is abundant in content. It cannot be synthesized by the animal body itself and can only be obtained from food. It is a rare amino acid. Ergothioneine plays a unique role in the synthesis of cells in the body and has strong antioxidant properties. With the continuous deepening of the research on antioxidants, ergothioneine, as a natural bioactive substance, has begun to attract widespread attention. There have been some related reports abroad, while domestic research is still in the exploratory stage.

[0003] Ergothioneine, whose scientific name is 2-mercapto-L-histidine trimethyl inner salt, was first isolated from ergot (a sclerotium of a fungus that parasitizes on the plant Avena sterilis) by Tanret C in 1909. Subsequently, ergothioneine was also found in animal blood, which aroused great interest. Since then, ergothioneine has been found in animal semen and various tissues, especially in the liver and lungs. Further studies have shown that many microorganisms such as fungi and actinomycetes can synthesize ergothioneine, but bacteria cannot synthesize this compound. Among edible fungi, the two varieties of brown double-spore mushroom, Crimini mushroom when the cap is not opened, and Portabella mushroom after the cap is opened, have the highest content of ergothioneine, followed by shiitake mushroom, and delicious boletus, volvariella and other edible fungi can produce the precursor histidine trimethyl inner salt required for the synthesis of ergothioneine. Ergothioneine is a white crystal with good water solubility, and the solubility limit at room temperature is 0.9 mol / L. It is quite stable under strong alkaline conditions. It is an excellent divalent metal chelator, especially for copper and zinc. These special properties make it significantly different from other biological thiols.

[0004] Current studies have shown that ergothioneine has many physiological functions such as free radical scavenging, anti-inflammatory, maintaining DNA biosynthesis, normal cell growth, and cell immunity.

[0005] In the prior art, for example, Chinese Patent Application CN115944548 discloses an oil control composition for improving the light stability of ergothioneine and its application. The oil control composition combines ergothioneine with glucosyl rutin and extract of leucogene eriocaulon to significantly improve the light stability of ergothioneine. The composition has good oil control efficacy and can be used to prepare cosmetics with oil control efficacy. The mass ratio of the three components is ergothioneine: glucosyl rutin: extract of leucogene eriocaulon = 1:2:17.

[0006] For example, Chinese patent application CN114681451 discloses a use of hyaluronic acid or a salt thereof and / or trehalose in stabilizing ergothioneine, an additive for stabilizing ergothioneine, and a composition. The additive for stabilizing ergothioneine comprises hyaluronic acid or a salt thereof and trehalose, and the mass ratio of hyaluronic acid or a salt thereof to trehalose is 1:19-1:10. The molecular weight of hyaluronic acid or a salt thereof is 3KDa-30KDa, preferably 3KDa-10KDa.

[0007] For example, Chinese patent application CN115363980 discloses a composition containing ergothioneine and persimmon fruit extract, which contains 0.005-10% ergothioneine and 0.05-5% persimmon (DIOSPYROS KAKI) fruit extract. In view of the odor generated by ergothioneine during application, it is found that the persimmon (DIOSPYROS KAKI) fruit extract can stabilize ergothioneine, reduce the generation of odor, expand its application, and provide a better experience for consumers; in the case of using a pH regulator at the same time, the deodorizing effect is more obvious. In addition, the persimmon (DIOSPYROS KAKI) fruit extract has anti-wrinkle effect and can synergistically resist aging with ergothioneine.

[0008] The above prior art uses a single component or composition to investigate its effect on the light stability of ergothioneine, and has made considerable progress. The stability of ergothioneine in the application formula system has not been studied in the prior art.

[0009] The present application unexpectedly found that the use of a specific emulsion system not only stabilizes ergothioneine, but also greatly breaks through the stable expression of ergothioneine content, and further quantifies the contribution of the emulsion system to the stability of ergothioneine. SUMMARY

[0010] The purpose of the present application is to provide an emulsion for stabilizing the content of ergothioneine, which is prepared according to the provided formula and preparation method, and can not only stabilize the stability of ergothioneine under light, but also ensure the active content of ergothioneine.

[0011] The present application provides an emulsion system capable of stabilizing ergothioneine, which comprises an aqueous phase composed of a polyacrylic acid polymer and an aqueous carrier, an oil phase composed of cetylstearyl alcohol, hexadecyl / octadecyl glycoside and isopropyl palmitate, and triethanolamine, wherein the weight ratio of the aqueous phase, the oil phase and triethanolamine is 80-95:8-10:0.01-0.5, and the content of triethanolamine in the emulsion system is at least 0.1wt%.

[0012] In a preferred embodiment, the weight ratio of the water phase, the oil phase and the triethanolamine is 80.39-90.95:8-9:0.01-0.4;

[0013] In a preferred embodiment, the content of the triethanolamine in the emulsifying system is 0.1-0.5wt%.

[0014] In a preferred embodiment, the weight ratio of the water phase, the oil phase and the triethanolamine is 80.39-90.95:8-9:0.01-0.1;

[0015] In a preferred embodiment, the content of the triethanolamine in the emulsifying system is 0.1-0.4wt%.

[0016] In a preferred embodiment, the weight ratio of the polyacrylic polymer and the aqueous carrier is 0.1-0.5:80.29-90.45.

[0017] Preferably, the polyacrylic polymer is polyacrylic acid-30 polymer or polyacrylic acid polymer TR-1.

[0018] Preferably, the aqueous carrier is selected from at least one of deionized water, small molecule water, glacial water.

[0019] In a preferred embodiment, the weight ratio of the cetyl stearyl alcohol, hexadecyl / octadecyl glycoside and isopropyl palmitate is 1-2:2:4-6.

[0020] In a preferred embodiment, the emulsifying system further comprises a PH adjuster.

[0021] Preferably, the PH adjuster is citric acid or tromethamine.

[0022] In a preferred embodiment, the PH value of the emulsifying system is 4-8.

[0023] In a preferred embodiment, the emulsifying system can stably carry 0.01wt%-5wt% of ergothioneine.

[0024] Preferably, the emulsifying system can stably carry 0.01wt%-2wt% of ergothioneine.

[0025] In a preferred embodiment, the viscosity of the emulsifying system is 9000-60000 m.pas.

[0026] Preferably, the viscosity of the emulsifying system is 9600-56000 m.pas.

