Preparation method of olive oil unsaponifiables and application thereof in skin care products

Abstract

This invention discloses an unsaponifiable extract of olive oil, its preparation method, and its applications. The preparation method includes: S1, raw material pretreatment: after drying and pulverizing olive pulp, antioxidants and a compound enzyme are added for enzymatic hydrolysis, followed by ultrasonic cell disruption; S2, solvent extraction: extraction is performed using a hexane-anhydrous ethanol compound solvent to obtain an extract; S3, multi-stage saponification: the extract undergoes two stages of saponification sequentially, and the organic phase is collected; S4, purification and refining: after dehydration and concentration, the organic phase is dissolved in squalane and filtered to obtain a refined concentrate. This invention achieves efficient and gentle extraction of high-value components such as squalene and phytosterols through low-temperature processing, enzyme-ultrasonic synergistic cell disruption, and gradient saponification. The obtained product can be directly used in cosmetics, exhibiting excellent anti-aging and skin repair effects, and the formulation is extremely simple, gentle, and non-irritating, showing promising application prospects.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, specifically to a method for preparing unsaponifiable olive oil and its application in skincare products. Background Technology

[0002] Skin aging is a complex pathophysiological process driven by both intrinsic genetic programs (chronological aging) and external environmental stresses. Photoaging is a major extrinsic cause of skin aging, accounting for over 80% of environmental aging factors. Prolonged exposure to ultraviolet radiation (especially UVA and UVB) leads to excessive production of reactive oxygen species (ROS) in the skin, triggering oxidative stress. This not only directly damages cellular DNA but also activates the mitogen-activated protein kinase (MAPK) pathway, upregulating transcription factors AP-1 and NF-κB. The consequences are twofold: on the one hand, it inhibits the synthesis of type I and type III collagen and elastin; on the other hand, it significantly induces the expression of matrix metalloproteinases (MMPs), accelerating the degradation of the extracellular matrix (ECM), ultimately leading to deep wrinkles, sagging, pigmentation, and impaired skin barrier function.

[0003] For photoaging, current anti-aging strategies mainly focus on anti-oxidation, inhibiting MMPs activity, and promoting collagen regeneration. However, mainstream high-efficiency anti-aging ingredients on the market (such as all-trans retinoic acid and high-concentration acids) often have strong skin irritation and can easily cause adverse reactions such as erythema and peeling, limiting their application in sensitive skin and people with poor tolerance.

[0004] Therefore, developing natural plant-derived active ingredients that combine the dual benefits of "powerful antioxidant / anti-aging" and "gentle barrier repair" has become a research hotspot in the cosmetics field.

[0005] Olive oil unsaponifiables (OOUs) are natural active compounds in olive oil that cannot be saponified with lye and are soluble in ether. They comprise only 1%–2% of the total olive oil content, yet concentrate most of the bioactive essence of the olive fruit. Their main components include squalene (a natural moisturizer), phytosterols (anti-inflammatory and repairing), polyphenolic compounds (such as hydroxytyrosol and oleuropein, powerful antioxidants), and tocopherols. Studies have shown that olive unsaponifiables have significant advantages in scavenging free radicals, inhibiting lipid peroxidation, regulating skin microecology, and promoting collagen synthesis, making them an ideal natural anti-aging ingredient.

[0006] In the existing technology, several technical solutions have been disclosed that focus on the extraction or purification of unsaponifiables. For example, Chinese patent document CN101597204B (application number: CN200910069670.8) discloses a "Method for Extracting High-Purity Squalene from Olive Oil". This method directly uses commercially available olive oil as the starting material and employs a process route combining two-stage molecular distillation and silica gel column chromatography to further separate and purify high-purity squalene from unsaponifiables. Although this method can obtain a high-purity single component, its process is complex, requires sophisticated equipment, and consumes a large amount of energy. Furthermore, its starting material is already olive oil obtained through processes such as pressing, and it does not solve the problem of high-value utilization of by-products such as pomace in the olive oil industry chain. Essentially, it still belongs to the deep processing of olive oil, resulting in low raw material utilization efficiency.

