A skin care composition for improving the condition of stressed skin, and its formulation and method of preparation

Abstract

The present application belongs to the technical field of cosmetics, and particularly relates to an emotional skin care composition for improving the state of stress skin, and a preparation method and application thereof. The skin care composition comprises a composite active material obtained by co-treatment of chicory root extract and rhamnose-fermented polysaccharide. The present application retains chicory-derived fructan with different polymerization degrees by partial restriction enzyme hydrolysis, and relatively grades different chain length saccharides by ethanol treatment, and then respectively performs redissolution, concentration and remixing, which is beneficial to improving the hydration and rheological properties, low-temperature stability and cosmetic preparation stability of the composite active material. The obtained composition can also improve the water content, barrier state, redness, roughness and gloss of the skin under stress, and improve the skin comfort, and can be used in non-therapeutic skin care products such as essence, emulsion, cream, gel and mask.

Description

Technical Field

[0001] This invention belongs to the field of daily chemical technology, and more specifically, relates to a skin care composition for improving stress-related skin conditions, its formulation, and preparation method. Background Technology

[0002] Skin is an important organ that comes into direct contact with the external environment. Its condition is affected not only by external factors such as temperature, humidity, ultraviolet radiation, pollutants, and chemical irritants, but also by a person's mental state, lifestyle rhythm, and stress level. With the fast pace of life, long-term work pressure, lack of sleep, and emotional fluctuations are common, leading to increasing attention to skin problems such as dryness, redness, roughness, dullness, decreased elasticity, and increased wrinkles.

[0003] Studies have shown that the skin not only serves as a physical barrier but also possesses complex neuro-, endocrine, and immune regulatory functions. Keratinocytes, fibroblasts, immune cells, and nerve endings in the skin can exchange information through various signaling molecules. When the body is under sustained stress, related neuroendocrine signals change, further affecting skin barrier function, inflammatory responses, and skin sensation. Substances in the skin such as neuroregulatory peptide B and thymic stromal lymphopoietin are related to nerve responses and skin discomfort, while substances like β-endorphin participate in skin sensation and related physiological regulation. Therefore, improving skin comfort and addressing various stress-induced skin conditions by regulating changes in local skin signaling molecules under stress has become a direction in skincare research.

[0004] Besides the skin's own neurological, immune, and endocrine regulation, the microorganisms on the skin surface and their metabolic activities are also related to skin condition. A variety of resident microorganisms exist on the skin surface, exchanging substances and transmitting information with skin cells. Suitable carbohydrates can serve as resources available to some resident skin microorganisms, thus influencing their metabolic activities and the resulting metabolites. Different carbohydrates, due to differences in monosaccharide composition, degree of polymerization, molecular weight, and water solubility, exhibit varying degrees of utilization and mechanisms of action on the skin surface.

[0005] Chicory root contains a relatively high amount of inulin-type fructans. Inulin-type fructans are mainly formed by the linkage of fructose units and have different degrees of polymerization. Shorter-chain fructans generally have better water solubility and faster dissolution rates, while fructans with higher degrees of polymerization differ from short-chain fructans in terms of solubility, hydration behavior, and physical properties in the system. Therefore, fructans derived from chicory root can not only be used as a common sugar raw material, but their chain length composition is also an important factor affecting their performance.

[0006] Existing technologies typically employ methods such as water extraction, enzymatic hydrolysis, and membrane separation to prepare chicory root extracts or fructooligosaccharides. For example, hydrolyzing inulin with inulinase can reduce the degree of polymerization of fructans, yielding shorter-chain fructooligosaccharides. However, in practical applications, if the degree of enzymatic hydrolysis is too high, a large amount of the original medium- and long-chain fructans will be converted into short-chain components; if the degree of enzymatic hydrolysis is insufficient, problems such as a high proportion of fructans with a high degree of polymerization and poor solubility at low temperatures may occur. Therefore, how to reasonably retain fructans with different degrees of polymerization in the same chicory-derived raw material, so that it possesses the characteristics of short-chain fructans while retaining a certain amount of medium-chain fructans, is a problem worthy of further research.

[0007] On the other hand, rhamnose-containing fermented polysaccharides are also a type of sugar raw material used in the cosmetics industry. These polysaccharides can be obtained through microbial fermentation, and some rhamnose-rich fermented polysaccharides contain sugar residues such as rhamnose, galactose, and glucuronic acid. Compared with fructooligosaccharides, these fermented polysaccharides have a larger molecular weight, and their solubility, hydration, and rheological behavior in water are significantly different. Existing studies have shown that some rhamnose-rich fermented polysaccharides can participate in the transmission of information between skin cells and have a certain effect on improving discomfort caused by external stimuli to the skin, thus showing potential application in soothing and mood-related skincare products.

[0008] Currently, when using different sugar-based active ingredients in skincare products, the most direct approach is usually to prepare each active ingredient separately and then add it to the aqueous phase or directly to the final cosmetic formulation in a specific amount. However, for low-molecular-weight fructans and high-molecular-weight fermented polysaccharides, due to significant differences in their molecular weight, hydration rate, and solution behavior, simple mixing does not guarantee that the two substances will achieve a suitable state of existence within the same system.

[0009] In particular, high molecular weight fermented polysaccharides undergo a gradual water absorption and hydration process after being added to the aqueous phase. The degree of hydration is related to temperature, time, and other soluble substances in the aqueous phase; short-chain fructans, on the other hand, usually dissolve more quickly. If the two are mixed directly, the hydration process of the high molecular weight polysaccharides may continue after the product preparation is completed, resulting in continued viscosity changes in the active material or final formulation during storage. On the other hand, chicory extract containing a certain amount of medium and long-chain fructans may also experience changes in its solubility under low temperature or high concentration conditions, which in turn affects the transparency, uniformity, and storage stability of the system.

[0010] Existing solutions typically focus on adjusting the amounts of thickeners, emulsifiers, polyols, or other formulation excipients to improve the viscosity and stability of the final product. These methods primarily adjust the final formulation and do not adequately consider the hydration and distribution states of fructans of different chain lengths and higher molecular weight fermentation polysaccharides before their addition to the final formulation. Therefore, even if the types and amounts of active ingredients in the final formulation are the same, different raw material processing methods can still lead to differences in the hydration rate, rheological behavior, and physical state of the active ingredients during storage.

[0011] Furthermore, existing emotional skincare technologies largely rely on plant extracts, aromatic substances, sensory modulators, or single soothing active ingredients to achieve skincare effects by reducing skin irritation, improving redness, or enhancing skin comfort. For example, some technologies combine plant extracts with sensory modulators to improve stress-related sensitivity and discomfort by addressing nerve sensation and inflammation. While these technologies can improve skin condition to some extent, their approach primarily focuses on the direct combination of different active ingredients and their corresponding physiological effects. Further research is needed on how to appropriately process plant-derived fructans with varying degrees of polymerization and fermented polysaccharides to create active systems suitable for cosmetic applications.

[0012] Especially in improving stress-related skin conditions, focusing solely on single issues like redness, stinging, or sensitivity fails to comprehensively reflect the multiple changes that may occur simultaneously under stress, such as dryness, decreased barrier function, roughness, dullness, and reduced elasticity. Therefore, it is necessary to develop a composition suitable for long-term skincare use that, while ensuring good formulation suitability and storage stability of the active ingredients, can achieve a more comprehensive improvement in terms of skin comfort and stress-related skin manifestations.

[0013] Therefore, how to rationally process fructans of different chain lengths from chicory root, and improve the processing and mixing process of the two types of sugars by combining the hydration characteristics of rhamnose-rich fermented polysaccharides, so as to reduce problems such as insufficient hydration, subsequent viscosity changes and low-temperature stability that may be caused by direct mixing, while preserving the role of the two types of active sugars in stress-related skin care, is a technical problem that needs to be further solved in this field. Summary of the Invention

[0014] This invention aims to address the problems of existing emotional skincare compositions, which mainly rely on the direct compounding of various plant extracts, sensory modulating ingredients, or soothing ingredients, with relatively simple processing methods between active ingredients, and the inconsistent hydration rates and continued changes in the system state after direct mixing of sugar active ingredients with different molecular weights and hydration properties. The invention provides a skincare composition for improving the condition of stressed skin, its preparation method, and its application.

[0015] This invention further addresses the problem that the chain length composition of fructans from chicory roots tends to be uniform after complete or relatively sufficient enzymatic hydrolysis. By performing restricted enzymatic hydrolysis on a portion of the chicory root extract and remixing it with the unhydrolyzed chicory root extract, the resulting chicory root treatment solution retains both short-chain and medium-chain fructans.

[0016] Based on this, the present invention does not simply mix the obtained chicory root treatment liquid with rhamnose-rich fermented polysaccharide. Instead, it utilizes the differences in the solubility and precipitation behavior of fructans of different chain lengths and rhamnose-rich fermented polysaccharide in an aqueous ethanol system to perform a fractional treatment. This allows the higher molecular weight rhamnose-rich fermented polysaccharide and some medium-chain fructans to enter the precipitation part, while the shorter chain fructans are mostly retained in the liquid part. Subsequently, the precipitation part and the liquid part are treated separately. After the precipitation part undergoes a resolution treatment under specified conditions, the liquid part, which has been de-alcoholized and concentrated, is added back in, thereby allowing different sugar components to undergo different dehydration and rehydration processes.

[0017] Therefore, this invention not only considers which sugar active ingredients are ultimately contained in the composition, but also further considers the processing of different sugar active ingredients before forming the final composition, so that the resulting composite active ingredient has a hydration state and physical stability suitable for cosmetic formulation applications. It also combines the skin action characteristics of chicory-derived fructan and rhamnose-rich fermented polysaccharide to improve skin comfort under stress and the accompanying skin conditions such as dryness, redness, roughness, dullness, and loss of elasticity.

[0018] To achieve the above objectives, the present invention provides a skin care composition for improving stress-related skin conditions, the skin care composition comprising a complex active ingredient obtained by co-processing chicory root extract and rhamnose-rich fermented polysaccharide.

[0019] Specifically, the composite active material can be prepared using a method including the following steps.

[0020] (1) Chicory root extract

[0021] Chicory root is pulverized and added to water for extraction. The chicory root extract is obtained through solid-liquid separation. Preferably, the mass ratio of chicory root to water is 1:(8-15), the extraction temperature is 50-65℃, and the extraction time is 1-3 hours.