[0027] The present application also provides a method for preparing the emulsifying system, the method comprising the following steps:

[0028] 1) Take 0.1-0.5 parts by mass of polyacrylic acid-30 polymer and 50-80 parts by mass of deionized water, heat to 80-85℃ and mix and stir until completely dissolved to obtain an aqueous phase;

[0029] 2) Take 1-2 parts by mass of cetyl stearyl alcohol and 1-2 parts by mass of hexadecyl / octadecyl glycoside and add to 4-6 parts by mass of isopropyl palmitate, heat to 80-85℃ until completely melted to obtain an oil phase;

[0030] 3) At 80-85℃, slowly add the oil phase of step 2) to the aqueous phase of step 1) and homogenize until uniform and free of particles; after the temperature is reduced to 55-60℃, add 0.1-0.4 parts by mass of a 10wt% aqueous solution of triethanolamine to neutralize and stir to react, then add 0.01-2 parts by mass of ergothioneine and citric acid or tromethamine to adjust the pH to 4-8, and add deionized water to make up to 100 parts by mass, and continue to stir until uniform.

[0031] In a preferred embodiment, the stirring rate in step 1) is 100-500 rpm; the homogenization conditions in step 3) are 2500-3000 rpm for 10-15 min.

[0032] The present application also provides the use of the emulsion system in skin external agents.

[0033] In a preferred embodiment, the skin external agent can be used in the field of cosmetics.

[0034] In a preferred embodiment, the skin external agent is selected from the group consisting of emulsions, creams, masks, etc.

[0035] The present application has the following advantages:

[0036] The present application forms an emulsion system by optimizing the combination of an aqueous phase composed of a polyacrylic acid polymer and an aqueous carrier, an oil phase composed of cetyl stearyl alcohol, hexadecyl / octadecyl glycoside and isopropyl palmitate, and triethanolamine. This system not only can stably carry 0.1-5wt% ergothioneine, but also has moderate viscosity (e.g. 9000-60000 m.pas), and can successfully pass the high and low temperature stability tests described in the test examples. In addition, by testing the ergothioneine content in the emulsion containing ergothioneine after the high and low temperature stability test, it is found that the ergothioneine content in the system is ≥90%, which indicates that the emulsion system containing ergothioneine described in the present application can successfully stably carry an effective amount of ergothioneine, i.e. the stability of the emulsion system to ergothioneine fully meets the application requirements in the field of skin external agents.

[0037] In addition, the present application finds that using a specific emulsifying system can not only stabilize the carrying of an effective amount of ergothioneine, but also greatly break through the stable expression of the content of ergothioneine, and further quantify the contribution of the emulsifying system to the stability of ergothioneine. DETAILED DESCRIPTION

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. For the purposes of the present application, the following terms are defined below.

[0039] For the sake of providing a more concise description, some quantitative expressions given herein are not modified with the term "about". It should be understood that, whether or not the term "about" is expressly used, each quantity given herein is intended to refer to the actual given value, and is also intended to refer to the approximation that would be made by one of ordinary skill in the art given the experimental and / or measurement conditions, including the approximation that would be made by one of ordinary skill in the art to account for the inherent inexactitude in experimentally and / or measurably determining values.

[0040] For the sake of providing a more concise description, some quantitative expressions given herein are recited as a range from about X amount to about Y amount. It should be understood that when a range is recited, the range is not limited to the recited upper and lower values, but is intended to include the entire range of values between about X amount and about Y amount, or any amount therebetween.

[0041] Rotational speed

[0042] The rotational speed described in the present application has the unit of rpm or revolutions per minute, both of which have the same meaning.

[0043] Polyacrylic polymers

[0044] The polyacrylic polymer described in the present application is a polymer containing acrylic acid (ester) modified groups, for example, the polyacrylic polymer is polyacrylic acid-30 polymer, polyacrylic acid polymer TR-1, etc.

[0045] Aqueous carrier

[0046] The aqueous carrier described in the present application is selected from at least one of deionized water, small molecule water, glacial water.

[0047] Hereinafter, the technical solutions of the present application will be described in detail through preferred embodiments, but the scope of the present application is not limited to these embodiments, which are intended to illustrate the technical solutions of the present application rather than limit the scope of the present application. The test methods in the following examples are not specified, which are usually carried out according to the conventional conditions, or according to the conditions recommended by the manufacturers. Unless otherwise specified, all percentages and parts are by weight.

[0048] The experimental materials used in the embodiments of the present application are as follows:

[0049] Cetyl Stearyl Alcohol, trade name Emercol C16-18 30:70 MY, purchased from Emery Oleochemicals (M) Sdn Bhd

[0050] Ergothioneine, trade name Ergothioneine, purchased from Shanghai Ergothioneine Biotech Group Co., Ltd.

[0051] Isopropyl Palmitate, trade name Isopropyl Palmitate 1517, purchased from Torkem Palmitate (Zhangjiagang) Co., Ltd.

[0052] Cetearyl Glucoside, trade name Montanov 68 MB, purchased from SEPPIC S.A.

[0053] Polyacrylic Acid-30 Polymer, trade name STABYLEN 30, purchased from 3V SIGMA S.p.A.

[0054] Triethanolamine, trade name Triethanolamine 99%, purchased from Dow Chemical (Shanghai) Co., Ltd.

[0055] Acrylates / Dimethicone Copolymer, trade name Aristoflex AVC, purchased from Clariant Chemicals (Shanghai) Co., Ltd.

[0056] Sodium Hyaluronate, trade name Hybloom™ Sodium Hyaluronate (HA-TG), purchased from Huaxi Biotech Co., Ltd.

[0057] Tromethamine, trade name TRIS AMINO™ ULTRA PC, Tromethamine, purchased from Yalu Chemical Trading (Shanghai) Co., Ltd.

[0058] Citric Acid, trade name Citric Acid Monohydrate, purchased from Shandong Yingxuan Industry Co., Ltd.

[0059] White Oil 26#, trade name Mineral Oil Oliba 13C, purchased from Tianjin Kaiwei Yongli Joint Chemical Co., Ltd.

[0060] Xanthan Gum, trade name GRINDSTED ® Xanthan 200, purchased from Danisco France SAS Usine de Melle

[0061] PEG-20 methyl glucose sesqui-stearate, trade name Glucamate™ SSE-20 emulsifier, purchased from Lubrizol Advanced Materials, Inc.

[0062] Deionized water, self-made

[0063] Formic acid, analytical pure, purchased from Sinopharm Chemical Reagent Co., Ltd.

[0064] Methanol, chromatographically pure, purchased from Merck KGAA

[0065] Acetonitrile, chromatographically pure, purchased from Sinopharm Chemical Reagent Co., Ltd.

[0066] High performance liquid chromatograph, Agilent

[0067] Constant temperature box, Friocell

[0068] Example 1: Preparation of a gel containing ergothioneine

[0069] 0.1 parts by mass of ergothioneine and 90 parts by mass of deionized water were mixed and dissolved, and mixed and stirred at a rotation speed of 100 rpm at room temperature until completely dissolved; 0.8 parts by mass of dimethylammonium propylene glycol taurate / VP copolymer was added, and homogenized at 2500-3000 rpm at room temperature for 5-6 minutes until uniform and free of particles; citric acid or tromethamine was added to adjust the pH value to 6, deionized water was added to make up to 100, and mixed and stirred at a rotation speed of 100 rpm for 10-20 minutes; then, it was left to stand at room temperature for 12 hours, and was ready for use.

