Lycopene-rich impregnated vegetable oil and method for preparing same

Abstract

The invention relates to an impregnated vegetable oil which contains lycopene and terpenes from natural sources, characterised by having high stability and good biological properties. The impregnated vegetable oil is enriched with lycopene by two methods: a direct solubilisation method, in which the impregnated, flavoured and / or fragranced vegetable oil, containing terpenes, constitutes a green solvent with a high solubilising power for extracting and / or solubilising lycopene; and an indirect dissolution method, in which a pure and stable lycopene extract containing terpenes, obtained by selective extraction of tomato peels with the ethanol-terpene solvent, constitutes the agent for impregnating the vegetable oil. The lycopene-rich impregnated vegetable oil of the present invention has improved bioavailability and nutritional attributes, natural antioxidant capacity, a special odour and flavour, a long shelf life, and is useful for manufacturing nutraceutical, cosmetic, pharmaceutical or food products.

Description

Lycopene-rich impregnated vegetable oil and its preparation method DESCRIPTION [1] TECHNICAL FIELD [2] The present invention relates to the field of manufacturing lycopene-enriched formulations. More specifically, it relates to a flavored and / or perfumed vegetable oil, rich in lycopene extracted from natural raw materials. This oil has excellent properties and functionalities, and is intended for use in the manufacture of nutraceutical, cosmetic, pharmaceutical or food products. [3] OBJECTIVE OF THE INVENTION [4] The objective of the invention is to prepare an impregnated vegetable oil rich in lycopene, with increased stability, solubility, bioactivity and bioavailability. This objective is achieved through an innovative process that proposes two approaches: the first method consists of directly solubilizing tomato peels in a flavored and / or perfumed vegetable oil, enriched with Magnoliophyte plants and an essential oil derived from these plants; the second method involves the indirect dissolution of a pure and stable lycopene extract, combining lycopene and certain terpenes from an ethano-terpene extraction solvent, in a vegetable oil. [5] STATE OF THE ART [6] Lycopene is a linear carotenoid with the molecular formula C 40 H 56. Thanks to its 11 conjugated double bonds, it has a bright red color and is the most powerful antioxidant among carotenoids. Its high capacity to neutralize free radicals, associated with cardiovascular diseases and various cancers, highlights its crucial role in reducing the risks associated with oxidative stress. [7] Tomatoes, due to their high lycopene content, are the main source of this carotenoid. The highest concentration is found in the peel, which contains five times more lycopene than the rest of the fruit. Therefore, tomato peels are widely used in lycopene extraction processes, especially those from the tomato processing industry, which generate a large amount of by-products. [8] Several industries use tomato peels as a source for the production of natural lycopene for human consumption. Among them are Chenguang Biotech Group Co. Ltd (US11499052B2) and OmniActive Health Technologies Ltd (AU2012311207B2), which have filed patent applications for their methods for producing natural lycopene in crystalline form. [9] Crystalline lycopene, with its structured molecular arrangement, presents a solid form that can pose problems for digestion and absorption, thus reducing its bioavailability and physiological effectiveness. The bioavailability of lycopene is influenced by various factors, such as the food matrix, its solubility in lipids, and its interactions with other compounds. Since lycopene is lipophilic, its bioavailability is enhanced when ingested with lipids. This improvement is particularly marked in an oily matrix, which favors both its absorption and biological activity.

[0010] The association of lycopene with oil is the result of innovative methods using either oil as a solvent for lycopene extraction or the direct incorporation of lycopene into oily matrices.

[0011] Numerous patents and studies have explored the use of oil as a solvent to extract lycopene. Chinese patent application CN 102210357A (Henan University of Technology) describes a method for preparing lycopene-enriched vegetable oil. This method involves cleaning and peeling fresh tomatoes, crushing the peels with a stirrer, and mixing the peels with vegetable oil in a specific ratio. The mixture is then extracted at a constant temperature in the dark to obtain an oil containing between 24 and 95 pg / mL of lycopene.

[0012] Patent KR 100692253B 1 describes a method for the direct extraction of lycopene from tomatoes using vegetable oil. The technique involves selecting a deep red tomato and grinding it to obtain a homogeneous tomato solution. A combination of this liquid and vegetable oil in a mass ratio of 2:1 is prepared. The mixture is loaded into a pressing machine rotating at 200-500 rpm and compressed. Then, a leaching process is carried out under different temperature and time conditions, ranging from 1 to 10 hours at an ambient temperature of 25-35°C, 15 to 24 hours at an elevated temperature of about 60-70°C, and 5 to 10 days at a temperature of 25°C. This leaching process achieves a lycopene recovery rate of about 5 to 20%. Moreover, US patent document US 7557146B2 describes a simple methodology that uses vegetable oil as an extractant lipid to obtain a lycopene-enriched formulation.This process involves combining tomato paste with vegetable oil in a ratio. mass ratio of 1:1 and subjecting the mixture to a speed of 4500 rpm for 5 minutes. Centrifugation at 4000 rpm then separates the two phases.