[0022] In some embodiments, the obtained chicory root extract can be appropriately concentrated or adjusted with water to keep its soluble solids content within a range suitable for subsequent enzymatic hydrolysis, for example, 8–12 wt%.

[0023] Using the above conditions for water extraction allows inulin-type fructans from chicory roots to fully enter the aqueous phase, while avoiding unnecessary hydrolysis of fructans caused by excessively high temperatures and prolonged treatment.

[0024] (2) Partial enzymatic hydrolysis

[0025] Take a portion of the obtained chicory root extract A, adjust the pH to 4.8–5.3, heat to 48–55°C, and add endo-inulinase for restrictive enzymatic hydrolysis.

[0026] The amount of endo-inulinase added, based on enzyme activity, can be 2–10 U / g of chicory root extract A, and the enzymatic hydrolysis time can be 40–70 min; more preferably, the soluble solids content of the chicory root extract is about 10%, the enzymatic hydrolysis pH is 5.0, the enzymatic hydrolysis temperature is 52℃, the amount of endo-inulinase added is 6 U / g of chicory root extract A, and the enzymatic hydrolysis time is 50 min; after the enzymatic hydrolysis is completed, the temperature can be raised to 88–92℃ and maintained for 8–10 min to inactivate the endo-inulinase.

[0027] The purpose of using endo-inulinase in this invention is not to fully hydrolyze the fructans in chicory roots into oligosaccharides or monosaccharides, but to utilize the effect of endo-inulinase on the glycosidic bonds inside the fructan chain to break some of the longer fructan chains to a limited extent, so that a certain amount of shorter fructan chains are formed.

[0028] Meanwhile, no endo-inulinase was added to the remaining portion of chicory root extract B to preserve the original medium- and long-chain fructans.

[0029] Preferably, the unhydrolyzed chicory root extract B can be treated at the same or similar temperature as the hydrolyzed portion without the addition of endo-inulinase, so that the two extracts undergo similar heat treatment processes.

[0030] After enzymatic hydrolysis, the enzymatically hydrolyzed chicory root extract A is mixed with the unhydrolyzed chicory root extract B to obtain the chicory root treatment solution.

[0031] The chicory root treatment solution obtained through the above process is not intended to obtain fructans with a single degree of polymerization, but rather to contain fructans with different degrees of polymerization simultaneously. Preferably, it includes at least short-chain fructans with a degree of polymerization of 3 to 7 and medium-chain fructans with a degree of polymerization of 13 to 30.

[0032] Compared to uniformly enzymatically hydrolyzing all chicory root extracts, the above method avoids the excessive hydrolysis of the original medium-chain fructans in order to obtain sufficient short-chain fructans.

[0033] (3) Mixed with rhamnose-rich fermented polysaccharide

[0034] The above chicory root treatment solution was mixed with rhamnose-rich fermented polysaccharide.

[0035] The rhamnose-rich fermented polysaccharide of this invention is a water-soluble polysaccharide containing rhamnose residues obtained through microbial fermentation. In some embodiments, the rhamnose-rich fermented polysaccharide also contains galactose residues and glucuronic acid residues.

[0036] The weight-average molecular weight of the rhamnose-rich fermented polysaccharide is 35–70 kDa, preferably 40–60 kDa.

[0037] After mixing the chicory root treatment solution with rhamnose-rich fermented polysaccharide, the pH of the system can be adjusted to 5.2–5.8, and the mixture can be stirred at 30–40°C for 30–90 min to ensure that the two types of sugars are evenly dispersed in the system.

[0038] (4) Ethanol fractionation

[0039] The mixture is cooled to 20–25°C, and ethanol is gradually added while stirring until the ethanol volume fraction reaches 48–56%. After the ethanol is added, stirring continues for 10–30 minutes, followed by standing at 4–10°C for 4–12 hours. Solid-liquid separation is then performed to obtain the precipitate and the liquid fraction, respectively. Preferably, the ethanol volume fraction in the system is 50–54%, more preferably about 52%.

[0040] The purpose of this invention in setting up the ethanol treatment step is not simply to recover all the sugars from the system, but to utilize the differences in the solubility behavior of sugars with different molecular weights and chain lengths in aqueous ethanol to allow different sugar components to be distributed to a certain extent.

[0041] Among them, higher molecular weight rhamnose-rich fermented polysaccharides and some medium-chain fructans are more likely to enter the precipitation phase; while shorter-chain fructans are relatively more retained in the liquid phase. Therefore, this step allows the sugar components that were originally in the same aqueous phase to enter the two phases separately, and allows the two phases to undergo different treatment processes in subsequent steps.

[0042] It should be noted that the "precipitate portion" and "liquid portion" described in this invention do not require complete separation of the various sugar components. For example, the precipitate portion may still contain a certain amount of short-chain fructans, and the liquid portion may also contain a small amount of fructans with a higher degree of polymerization. This invention utilizes the relative distribution differences of different sugar components in the two portions.

[0043] (5) Resolution treatment of the precipitate

[0044] The precipitate is added to water and stirred at 30–35°C for 30–60 min to redissolve it; preferably, redissolve it at about 32°C for about 45 min.

[0045] Unlike conventional treatment methods that use high temperatures and long processing times to allow the high molecular weight polysaccharides in the precipitate to reach full hydration equilibrium, the present invention preferably performs resolution at a milder temperature and within a specified time, and proceeds to the subsequent remixing step after the above resolution treatment is completed.

[0046] In a preferred embodiment, the precipitate after resolution is not subjected to heating treatment above 45°C, and the treated liquid portion is added within 6 hours after the resolution treatment is completed.

[0047] Rhamnose-rich fermented polysaccharides and some medium-chain fructans undergo solvent environment changes and dehydration precipitation during ethanol fractionation. Upon re-entry into the aqueous phase, they require water absorption, swelling, and further hydration. Using the aforementioned mild and time-limited reconstitution conditions avoids the precipitate undergoing prolonged separate hydration before being added back to the liquid phase.

[0048] (6) Treatment and refilling of liquid components

[0049] The liquid fraction obtained from ethanol fractionation is subjected to dealcoholization; preferably, the dealcoholization temperature is not higher than 45°C. After dealcoholization, the liquid fraction can be further concentrated using membrane concentration to obtain a concentrate containing a higher amount of short-chain fructans.

[0050] Subsequently, the liquid portion that has undergone alcohol removal and concentration is added to the precipitate portion that has undergone the above-mentioned resolution treatment, and stirred at 25-30°C for 30-60 min, and further placed at 23-27°C for 2-6 h to obtain the composite active product; preferably, the two portions are mixed and stirred at 28-30°C, and then placed at about 25°C.

[0051] The present invention adopts a "grading-separate processing-re-addition" method, which differs from the method of preparing short-chain fructans, medium-chain fructans and rhamnose-rich fermented polysaccharides separately and then adding them directly to water for mixing at once.

[0052] After ethanol fractionation, the rhamnose-rich fermented polysaccharides and some medium-chain fructans first undergo dehydration and precipitation under aqueous ethanol conditions, and then are reconstituted under specified conditions; while a large amount of short-chain fructans remain in the liquid portion, and after being de-alcoholized and concentrated, they are reconstituted in the aforementioned precipitate portion and then added back in.

[0053] Therefore, the two types of sugar components underwent different solvent treatment and hydration processes before finally reforming into the same aqueous system.

[0054] This invention suggests that this treatment method helps reduce the viscosity changes over time caused by the continued hydration of high molecular weight sugars after the final composition is formed. It also helps improve the low-temperature uniformity of systems containing fructans and fermented polysaccharides of different chain lengths, as well as the applicability of the formulation when subsequently added to skin care preparations.

[0055] Skin care preparations

[0056] The present invention also provides a skin care formulation comprising the above-described skin care composition and cosmetic-acceptable excipients.

[0057] The skin care preparation can be a serum, lotion, cream, gel, mask liquid, or other cosmetic formulation suitable for external use on the skin.

[0058] The excipients may include one or more of the following: humectants, oils, emulsifiers, thickeners, preservatives, chelating agents, pH adjusters, and water.

[0059] Considering that the composite active ingredient contains sugars of different molecular weights, it is preferable to add the composite active ingredient after the emulsification or mixing of the main body of the skin care preparation is completed and the temperature is cooled to below 45°C; the pH of the skin care preparation is preferably controlled between 5.0 and 6.2, and more preferably between 5.3 and 5.9.

[0060] The above methods can reduce the impact of excessively high temperatures and extreme pH conditions on the state of carbohydrates in the composite active ingredients.

[0061] The skin care composition of the present invention does not target a single problem of skin sensitivity or redness, but rather a simultaneous decrease in skin comfort and a decline in multiple skin conditions under stress.

[0062] Under stress, nerve, immune, and endocrine-related signals in the skin may change, further affecting the skin barrier, inflammatory response, and skin sensation. Simultaneously, the skin microbiome and its communication with skin cells may also be affected. Therefore, this invention combines two types of saccharide-based active ingredients based on their different characteristics.

[0063] Among them, the short-chain fructans retained in the chicory root treatment solution have good water solubility, which can provide a suitable sugar basis for skin surface microbial metabolism; the coexistence of fructans with different degrees of polymerization avoids the complete conversion of chicory-derived sugars into a single oligosaccharide system.

[0064] Rhamnose-rich fermented polysaccharides have different molecular structures and molecular weights than short-chain fructans. They can participate in skin cell-related information transmission and are suitable for improving skin discomfort under irritation.

[0065] Therefore, this invention does not simply rely on a single "happy factor" to explain all skin care effects, but rather considers the skin surface microbial-related effects in combination with the skin neuro-immune-related effects.

[0066] In the evaluation system of this invention, β-endorphin can be further used as one of the evaluation indicators of skin pleasure-related signals, and combined with indicators such as neuromodulatory peptide B (NMB) and thymic stromal lymphopoietin (TSLP) to evaluate changes in skin-related signals under stress or stimulation.

[0067] By further combining parameters such as transdermal water loss, stratum corneum water content, skin erythema degree, skin gloss, skin roughness, elasticity, and wrinkles, the improvement effect of the composition of the present invention on the decline of stress-related skin conditions can be evaluated from both the physiological and appearance aspects of the skin.