[0007] Another type of existing technology focuses on the direct extraction of unsaponifiables from plant raw materials. For example, Chinese patent document CN102648271B (application number: CN201080047939.3) discloses a "Method for Extracting Unsaponifiables from Renewable Raw Materials". This method aims to extract unsaponifiables from renewable raw materials such as oilseed fruits (e.g., olives) and seeds. Its core steps include: after dehydrating and conditioning the raw materials, reactive grinding is carried out in the presence of lower alcohols and catalysts to obtain a liquid phase containing fatty acid alkyl esters and unsaponifiables, followed by saponification and extraction of the liquid phase. Although this method achieves one-step extraction from fruit to unsaponifiables, its process involves reactive grinding and transesterification reactions, which are relatively vigorous. Furthermore, the lower alcohols and catalysts used may adversely affect the purity of subsequent active products and the retention of heat-sensitive components (such as squalene and polyphenols).

[0008] Based on this, a new process for extracting unsaponifiables from olives that utilizes the whole fruit, protects against low temperatures, and retains high activity has been developed. This process has significant industrial value and application prospects for improving raw material utilization, reducing production costs, and preparing highly effective anti-aging skincare products. Summary of the Invention

[0009] This invention provides an unsaponifiable olive oil and its application in the preparation of minimalist cosmetics that combine anti-aging and repairing effects.

[0010] To achieve the above objectives, the present invention adopts the following technical solution: An unsaponifiable matter from olive oil is prepared by extraction using the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into small pieces of 0.5-1cm. Place the pulp pieces in a hot air drying oven at 50-55℃ and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 40-60 mesh sieve to obtain olive pulp powder.

[0011] Antioxidant and deionized water were added to the fruit pulp powder at a ratio of 1:2 g / mL, and the mixture was stirred at low speed for 30 minutes to fully wet the pulp. Then, a complex enzyme preparation consisting of cellulase and pectinase was added, and the pH of the system was adjusted to 5.0-5.5. The mixture was then enzymatically hydrolyzed at 45°C for 1 hour. After the enzymatic hydrolysis was completed, the temperature was raised to 60°C and kept at that temperature for 10 minutes to inactivate the enzyme activity. After the temperature was lowered to 30°C, the mixture was subjected to ultrasonic cell disruption at 200W for 15 minutes.

[0012] Step S1 achieves efficient cell wall disruption and gentle release of unsaponifiables through a synergistic process of low-temperature drying, compound enzymatic hydrolysis, and ultrasonic cell wall disruption. Specifically: low-temperature drying at ≤55℃ avoids the thermal decomposition of heat-sensitive components such as squalene, tocopherol, and polyphenols; cellulase and pectinase work together to soften the cell wall structure, and combined with ultrasonic physical disruption, significantly improving the dissolution efficiency of cell contents; the addition of antioxidants effectively inhibits the oxidative loss of unsaponifiables during pretreatment.

[0013] Preferably, the antioxidant in step S1 is ascorbic acid, and the amount added is 0.02% to 0.03% of the weight of olive pulp powder.

[0014] Preferably, in step S1, the mass ratio of cellulase to pectinase in the compound enzyme is 2:1, and the total amount added is 0.5% to 0.8% of the weight of the fruit pulp powder.

[0015] S2, Solvent extraction The broken olive pulp powder was transferred to an extraction tank, and a hexane-anhydrous ethanol composite solvent was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was stirred and extracted at 35℃ for 2 hours at a speed of 80 rpm under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh composite solvent was added to the filter residue, and the extraction was repeated once. The supernatants from both extractions were combined to obtain the crude extract.

[0016] Step S2 employs a hexane-ethanol composite solvent system, utilizing its non-polar and weakly polar solubility properties to selectively extract fat-soluble unsaponifiables (such as squalene, sterols, and tocopherols) from the fruit pulp, while retaining some triglycerides and polar impurities, effectively improving the extraction rate of unsaponifiables. This method directly extracts unsaponifiables from the fruit pulp, avoiding the resource waste of traditional processes that require first extracting large quantities of olive oil and then performing saponification, significantly improving raw material utilization efficiency; the extracted pomace can still be used for oil extraction or energy utilization.

[0017] Preferably, in step S2, the volume ratio of n-hexane to anhydrous ethanol in the n-hexane-anhydrous ethanol composite solvent is 2 to 3:1.

[0018] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.0-9.5. Saponification was carried out at constant temperature for 1.5h to convert free fatty acids into water-soluble fatty acid potassium soaps. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15min. The mixture was allowed to stand for 30min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50°C, add a higher concentration of KOH-ethanol solution, adjust the pH to 11.0-11.5, and saponify at a constant temperature for 2.5 h under nitrogen protection; after saponification, cool to 25°C, add an equal volume of deionized water and shake and wash for 10 min, let stand for 20 min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0019] Step S3 employs a gradient saponification strategy, gradually removing lipid impurities and enriching the target unsaponifiables through two stages of saponification conditions with different intensities. The first stage of saponification is mild, primarily removing free fatty acids; the second stage, under strongly alkaline conditions, thoroughly hydrolyzes residual glycerides, ensuring a high enrichment of unsaponifiables in the organic phase.