[0070] Example 2: Preparation of a gel containing ergothioneine

[0071] 0.1 parts by mass of ergothioneine and 90 parts by mass of deionized water were mixed and dissolved, and mixed and stirred at a rotation speed of 100 rpm at room temperature until completely dissolved; 0.8 parts by mass of dimethylammonium propylene glycol taurate / VP copolymer was added, and homogenized at 2500-3000 rpm at room temperature for 5-6 minutes until uniform and free of particles; citric acid or tromethamine was added to adjust the pH value to 4, deionized water was added to make up to 100, and mixed and stirred at a rotation speed of 100 rpm for 10-20 minutes; then, it was left to stand at room temperature for 12 hours, and was ready for use.

[0072] Example 3: Preparation of a gel containing ergothioneine

[0073] Take 0.1 parts by mass of ergothioneine and 90 parts by mass of deionized water to mix and dissolve, mix and stir at room temperature at a rotation speed of 100 rpm until completely dissolved; add 0.8 parts by mass of dimethyl ammonium taurate / acrylic acid / VP copolymer, homogenize at room temperature at 2500-3000 rpm, after homogenizing for 5-6 minutes, it is uniform and no particles; add citric acid or tromethamine to adjust the pH value to 8, add deionized water to make up to 100, continue to mix and stir at a rotation speed of 100 rpm for 10-20 minutes; then stand at room temperature for 12 hours, ready for use.

[0074] Example 4: Preparation of gel containing ergothioneine

[0075] Take 0.01 parts by mass of ergothioneine and 90 parts by mass of deionized water to mix and dissolve, mix and stir at room temperature at a rotation speed of 100 rpm until completely dissolved; add 0.8 parts by mass of dimethyl ammonium taurate / acrylic acid / VP copolymer, homogenize at room temperature at 2500-3000 rpm, after homogenizing for 5-6 minutes, it is uniform and no particles; add citric acid or tromethamine to adjust the pH value to 6, add deionized water to make up to 100, continue to mix and stir at a rotation speed of 100 rpm for 10-20 minutes; then stand at room temperature for 12 hours, ready for use.

[0076] Test Example 1: Viscosity test and high and low temperature stability test

[0077] Test sample: After measuring the viscosity of the gel containing ergothioneine prepared in Examples 1-4, the high and low temperature stability was investigated.

[0078] Use BROOKFIELD RVDVC digital viscometer (with RV type, T type rotor), sample test temperature: 25℃, constant temperature 1h, sample test container: try to use the same specification beaker (180ml bottle) when testing different batches of samples; reading recording time: LV / RV type rotor 60s; T type rotor 90s; Before sample determination, confirm that there is no bubble interference. According to the standard, select the corresponding rotor and rotation speed. After taking the sample out of the constant temperature tank, immediately measure its viscosity with the rotary viscometer. When selecting LV and RV rotors, insert the rotor obliquely into the sample, so that the liquid surface is just in the recess of the rotor, and keep the rotor wall free of bubbles; when selecting T type rotor, make the rotor just close to the liquid surface, and select a certain up and down movement distance, so that the rotor has enough up and down space and will not damage the beaker bottom. After the reading is stable (or after the standard time), record the viscosity result.

[0079] The high and low temperature stability evaluation method is as follows: place the gel containing ergothioneine prepared in Examples 1-4 in a constant temperature oven with high temperature (48℃), room temperature (25℃) and low temperature (4℃) stability, and observe for 30 days. Observe whether there is particle solid precipitation, jelly, delamination, etc.

[0080] If the sample is normal without any change before and after a certain time within 30 days, it is indicated as "pass", the appearance of particles is indicated as "precipitation", the solidification of the material body is indicated as "jelly", and the appearance of two layers of liquid is indicated as "layering". The results are shown in the following table.

[0081] Conclusion: The gel containing ergothioneine is prepared, and the stability of ergothioneine in the gel thickening system with different pH values is investigated.

[0082] From the above experimental results, it can be seen that the gel containing ergothioneine has a moderate viscosity (e.g. 9000-60000 m.pas), which meets the viscosity range of consumers using the gel dosage form, and the dosage form performs well in terms of high and low temperature stability.

[0083] Test Example 2: Ergothioneine Content Test

[0084] Test sample: The gel containing ergothioneine prepared according to Examples 1-4 after passing the high and low temperature stability test in Test Example 1

[0085] Sample extraction: The sample is directly taken and dissolved in a 0.1 wt% formic acid methanol solution after filtering the membrane, and then measured.

[0086] Sample test: High performance liquid chromatograph (with ultraviolet detector), high performance liquid chromatograph conditions: chromatograph column:

[0087] HILIC chromatograph column (2.1 mm x 100 mm, 1.7 um); mobile phase: 0.1% formic acid acetonitrile solution-0.1% formic acid aqueous solution (88+12, V / V); flow rate: 0.4 Ml / min; column temperature: 40°C; wavelength: 262 nm; injection volume: 3 microliters.

[0088] A series of standard samples with ergothioneine concentrations of 1.0 mg / L, 2.0 mg / L, 5.0 mg / L, 10.0 mg / L, 20.0 mg / L, and 50.0 mg / L are tested according to the above chromatograph conditions. The standard curve of ergothioneine is drawn with the mass concentration of the standard working solution as the abscissa and the peak area as the ordinate. Qualitative analysis is performed according to the retention time, and the relative deviation of the retention time of the sample and the standard sample with similar concentration is not more than 2%, and quantitative analysis is performed by external standard method.

[0089] The response value of ergothioneine in the test sample solution should be within the range of the standard curve. If it exceeds the range of 1.0 mg / L-50.0 mg / L of the standard working curve, it should be diluted and analyzed again.

[0090] Recovery % = ergothioneine test content / ergothioneine added amount * 100%

[0091]

[0092] Conclusion: Prepare the gel containing ergothioneine, and investigate the effective content of ergothioneine in the gel thickening system of different PH values after light treatment. Considering the test deviation and addition loss, it is generally considered that the effective concentration of active substances is more than 90%, and less than 90% is considered to have the risk of decomposition and deterioration.