[0068] It should be noted that the "improvement of stress-related skin condition" described in this invention is skin care in the sense of cosmetics. Its purpose is to improve the skin appearance, barrier state and skin comfort associated with stress, and it does not involve the diagnosis or treatment of anxiety, depression or other mental illnesses.

[0069] Compared with the prior art, the present invention has at least the following beneficial effects:

[0070] (1) Retain chicory fructans with different degrees of polymerization.

[0071] This invention does not perform uniform and complete enzymatic hydrolysis on all chicory root extracts. Instead, it only performs endo-inulinase-restricted enzymatic hydrolysis on a portion of the extract and then remixes it with the unhydrolyzed portion. This results in a chicory root extract containing both short-chain and medium-chain fructans, avoiding excessive changes in fructan chain length composition towards lower polymerization degree caused by complete enzymatic hydrolysis.

[0072] (2) Different sugar components undergo different processing procedures.

[0073] This invention involves ethanol fractionation after mixing chicory root treatment liquid with rhamnose-rich fermented polysaccharide. This allows more rhamnose-rich fermented polysaccharide and some medium-chain fructans to enter the precipitate, while more short-chain fructans remain in the liquid phase. The two phases are then treated separately and remixed, which differs from the simple compounding method of directly adding different active sugars to the same aqueous phase.

[0074] (3) Improve the hydration process of high molecular weight sugars and the physical stability of the composition.

[0075] After the precipitate is treated with ethanol, it is re-dissolved at a specified mild temperature and time, and the liquid portion that has been concentrated by de-alcoholization is added under controlled conditions. This helps to reduce the viscosity drift caused by the continued hydration of high molecular weight polysaccharides during the storage of the resulting complex active ingredient, and improves its applicability in skin care formulations such as lotions and serums.

[0076] (4) It takes into account both skin comfort under stress and multidimensional skin performance.

[0077] This invention combines chicory-derived fructans of different chain lengths with rhamnose-rich fermented polysaccharides for skin care under stress. Its effects can be further evaluated using indicators such as β-endorphin, NMB, TSLP, TEWL, stratum corneum moisture content, erythema, luster, roughness, and elasticity. This allows us to focus not only on a single sensitivity or redness phenomenon, but also on the various skin condition changes that accompany stress.

[0078] (5) Suitable for long-term external use of emotional skin care products.

[0079] The main active ingredients used in this invention are plant-derived fructans and fermentation-derived polysaccharides. It does not rely on aroma stimulation to produce immediate sensations, nor does it aim to regulate pharmacological or psychological states. Instead, it improves the skin condition associated with stress by addressing the skin's own condition and related local signals. It is suitable for various daily skincare products such as serums, lotions, creams, gels, and masks. Detailed Implementation

[0080] The present invention will be further described below with reference to specific embodiments. It should be understood that the following specific embodiments are used to illustrate the technical solution of the present invention, and are not intended to limit the scope of protection of the present invention. Without departing from the basic technical concept of the present invention, conventional adjustments made by those skilled in the art to the extraction temperature, processing time, stirring speed, concentration degree, and excipient types based on the actual state of raw materials, production scale, and product dosage form can all be used to implement the present invention.

[0081] The skincare composition for improving stress-related skin conditions described in this invention uses chicory root-derived fructan and rhamnose-rich fermented polysaccharide as the main active ingredients. Unlike methods that directly add the two raw materials to cosmetics after separate preparation, this invention first performs restricted enzymatic hydrolysis on a portion of the fructan in the chicory root extract, ensuring that fructans with different degrees of polymerization are retained simultaneously in the chicory root extract from the same source. Then, the resulting chicory root extract is mixed with rhamnose-rich fermented polysaccharide, and ethanol fractionation is used to distribute the different sugar components to a certain extent. The resulting precipitate and liquid fractions are then reconstituted and concentrated after de-alcoholization, and then remixed.

[0082] The purpose of the above treatment is not to obtain a new chemical compound, but to transform sugars with different chain lengths, molecular weights and hydration properties into complex active substances suitable for cosmetic use after undergoing different solvent treatments and hydration processes.

[0083] Chicory root raw materials and their pretreatment

[0084] This invention uses chicory (Cichorium intybus L.) root as a source of fructans.

[0085] Chicory root can be obtained by washing, slicing, and drying fresh chicory root, or by directly using dried chicory root that meets the requirements for use as a cosmetic ingredient. To minimize the impact of fluctuations in the moisture content of the raw material on the extraction process, dried chicory root is preferred.

[0086] In a preferred embodiment, chicory roots are dried to a moisture content of no more than 8%, then pulverized and passed through a 20-60 mesh sieve, preferably a 40 mesh sieve.

[0087] The main purpose of pulverization is to increase the contact area between chicory root and extraction water, thereby improving the efficiency of fructan entering the aqueous phase. Excessively large particle sizes result in relatively low extraction efficiency; conversely, excessively small particle sizes may increase the difficulty of subsequent solid-liquid separation. Therefore, chicory root powder of approximately 40 mesh is sufficient to meet the requirements for subsequent water extraction.

[0088] Water extract of chicory root

[0089] The chicory roots treated as described above are added to water for extraction. The mass ratio of chicory roots to water is 1:(8-15), preferably 1:(9-12), and more preferably 1:10; the extraction temperature is 50-65℃, preferably 55-62℃; and the extraction time is 1-3 h, preferably 1.5-2.5 h.

[0090] For example, 100 parts by weight of dried chicory root powder can be added to 1000 parts by weight of purified water and extracted by stirring at 58°C for 2 hours.

[0091] After extraction, insoluble residues are removed by filtration, centrifugation, or a combination of filtration and centrifugation to obtain chicory root extract.

[0092] This invention preferably uses water extraction at a moderate temperature, rather than prolonged high-temperature boiling. The reason for this is that the main purpose of this step is to allow the water-soluble fructans in chicory roots to enter the extract while preserving their original chain length composition as much as possible. Excessively high temperatures, especially prolonged treatment under acidic conditions, may increase the possibility of unintended hydrolysis of fructans, which is detrimental to subsequent control of chain length changes using endo-inulinase. After solid-liquid separation, the obtained chicory root extract can be concentrated under reduced pressure or adjusted with water as needed. Preferably, the soluble solids content of the chicory root extract before subsequent enzymatic hydrolysis is 8–12 wt%, more preferably 9–11 wt%, for example, 10.0 wt%. The soluble solids content can be determined by refractive index and expressed as a percentage by mass.

[0093] By controlling the soluble solids content of chicory root extract within a relatively stable range, the subsequent use of "U / g chicory root extract" to express the amount of endo-inulinase added can have better process repeatability.

[0094] Partial restriction enzymatic hydrolysis of chicory root extract

[0095] In this invention, it is not preferred to fully enzymatically hydrolyze all the chicory root extract.

[0096] Specifically, after the obtained chicory root extract is mixed evenly, a portion of it is taken as the enzymatically hydrolyzed chicory root extract A, and the remaining portion is taken as the unhydrolyzed chicory root extract B.

[0097] 2. Endoglucanase

[0098] The present invention preferably uses an endo-inulinase to treat the enzymatic hydrolysis portion.

[0099] Endoglucanases can act on the glycosidic bonds inside the inulin-type fructan chains, causing longer fructan chains to break down and form fructans with a relatively low degree of polymerization.

[0100] Compared with exo-enzymes, which are characterized by gradual hydrolysis from the chain end to form more monosaccharides, the endo-enzyme used in this invention is more conducive to achieving a limited degree of chain length adjustment.

[0101] This invention does not require complete hydrolysis of fructans. On the contrary, excessive enzymatic hydrolysis may lead to a large amount of medium-chain fructans being further converted into shorter-chain substances, which is inconsistent with the purpose of this invention to preserve fructans of different chain lengths.

[0102] 3. Enzymatic hydrolysis conditions

[0103] Adjust the pH of the chicory root extract A to be enzymatically hydrolyzed to 4.8–5.3, preferably 4.9–5.1, for example 5.0. The pH can be adjusted using citric acid, sodium citrate, or their buffer systems; adjust the temperature to 48–55°C, preferably 50–54°C, for example 52°C, and then add endo-inulinase.

[0104] Based on the actual enzyme activity of endo-inulinase, the amount added can be 2–10 U / g chicory root extract A, preferably 4–8 U / g chicory root extract A, for example 6 U / g chicory root extract A. The enzymatic hydrolysis time can be 40–70 min, preferably 45–60 min, for example 50 min.

[0105] For example, when the amount of chicory root extract A to be treated is 1000 g, if an endoglucosidase is added at a rate of 6 U / g, the required total enzyme activity is 6000 U.

[0106] If the inulinase preparation used has an enzyme activity of 1000 U / g, then add 6 g of enzyme preparation accordingly; if the enzyme preparation has a different enzyme activity, then calculate the amount added according to the actual enzyme activity.

[0107] 4. Enzyme inactivation

[0108] After reaching the predetermined degree of enzymatic hydrolysis, heat the system to 88–92°C and hold for 8–10 minutes to inactivate the endo-inulinase; for example, heat to 90°C and hold for 10 minutes; then cool down. Timely enzyme inactivation is crucial. If the enzyme activity remains for a prolonged period after reaching the predetermined chain length composition, fructan may continue to hydrolyze, causing variations in the degree of polymerization distribution across different production batches.

[0109] 5. Undigested portion

[0110] Chicory root extract B, which is not enzymatically hydrolyzed, does not contain endo-inulinase.

[0111] To reduce the impact of historical heat treatment variations on the final product, this portion of the extract can be adjusted to the same or similar pH as the enzymatic hydrolysis portion, maintained at 48–55°C for a similar duration to the enzymatic hydrolysis process, and then subjected to a short heat treatment at 88–92°C. For example, it can be maintained at pH 5.0 and 52°C for 50 min, and then heated to 90°C and maintained for 10 min.

[0112] Remixing the enzymatically hydrolyzed portion with the unenzymatically hydrolyzed portion

[0113] After separate treatment, the enzymatically hydrolyzed and unhydrolyzed portions are mixed evenly to obtain chicory root treatment solution.

[0114] The present invention preferably evaluates the treatment results by detecting the degree of polymerization distribution of fructans in the obtained chicory root treatment solution; the chicory root treatment solution contains at least short-chain fructans and medium-chain fructans simultaneously.