[0013] Although lycopene is fat-soluble, its solubility in vegetable oils may be limited, resulting in incomplete dissolution in some types of oils (Kubola et al., Food Res Int., 50:664-669 (2013)). Heating may improve its solubility.

[0014] The production of lycopene-enriched oil matrices relies on the acquisition of lycopene-rich extracts through various lycopene extraction methods. JP 2743247B2 describes a method for producing lycopene oil by dissolving a lycopene extract in vegetable oil. Tomato by-product peels from the tomato juice production process, crushed and treated with a plant cell-disrupting enzyme, are subjected to dehydration with 90% ethanol, then extracted with supercritical carbon dioxide, and the lycopene extract is dissolved in vegetable oil with a recovery rate of 84.6%.

[0015] Similarly, patent application CN 10114863 IA describes a process for preparing lycopene-enriched vegetable oil from tomato pomace using supercritical carbon dioxide. The process involves grinding, dehydrating, and treating the tomato pomace with ethanol to destroy the cell walls. The treated pomace is then freeze-dried and pulverized to a size between 0.20 and 0.45 mm. The resulting tomato pomace is extracted using supercritical carbon dioxide, and the resulting extract is absorbed with vegetable oil.

[0016] Patent application KR 20130095492A describes a method for preparing a colored vegetable oil comprising lycopene as a natural pigment. The method involves extracting the essential oil from tomatoes and then blending it with a vegetable oil. The tomatoes are freeze-dried and crushed, then subjected to supercritical carbon dioxide treatment at 30-60°C for 100 hours at 100-300 bar to extract the essential oils, including lycopene, and 95% ethanol was used as a co-solvent. The essential oils are then blended with edible oils at a ratio of 1 and 5%, and stirred to prepare a colored oil containing 50-1500 μg of lycopene.

[0017] Patent application CN 105595358A discloses a method for extracting lycopene from tomatoes for incorporation into edible oils. The method involves weighing fresh tomatoes, cleaning them, chopping them, adding sodium hydroxide to regulate the pH to 8, heating them in a thermostatically controlled water bath, stirring them during the water bath, and adding absolute ethyl alcohol to dehydrate them after the water bath. After Dehydration, the circulating water type multiplex vacuum pump provides suction filtration to prepare crude lycopene extract. The edible oil is added to the crude lycopene extract, the mixture is stirred sufficiently, sealed to be stored away from light and heated in a water bath.

[0018] Rather than extracting lycopene and incorporating it into vegetable oils, patent application CN 103843914A proposes adding lycopene directly to tea oil as an antioxidant. This method involves adding 26.86 mg of lycopene to tea oil and then stirring the mixture at 62.61 °C for 8.06 hours, resulting in a concentration of 30.42 μg / mL.

[0019] Vegetable oils can degrade through oxidation when exposed to light and oxygen, causing them to turn rancid and their quality to deteriorate (Henry., Int J Innov Res Sci Eng Technol., 5: 6609-16 (2016)). Adding natural antioxidants such as lycopene can protect oils and extend their shelf life.

[0020] Patent application CN 105875865A proposes a sesame oil enriched with lycopene and added with a composite essential oil as a natural antioxidant. The process begins with the extraction of lycopene. Crushed tomatoes are extracted 2 to 3 times with an extraction solution in a mass ratio of 2:7, at a temperature of 50 to 55°C for a stirring time of 30 to 40 min. The crude lycopene extract is then concentrated into a liquor by absorption of the macroporous resin. The composite essential oil contains essential oils prepared using the supercritical CO2 extraction technique. Lycopene, the composite essential oil, and sesame oil are mixed in different portions by weight, and the mixture is slowly stirred at a temperature of 40 to 55°C. Thanks to the addition of lycopene and composite essential oil, sesame oil has no particular scent and is 40 to 60% more resistant to oxidation.

[0021] Lycopene is particularly unstable due to its highly unsaturated structure. Its degradation involves complex processes such as oxidation and trans-cis isomerization, influenced by light, heat, and oxygen (Zuorro et al., Orient. J. Chem., 34: 2229 (2018)). The stability of lycopene in vegetable oils may be low due to their fatty acid and antioxidant composition. The addition of other antioxidants may help protect and stabilize lycopene.