[0093] The gel containing ergothioneine prepared by examples 1-4 passed the high and low temperature stability test described in test example 1, that is, after the high and low temperature stability test, the content of ergothioneine in the gel containing ergothioneine prepared by example 1 decreased to 0.068 mass parts, and 68% of ergothioneine could be effectively recovered relative to the original 0.1 mass parts of the added amount; the content of ergothioneine in the gel containing ergothioneine prepared by example 2 decreased to 0.071 mass parts, and 71% of ergothioneine could be effectively recovered relative to the original 0.1 mass parts of the added amount; the content of ergothioneine in the gel containing ergothioneine prepared by example 3 decreased to 0.067 mass parts, and 67% of ergothioneine could be effectively recovered relative to the original 0.1 mass parts of the added amount; the content of ergothioneine in the gel containing ergothioneine prepared by example 4 decreased to 0.0063 mass parts, and 63% of ergothioneine could be effectively recovered relative to the original 0.01 mass parts of the added amount; after one month of high and low temperature stability test, the test content of ergothioneine in the gel containing ergothioneine prepared by examples 1-4 with different PH values was less than 90% of the added amount, indicating that ergothioneine was unstable in the gel system of examples 1-4, and the content and efficacy were decreased due to photodecomposition, thereby indicating that ergothioneine could not be stabilized in the gel system.

[0094] Example 5: Preparation of gel containing ergothioneine

[0095] 0.1 mass parts of ergothioneine and 90 mass parts of deionized water were mixed and dissolved, and mixed and stirred at room temperature at a rotation speed of 100 rpm until completely dissolved; 0.8 mass parts of dimethyl ammonium acrylate taurate / VP copolymer and 0.1% sodium hyaluronate were added, and homogenized at 2500-3000 revolutions per minute at room temperature for 5-6 minutes until uniform and no particles; citric acid or tromethamine was added to adjust the PH value to 6, deionized water was added to make up to 100, and mixed and stirred at a rotation speed of 100 rpm for 10-20 minutes; then it was placed at room temperature for 12 hours and ready for use.

[0096] Example 6: Preparation of a gel containing ergothioneine

[0097] 0.1 parts by mass of ergothioneine and 90 parts by mass of deionized water were mixed and dissolved, and mixed and stirred at room temperature at 100 rpm until completely dissolved; 0.8 parts by mass of dimethylammonium propylene glycol taurate / VP copolymer and 0.1% sodium hyaluronate were added, and homogenized at 2500-3000 rpm at room temperature for 5-6 minutes until uniform and free of particles; citric acid or tromethamine was added to adjust the pH to 4, deionized water was added to make up to 100, and mixed and stirred at 100 rpm for 10-20 minutes; and then left to stand at room temperature for 12 hours.

[0098] Example 7: Preparation of a gel containing ergothioneine

[0099] 0.1 parts by mass of ergothioneine and 90 parts by mass of deionized water were mixed and dissolved, and mixed and stirred at room temperature at 100 rpm until completely dissolved; 0.8 parts by mass of dimethylammonium propylene glycol taurate / VP copolymer and 0.1% sodium hyaluronate were added, and homogenized at 2500-3000 rpm at room temperature for 5-6 minutes until uniform and free of particles; citric acid or tromethamine was added to adjust the pH to 8, deionized water was added to make up to 100, and mixed and stirred at 100 rpm for 10-20 minutes; and then left to stand at room temperature for 12 hours.

[0100] Example 8: Preparation of a gel containing ergothioneine

[0101] 0.01 parts by mass of ergothioneine and 90 parts by mass of deionized water were mixed and dissolved, and mixed and stirred at room temperature at 100 rpm until completely dissolved; 0.8 parts by mass of dimethylammonium propylene glycol taurate / VP copolymer and 0.1% sodium hyaluronate were added, and homogenized at 2500-3000 rpm at room temperature for 5-6 minutes until uniform and free of particles; citric acid or tromethamine was added to adjust the pH to 6, deionized water was added to make up to 100, and mixed and stirred at 100 rpm for 10-20 minutes; and then left to stand at room temperature for 12 hours.

[0102] Test Example 3: Viscosity test and high and low temperature stability test

[0103] Test sample: The system viscosity of the gel containing ergothioneine prepared in Examples 5-8 was measured, and high and low temperature stability was investigated.

[0104] Using BROOKFIELD RVDVC digital viscometer (attached RV type, T type rotor), sample test temperature: 25℃, constant temperature 1h, sample test container: the same specification beaker (180ml bottle) as possible when testing different batches of samples; reading record time: LV / RV type rotor 60s; T type rotor 90s; The sample should be confirmed without bubble interference before determination. According to the standard, select the corresponding rotor and speed. After taking the sample out of the constant temperature tank, immediately measure its viscosity with the rotary viscometer. When selecting LV and RV rotors, insert the rotor obliquely into the sample, so that the liquid surface is just in the recess of the rotor, and keep the rotor wall without bubbles; when selecting T type rotor, make the rotor just close to the liquid surface, and select a certain up and down movement distance, so that the rotor has enough up and down space and will not damage the beaker bottom. Record the viscosity result after the reading is stable (or after the standard time).

[0105] The high and low temperature stability evaluation method is as follows: the ergot alkaloids-containing gel prepared in examples 5-8 is placed in a constant temperature box at high temperature (48℃), room temperature (25℃) and low temperature (4℃) for 30 days.

[0106] Observe whether the sample has particle solid precipitation, jelly, delamination and other conditions. If the sample has no change before and after a certain time within 30 days, it is indicated that "pass", the appearance of particles indicates "precipitation", the solidification of the material body indicates "jelly", and the appearance of upper and lower layers of liquid indicates "delamination". The results are shown in the following table.

[0107]

[0108] Conclusion: The ergot alkaloids-containing gel is prepared, and the stability of ergot alkaloids in the sodium hyaluronate gel thickening system at different pH values is investigated.

[0109] From the above experimental results, it can be seen that the viscosity of the ergot alkaloids-containing gel is moderate (for example, 9000-60000 m.pas), which meets the viscosity range of consumers using the gel type, and the dosage form performs well in high and low temperature stability investigation.

[0110] Test example 4: ergot alkaloids content test

[0111] Test sample: the ergot alkaloids-containing gel prepared in examples 5-8 which has passed the high and low temperature stability test in test example 3

[0112] Sample extraction: directly take the sample, dissolve the filter membrane with 0.1wt% formic acid methanol solution, and then determine.

[0113] Sample test: high performance liquid chromatograph (with ultraviolet detector), high performance liquid chromatography conditions: chromatographic column:

[0114] HILIC column (2.1 mm x 100 mm, 1.7 um); mobile phase: 0.1% formic acid acetonitrile solution-0.1% formic acid aqueous solution (88+12, V / V); flow rate: 0.4 Ml / min; column temperature: 40℃; wavelength: 262 nm; injection volume: 3 microliters. Ergothioneine standard samples with concentrations of 1.0 mg / L, 2.0 mg / L, 5.0 mg / L, 10.0 mg / L, 20.0 mg / L, 50.0 mg / L were used. The standard curve of ergothioneine was drawn with the mass concentration of the standard working solution as the abscissa and the peak area as the ordinate under the above chromatographic conditions. Qualitative analysis was performed according to the retention time, and the relative deviation of the retention time of the sample and the standard sample at a similar concentration was not more than 2%, and quantitative analysis was performed by the external standard method.