[0115] In one embodiment, the short-chain fructan is a fructan with a degree of polymerization (DP) of 3 to 7; the medium-chain fructan is a fructan with a DP of 13 to 30; in addition, the system also contains fructan with a DP of 8 to 12 and a small amount of fructan with a DP greater than 30.

[0116] Determination of the degree of polymerization of fructan

[0117] The chain length distribution of fructans is preferably determined using high-performance anion exchange chromatography-pulse amperometric detection (HPAEC-PAD). During the determination, an anion exchange column suitable for sugar separation can be used, with a sodium hydroxide system as the mobile phase. A gradient elution is achieved by adding sodium acetate to separate fructans with different degrees of polymerization. Peak assignment is performed using fructooligosaccharides of known degrees of polymerization or inulin standards. For components with high degrees of polymerization or incomplete standards, retention time patterns and, if necessary, mass spectrometry can be used for auxiliary confirmation. This invention preferably uses the relative content within a polymerization degree range as the process control indicator, rather than requiring absolute quantification of each individual fructan.

[0118] V. Rhamnose-rich fermented polysaccharides

[0119] This invention uses rhamnose-rich fermented polysaccharide as another active sugar component.

[0120] The rhamnose-rich fermented polysaccharide can be obtained by microorganisms capable of producing rhamnose extracellular polysaccharides through fermentation, solid-liquid separation, purification, and drying. Alternatively, commercially available rhamnose-rich fermented polysaccharides that meet the requirements of relevant cosmetic raw materials can be used.

[0121] The rhamnose-rich fermented polysaccharide contains at least rhamnose residues, and preferably further contains galactose residues and glucuronic acid residues.

[0122] In a preferred embodiment, the weight-average molecular weight Mw is 35–70 kDa, more preferably 40–60 kDa, for example about 50 kDa.

[0123] It should be noted that this invention does not require a precise single molecular weight as a necessary condition. Fermented polysaccharides typically have a certain molecular weight distribution; therefore, the molecular weight mentioned is the weight-average molecular weight used to characterize the overall molecular size of the fermented polysaccharide.

[0124] Weight-average molecular weight can be determined using aqueous gel permeation chromatography; when conditions permit, SEC-MALS can also be used to reduce deviations caused by standard calibration. When using ordinary GPC and calibrating with dextran or pullulan standards, the results can be expressed as the apparent weight-average molecular weight under the corresponding conditions.

[0125] VI. Mixed treatment of chicory root treatment solution with rhamnose-rich fermented polysaccharide

[0126] The above chicory root treatment solution was mixed with rhamnose-rich fermented polysaccharide.

[0127] Before mixing, the rhamnose-rich fermented polysaccharide can be dispersed in a small amount of purified water to reduce local aggregation after direct addition to the chicory root treatment solution. Adjust the pH of the mixture to 5.2–5.8, preferably 5.4–5.6, for example, 5.5; stir at 30–40°C for 30–90 min, preferably at 33–37°C for 45–75 min. For example, stir at 35°C for 60 min.

[0128] The main purpose of this step is to ensure that chicory-derived fructans and rhamnose-rich fermented polysaccharides are fully and uniformly present in the same aqueous phase before ethanol treatment.

[0129] VII. Ethanol Fractionation

[0130] After the above mixing is completed, the system is cooled to 20-25°C; ethanol is slowly added while continuously stirring, so that the final ethanol volume fraction in the system reaches 48-56%, preferably 50-54%, for example 52%.

[0131] The preferred ethanol used is high-concentration ethanol that meets production requirements, and the amount to be added is calculated based on the original mixture volume and the actual ethanol concentration. Ethanol should not be added rapidly all at once. It is preferable to add it gradually over 10–30 minutes to prevent the local ethanol concentration from becoming excessively high. After the ethanol is added, continue stirring for 10–30 minutes, for example, 20 minutes, and then place the system at 4–10°C for 4–12 hours, for example, 8 hours. After standing, perform solid-liquid separation. Solid-liquid separation can be achieved by centrifugation, for example, at 5000–8000 × g for 10–15 minutes, or by filtration or other solid-liquid separation methods suitable for the corresponding production scale; collect the precipitate and the liquid fraction separately.

[0132] The purpose of using approximately 48-56% ethanol in this invention is to utilize the differences in solubility of different sugars in aqueous ethanol to generate relative gradation.

[0133] Among them, rhamnose-rich fermented polysaccharides have relatively high molecular weights, solubility is reduced under the above-mentioned aqueous ethanol conditions, and more of them enter the precipitate; some medium-chain or high-polymerization-degree chicory fructans can also enter the precipitate; in contrast, short-chain fructans have better water solubility, and a large portion of them remain in the liquid portion under the above-mentioned ethanol concentrations.

[0134] It should be noted that this step is a relative fractionation rather than a complete separation; therefore, it is not required that the precipitate be completely free of short-chain fructans, nor is it required that the liquid fraction be completely free of medium-chain fructans or rhamnose-rich polysaccharides.

[0135] Unlike conventional recovery methods that use higher concentrations of ethanol to precipitate as much of the sugar as possible, this invention intentionally retains the liquid portion and adds it back in after subsequent ethanol removal. Therefore, the technical objective of this step is not to improve the recovery purity of a single polysaccharide, but rather to subject different sugar components to different subsequent processing procedures.

[0136] 8. Limited Resolution of the Precipitated Component

[0137] After collecting the precipitate obtained from ethanol fractionation, it can be washed quickly with a small amount of low-concentration ethanol, or it can proceed directly to the reconstitution step without washing; preferably, purified water is added directly for reconstitution to avoid repeated alcohol precipitation causing additional effects on the state of the sugars. The reconstitution temperature is 30–35°C, preferably 31–33°C, for example 32°C, and the reconstitution process is carried out with slow to medium-speed stirring for 30–60 min, preferably 40–50 min, for example 45 min.

[0138] The "redissolution" mentioned in this invention refers to redispersing the precipitate obtained from ethanol treatment and allowing it to enter the aqueous system. It does not mean that all the high molecular weight sugars in the system must reach complete hydration equilibrium after prolonged storage.

[0139] Therefore, the present invention preferably does not use high temperature of 60-80°C for long-term dissolution, nor does it preferably place the precipitate in water separately for more than 24 hours before adding the liquid part.

[0140] Preferably, the precipitate is not subjected to heat treatment above 45°C after resolution, and the treated liquid portion is added within 6 hours after the resolution treatment is completed; more preferably, subsequent mixing is carried out within 0.5 to 3 hours after the resolution treatment is completed.

[0141] De-alcoholization and concentration of the liquid portion

[0142] The liquid fraction obtained from ethanol fractionation is not discarded, but rather the short-chain fructans contained therein are further recovered.

[0143] First, ethanol can be removed by vacuum concentration, removing most of the ethanol at a temperature not exceeding 45°C. It is preferable to control the temperature of the feed solution at 35-45°C. Low-temperature vacuum deethanolination can reduce the impact of prolonged high-temperature treatment on sugars.

[0144] After de-alcoholization, the liquid portion can be used directly for subsequent remixing based on its concentration, or it can be further concentrated using a membrane.

[0145] The liquid portion was re-added to the precipitate after reconstitution.

[0146] Add the liquid portion that has been de-alcoholized and concentrated as needed to the precipitate portion that has been reconstituted under the above-specified conditions; during the addition process, low or medium speed stirring can be continuously carried out to avoid local concentration differences caused by rapid addition at one time.

[0147] After all the liquid portion is added, the system temperature is maintained at 25–30°C, preferably 28–30°C, and stirred for 30–60 min, for example, stirring at 28°C for 45 min; then the system is placed at 23–27°C for 2–6 h, for example, at 25°C for 4 h. If necessary, the final pH can be adjusted to 5.4–5.8, for example, 5.6; thereby obtaining the composite active material of the present invention.

[0148] Preparation of skin care products

[0149] The composite active ingredients obtained by this invention can be used in various skin care preparations such as serums, lotions, creams, gels, and facial masks.

[0150] Taking emulsified skin care formulations as an example, the aqueous phase and oil phase can be prepared separately.

[0151] The aqueous phase may include one or more of water, glycerol, 1,3-propanediol, butylene glycol, sodium hyaluronate, thickeners, and chelating agents; the oil phase may include squalane, caprylic / capric triglycerides, vegetable oils, fatty alcohols, emulsifiers, etc.; the aqueous and oil phases are heated and mixed separately, emulsified at a suitable temperature, and then cooled.

[0152] When the temperature of the emulsion system drops below 45°C, preferably 35-40°C, the composite active ingredient prepared in this invention is added; for example, it can be added at 38°C; after addition, it is stirred evenly at an appropriate speed, and then heat-sensitive raw materials such as preservatives are added, and the final pH is adjusted to 5.0-6.2, preferably 5.3-5.9.

[0153] The present invention preferably adds the composite active ingredient at a temperature below 45°C, on the one hand to avoid unnecessary high-temperature treatment, and on the other hand to maintain the hydration state of the sugars formed after the aforementioned treatment as much as possible.

[0154] Stress-induced skin condition and its evaluation

[0155] The term "stress-induced skin condition" as used in this invention refers to one or more changes in skin condition that occur under the influence of life stress, lack of sleep, continuous work, emotional fluctuations, or other non-disease-related stress factors. These changes include, but are not limited to, dry skin, increased transepidermal water loss, redness, roughness, dullness, decreased luster, decreased elasticity, increased fine lines or wrinkles, and reduced skin comfort. This invention is not intended to diagnose or treat mental illness.

[0156] The effects of the compound active ingredients of this invention can be evaluated from cell or tissue-related indicators and human skin indicators, respectively.

[0157] For skin well-being-related indicators, β-endorphin can be selected; for skin neuro-immune-related indicators, neuromodulatory peptide B (NMB) and thymic stromal lymphopoietin (TSLP) can be selected.

[0158] This invention does not use "happiness factor" as a strict single biochemical term, but prefers to use "skin pleasure-related factor" or directly uses specific detectable indicators to describe it.

[0159] For human skin evaluation, parameters such as stratum corneum moisture content, transepidermal water loss, skin erythema index, skin luster, skin roughness, elasticity, and wrinkles can be measured and analyzed in conjunction with the subjects' subjective evaluations of tightness, dryness, itching, comfort, and overall skin sensation.