[0022] Patent CN 102204595B proposes adding vitamin E as an antioxidant to a vegetable oil enriched with lycopene to improve the synergy and bio-activity of the lycopene. The functional edible oil contains between 12 and 200 mg of lycopene and 2 to 20 mg of vitamin E to enhance its stability and antioxidant properties. Similarly, patent CN 102934704B describes an antioxidant edible oil characterized by the fact that each kilogram contains 0.16 g of lycopene, 0.12-0.20 g of astaxanthin and a vitamin E content not exceeding 0.20 g, in order to overcome the defect of single lycopene and improve health care functions such as anti-oxidation and free radical scavenging in the human body.

[0023] The present invention provides two alternatives to the prior art approaches mentioned above, which combine, on the one hand, the preparation of matrices of vegetable oils enriched with lycopene by dissolving a stable extract of lycopene rich in natural antioxidants of plant origin and, on the other hand, the use of a vegetable oil as an extraction solvent with high solubilizing power for lycopene, by a process of impregnation of the vegetable oil allowing it to be enriched with natural antioxidants which improve the solubility of lycopene, protect it and act in synergy with it to reinforce its bioactivity and stability.

[0024] STATEMENT OF THE INVENTION

[0025] The main objective of the present invention is to prepare an impregnated vegetable oil rich in lycopene, characterized by increased stability, solubility, bioactivity and bioavailability.

[0026] The objective of the invention is achieved by means of a process which comprises a method of direct solubilization of tomato peels in a vegetable oil impregnated, flavored and / or perfumed, with Magnoliophyte plants and an essential oil derived from one of these plants, or a method of indirect dissolution of the pure and stable extract of lycopene, composed of lycopene and certain terpenes derived from the ethano-terpene extraction solvent, in a vegetable oil.

[0027] The said process for preparing impregnated vegetable oil rich in lycopene comprises the following steps: Direct solubilization: Vegetable oil is flavored with plant material having a moisture content of 5% and a size of 1 to 2 cm, by means of microwave-assisted infusion, in a mass ratio of 10% (w / w). The flavored oil is recovered by filtration through a suitable filter to remove as much plant residue as possible. The vegetable oil is perfumed with essential oil at a concentration of 1% (w / w), homogenized under mechanical stirring using a three-blade propeller stirrer at room temperature. Dried tomato peels are finely ground to obtain particles smaller than 0.5 mm. The peels are then macerated with the impregnated, flavored and / or perfumed vegetable oil. The interaction is carried out at a temperature of 60°C for 40 minutes, under reduced light conditions. The impregnated vegetable oil enriched with lycopene is recovered by a filtration method, using a suitable filter. Indirect dissolution: The vegetable oil is impregnated and enriched with lycopene by mixing with pure and stable lycopene extract at a concentration of 2 to 40 mg / 100 g and by magnetic stirring for 40 minutes at a temperature of 60°C.

[0028] BRIEF DESCRIPTION OF THE FIGURES

[0029] Figure 1. Chromatographic profile of rosemary impregnated oil / lycopene-rich rosemary essential oil.

[0030] Figure 2. Chromatographic profile of peppermint oil / lycopene-rich peppermint essential oil.

[0031] Figure 3. Chromatographic profile of thyme-infused oil / lycopene-rich thyme essential oil.

[0032] Figure 4. Chromatographic profile of verbena-infused oil / lycopene-rich verbena essential oil.

[0033] Figure 5. Chromatographic profile of coriander-infused oil / lycopene-rich coriander essential oil.

[0034] DETAILED DESCRIPTION OF THE INVENTION

[0035] The invention relates to a novel impregnated oil matrix rich in lycopene and its preparation method using a direct solubilization method or an indirect dissolution method.

[0036] The oily matrix according to the invention is a natural oil of food and / or pharmaceutical quality, or a combination thereof.

[0037] In a first embodiment, the oily matrix is chosen according to its oxidative stability and its sensory characteristics.

[0038] The term "oxidative stability of oil" refers to the measure of the time required for oxidative deterioration to occur.

[0039] In a preferred embodiment, the oily matrix is a vegetable oil with a low natural antioxidant content.

[0040] The word "antioxidant" refers to a substance that has the ability to delay oxidation by inhibiting the initiation or propagation of oxidation chain reactions caused by free radicals.

[0041] Many vegetable oils naturally contain antioxidants, such as polyphenols, tocols (α-, β-, γ-, and δ-tocols (tocopherol and tocotrienol)), and sterols, including campesterol, stigmasterol, β-sitosterol, and avenasterol (Liu et al., Eur. J. Lipid Sci. Technol., 122: 1900079 (2020)).

[0042] The oxidative stability of vegetable oils is influenced by their unsaturated fatty acid composition and their natural antioxidant content. The autoxidation of unsaturated fats transforms them into free radicals, which readily oxidize to produce hydroperoxides and organic compounds, such as aldehydes, ketones, or acids, which give rise to undesirable odors and flavors. Whereas, natural antioxidants act either by inhibiting the formation of free alkyl radicals during the initiation phase or by interrupting the oxidation process by reacting preferentially with the fatty radical to form a stable radical that does not react rapidly with oxygen (Aluyor et al., Afr. J. Biotechnol., 7 (2008)).