[0115] The response value of ergothioneine in the test sample solution should be within the range of the standard curve. If it exceeds the range of 1.0 mg / L-50.0 mg / L of the standard working curve, it should be diluted and analyzed again.

[0116] Recovery rate%=ergothioneine test content / ergothioneine added amount*100%

[0117]

[0118] Conclusion: The gel containing ergothioneine was prepared, and the effective content of ergothioneine in the sodium hyaluronate gel thickening system at different pH values after light treatment was investigated. Considering the test deviation and addition loss, it is generally considered that the effective concentration of active substances is above 90% recovery rate, and below 90% is considered to have the risk of decomposition and deterioration.

[0119] The ergothioneine-containing gel prepared in Examples 5-8 passed the high and low temperature stability test described in Test Example 3, i.e. after the high and low temperature stability test, the content of ergothioneine in the ergothioneine-containing gel prepared in Example 5 decreased to 0.059 parts by mass, 59% of the original added amount of 0.1 parts by mass of ergothioneine can be effectively recovered; the content of ergothioneine in the ergothioneine-containing gel prepared in Example 6 decreased to 0.064 parts by mass, 64% of the original added amount of 0.1 parts by mass of ergothioneine can be effectively recovered; the content of ergothioneine in the ergothioneine-containing gel prepared in Example 7 decreased to 0.056 parts by mass, 56% of the original added amount of 0.1 parts by mass of ergothioneine can be effectively recovered; the content of ergothioneine in the ergothioneine-containing gel prepared in Example 8 decreased to 0.006 parts by mass, 60% of the original added amount of 0.01 parts by mass of ergothioneine can be effectively recovered; after the high and low temperature stability test for one month, the test content of ergothioneine in the ergothioneine-containing gels prepared in Examples 5-8 with different pH values were all less than 90% of the added amount, indicating that ergothioneine was unstable in the gel systems of Examples 5-8, and the content and efficacy of ergothioneine were decreased due to photodegradation, thereby indicating that ergothioneine cannot be stabilized in the gel system.

[0120] Example 9: Preparation of an ergothioneine-containing emulsion

[0121] The water phase: 0.1 parts by mass of polyacrylic acid-30 polymer, 0.5 parts by mass of PEG-20 methylglucose sesquistearate, 0.1 parts by mass of xanthan gum and 80 parts by mass of deionized water were heated to 80-85°C and mixed, and mixed and stirred at a speed of 100 rpm until completely dissolved;

[0122] The oil phase: 2 parts by mass of cetylstearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glucoside were added to 3 parts by mass of 26# white oil and 4 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0123] The oil phase was slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm for 10-15 minutes until uniform and no particles were formed; when the temperature decreased to 55-60°C, 0.1 parts by mass of triethanolamine (10 wt% aqueous solution) was added, and after neutralization and stirring reaction, 0.01 parts by mass of ergothioneine was added and stirred to dissolve. Then, citric acid or tromethamine was added to adjust the pH value to 6, and deionized water was added to make up to 100, and mixed and stirred at a speed of 100 rpm for 10-20 minutes; then, it was placed at room temperature for 12 hours and ready for use.

[0124] Example 10: Preparation of an ergothioneine-containing emulsion

[0125] Water phase: 0.1 parts by mass of polyacrylic acid-30 polymer, 0.5 parts by mass of PEG-20 methyl glucose sesqui stearate, 0.1 parts by mass of xanthan gum and 80 parts by mass of deionized water were heated to 80-85°C and mixed, and mixed and stirred at a rotation speed of 100 rpm until completely dissolved;

[0126] Oil phase: 2 parts by mass of cetyl stearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glucoside were added to 3 parts by mass of 26# white oil and 4 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0127] The oil phase was slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm, and after homogenization for 10-15 minutes, it was evenly and non-particulate. When the temperature dropped to 55-60°C, 0.1 parts by mass of triethanolamine (10wt% aqueous solution) was added, and after neutralization and stirring reaction, 0.1 parts by mass of ergothioneine was added and stirred to dissolve. Then, citric acid or tromethamine was added to adjust the pH value to 6, deionized water was added to make up to 100, and mixed and stirred at a rotation speed of 100 rpm for 10-20 minutes; then, it was left to stand at room temperature for 12 hours, ready for use.

[0128] Example 11: Preparation of emulsion containing ergothioneine

[0129] Water phase: 0.1 parts by mass of polyacrylic acid-30 polymer, 0.5 parts by mass of PEG-20 methyl glucose sesqui stearate, 0.1 parts by mass of xanthan gum and 80 parts by mass of deionized water were heated to 80-85°C and mixed, and mixed and stirred at a rotation speed of 100 rpm until completely dissolved;

[0130] Oil phase: 2 parts by mass of cetyl stearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glucoside were added to 3 parts by mass of 26# white oil and 4 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0131] The oil phase was slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm, and after homogenization for 10-15 minutes, it was evenly and non-particulate. When the temperature dropped to 55-60°C, 0.1 parts by mass of triethanolamine (10wt% aqueous solution) was added, and after neutralization and stirring reaction, 0.1 parts by mass of ergothioneine was added and stirred to dissolve. Then, citric acid or tromethamine was added to adjust the pH value to 6, deionized water was added to make up to 100, and mixed and stirred at a rotation speed of 100 rpm for 10-20 minutes; then, it was left to stand at room temperature for 12 hours, ready for use.

[0132] Example 11: Preparation of emulsion containing ergothioneine

[0133] Water phase: 0.1 parts by mass of polyacrylic acid-30 polymer, 0.5 parts by mass of PEG-20 methyl glucose sesqui stearate, 0.1 parts by mass of xanthan gum and 80 parts by mass of deionized water were heated to 80-85°C and mixed, and mixed and stirred at a speed of 100 rpm until completely dissolved;

[0134] Oil phase: 2 parts by mass of cetyl stearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside were added to 3 parts by mass of 26# white oil and 4 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0135] The oil phase was slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm, and after homogenization for 10-15 minutes, it was evenly and no particles; when the temperature dropped to 55-60°C, 0.1 parts by mass of triethanolamine (10wt% aqueous solution) was added, and after neutralization and stirring reaction, 1 part by mass of ergothioneine was added and stirred to dissolve. Then, citric acid or tromethamine was added to adjust the pH value to 6, deionized water was added to make up to 100, and mixed and stirred at a speed of 100 rpm for 10-20 minutes; then, it was placed at room temperature for 12 hours and ready for use.