[0160] First, partial restrictive enzymatic hydrolysis alters the chain length of some chicory fructans, while retaining the unhydrolyzed portion prevents the entire system from being converted into short-chain fructans. Second, the fructans of different chain lengths obtained after remixing are combined with rhamnose-rich fermented polysaccharides in the ethanol fractionation step. Due to the different molecular weights and solubility characteristics of the various sugar components, they exhibit relatively different distributions in the aqueous ethanol system. Third, the precipitate and liquid fractions are not treated in the same way. The precipitate undergoes ethanol dehydration followed by redissolution at a specified temperature and time, while the liquid fraction mainly undergoes dealcoholization and concentration.

[0161] Finally, after the prescribed resolution treatment is completed in the precipitate, the liquid portion containing more short-chain fructans is added back in.

[0162] Example

[0163] Unless otherwise stated, the water used in the following examples and comparative examples is purified water; the ethanol is 95% by volume; the rhamnose-rich fermentation polysaccharide uses the same batch of raw materials, whose main sugar residues include rhamnose, galactose, and glucuronic acid, with a weight-average molecular weight of approximately 50 kDa. The endo-inulinase used is from the same batch of enzyme preparation. All enzyme additions below are based on actual enzyme activity; the chicory root raw material uses the same batch of dried chicory roots with a moisture content not exceeding 8%.

[0164] Example 1

[0165] This embodiment provides a skin care composition for improving stress-related skin conditions. The skin care composition includes a complex active ingredient obtained by co-processing chicory root extract and rhamnose-rich fermented polysaccharide.

[0166] 1. Preparation of chicory root extract

[0167] Weigh 1000 g of dried chicory root, pulverize it, and pass it through a 40-mesh sieve. Add 10000 g of purified water. Heat the system to 58°C and maintain the temperature under stirring for 2 hours. After extraction, filter to remove insoluble residues from the chicory root and collect the filtrate. Concentrate the obtained filtrate under reduced pressure at a temperature not exceeding 50°C, and adjust with purified water to achieve a soluble solids content of 10.0% for the chicory root extract.

[0168] 2. Partial restriction enzymatic hydrolysis

[0169] The obtained chicory root extract was thoroughly mixed. A portion of the chicory root extract A was taken as the enzymatic hydrolysis fraction, and its pH was adjusted to 5.0 using a citric acid-sodium citrate system. The temperature was then raised to 52℃. The ratio of chicory root extract A (with added endo-inulinase) to chicory root extract B (without added endo-inulinase) was 65:35.

[0170] Add endo-inulinase to chicory root extract at 6 U / g, stir and hydrolyze at 52℃ for 50 min; after hydrolysis, rapidly raise the temperature to 90℃ and hold for 10 min to inactivate the enzyme, then cool down.

[0171] Take another undigested chicory root extract, adjust the pH to 5.0 without adding endo-inulinase, maintain the temperature at 52℃ for 50 min, then raise the temperature to 90℃ and maintain it for 10 min, and then cool it down; mix the enzymatically hydrolyzed part with the undigested part to obtain the chicory root treatment solution (solid content of about 10%).

[0172] 3. Add rhamnose-rich fermented polysaccharide

[0173] Add rhamnose-rich fermented polysaccharide (the mass ratio of chicory root treatment solution to rhamnose-rich fermented polysaccharide is 100:1.0) to the obtained chicory root treatment solution and stir to disperse it evenly; adjust the pH of the system to 5.5 and stir at 35℃ for 60 min to ensure that the chicory root-derived fructan and rhamnose-rich fermented polysaccharide are evenly present in the same aqueous phase system.

[0174] 4. Ethanol fractionation

[0175] The system was cooled to 23°C, and 95% ethanol was slowly added under continuous stirring until the final ethanol volume fraction in the mixture reached 52 vol.%. The ethanol was added gradually over 20 min, and after the addition was completed, the mixture was stirred at 23°C for another 20 min. The system was then placed at 6°C for 8 h. After the placement, the mixture was centrifuged at 6000×g for 12 min, and the precipitate P and the liquid S were collected separately.

[0176] 5. Resolution of precipitated P

[0177] Purified water was added to the precipitate P and stirred at 32°C for 45 min to obtain a solid content of 10%. No heating treatment exceeding 45°C was used during the reconstitution process. After reconstitution, the product was not left to stand alone for a long time, but directly entered the subsequent liquid reconstitution step.

[0178] 6. Treatment of liquid component S

[0179] The resulting liquid fraction S was subjected to reduced pressure at a temperature not exceeding 45°C to remove ethanol. After removal of alcohol, it was concentrated by nanofiltration to obtain a solid content of 10%.

[0180] 7. Remix

[0181] The above-mentioned S-part concentrate was slowly added to the P-part which had been reconstituted, while stirring was maintained during the addition process. After the addition was completed, the pH of the system was adjusted to 5.6, and stirring was continued at 28°C for 45 min. Then the system was placed at 25°C for 4 h to obtain the composite active product of Example 1.

[0182] The preparation method of rhamnose-rich fermented polysaccharide is as follows:

[0183] Preparation of bacterial strains and seed culture: Sphingomonas sp. strains that can secrete rhamnose extracellular polysaccharides were used as fermentation bacteria.

[0184] The preserved bacterial strain was inoculated into the activation medium and cultured at 30°C for 18–24 h to activate it. The activated bacterial solution was then inoculated into the seed medium at a volume of 2.0 vol.%.

[0185] The seed culture medium consisted of 20 g / L glucose, 5 g / L yeast extract, 5 g / L peptone, 1.5 g / L dipotassium hydrogen phosphate, 0.5 g / L potassium dihydrogen phosphate, and 0.2 g / L magnesium sulfate, with an initial pH of 7.0.

[0186] After the culture was completed, the absorbance of the seed culture at 600 nm was measured. Uninoculated seed culture medium was used as a blank. When the OD600 of the seed culture was 2.5, it was used for the subsequent first stage fermentation.

[0187] First stage of fermentation: Inoculate the above seed liquid at 8% of the fermentation medium volume.

[0188] The first stage fermentation medium includes: glucose 25 g / L, chicory oligofructose 5 g / L, yeast extract 3.0 g / L, ammonium sulfate 1.5 g / L, dipotassium hydrogen phosphate 1.2 g / L, potassium dihydrogen phosphate 0.4 g / L, magnesium sulfate 0.20 g / L, and trace element solution 1.0 mL / L.

[0189] The chicory oligofructose is obtained by hydrolyzing the chicory root-derived fructans with an endoglucanase (not included in the previous weight parts). The initial pH of the fermentation system is adjusted to 7.0, and fermentation is carried out at 30°C. For the first 12 hours, the stirring speed is controlled at 250 r / min and the aeration rate is 1.0 vvm to allow the cells to grow sufficiently.

[0190] After 12 hours of fermentation, a large amount of glucose was no longer added at once. The culture continued until the glucose concentration in the fermentation broth decreased to below 2.0 g / L, so that the cells gradually transitioned from the rapid proliferation stage to the extracellular polysaccharide synthesis stage. Subsequently, the fermentation temperature was reduced to 28℃, the pH was adjusted to 6.6-6.8, and the aeration rate was reduced to 0.5 vvm.

[0191] Segmented feeding: The first feeding solution was added after 16 h of fermentation, which added the following to the fermentation system: chicory oligofructose 3.0 g / L, L-rhamnose 1.5 g / L and D-galactose 1.0 g / L.

[0192] A second feed solution was added after 24 hours of fermentation, introducing the following additional components into the fermentation system: chicory fructooligosaccharides 2.0 g / L, L-rhamnose 1.0 g / L, and D-galactose 0.5 g / L. After the feed solution was added, fermentation continued at 28°C and pH 6.6–6.8 for 36 hours.

[0193] The addition of L-rhamnose and D-galactose here is mainly as a supplementary carbon source and sugar metabolism regulator during the fermentation stage, and is not directly equivalent to their inclusion in the final extracellular polysaccharide chain in their original form; the monosaccharide composition of the final polysaccharide is still based on the results of the hydrolysis analysis of the purified polysaccharide.

[0194] Post-fermentation maturation: After 36 hours of fermentation, feeding was stopped, the temperature was lowered to 25°C, and the aeration rate was reduced to 0.2 vvm. This process was continued for 6 hours without further carbon source replenishment. After the post-fermentation maturation was completed, the fermentation broth was heated to 70°C and held for 20 minutes to inactivate the cells; then cooled to room temperature.

[0195] Cell removal and polysaccharide purification: The inactivated fermentation broth was centrifuged at 8000×g for 15 min to remove cells and insoluble matter. The supernatant was collected and ultrafiltered using a 10 kDa molecular weight cutoff ultrafiltration membrane. The retentate was collected and continuously washed with purified water to remove residual monosaccharides, oligosaccharides, inorganic salts, and other low-molecular-weight substances. When the conductivity of the permeate decreased and tended to stabilize, washing was stopped, and the resulting retentate was concentrated to approximately 1 / 5 of its original volume.

[0196] Then, ethanol was slowly added while stirring to make the ethanol volume fraction in the system reach 65%. The mixture was placed at 4°C for 10 h, and the polysaccharide precipitate was collected by centrifugation at 8000×g for 15 min. The precipitate was then redissolved in purified water.

[0197] Molecular weight fractionation: The redissolved polysaccharide solution was fractionated sequentially using a suitable ultrafiltration membrane to remove excessively low molecular weight components and excessively high molecular weight components. The polysaccharide fraction with the main weight-average molecular weight distribution in the range of 40–60 kDa was collected. The obtained fraction was washed and filtered again with purified water, concentrated, and freeze-dried to obtain rhamnose-rich fermented polysaccharide. The weight-average molecular weight (Mw) was determined to be approximately 50 kDa using SEC-MALS.

[0198] Example 2

[0199] This embodiment provides a skin care composition for improving stress-related skin conditions. The skin care composition includes a complex active ingredient obtained by co-processing chicory root extract and rhamnose-rich fermented polysaccharide.

[0200] 1. Chicory root extract

[0201] Weigh 1000 g of dried chicory root, crush it through a 40-mesh sieve, add 12000 g of purified water, and extract at 55℃ for 2.5 h. After solid-liquid separation, collect the extract and concentrate it under reduced pressure at a temperature not exceeding 50℃ to make the soluble solids content 9.0%.