[0020] Preferably, the concentration of the primary saponification KOH-ethanol solution in step S3 is 0.3-0.5 mol / L.

[0021] Preferably, the concentration of the secondary saponification KOH-ethanol solution in step S3 is 1-2 mol / L.

[0022] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate to dissolve it, and the concentrate was filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0023] Step S4 further enhances the purity and stability of the product through dehydration, low-temperature concentration, and membrane filtration. Anhydrous sodium sulfate effectively removes trace amounts of moisture, preventing hydrolysis or oxidation during subsequent concentration; low-temperature concentration under reduced pressure avoids loss of heat-sensitive components; and squalane is selected as the carrier solvent, exhibiting good compatibility with unsaponifiables and possessing excellent skincare properties, making it suitable for direct use in cosmetic formulations.

[0024] Preferably, the amount of anhydrous sodium sulfate used in step S4 is 3% to 5% of the weight of the upper n-hexane phase.

[0025] Preferably, the amount of squalane used in step S4 is 5 to 10 times the weight of the concentrate.

[0026] The unsaponifiable matter in olive oil described in this invention can be directly added to cosmetic formulations as an active ingredient. The addition amount is 0.1-10%.

[0027] The present invention also discloses a minimally formulated olive oil moisturizing essence, comprising the following components in weight percentage: 0.1-10% squalane, 0.1-10% unsaponifiables of olive oil prepared in Example 1, and the balance of olive oil.

[0028] The beneficial effects of the technical solution provided by the embodiments of the present invention are as follows: 1. Gentle preparation throughout the entire process, effectively protecting active ingredients. Low-temperature drying (≤55℃), vacuum concentration (45℃), and nitrogen protection throughout the process are used to avoid the degradation and loss of heat-sensitive and easily oxidized components such as squalene, tocopherol, and polyphenols, thus preserving the natural activity of unsaponifiables in olive oil to the greatest extent.

[0029] 2. Enzyme-ultrasound synergistic cell disruption significantly improves extraction efficiency. By combining cell wall softening with complex enzymes (cellulase / pectinase) with ultrasonic physical disruption, the dissolution rate of unsaponifiables within cells is significantly improved, resulting in a higher extraction rate and shorter extraction time compared to traditional single extraction methods.

[0030] 3. Extracted directly from the fruit pulp, making fuller use of raw materials. This process eliminates the traditional steps of pressing oil and then saponifying it, directly extracting unsaponifiables from the olive pulp, significantly improving resource utilization efficiency. The remaining pomace can still be further pressed for oil or used for energy, making it green and environmentally friendly.

[0031] 4. Gradient saponification removes impurities, resulting in high product purity. A two-stage gradient saponification strategy is adopted: the first stage is mild saponification to remove free fatty acids, and the second stage is strong alkaline saponification to completely hydrolyze residual glycerides, effectively enriching target unsaponifiables such as squalene, phytosterols, and tocopherols, resulting in high purity and few impurities.

[0032] 5. Combining dehydration and membrane filtration results in strong stability. Anhydrous sodium sulfate removes trace amounts of moisture through dehydration, preventing subsequent hydrolysis or oxidation; 0.22μm microfiltration removes minute impurities, extending product shelf life and ensuring the stability and safety of cosmetic formulations.

[0033] 6. Squalane, as a carrier, can be directly adapted to cosmetics. Squalane was chosen as the final carrier solvent because it has good compatibility with unsaponifiables of olive oil. Squalane itself has excellent skin care properties, and the resulting concentrate can be directly added to cosmetic formulations, simplifying the production process and facilitating industrial application. Attached Figure Description

[0034] Figure 1 Example 1: Gas chromatogram of the sample; Figure 2 Gas chromatogram of squalene standard. Detailed Implementation

[0035] The embodiments described in this specification are for illustrative purposes only and do not limit the scope of protection of this invention. The scope of protection of this invention is defined only by the claims, and any omissions, substitutions, or modifications made based on the embodiments disclosed in this invention will fall within the scope of protection of this invention.

[0036] To enhance understanding of the present invention, the present invention will be further described in detail below with reference to embodiments. These embodiments are only used to explain the present invention and do not constitute a limitation on the scope of protection of the present invention.