[0136] Example 13: Preparation of emulsion containing ergothioneine

[0137] Water phase: 0.1 parts by mass of polyacrylic acid-30 polymer, 0.5 parts by mass of PEG-20 methyl glucose sesqui stearate, 0.1 parts by mass of xanthan gum and 80 parts by mass of deionized water were heated to 80-85°C and mixed, and mixed and stirred at a speed of 100 rpm until completely dissolved;

[0138] Oil phase: 2 parts by mass of cetyl stearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside were added to 3 parts by mass of 26# white oil and 4 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0139] The oil phase was slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm, and after homogenization for 10-15 minutes, it was evenly and no particles; when the temperature dropped to 55-60°C, 0.1 parts by mass of triethanolamine (10wt% aqueous solution) was added, and after neutralization and stirring reaction, 1 part by mass of ergothioneine was added and stirred to dissolve. Then, citric acid or tromethamine was added to adjust the pH value to 6, deionized water was added to make up to 100, and mixed and stirred at a speed of 100 rpm for 10-20 minutes; then, it was placed at room temperature for 12 hours and ready for use.

[0140] Test Example 5: Viscosity test and high and low temperature stability test

[0141] Test sample: After measuring the viscosity of the emulsion containing ergothioneine prepared in Examples 9-13, the high and low temperature stability was investigated.

[0142] Using BROOKFIELD RVDVC digital viscometer (with RV type, T type rotor), sample test temperature: 25℃, constant temperature 1h, sample test container: the same specification beaker (180ml bottle) as possible when testing different batches of samples; reading record time: LV / RV type rotor 60s; T type rotor 90s; The sample should be confirmed without bubble interference before determination. According to the standard, select the corresponding rotor and speed. After taking the sample out of the constant temperature tank, immediately determine its viscosity with the rotary viscometer. When selecting LV and RV rotors, insert the rotor obliquely into the sample, so that the liquid surface is just in the recess of the rotor, and keep the rotor wall without bubbles; when selecting T type rotor, make the rotor just close to the liquid surface, and select a certain up and down movement distance, so that the rotor has enough up and down space and will not damage the beaker bottom. Record the viscosity result after the reading is stable (or after the standard time).

[0143] The high and low temperature stability evaluation method is as follows: the emulsion containing ergothioneine prepared in examples 5-8 is placed in a constant temperature box at high temperature (48℃), room temperature (25℃) and low temperature (4℃) for 30 days. Observe whether the sample has particle solid precipitation, jelly, delamination and other conditions. If the sample has no change before and after a certain time within 30 days, it is "passed"; if the sample has particle phenomenon, it is "precipitated"; if the sample has solidification phenomenon, it is "jelly"; if the sample has upper and lower liquid phenomenon, it is "delamination". The results are shown in the following table.

[0144] Conclusion: The emulsion containing ergothioneine is prepared, and the viscosity and stability of the emulsion formula of the emulsified system containing different content of ergothioneine are investigated. Through the above experimental results, it can be obtained that the viscosity of the emulsion formula of the emulsified system containing different content of ergothioneine is moderate (for example, 9000-60000 m.pas), and when the content of ergothioneine increases, the viscosity of the emulsion decreases slightly, but it meets the viscosity range of the emulsion dosage form used by consumers, and at the same time, the dosage form shows good high and low temperature stability.

[0145] Test example 6: ergothioneine content test

[0146] Test sample: the emulsion containing ergothioneine prepared in examples 9-13 which passed the high and low temperature stability test in test example 5

[0147] Sample extraction: directly take the sample, dissolve the filter membrane with 0.1 wt % formic acid methanol solution, and then determine.

[0148] Sample test: high performance liquid chromatograph (with ultraviolet detector), high performance liquid chromatograph conditions: chromatographic column:

[0149] HILIC column (2.1 mm x 100 mm, 1.7 um); mobile phase: 0.1% formic acid acetonitrile solution-0.1% formic acid aqueous solution (88+12, V / V); flow rate: 0.4 Ml / min; column temperature: 40°C; wavelength: 262 nm; injection volume: 3 microliters. Ergothioneine standard samples with concentrations of 1.0 mg / L, 2.0 mg / L, 5.0 mg / L, 10.0 mg / L, 20.0 mg / L, and 50.0 mg / L were used. The standard curve of ergothioneine was prepared by taking the mass concentration of the standard working solution as the abscissa and the peak area as the ordinate under the above chromatographic conditions. Qualitative analysis was performed according to the retention time, and the relative deviation of the retention time of the sample and the standard sample at a similar concentration was not more than 2%, and quantitative analysis was performed by the external standard method.

[0150] The response value of ergothioneine in the sample to be tested should be within the range of the standard curve. If it exceeds the range of 1.0 mg / L-50.0 mg / L of the standard working curve, it should be diluted and then analyzed.

[0151] Recovery rate = (ergothioneine test content / ergothioneine added amount) * 100%

[0152]

[0153] Conclusion: Emulsions containing ergothioneine were prepared, and the effective content of ergothioneine in emulsion formulations with different ergothioneine contents after light treatment was investigated. Considering the test deviation and addition loss, it is generally considered that a recovery rate of more than 90% is the effective concentration of active substances, and less than 90% is considered to have the risk of decomposition and deterioration.

[0154] The ergothioneine-containing emulsions prepared in Examples 9-13 passed the high and low temperature stability test described in Test Example 5, i.e. after the high and low temperature stability test, the content of ergothioneine in the ergothioneine-containing emulsion prepared in Example 9 decreased to 0.0065 parts by mass, 65% of the ergothioneine could be effectively recovered relative to the original added amount of 0.01 parts by mass; the content of ergothioneine in the ergothioneine-containing emulsion prepared in Example 10 decreased to 0.081 parts by mass, 81% of the ergothioneine could be effectively recovered relative to the original added amount of 0.1 parts by mass; the content of ergothioneine in the ergothioneine-containing emulsion prepared in Example 11 decreased to 0.41 parts by mass, 82% of the ergothioneine could be effectively recovered relative to the original added amount of 0.5 parts by mass; the content of ergothioneine in the ergothioneine-containing emulsion prepared in Example 12 decreased to 0.82 parts by mass, 82% of the ergothioneine could be effectively recovered relative to the original added amount of 1 parts by mass; the content of ergothioneine in the ergothioneine-containing emulsion prepared in Example 13 decreased to 1.8 parts by mass, 90% of the ergothioneine could be effectively recovered relative to the original added amount of 2 parts by mass; after the high and low temperature stability test for one month, the test content of ergothioneine in the ergothioneine-containing emulsions prepared in Examples 9-12 was less than 90% of the added amount, indicating that ergothioneine was unstable in the emulsion system of Examples 9-12, and the content and efficacy of ergothioneine decreased due to photodegradation, thereby indicating that ergothioneine could not be stably present in the emulsion system.