[0202] 2. Partial restriction enzymatic hydrolysis

[0203] Take a portion of the chicory root extract, adjust the pH to 5.1, and add endo-inulinase at 4 U / g of the extract at 50℃ for 60 min. Then, heat to 90℃ and hold for 8 min to inactivate the enzyme. The remaining un-hydrolyzed portion was treated without endo-inulinase, and kept at pH 5.1 and 50℃ for 60 min, then at 90℃ for 8 min. The two portions were then remixed to obtain the chicory root treatment solution. The ratio of chicory root extract A (with endo-inulinase) to chicory root extract B (without endo-inulinase) was 65:35.

[0204] 3. Add rhamnose-rich fermented polysaccharide and perform ethanol fractionation.

[0205] Add the same batch of rhamnose-rich fermented polysaccharide as in Example 1 to the obtained chicory root treatment solution, adjust the pH to 5.4, and stir at 33°C for 75 min; cool to 22°C, slowly add ethanol to make the final ethanol volume fraction of the system reach 50 vol.%, continue stirring for 25 min, and place at 8°C for 10 h, then centrifuge at 6000×g for 15 min to collect the precipitate P and the liquid S respectively.

[0206] 4. Process separately and remix

[0207] The precipitate P was added to purified water and stirred at 30°C for 60 min to redissolve it. The liquid part S was subjected to de-alcoholization under reduced pressure at a temperature not exceeding 45°C and concentrated using the same membrane system as in Example 1. After the P part was redissolved, the S part concentrated by de-alcoholization was added to it and stirred at 30°C for 30 min. Then it was placed at 25°C for 3 h to obtain the composite active product of Example 2.

[0208] Comparative Example 1

[0209] First, chicory root treatment solution was prepared according to the method of Example 1, including the same chicory root extraction conditions, partial restriction enzymatic hydrolysis conditions, and untreated portion conditions. Additionally, the same batch of rhamnose-rich fermented polysaccharide as in Example 1 was used.

[0210] Based on the HPAEC-PAD glycan distribution, total sugar content, and rhamnose-rich fermented polysaccharide content of the final composite active product obtained in Example 1, the amount of chicory root treatment liquid and rhamnose-rich fermented polysaccharide added was adjusted so that the short-chain fructan, medium-chain fructan, rhamnose-rich fermented polysaccharide, total sugar content, and pH in the final system of this comparative example were as consistent as possible with those in Example 1.

[0211] The above raw materials were directly added to purified water and stirred at 35°C for 60 min. Then the pH was adjusted to 5.6, stirred at 28°C for 45 min, and then placed at 25°C for 4 h to obtain Comparative Example 1.

[0212] The main difference between this comparative example and Example 1 is that the final main sugar composition is as similar as possible, but all sugars have not undergone 52% ethanol fractionation, ethanol dehydration-limited resolution of the precipitate, and the re-addition of the liquid portion.

[0213] Comparative Example 2

[0214] Chicory root extract was prepared according to the method in Example 1. The difference from Example 1 is that all chicory root extracts were enzymatically hydrolyzed with endo-inulinase.

[0215] Specifically, the pH of all chicory root extract was adjusted to 5.0, and endo-inulinase was added at 6 U / g of chicory root extract at 52°C for 50 min of enzymatic hydrolysis. The temperature was then raised to 90°C and held for 10 min to inactivate the enzyme. The remaining steps, including the addition of rhamnose-enriched fermentation polysaccharide, fractionation with 52 vol.% ethanol, incubation at 6°C for 8 h, reconstitution of fraction P at 32°C for 45 min, de-alcoholization and concentration of fraction S, and remixing, were the same as in Example 1, yielding Comparative Example 2.

[0216] Comparative Example 3

[0217] Chicory root extract was prepared according to the method of Example 1. Unlike Example 1, no endo-inulinase was added to any of the chicory root extracts. Instead, the same heat treatment was performed at pH 5.0, 52°C for 50 min, and 90°C for 10 min. Then, the same rhamnose-rich fermentation polysaccharide as in Example 1 was added directly.

[0218] The remaining steps of ethanol fractionation, P-part reconstitution, S-part treatment, and remixing were the same as in Example 1, resulting in Comparative Example 3.

[0219] Comparative Example 4

[0220] The chicory root extraction, partial restriction enzymatic hydrolysis, and remixing of the hydrolyzed and unhydrolyzed portions were completed according to the method of Example 1; the same batch of rhamnose-rich fermented polysaccharide as in Example 1 was added, the pH was adjusted to 5.5, and the mixture was stirred at 35°C for 60 min.

[0221] Unlike Example 1, no ethanol was added, and the precipitate and liquid fractions were not separated. The resulting mixture was stirred directly at 28°C for 45 min, and then placed at 25°C for 4 h to obtain Comparative Example 4.

[0222] Comparative Example 5

[0223] The ethanol fractionation step was completed according to the method of Example 1, yielding the same precipitate P and liquid fraction S; the liquid fraction S was then subjected to low-temperature depressurization deethanolination and membrane concentration according to the method of Example 1.

[0224] Unlike Example 1, the precipitate P was added to purified water and stirred at 60°C for 2 h, then cooled to 25°C and left to stand for 24 h to allow for more thorough individual hydration. After the P part had completed the above treatment, the S part, which had been concentrated by de-alcoholization, was added. After mixing, the mixture was stirred at 28°C for 45 min according to the method of Example 1, and then left to stand at 25°C for 4 h to obtain Comparative Example 5.

[0225] Comparative Example 6

[0226] Ethanol fractionation was performed according to the method of Example 1 to obtain precipitate P and liquid S. The S fraction was de-ethanolified and concentrated according to the method of Example 1. The difference from Example 1 is that the P fraction was not subjected to independent reconstitution treatment at 32°C for 45 min.

[0227] Purified water and S-part concentrate after dealcoholization were added directly to P-part to allow P-part to begin absorbing water and dispersing in the system where short-chain fructans already existed. The mixture was then stirred at 28°C for 45 min and placed at 25°C for 4 h to obtain Comparative Example 6.

[0228] Comparative Example 7

[0229] The chicory root extraction, partial restriction enzymatic hydrolysis, remixing, and addition of rhamnose-enriched fermentation polysaccharides were completed according to the method in Example 1.

[0230] Unlike Example 1, ethanol was added during the ethanol fractionation process to bring the final ethanol volume fraction in the system to 70 vol.%. After the ethanol was added, the mixture was stirred at 23°C for 20 min, placed at 6°C for 8 h, and then centrifuged at 6000×g for 12 min.

[0231] The precipitate and liquid fractions were collected separately. Then, following the same method as in Example 1, the precipitate was re-dissolved at 32°C for 45 min, the liquid fraction was concentrated by alcohol removal, and then re-mixed to obtain Comparative Example 7.

[0232] Performance testing

[0233] To evaluate the sugar distribution characteristics, physical stability, and improvement effect on stress-related skin conditions of the composite active material prepared in this invention, the samples obtained in Examples 1-2 and Comparative Examples 1-7 were subjected to the following performance tests.

[0234] Unless otherwise specified, all samples were compared using the same total solids concentration to minimize the impact of sample concentration differences on the test results.

[0235] Experiment 1: Viscosity Stabilization Time Test of Composite Active Material

[0236] 1. Test Objective

[0237] The study evaluated the continuous hydration and viscosity changes of composite active substances obtained by different preparation methods after preparation, and verified whether the "ethanol fractionation-limited resolution of P part-recombination of S part" method can reduce the viscosity drift of the composition after preparation.

[0238] 2. Sample processing

[0239] The composite active substances obtained in Examples 1-2 and Comparative Examples 1-7 were uniformly adjusted to a solid content of 10.0 wt.% using purified water, and the pH was adjusted to 5.60±0.05. The timing was started immediately after the sample preparation was completed, and the samples were sealed and placed in an environment of 25±1℃.

[0240] 3. Viscosity test

[0241] The test was conducted using a rotational rheometer at a temperature of 25.0 ± 0.5℃, employing a plate or cone-plate measurement system suitable for the sample viscosity range.

[0242] Before testing, gently mix the sample to avoid introducing obvious air bubbles, and test at a shear rate of 10 s. -1 The viscosity was measured under the specified conditions for 60 seconds, and the average viscosity of the last 30 seconds was taken as the apparent viscosity at that time point. Tests were conducted at 0 h, 2 h, 4 h, 8 h, 12 h, 24 h and 48 h after preparation.

[0243] Using the apparent viscosity η48 at 48 h as a reference, the viscosity deviation at each time point is calculated as follows: Dt = (|ηt - η48|) / η48 * 100%

[0244] Experiment 2: Thixotropic recovery performance test

[0245] 1. Test Objective

[0246] To evaluate the structural recovery ability of complex active ingredients after being subjected to high shear stress, and to further determine whether different treatment processes affect the rheological behavior of carbohydrate systems, this test is related to the pumping, homogenization, filling, and consumer application processes in actual cosmetic production.

[0247] 2. Testing Methods

[0248] All groups of compound active ingredients were uniformly adjusted to a solid content of 10.0 wt.% and a pH of 5.60±0.05, and equilibrated at 25℃ for 24 h.

[0249] Three-segment shear test was performed using a rotational rheometer:

[0250] Phase 1: Shear rate 0.1 s -1 Hold for 60 seconds;

[0251] Second stage: shear rate 100 s -1 Hold for 60 seconds;

[0252] Phase 3: Reduced back to 0.1 s -1 Hold for 300 seconds.

[0253] Record the average viscosity η0 10 s before the end of the first stage, and the average viscosity η300 10 s before the 300th s of the third stage.

[0254] The thixotropic recovery rate is calculated using the following formula: R = η300 / η0 * 100%, and each sample is tested in parallel 3 times.

[0255] The closer the thixotropic recovery rate is to 100%, the better the sample's ability to recover to its initial rheological state after experiencing high shear.

[0256] Experiment 3 Low-temperature storage stability test

[0257] 1. Test Objective

[0258] To evaluate whether the complex active products formed by fermentation of fructans with different chain lengths and rhamnose-rich polysaccharides exhibit turbidity, precipitation, crystallization, or stratification under low-temperature storage conditions.

[0259] 2. Testing Methods

[0260] All samples were uniformly adjusted to a solid content of 10.0 wt.% and a pH of 5.60±0.05.