[0037] Example 1 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 0.5cm pieces. Place the pulp pieces in a 55℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 40-mesh sieve to obtain olive pulp powder.

[0038] Add ascorbic acid to the olive pulp powder at a rate of 0.03% of the olive pulp powder weight; then add deionized water at a material-to-liquid ratio of 1:2 g / mL, and stir at low speed for 30 minutes to fully wet the pulp powder; subsequently add a compound enzyme preparation composed of cellulase and pectinase, wherein the mass ratio of cellulase to pectinase in the compound enzyme is 2:1, and the total addition amount is 0.5% of the olive pulp powder weight; adjust the pH of the system to 5.5, and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and keep it at 10 minutes to inactivate the enzyme activity, and after cooling to 30℃, use 200W ultrasonic wave to break the cell wall for 15 minutes.

[0039] S2, Solvent extraction The crushed olive pulp powder was transferred to an extraction tank, and a hexane-anhydrous ethanol composite solvent with a volume ratio of 2:1 (hexane to anhydrous ethanol) was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was extracted at a constant temperature of 35℃ with stirring for 2 hours at a speed of 80 rpm, under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh composite solvent was added to the filter residue, and the extraction was repeated once. The supernatants from both extractions were combined to obtain the crude extract.

[0040] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.5 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.0. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1 mol / L KOH-ethanol solution, adjust pH to 11.0, and saponify at a constant temperature for 2.5 h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10 min, let stand for 20 min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0041] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 5% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 5 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0042] Example 2 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 1cm pieces. Place the pulp pieces in a 50℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 60-mesh sieve to obtain olive pulp powder.

[0043] Add ascorbic acid to the olive pulp powder at a weight of 0.02%; then add deionized water at a ratio of 1:2 g / mL and stir at low speed for 30 minutes to fully wet the pulp powder; subsequently add a compound enzyme preparation composed of cellulase and pectinase at a mass ratio of 2:1, with a total addition amount of 0.8% of the olive pulp powder weight; adjust the pH of the system to 5.0 and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and incubate for 10 minutes to inactivate the enzyme activity; after cooling to 30℃, use 200W ultrasonic wave to break up the cell walls for 15 minutes.

[0044] S2, Solvent extraction The crushed olive pulp powder was transferred to an extraction tank, and a hexane-anhydrous ethanol composite solvent with a volume ratio of 3:1 (hexane to anhydrous ethanol) was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was extracted at a constant temperature of 35℃ with stirring for 2 hours at a speed of 80 rpm, under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh composite solvent was added to the filter residue, and the extraction was repeated once. The supernatants from both extractions were combined to obtain the crude extract.

[0045] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.3 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.5. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 2 mol / L KOH-ethanol solution, adjust pH to 11.5, and saponify at a constant temperature for 2.5 h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10 min, let stand for 20 min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0046] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 3% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 10 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0047] Example 3 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 0.8cm pieces. Place the pulp pieces in a 53℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 50-mesh sieve to obtain olive pulp powder.

[0048] Add ascorbic acid to the olive pulp powder at a weight of 0.02%; then add deionized water at a ratio of 1:2 g / mL and stir at low speed for 30 minutes to fully wet the pulp powder; subsequently add a compound enzyme preparation composed of cellulase and pectinase at a mass ratio of 2:1, with a total addition amount of 0.6% of the olive pulp powder weight; adjust the pH of the system to 5.2 and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and incubate for 10 minutes to inactivate the enzyme activity; after cooling to 30℃, use 200W ultrasonic wave to break up the cell walls for 15 minutes.

[0049] S2, Solvent extraction The crushed olive pulp powder was transferred to an extraction tank, and a hexane-anhydrous ethanol composite solvent with a volume ratio of 2.5:1 (hexane to anhydrous ethanol) was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was extracted at a constant temperature of 35℃ with stirring for 2 hours at a speed of 80 rpm, under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh composite solvent was added to the filter residue, and the extraction was repeated once. The supernatants from both extractions were combined to obtain the crude extract.