[0155] Furthermore, it was indicated that ergothioneine with a content of less than 2% could not be stably present in the emulsion. Compared with the hydrogel system of Examples 1-8, the emulsion system of Examples 9-12 containing ergothioneine with different contents had certain improvement in stabilizing the content of ergothioneine, but could not meet the stabilization of low-concentration ergothioneine (e.g. < 2 wt%).

[0156] Example 14: Preparation of an ergothioneine-containing emulsion

[0157] The water phase: 0.5 parts by mass of polyacrylic acid-30 polymer and 80 parts by mass of deionized water were weighed and heated to 80-85°C and mixed, and mixed and stirred at a rotation speed of 100 rpm until completely dissolved;

[0158] The oil phase: 1 part by mass of cetylstearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside were added to 6 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0159] The oil phase is slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm for 10-15 minutes until uniform and free of particles. When the temperature drops to 55-60°C, 0.4 parts by mass of triethanolamine (10 wt% aqueous solution) is added, and after neutralization and stirring, 0.01 parts by mass of ergothioneine is added and dissolved by stirring. Then, citric acid or tromethamine is added to adjust the pH to 6, deionized water is added to make up to 100, and mixing and stirring are continued at 100 rpm for 10-20 minutes. Then, it is left to stand at room temperature for 12 hours and is ready for use.

[0160] Example 15: Preparation of an emulsion containing ergothioneine

[0161] Water phase: 0.5 parts by mass of polyacrylic acid-30 polymer and 80 parts by mass of deionized water are heated to 80-85°C and mixed and stirred at 100 rpm until completely dissolved.

[0162] Oil phase: 1 part by mass of cetylstearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside are added to 6 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0163] The oil phase is slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm for 10-15 minutes until uniform and free of particles. When the temperature drops to 55-60°C, 0.4 parts by mass of triethanolamine (10 wt% aqueous solution) is added, and after neutralization and stirring, 0.01 parts by mass of ergothioneine is added and dissolved by stirring. Then, citric acid or tromethamine is added to adjust the pH to 6, deionized water is added to make up to 100, and mixing and stirring are continued at 100 rpm for 10-20 minutes. Then, it is left to stand at room temperature for 12 hours and is ready for use.

[0164] Example 16: Preparation of an emulsion containing ergothioneine

[0165] Water phase: 0.5 parts by mass of polyacrylic acid-30 polymer and 80 parts by mass of deionized water are heated to 80-85°C and mixed and stirred at 100 rpm until completely dissolved.

[0166] Oil phase: 1 part by mass of cetylstearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside are added to 6 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0167] The oil phase is slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm for 10-15 minutes until uniform and free of particles. When the temperature drops to 55-60°C, 0.4 parts by mass of triethanolamine (10 wt% aqueous solution) is added, and after neutralization and stirring, 0.5 parts by mass of ergothioneine is added and dissolved by stirring. Then, citric acid or tromethamine is added to adjust the pH to 6, deionized water is added to make up to 100, and mixing and stirring are continued at 100 rpm for 10-20 minutes. Then, it is left to stand at room temperature for 12 hours and is ready for use.

[0168] Example 17: Preparation of an emulsion containing ergothioneine

[0169] Water phase: 0.5 parts by mass of polyacrylic acid-30 polymer and 80 parts by mass of deionized water are heated to 80-85°C and mixed and stirred at 100 rpm until completely dissolved.

[0170] Oil phase: 1 part by mass of cetylstearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside are added to 6 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0171] The oil phase is slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm for 10-15 minutes until uniform and free of particles. When the temperature drops to 55-60°C, 0.4 parts by mass of triethanolamine (10 wt% aqueous solution) is added, and after neutralization and stirring, 0.5 parts by mass of ergothioneine is added and dissolved by stirring. Then, citric acid or tromethamine is added to adjust the pH to 6, deionized water is added to make up to 100, and mixing and stirring are continued at 100 rpm for 10-20 minutes. Then, it is left to stand at room temperature for 12 hours and is ready for use.

[0172] Example 18: Preparation of an emulsion containing ergothioneine

[0173] Water phase: 0.5 parts by mass of polyacrylic acid-30 polymer and 80 parts by mass of deionized water are heated to 80-85°C and mixed and stirred at 100 rpm until completely dissolved.

[0174] Oil phase: 1 part by mass of cetylstearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside are added to 6 parts by mass of isopropyl palmitate, and placed in a 85°C water bath to completely melt.

[0175] The oil phase is slowly added to the water phase at 80-85°C and homogenized at 2500-3000 rpm for 10-15 minutes until it is uniform and free of particles; when the temperature drops to 55-60°C, 0.4 parts by mass of triethanolamine (10% aqueous solution) is added, and after neutralization and stirring, 1 part by mass of ergothioneine is added and dissolved by stirring. Then, citric acid or tromethamine is added to adjust the pH to 6, deionized water is added to make up to 100, and mixing and stirring are continued at 100 rpm for 10-20 minutes; then, the sample is left to stand at room temperature for 12 hours and is ready for use.

[0176] Test Example 7: Product viscosity test and high and low temperature stability test

[0177] Test sample: After measuring the viscosity of the emulsion containing ergothioneine prepared in Examples 14-18, the high and low temperature stability of the system was investigated.

[0178] A BROOKFIELD RVDVC digital viscometer (RV type, T type rotor) was used, the sample test temperature was 25°C, the constant temperature was 1 hour, the sample test container was the same size beaker (180 ml bottle) as used in the test of different batches of samples; the reading recording time was LV / RV type rotor 60s; T type rotor 90s; before the sample was measured, it should be confirmed that there was no bubble interference. According to the standard, the corresponding rotor and speed were selected. After the sample to be tested was taken out of the constant temperature tank, its viscosity was immediately measured with a rotary viscometer. When selecting LV and RV rotors, the rotors were inserted obliquely into the sample, with the liquid surface just at the recess of the rotor, and the rotor wall was kept free of bubbles; when selecting T type rotor, the rotor was just close to the liquid surface, and a certain up and down movement distance was selected, so that the rotor had enough up and down space and would not damage the beaker bottom. After the reading was stable (or after the standard time), the viscosity result was recorded.

[0179] The high and low temperature stability evaluation method is as follows: the emulsion containing ergothioneine prepared in Examples 14-18 is placed in a constant temperature oven at high temperature (48°C), room temperature (25°C), and low temperature (4°C) for 30 days. Whether the sample has particles, solidification, stratification, and other conditions is observed.

[0180] If the sample is normal without change before and after a certain time within 30 days, it is indicated as "pass", the appearance of particles is indicated as "precipitation", the solidification of the material body is indicated as "jelly", and the appearance of upper and lower layers of liquid is indicated as "stratification". The results are shown in the following table.

[0181]

[0182] Conclusion: The emulsion containing ergothioneine is prepared, and the viscosity and stability of the emulsion formula of the emulsified system containing different contents of ergothioneine are investigated.