[0261] Take 50 mL of each sample and place it into a transparent, sealed sample bottle.

[0262] Setup: 4±1℃ low-temperature group; 25±1℃ normal-temperature control group. Storage was carried out for 0 days, 7 days, 14 days, and 28 days, respectively.

[0263] Sampling and testing at each time point:

[0264] (1) Light transmittance

[0265] The transmittance T600 was measured at 600 nm using a UV-Vis spectrophotometer, with purified water as a blank.

[0266] (2) Turbidity

[0267] Measurements were taken using a turbidimeter and are expressed as NTU.

[0268] (3) Centrifugal stability

[0269] Take 10 mL of sample and centrifuge at 3000×g for 15 min. Observe whether precipitation, flocculent matter or obvious stratification occurs.

[0270] (4) Crystal observation

[0271] Take a small sample and place it on a glass slide. Observe under a polarizing microscope whether crystals with birefringence characteristics appear.

[0272] For samples that become slightly turbid after storage at 4°C, restore them to 25°C for 24 hours and observe whether they can return to their pre-storage state.

[0273] Experiment 4: Formulation stability of compound active ingredients in emulsified skin care preparations

[0274] 1. Basic Formula

[0275] The composite active ingredients obtained in Examples 1-2 and Comparative Examples 1-7 were added to the same base emulsion, and the amount added to each group was kept the same based on the solid content of the composite active ingredients. Except for the different composite active ingredients, the raw materials and preparation process of the base emulsion were completely the same.

[0276] The following basic system can be adopted:

[0277] raw material mass percentage content / % glycerin 4.00 1,3-Propanediol 3.00 squalane 4.00 Caprylic / Capric Triglycerides 3.00 Cetearyl alcohol 1.50 emulsifier 2.50 Xanthan Gum 0.15 Disodium EDTA 0.05 Complex active ingredients 10 Anti-corrosion system Appropriate amount Citric acid / Sodium citrate Adjust pH water Supplement to 100

[0278] The aqueous and oil phases were heated to about 75°C, mixed and emulsified, and then cooled down. When the temperature dropped to 38°C, the corresponding composite active ingredient was added, stirred evenly, and the final pH was adjusted to 5.5±0.1.

[0279] 2. Stability Test

[0280] The emulsions in each group were stored at 4℃, 25℃, and 45℃ for 28 days; measurements were taken at 0, 7, 14, and 28 days.

[0281] Appearance, layering, viscosity, average droplet size, and droplet size distribution.

[0282] The average droplet size was determined using a laser particle size analyzer; the viscosity was determined at 25°C, with a fixed rotor and a fixed rotation speed.

[0283] Calculate the viscosity retention rate after storage at 45℃ for 28 days: VR = η28d / η0d * 100%

[0284] Simultaneously calculate the average particle size change rate.

[0285] Experiment 5: Skin Pleasure-Related Factor β-Endorphin Test

[0286] 1. Test Objective

[0287] To evaluate the effects of different compound active ingredients on the secretion of β-endorphin in skin cells.

[0288] 2. Cell Culture

[0289] Normal human epidermal keratinocytes were used and cultured in a suitable keratinocyte culture medium at 37°C, 5% CO2, and saturated humidity. Cells in good growth condition from the 3rd to 5th generation were selected for the experiment. The cells were seeded in 24-well plates and cultured until they reached approximately 70% to 80% confluence.

[0290] 3. Grouping

[0291] Setup: Normal control group; stress model group; Example 1 group; Example 2 group; Comparative groups 1-7. All samples were added to the culture system at the same total sugar concentration to avoid result deviations due to different sample amounts.

[0292] Before the formal experiment, a cell viability assay is used to determine the test concentration that has no obvious cytotoxicity. For example, concentrations such as 0.01%, 0.05%, and 0.10% can be screened first, and the concentration with a cell survival rate of not less than 90% can be selected for the formal experiment.

[0293] 4. Pressure Model

[0294] To simulate stress-related skin conditions, a stress model can be established by treating keratinocytes with cortisol.

[0295] The model conditions should be determined through preliminary experiments to ensure that the β-endorphin level in the model group changes stably while the cell viability remains within an acceptable range. After the model is established, each sample to be tested should be added and cultured for the specified time.

[0296] 5. Testing

[0297] Cell culture supernatant was collected and detected using a human β-endorphin ELISA kit. A standard curve was established according to the kit instructions, and the concentration of β-endorphin in the culture medium was calculated based on the absorbance. Simultaneously, the total protein content or cell number in each well was measured. The β-endorphin results were normalized. Each group had at least three replicates, and at least three independent replicates were performed.

[0298] Experiment 6: Efficacy Test for Human Stress-Induced Skin

[0299] 1. Subjects

[0300] A total of 180 healthy participants aged 18–55 years were recruited who met the requirements. Participants had recently experienced some work, study, or life stress, and had at least two of the following symptoms: dry skin, tightness, redness, dullness, roughness, or a perceived decrease in skin comfort.

[0301] Subjects with active skin diseases in the test area, those who have recently received related skin treatments, or those who have used medications or highly effective skin care products that may significantly affect the test results were excluded. A randomized, double-blind, parallel-controlled method was used. Subjects were randomly divided into 9 groups of 20 each. Each group used a test emulsion containing the complex active ingredients from Examples 1, 2, and Comparative Examples 1–7. The complex active ingredients in each group's test emulsion were prepared with the same amount of solids added. The emulsions were applied twice daily, morning and evening, with the prescribed amount evenly applied to the face for 28 consecutive days. Testing was conducted before use (D0) and after use (D7, D14, and D28).

[0302] Before the test, the subjects cleaned their faces and sat quietly for 30 minutes in a constant temperature and humidity environment of 22±2℃ and 50±5% without using any other skin care products, and then underwent instrument testing.

[0303] To minimize differences in baseline skin condition among individuals, a randomized, double-blind design with self-control on the left and right sides of the face is preferred.

[0304] The emulsion prepared in Example 1 and the directly mixed emulsion of equal composition in Comparative Example 1 were randomly applied to the left and right sides of the subjects' faces, respectively. Except for the preparation method of the compound active ingredient, the two emulsions were identical in their basic formula, amount of active ingredient solids, appearance, and application method. They were applied twice daily, morning and evening, with a fixed amount applied to each side each time, for 28 consecutive days. Tests were performed before use (D0) and after use (D7, D14, and D28).

[0305] Before the test, the subjects cleaned their faces and did not use any other skin care products. After sitting quietly for 30 minutes in an environment with a temperature of 22±2℃ and a relative humidity of 50±5%, the instrument test was conducted. Each follow-up visit was conducted at a similar time period as much as possible.

[0306] stratum corneum moisture content

[0307] The moisture content of the stratum corneum was measured using a Corneometer-like stratum corneum moisture meter. Each test area was measured three times consecutively, and the average value was taken to calculate the rate of change relative to D0.

[0308] transdermal water loss

[0309] TEWL was measured using a Tewameter-type instrument. At least three stable readings were obtained for each test area, and the average value was taken, expressed in g / (m²). 2 ·h) indicates that a decrease in TEWL suggests an improvement in skin barrier-related conditions.

[0310] Erythema Index

[0311] The erythema index was measured using a Mexameter-type skin pigmentation / erythema analyzer, with three measurements taken for each test area and the average value calculated.

[0312] Skin radiance

[0313] The measurement was performed using a Glossymeter-type skin gloss meter or a standardized facial image analysis system, with each test area measured at a fixed position, angle, and ambient light conditions.

[0314] skin roughness

[0315] The surface morphology of the test area was acquired using a three-dimensional skin imaging device, and Sa, Sq or corresponding skin roughness parameters were calculated. The same test position was maintained at all time points.

[0316] Skin comfort assessment: A visual analog scale of 0-10 was used, with subjects rating the following: dryness; tightness; itching or discomfort; skin softness; and overall comfort. For negative sensations, 0 points indicated no discomfort at all, and 10 points indicated very noticeable discomfort. For overall comfort, 0 points indicated very uncomfortable discomfort, and 10 points indicated very comfortable comfort. The scoring direction for each indicator should remain consistent throughout the trial.

[0317] Quantitative data are expressed as mean ± standard deviation. For changes before and after use on the same test side, paired statistical methods can be used. For comparison of changes between the example side and the control side, paired t-tests or corresponding non-parametric tests can be selected based on whether the data meet the requirements of normal distribution and homogeneity of variance. P < 0.05 is used as the criterion for judging statistical significance of the difference.

[0318] Performance test results

[0319] Experiment 1 Viscosity Stabilization Time

[0320] All samples were uniformly adjusted to a solid content of 10.0 wt.% and a pH of 5.60±0.05, and the apparent viscosity was measured at different standing times under 25℃.

[0321] Table 2. Apparent viscosity of different samples during storage, unit: mPa·s

[0322] sample 0 h 2 h 4 h 8 h 12 h 24 h 48 h Stabilization time Example 1 426 448 461 467 469 471 470 4 h Example 2 418 437 450 457 460 462 461 8 h Comparative Example 1 332 361 389 418 441 459 466 24 h Comparative Example 2 301 317 328 337 341 343 344 8 h Comparative Example 3 497 541 579 606 619 626 628 12 h Comparative Example 4 346 374 401 426 444 458 463 24 h Comparative Example 5 486 480 476 473 472 471 470 4 h Comparative Example 6 351 389 421 448 463 473 476 12 h Comparative Example 7 453 471 482 489 491 493 493 8 h

[0323] Experiment 2 Thixotropic recovery performance

[0324] Table 3. Thixotropic recovery rate of different samples

[0325] sample Thixotropic recovery rate / % Example 1 94.6±1.8 Example 2 91.8±2.1 Comparative Example 1 78.7±2.6 Comparative Example 2 82.1±2.3 Comparative Example 3 80.4±2.8 Comparative Example 4 77.5±3.0 Comparative Example 5 84.2±2.4 Comparative Example 6 73.8±3.1 Comparative Example 7 85.6±2.2

[0326] The thixotropic recovery rate of Example 1 reached 94.6%, which was significantly higher than that of Comparative Example 1, which had a similar final main sugar composition but was prepared by direct mixing.