[0050] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.4 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.3. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1.5mol / L KOH-ethanol solution dropwise, adjust the pH to 11.5, and saponify at a constant temperature for 2.5h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10min, let stand for 20min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0051] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 4% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 8 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0052] Comparative Example 1 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Take 1 kg of commercially available extra virgin olive oil and place it in a constant temperature reaction vessel. Slowly add 0.4 mol / L KOH-ethanol solution while stirring at 40℃ to adjust the pH of the system to 9.3. Saponify at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, cool to room temperature, add an equal volume of n-hexane and shake to extract for 15 min. Let stand for 30 min to separate the layers, discard the lower saponified aqueous phase, and collect the upper n-hexane phase. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1.5mol / L KOH-ethanol solution dropwise, adjust the pH to 11.5, and saponify at a constant temperature for 2.5h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10min, let stand for 20min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0053] S2, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 4% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 8 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0054] Comparative Example 2 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 0.8cm pieces. Place the pulp pieces in a 53℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 50-mesh sieve to obtain olive pulp powder.

[0055] Add ascorbic acid to the olive pulp powder at a weight of 0.02%; then add deionized water at a ratio of 1:2 g / mL and stir at low speed for 30 minutes to fully wet the pulp powder; then add cellulase preparation at a weight of 0.6% of the pulp powder; adjust the pH of the system to 5.2 and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and keep it at 10 minutes to inactivate the enzyme activity; after cooling to 30℃, use 200W ultrasonic wave to break up the cell walls for 15 minutes.

[0056] S2, Solvent extraction The crushed olive pulp powder was transferred to an extraction tank, and a hexane-anhydrous ethanol composite solvent with a volume ratio of 2.5:1 (hexane to anhydrous ethanol) was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was extracted at a constant temperature of 35℃ with stirring for 2 hours at a speed of 80 rpm, under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh composite solvent was added to the filter residue, and the extraction was repeated once. The supernatants from both extractions were combined to obtain the crude extract.

[0057] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.4 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.3. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1.5mol / L KOH-ethanol solution dropwise, adjust the pH to 11.5, and saponify at a constant temperature for 2.5h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10min, let stand for 20min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0058] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 4% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 8 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0059] Comparative Example 3 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 0.8cm pieces. Place the pulp pieces in a 53℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 50-mesh sieve to obtain olive pulp powder.

[0060] Add ascorbic acid to the olive pulp powder at a weight of 0.02%; then add deionized water at a ratio of 1:2 g / mL and stir at low speed for 30 minutes to fully wet the pulp powder; then add pectinase preparation at a weight of 0.6% of the pulp powder; adjust the pH of the system to 5.2 and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and keep it at 60℃ for 10 minutes to inactivate the enzyme activity; after cooling to 30℃, use 200W ultrasonic wave to break up the cell walls for 15 minutes.

[0061] S2, Solvent extraction The crushed olive pulp powder was transferred to an extraction tank, and a hexane-anhydrous ethanol composite solvent with a volume ratio of 2.5:1 (hexane to anhydrous ethanol) was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was extracted at a constant temperature of 35℃ with stirring for 2 hours at a speed of 80 rpm, under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh composite solvent was added to the filter residue, and the extraction was repeated once. The supernatants from both extractions were combined to obtain the crude extract.

[0062] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.4 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.3. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1.5mol / L KOH-ethanol solution dropwise, adjust the pH to 11.5, and saponify at a constant temperature for 2.5h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10min, let stand for 20min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0063] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 4% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 8 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0064] Comparative Example 4 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 0.8cm pieces. Place the pulp pieces in a 53℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 50-mesh sieve to obtain olive pulp powder.

[0065] Add ascorbic acid to the olive pulp powder at a weight of 0.02%; then add deionized water at a ratio of 1:2 g / mL and stir at low speed for 30 minutes to fully wet the pulp powder; subsequently add a compound enzyme preparation composed of cellulase and pectinase at a mass ratio of 2:1, with a total addition amount of 0.6% of the olive pulp powder weight; adjust the pH of the system to 5.2 and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and incubate for 10 minutes to inactivate the enzyme activity; after cooling to 30℃, use 200W ultrasonic wave to break up the cell walls for 15 minutes.

[0066] S2, Solvent extraction The broken olive pulp powder was transferred to an extraction tank, and n-hexane was added. The material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was stirred and extracted at a constant temperature of 35℃ for 2 hours at a speed of 80 r / min, with nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 r / min for 15 minutes, and the supernatant was collected. An equal volume of fresh compound solvent was added to the filter residue, and the extraction was repeated once. The supernatants from the two extractions were combined to obtain the crude extract.