[0183] From the above experimental results, it can be seen that the emulsion formula of the emulsion system containing different content of ergothioneine has moderate viscosity (e.g. 9000-60000 m.pas), and when the content of ergothioneine increases, the viscosity of the emulsion decreases slightly, but it meets the viscosity range of the emulsion dosage form (cream) used by consumers, and the dosage form performs well in high and low temperature stability test.

[0184] Test Example 8: Ergothioneine content test

[0185] Test sample: emulsion containing ergothioneine prepared by examples 14-18 after passing the high and low temperature stability test described in test example 7

[0186] Sample extraction: directly take sample and dissolve the filter membrane with 0.1% formic acid methanol solution for determination.

[0187] Sample test: high performance liquid chromatograph (with ultraviolet detector), high performance liquid chromatograph conditions: chromatographic column:

[0188] HILIC chromatographic column (2.1 mm x 100 mm, 1.7 um); mobile phase: 0.1% formic acid acetonitrile solution-0.1% formic acid aqueous solution (88+12, V / V); flow rate: 0.4 Ml / min; column temperature: 40℃; wavelength: 262 nm; injection volume: 3 microliters. Use ergothioneine concentration of 1.0, 2.0, 5.0, 10.0, 20.0, 50.0 mg / L series standard sample. Test according to the above chromatographic conditions, draw the standard curve of ergothioneine with the mass concentration of standard working solution as the abscissa and the peak area as the ordinate. Qualitative according to retention time, the relative deviation of sample and standard product retention time under similar concentration is not more than 2%, and quantitative by external standard method.

[0189] The response value of ergothioneine in the test sample solution should be within the range of the standard curve, and if it exceeds the range of 1.0 mg / L-50.0 mg / L of the standard working curve, it should be diluted and analyzed again.

[0190] Recovery rate%=ergothioneine test content / ergothioneine addition amount*100%

[0191]

[0192] Conclusion: gel containing ergothioneine is prepared, and the effective content of ergothioneine in emulsion formula containing different content of ergothioneine after light treatment is investigated. Considering the test deviation and addition loss, it is generally considered that the recovery rate above 90% is the effective concentration of active substance, and below 90% is considered to have the risk of decomposition and deterioration.

[0193] The ergothioneine-containing emulsions prepared in Examples 14-18 passed the high and low temperature stability test described in Test Example 7, i.e. after the high and low temperature stability test, the ergothioneine content in the ergothioneine-containing emulsion prepared in Example 14 was maintained at 0.01 parts by mass, 100% of the ergothioneine could be effectively recovered relative to the original 0.01 parts by mass of the added amount; the ergothioneine content in the ergothioneine-containing emulsion prepared in Example 15 was maintained at 0.1 parts by mass, 100% of the ergothioneine could be effectively recovered relative to the original 0.1 parts by mass of the added amount; the ergothioneine content in the ergothioneine-containing emulsion prepared in Example 16 was maintained at 0.2 parts by mass, 100% of the ergothioneine could be effectively recovered relative to the original 0.2 parts by mass of the added amount; the ergothioneine content in the ergothioneine-containing emulsion prepared in Example 17 decreased to 0.48 parts by mass, 96% of the ergothioneine could be effectively recovered relative to the original 0.5 parts by mass of the added amount; the ergothioneine content in the ergothioneine-containing emulsion prepared in Example 18 decreased to 0.92 parts by mass, 92% of the ergothioneine could be effectively recovered relative to the original 1 parts by mass of the added amount; the ergothioneine content in the ergothioneine-containing emulsions prepared in Examples 14-18 was higher than 90% of the added amount after the high and low temperature stability test for one month, and the ergothioneine content and efficacy of the ergothioneine-containing emulsion systems of Examples 14-18 were considered to be successfully stabilized. Since ergothioneine is expensive, the addition amount of a daily care formula is usually in the range of 0.01-1%, and the stability of the emulsion system to ergothioneine has completely met the application range.

Claims

1. A stable emulsion system for carrying ergothioneine, comprising: an aqueous phase consisting of a polyacrylic polymer and an aqueous carrier; an oil phase consisting of cetostearyl alcohol, hexadecyl / octadecyl glycoside and isopropyl palmitate; triethanolamine; and ergothioneine; wherein, the weight ratio of the aqueous phase, the oil phase and triethanolamine is 80.39-90.95:8-9:0.01-0.4; the content of the triethanolamine in the emulsion system is 0.1-0.5wt%; the weight ratio of the polyacrylic polymer and the aqueous carrier is 0.1-0.5:80.29-90.45; the polyacrylic polymer is STABYLEN 30; the aqueous carrier is selected from at least one of deionized water, glacial water; the weight ratio of the cetostearyl alcohol, hexadecyl / octadecyl glycoside and isopropyl palmitate is 1-2:2:4-6; the content of the ergothioneine in the emulsion system is 0.01wt%-2wt%; the pH value of the emulsion system is 4-8.

2. The emulsified system according to claim 1, characterized in that, the weight ratio of the aqueous phase, the oil phase and triethanolamine is 80.39-90.95:8-9:0.01-0.1, and the content of the triethanolamine in the emulsion system is 0.1-0.4wt%.

3. The emulsified system according to any one of claims 1-2, characterized in that, the viscosity of the emulsion system is 9600-56000m.pas.

4. A method for preparing the emulsion system of claim 3, comprising the following steps: 1) weighing 0.1-0.5 parts by mass of a polyacrylic polymer and 50-80 parts by mass of deionized water, heating to 80-85°C and mixing and stirring until completely dissolved to obtain an aqueous phase; 2) weighing 1-2 parts by mass of cetostearyl alcohol and 2 parts by mass of hexadecyl / octadecyl glycoside into 4-6 parts by mass of isopropyl palmitate, heating to 80-85°C until completely melted to obtain an oil phase; 3) at 80-85°C, slowly adding the oil phase of step 2) to the aqueous phase of step 1) and homogenizing until uniform and free of particles; after the temperature is reduced to 55-60°C, adding 0.1-0.4 parts by mass of a 10wt% aqueous solution of triethanolamine to neutralize and stir to react, then adding 0.01-2 parts by mass of ergothioneine and citric acid or tromethamine to adjust the pH value to 4-8, and adding deionized water to make up to 100 parts by mass, and continuing to stir until uniform.

5. The method of claim 4, wherein, In step 1), the mixing and stirring rate is 100-500rpm; in step 3), the homogenization conditions are 2500-3000rpm, and the homogenization time is 10-15min.

6. Use of the emulsion system of claim 3 in a skin external preparation for non-therapeutic purposes.

7. Use according to claim 6, wherein the skin external preparation for non-therapeutic purposes is used in the field of cosmetics.

8. Use according to claim 6, wherein the skin external preparation for non-therapeutic purposes is selected from the group consisting of emulsions, creams, and masks.

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