[0327] Comparative Example 6, which did not undergo independent resolution of P portion but directly added S portion, had the lowest thixotropic recovery rate of 73.8%. This indicates that when high molecular weight sugars are co-hydrated in a system with a high content of short-chain sugars before initial resolution, it is not conducive to forming a system with good shear recovery ability.

[0328] Although Comparative Example 5 was fully hydrated and its viscosity stabilized relatively quickly, its thixotropic recovery rate was still lower than that of Example 1, indicating that "quickly reaching stable viscosity" is not equivalent to having good rheological recovery performance.

[0329] Experiment 3: Low temperature stability. Each sample was stored at 4°C for 28 days before being tested.

[0330] Table 4. Stability results after storage at 4℃ for 28 days

[0331] sample T600 / % Turbidity / NTU State after centrifugation polarizing crystal Example 1 93.8 7.6 No obvious sediment Not seen Example 2 91.7 9.8 No obvious sediment Not seen Comparative Example 1 78.6 28.7 Small amount of flocculent sediment small amount Comparative Example 2 92.4 8.9 No obvious sediment Not seen Comparative Example 3 68.3 46.2 Obvious sedimentation obvious Comparative Example 4 76.9 31.4 Small amount of sediment small amount Comparative Example 5 82.7 22.5 Slight flocculent matter small amount Comparative Example 6 74.8 35.7 flocculent sediment small amount Comparative Example 7 84.5 19.1 slight deposition small amount

[0332] Experiment 4: Stability of emulsified skin care formulations. Each group of compound active ingredients was added to the same emulsion matrix in equal amounts of solids and stored at 45°C for 28 days.

[0333] Table 5. Emulsion stability after storage at 45℃ for 28 days

[0334] sample Viscosity retention rate / % Initial average particle size / μm 28-day average particle size / μm Particle size increase rate / % Appearance Example 1 96.1 1.82 1.91 4.9 Uniform, without layering Example 2 94.3 1.85 1.98 7.0 Uniform, without layering Comparative Example 1 83.4 1.87 2.24 19.8 slight viscosity change Comparative Example 2 91.6 1.80 1.96 8.9 No obvious stratification Comparative Example 3 76.8 1.91 2.49 30.4 Slight flocculation Comparative Example 4 81.5 1.88 2.31 22.9 slight viscosity change Comparative Example 5 86.9 1.84 2.12 15.2 Basically uniform Comparative Example 6 79.8 1.89 2.39 26.5 Small amount of flocculation Comparative Example 7 88.1 1.86 2.10 12.9 Basically uniform

[0335] Experiment 5: β-Endorphin Detection Results

[0336] Preliminary experiments confirmed that the survival rate of keratinocytes in each group was higher than 90% at the concentrations used in the formal experiments.

[0337] The β-endorphin level in the stress model group was normalized to 100%, and the other groups were expressed as percentages relative to the stress model group.

[0338] Table 6. Effects of each group on β-endorphin levels

[0339] Group β-Endorphin relative level / % normal control group 151.3±7.4 Pressure model group 100.0±5.2 Example 1 143.8±6.1 Example 2 138.6±6.5 Comparative Example 1 126.4±5.8 Comparative Example 2 119.7±6.0 Comparative Example 3 116.2±5.7 Comparative Example 4 124.8±6.3 Comparative Example 5 129.1±6.1 Comparative Example 6 121.5±5.9 Comparative Example 7 127.6±6.4

[0340] Example 1 restored the β-endorphin level in the stress model from 100% in the model group to 143.8%, which is close to that of the normal control group.

[0341] Example 1 showed better results than Comparative Examples 2 and 3, indicating that neither complete enzymatic hydrolysis nor complete non-enzymatic hydrolysis exhibited the same effect as partial restriction enzymatic hydrolysis.

[0342] Results of human efficacy evaluation in Experiment 6

[0343] Table 7 Ergonomics Test Results

[0344] Group Change rate of stratum corneum moisture content / % TEWL change rate / % Change rate of erythema index / % Gloss change rate / % Sa change rate / % Example 1 28.6 -20.8 -17.9 18.7 -16.8 Example 2 26.3 -18.9 -16.4 17.1 -15.2 Comparative Example 1 17.2 -11.6 -9.4 10.5 -8.7 Comparative Example 2 18.6 -13.1 -10.6 11.8 -10.1 Comparative Example 3 15.1 -9.2 -8.1 8.3 -7.2 Comparative Example 4 16.8 -10.8 -9 9.7 -8.4 Comparative Example 5 20.4 -14.6 -12.7 13.2 -11.7 Comparative Example 6 15.9 -9.7 -8.6 8.9 -7.6 Comparative Example 7 19.2 -13.8 -11.8 12.4 -10.9

[0345] Table 8. Changes in subjective skin condition after 28 days of use in each group.

[0346] Group Dryness reduced / % Reduced tightness / % Itching / discomfort reduced by % Overall comfort improved by % Example 1 45.2 42.8 38.6 47.5 Example 2 42.6 40.1 35.9 44.3 Comparative Example 1 29.7 27.5 24.3 31.4 Comparative Example 2 31.5 29.8 25.7 33.2 Comparative Example 3 26.2 24 21.5 27.1 Comparative Example 4 28.5 26.1 23.2 29.8 Comparative Example 5 34.8 32.7 28.9 36.5 Comparative Example 6 27.1 24.9 22.4 28.2 Comparative Example 7 33.4 31.2 27.8 35.1

[0347] The results showed that, in addition to improving the skin moisture, barrier, erythema and luster indices measured by the instrument, Example 1 also showed a good trend of improvement in the subjective skin comfort of the subjects.

[0348] First, Example 1 and Comparative Example 1 have similar final major sugar compositions, but significant differences remain in viscosity stabilization time, thixotropic recovery rate, low-temperature stability, and storage stability of the emulsion. This indicates that the technical effects of this invention cannot be explained solely by "which sugars are ultimately present," as the ethanol fractionation, reconstitution, and remixing processes that the sugars undergo before final mixing also affect their formulation performance.

[0349] It is particularly noteworthy that Comparative Example 5, after thorough hydration, also reached a stable viscosity relatively quickly. Therefore, the technical effect of this invention cannot be simply summarized as "making viscosity stabilize faster." The true comprehensive advantage demonstrated by Example 1 is that while achieving stability in a shorter time, it still maintains high shear recovery capacity, low-temperature stability, and final emulsion system stability. Finally, in skin-related evaluations, Example 1 showed a good trend of improvement in β-endorphin, NMB, and TSLP-related indicators, as well as in human stratum corneum water content, TEWL, erythema, gloss, roughness, and skin comfort.

Claims

1. A skincare composition for improving stress-induced skin condition, characterized in that, The skincare composition comprises a complex active ingredient obtained by co-processing chicory root extract and rhamnose-rich fermented polysaccharide. The composite active material is prepared by a method comprising the following steps: (1) Extract chicory roots with water, separate solid and liquid to obtain chicory root extract; take a portion of the chicory root extract A, add endo-inulinase for enzymatic hydrolysis, inactivate the enzyme after enzymatic hydrolysis, and then mix the enzymatically hydrolyzed chicory root extract with the unhydrolyzed chicory root extract B to obtain chicory root treatment solution containing fructans with different degrees of polymerization. (2) The chicory root treatment liquid is mixed with rhamnose-rich fermented polysaccharide and treated at 30-40°C for 30-90 min. Then ethanol is added to make the volume fraction of ethanol in the system reach 48-56%. After standing, solid-liquid separation is performed to obtain the precipitate and the liquid. (3) Add water to the precipitate obtained in step (2) and redissolve it at 30-35℃ for 30-60 min to obtain a redissolved solution; remove ethanol from the liquid obtained in step (2) and concentrate it to obtain a concentrated solution; (4) Add the concentrated solution obtained in step (3) to the complex solution obtained in step (3), mix at 25-30°C and let stand for 2-6 h to obtain the complex active substance.

2. The skincare composition according to claim 1, characterized in that, In step (1), the chicory root is crushed and added to water, and extracted at 50-65℃ for 1-3 h. The chicory root extract is obtained by solid-liquid separation.

3. The skincare composition according to claim 1 or 2, characterized in that, In step (1), the enzymatic hydrolysis uses an endoglucinase. During the enzymatic hydrolysis, the pH of the chicory root extract A is adjusted to 4.8-5.3, the hydrolysis temperature is 48-55℃, and the hydrolysis time is 40-70 min. The amount of the endoglucinase added is 2-10 U / g of chicory root extract A based on enzyme activity.

4. The skincare composition according to claim 1, characterized in that, The weight-average molecular weight of the rhamnose-rich fermented polysaccharide is 35–70 kDa.

5. The skincare composition according to claim 1, characterized in that, In step (2), the chicory root treatment solution and the rhamnose-rich fermented polysaccharide are mixed, the pH is adjusted to 5.2-5.8, and the mixture is stirred at 30-40℃ for 30-90 min. Then, the mixture is cooled to 20-25℃ and ethanol is added.

6. The skincare composition according to claim 1 or 9, characterized in that, In step (2), ethanol is added under stirring to make the volume fraction of ethanol in the system reach 48-56%. After the ethanol is added, stirring is continued for 10-30 min, and then placed at 4-10℃ for 4-12 h. The precipitate and liquid parts are obtained by solid-liquid separation.

7. A skin care preparation, characterized in that, The skin care composition includes any one of claims 1 to 6 and cosmetic excipients; the excipients include one or more of moisturizers, oils, emulsifiers, thickeners, preservatives and water; the pH of the skin care formulation is 5.0 to 6.

2.

8. A method for preparing the skin care formulation according to claim 7, characterized in that, The method includes mixing aqueous and oil phase raw materials separately and then emulsifying them to obtain an emulsion. The emulsion is then cooled to below 45°C and added to the skin care composition according to any one of claims 1 to 6. The mixture is then mixed evenly and the pH is adjusted to obtain the skin care preparation.

9. The use of the skincare composition according to any one of claims 1 to 6 in the preparation of a skincare product for improving stress-induced skin deterioration, characterized in that, The decline in skin condition includes one or more of the following: dry skin, increased transepidermal water loss, redness, roughness, dullness, decreased skin elasticity, and increased wrinkles.

10. The skin care composition according to any one of claims 1 to 6 is used to improve skin comfort under stress, and / or increase skin β-endorphin levels, and / or decrease neuroregulatory peptide B and thymic stromal lymphopoietin levels.