[0067] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.4 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.3. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1.5mol / L KOH-ethanol solution dropwise, adjust the pH to 11.5, and saponify at a constant temperature for 2.5h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10min, let stand for 20min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0068] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 4% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 8 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0069] Comparative Example 5 An unsaponifiable matter of olive oil is prepared by the following steps: S1. Raw material pretreatment Select ripe and plump olives, remove rotten fruit, impurities, and pits, quickly rinse with clean water to remove surface dirt, drain the water, and cut the pulp into 0.8cm pieces. Place the pulp pieces in a 53℃ hot air drying oven and dry until the moisture content is ≤8%; after cooling to room temperature, pulverize and pass through a 50-mesh sieve to obtain olive pulp powder.

[0070] Add ascorbic acid to the olive pulp powder at a weight of 0.02%; then add deionized water at a ratio of 1:2 g / mL and stir at low speed for 30 minutes to fully wet the pulp powder; subsequently add a compound enzyme preparation composed of cellulase and pectinase at a mass ratio of 2:1, with a total addition amount of 0.6% of the olive pulp powder weight; adjust the pH of the system to 5.2 and enzymatically hydrolyze at 45℃ for 1 hour; after enzymatic hydrolysis, raise the temperature to 60℃ and incubate for 10 minutes to inactivate the enzyme activity; after cooling to 30℃, use 200W ultrasonic wave to break up the cell walls for 15 minutes.

[0071] S2, Solvent extraction The broken olive pulp powder was transferred to an extraction tank, anhydrous ethanol was added, and the material-to-liquid ratio was controlled at 1:12 g / mL. The mixture was extracted at a constant temperature of 35℃ with stirring for 2 hours at a speed of 80 rpm, under nitrogen protection throughout the process. After extraction, the mixture was centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. An equal volume of fresh compound solvent was added to the filter residue, and the extraction was repeated once. The supernatants from the two extractions were combined to obtain the crude extract.

[0072] S3, Multi-stage saponification (1) Primary saponification: The crude extract was placed in a constant temperature reaction vessel, and 0.4 mol / L KOH-ethanol solution was slowly added dropwise under stirring at 40℃ to adjust the pH of the system to 9.3. Saponification was carried out at constant temperature for 1.5 h to convert free fatty acids into water-soluble fatty acid potassium soap. After saponification, the mixture was cooled to room temperature, and an equal volume of n-hexane was added for shaking extraction for 15 min. The mixture was allowed to stand for 30 min to separate into layers. The lower saponified aqueous phase was discarded, and the upper n-hexane phase was collected. (2) Secondary saponification: Add anhydrous ethanol to the upper n-hexane phase, heat to 50℃, add 1.5mol / L KOH-ethanol solution dropwise, adjust the pH to 11.5, and saponify at a constant temperature for 2.5h under nitrogen protection; after saponification, cool to 25℃, add an equal volume of deionized water and shake and wash for 10min, let stand for 20min to separate the layers, discard the lower aqueous phase, and retain the upper n-hexane phase.

[0073] S4, Purification and Refining (1) Dehydration treatment: Add anhydrous sodium sulfate to the upper n-hexane phase, wherein the amount of anhydrous sodium sulfate is 4% of the weight of the upper n-hexane phase; let stand for 2 hours to dehydrate, and remove anhydrous sodium sulfate by vacuum filtration to obtain purified extract; (2) Concentration and purification: The purified extract was placed in a rotary evaporator under reduced pressure and concentrated at 45°C and 0.09 MPa to remove n-hexane, resulting in a viscous concentrate. Squalane was added to the concentrate in an amount equal to 8 times its weight to dissolve it. The concentrate was then filtered through a 0.22 μm organic filter membrane to remove minor impurities, resulting in a purified unsaponifiable concentrate.

[0074] Experimental Example 1: Extraction Rate and Component Analysis of Unsaponifiables Olive oil is one of the plant-based oils with the highest squalene content, significantly higher than other common plant oils. This characteristic makes squalene a key chemical marker distinguishing olive oil from other oils. In terms of efficacy, squalene, with its strong antioxidant capacity, effectively protects skin lipid cells from free radical damage, maintaining skin elasticity and moisture. It is a core active ingredient in cosmetics for achieving anti-aging and barrier repair effects. However, as a highly unsaturated terpene compound containing six double bonds, squalene is extremely sensitive to heat and readily undergoes oxidative degradation. Based on the aforementioned source characteristics, efficacy value, and chemical instability, this application selects squalene as a key detection indicator to objectively evaluate the protective effect of the extraction process on the active ingredient and the efficacy potential of the final product.

[0075] The squalene content in the unsaponifiables of olive oils prepared in Examples 1-3 and Comparative Examples 1-5 was determined according to the method described in the literature Kang Heng, Fu Fan, Xue Peipei, et al. Analysis of squalene content in common edible vegetable oils [J]. Agricultural Products Processing, 2024, (11): 83-85.

[0076] Detection parameters: Initial temperature set at 180℃, then increased to 290℃ at a rate of 30℃ / min and held for 20 min; split ratio 10:1, injection port temperature 250℃, injection volume 2 μL. The sample solution was injected into the gas chromatograph for analysis, and the peak areas of squalene and squalane in the sample were recorded. The mass of squalene in the sample solution was calculated based on the peak area ratio of squalene and squalane in the test solution, the mass of the internal standard, and the correction factors for squalene and squalane.

[0077] Table 1 Comparison of squalene content in various test samples

[0078] The results showed that the extraction rates of squalene active ingredients in Examples 1-3 of this invention were significantly higher than those in the comparative examples. Although Comparative Example 1 (traditional saponification method) had a certain extraction rate, it was lower than that of this invention, indicating that direct extraction from the pulp combined with gradient saponification is more advantageous. The extraction rates of Comparative Examples 2-3 (different enzyme choices) and Comparative Examples 4-5 (different solvent choices) decreased significantly, proving that the choice of enzymatic hydrolysis and solvent has a significant impact on the extraction of active ingredients.

[0079] Experiment Example 2: Antioxidant Stability Test The unsaponifiable concentrates obtained in Example 1 and Comparative Example 1 were placed in a constant temperature incubator at 40°C for 30 days for accelerated storage, and the retention rate of squalene was determined. The results are shown in Table 2.

[0080]

[0081] Table 2 Accelerated stability test results The results showed that the squalene retention rate in Example 1 was significantly higher than that in Comparative Example 1 in the accelerated test, indicating that the nitrogen protection throughout the process, low temperature process and antioxidant addition strategy adopted in this invention effectively improved the oxidative stability of unsaponifiables.

[0082] Example 4 A minimally formulated olive oil moisturizing essence comprises the following components in weight percentages: 86% olive oil, 9% squalane, and 5% unsaponifiables of the olive oil prepared in Example 1.

[0083] Experimental Example 3: Evaluation of Cosmetic Efficacy Thirty volunteers aged 35-55 were recruited to conduct a volunteer trial efficacy evaluation test on the olive moisturizing essence oil with the minimal formula prepared in Example 4 of this invention.

[0084] Volunteer selection criteria: Suitable for both men and women, with skin problems such as wrinkles, dryness and tightness, and sensitive skin problems such as redness and stinging. No serious facial skin diseases (acne, eczema, dermatitis, etc.), no trauma, and no history of hormone dependence; I have not used any potent skincare products containing hormones, retinoic acid, fruit acids, or other strong effects in the past month, nor have I undergone any medical aesthetic procedures (such as photofacial rejuvenation or hyaluronic acid injections). Able to strictly follow testing specifications, participate in testing on time, complete daily usage records, and have no frequent absences; Voluntarily sign the informed consent form, understand the testing process and precautions, and declare any known allergens or have no history of allergies in advance.

[0085] Test method: Subjects strictly followed the instructions to use the minimally formulated olive moisturizing essence oil prepared according to Example 4 of this invention, once in the morning and once in the evening, for two consecutive months. During this period, they avoided using other skin care products and cosmetics, and avoided behaviors that could affect their skin condition, such as staying up late and excessive eye strain.

[0086] Evaluation method: Two months after use, volunteers completed a final questionnaire to comprehensively evaluate the improvement in skin problems. A 1-5 point rating scale (1 point = no improvement / very poor, 5 points = significant improvement / excellent) was used. Participants completed the evaluation through the questionnaire. The core indicators and scoring criteria are as follows: I. Dimensions of Sensitive Skin Repair 5 points: No redness or stinging on the skin, all sensitivity symptoms are completely relieved, and the skin barrier is stable; 4 points: Slight redness of the skin, significant improvement in sensitivity symptoms, no stinging or itching; 3 points: Skin redness and sensitivity have improved to some extent, with occasional slight stinging. 2 points: Skin redness and sensitivity show no significant improvement, with frequent stinging and itching; 1 point: Skin redness and increased sensitivity, with obvious discomfort or allergic reactions.

[0087] II. Anti-aging Dimensions 5 points: Fine lines on the face are significantly reduced and become shallower, the skin is firm and elastic, and the facial contours are improved; 4 points: Fine lines on the face have been reduced and become shallower, skin elasticity has improved, and there is no obvious sagging; 3 points: No obvious change in facial fine lines, and basically no improvement in skin elasticity; 2 points: Fine lines on the face have deepened slightly, and skin elasticity has decreased slightly; 1 point: Facial fine lines are significantly deepened, skin laxity worsens, and anti-aging effect is poor.

[0088] Table 3. Results of the Volunteer Questionnaire Survey

[0089] As shown in Table 3, after two months of using the minimally formulated olive oil moisturizing essence obtained in Example 4 of this invention, 86.7% of the volunteers experienced significant relief and repair of their sensitivity symptoms, and 83.3% of the volunteers felt an improved anti-aging effect. No adverse reactions were reported during the test, indicating that the product of this invention has excellent subjectively perceptible efficacy in repairing sensitive skin and anti-aging, and is gentle and non-irritating.

[0090] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0091] The above embodiments merely illustrate several implementation methods of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. An unsaponifiable matter of olive oil, obtained by the following preparation method: S1. Raw material pretreatment: After drying the olive pulp, crush it, add antioxidants and water to moisten it, then add compound enzyme preparation for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, inactivate it, and then perform ultrasonic cell wall breaking treatment to obtain cell wall broken pulp pulp. S2. Solvent extraction: Add n-hexane-anhydrous ethanol composite solvent to the cell wall broken fruit pulp for extraction, separate solid and liquid, and collect the extract. S3. Multi-stage saponification: The extract is subjected to primary and secondary saponification sequentially, and the organic phase is collected; the pH of the primary saponification is 9.0–9.5, and the pH of the secondary saponification is 11.0–11.

5. S4. Purification and refining: After dehydrating the organic phase, concentrate it, then add squalane to dissolve it, filter it, and obtain a concentrated solution of unsaponifiables in refined olive oil.

2. The method for preparing the unsaponifiable matter of olive oil according to claim 1, characterized in that, The drying process in step S1 involves hot air drying at 50–55°C until the moisture content is ≤8%; the pulverized material is then passed through a 40–60 mesh sieve.

3. The method for preparing unsaponifiable olive oil according to claim 1, characterized in that, The antioxidant mentioned in step S1 is ascorbic acid, and the amount added is 0.02%–0.03% of the weight of olive pulp powder.

4. The method for preparing the unsaponifiable matter of olive oil according to claim 1, characterized in that, The compound enzyme preparation mentioned in step S1 is composed of cellulase and pectinase in a mass ratio of 2:1, and the total amount added is 0.5%–0.8% of the weight of olive pulp powder.

5. The method for preparing the unsaponifiable matter of olive oil according to claim 1, characterized in that, The enzymatic hydrolysis conditions in step S1 are: pH 5.0–5.5, temperature 45℃, and time 1h; the ultrasonic cell disruption conditions are: power 200W and time 15min.

6. The method for preparing the unsaponifiable matter of olive oil according to claim 1, characterized in that, In step S2, the volume ratio of n-hexane to anhydrous ethanol in the n-hexane-anhydrous ethanol composite solvent is 2–3:1; the extraction conditions are: material-to-liquid ratio 1:12 g / mL, temperature 35℃, time 2h, and nitrogen gas is introduced for protection during the extraction process.

7. The method for preparing the unsaponifiable matter of olive oil according to claim 1, characterized in that, The primary saponification in step S3 is as follows: the pH is adjusted to 9.0–9.5 with 0.3–0.5 mol / L KOH-ethanol solution, and saponification is carried out at 40℃ for 1.5 h; the secondary saponification is as follows: the pH is adjusted to 11.0–11.5 with 1–2 mol / L KOH-ethanol solution, and saponification is carried out at 50℃ for 2.5 h, with nitrogen gas introduced for protection during the saponification process.

8. The method for preparing unsaponifiable olive oil according to claim 1, characterized in that, The dehydration in step S4 is performed by adding anhydrous sodium sulfate and allowing it to stand for dehydration, with the amount of anhydrous sodium sulfate being 3%–5% of the weight of the organic phase; the concentration is performed by vacuum concentration at a temperature of 45°C and a vacuum degree of 0.09 MPa; and the amount of squalane used is 5–10 times the weight of the concentrate.

9. A cosmetic composition, characterized in that, It contains the unsaponifiables of olive oil as described in any one of claims 1-8 and a cosmetically acceptable carrier.

10. A minimally formulated olive oil moisturizing essence, characterized in that, It comprises the following components in weight percentage: 0.1-10% squalane, 0.1-10% unsaponifiables of olive oil as described in any one of claims 1-8, and the balance being olive oil.

Images