[0043] In one embodiment, the vegetable oil is selected from at least one of the following oils: soybean oil, peanut oil, rapeseed oil, sunflower oil, sesame oil, macadamia oil, jojoba oil, avocado oil, grapeseed oil, coconut oil, flaxseed oil, safflower oil, hazelnut oil, walnut oil, wheat germ oil. . .

[0044] Compared to vegetable oils, olive oil is a notable exception. It contains a high concentration of tocopherols, which may have a synergistic effect with lycopene.

[0045] The process of the present invention consists in producing an impregnated, flavored and / or perfumed, and colored vegetable oil.

[0046] The term "impregnated oil" refers to an oil to which aromatic or perfume substances have been added in order to integrate them into its structure and thus endow it with specific characteristics in terms of flavor, aroma or fragrance.

[0047] In a first method of the invention, the vegetable oil is flavored and / or perfumed with a plant and an essential oil, and is colored by direct solubilization of lycopene.

[0048] In this document, the term "flavored oil" refers to a vegetable oil infused with a plant. The term "fragrance oil" refers to a vegetable oil blended with an essential oil.

[0049] In a preferred embodiment, the combination between a vegetable oil and a plant, as well as between a vegetable oil and an essential oil, is selected according to the sensory characteristics of each element. This selection aims for compatibility in order to obtain an odorous and tasty harmony.

[0050] The plant is a Magnoliophyte plant selected from the botanical class Magnoliopsida and Liliopsida.

[0051] The method according to the invention consists in impregnating the vegetable oil of a plant chosen from food plants, medicinal plants, aromatic plants and perfume plants, and of an essential oil obtained from one of these plants by hydrodistillation, supercritical carbon dioxide extraction, cold expression, enzymatic hydrolysis or by solvent extraction.

[0052] In a preferred embodiment, the plants are selected from Lamiaceae, Asteraceae, Rosaceae, Ericaceae, Apiaceae, Fabaceae, Schisandraceae, Myrtaceae, Rubiaceae, Rutaceae, Betulaceae, Brassicaceae, Monimiaceae, Malvaceae, Poaceae, Cupressaceae, Lauraceae, Pinaceae, Aquifoliaceae, Hamamelidaceae, Myristicaceae, Mimosaceae, Balsaminaceae, Myricaceae, Dipterocarpaceae, Cardiopteridaceae, Costaceae, Cistaceae, Winteraceae, Styracaceae, Papaveraceae, Acoraceae, Annonaceae, Caryophyllaceae, Amaranthaceae, Burseraceae, Amaryllidaceae, Geraniaceae, Zingiberaceae, Caprifoliaceae, Cannabaceae, Oleaceae, Liliaceae, Parmeliaceae, Iridaceae, Santalaceae, Asparagaceae, Anacardiaceae, Verbenaceae, Violaceae, Chenopodiaceae, Piperaceae, Theaceae and Tiliaceae.

[0053] Plants belonging to these botanical families are characterized by their composition of antioxidant molecules, such as flavonoids, phenolic acids, ascorbic acid, carotenoids, terpenes, organic acids, polyphenols, tannins, lignans, alkaloids, quinones, anthocyanins, tocopherols and coumarins.

[0054] Some of these antioxidant compounds, such as phenolic acids, carotenoids, terpenes, organic acids, and tocopherols, tend to be soluble in vegetable oils.

[0055] Terpenes are volatile organic compounds responsible for the smell and flavor of plants and the essential oils extracted from them.

[0056] The components of essential oils vary depending on the species and parts of the plant; they are chemically composed of terpenes, their oxygenated derivatives, terpenoids, which are esters of aromatic and aliphatic acids, phenolic compounds, and coumarins (Karpinsk., Biomolecules., 10: 103 (2020)).

[0057] Essential oils are found mainly in leaves, but also in flowers, buds, fruits, seeds, bark, wood or roots.

[0058] Terpenes, also known as isoprenes, are broadly classified into two categories: hydrocarbon-based terpenes, mainly mono-, sesqui-, and diterpenes, and their oxygenated derivatives including alcohols, oxides, aldehydes, ketones, phenols, acids, esters, and lactones (Moghaddam et al., In Soft Chem. Food Ferment., 379-419 (2017)).

[0059] According to one embodiment, the selected plants comprise hydrocarbon terpenes which may be: α-Pinene, β-Pinene, Limonene, Myrcene, Sabinene, α- Terpinene, γ-Terpinene, δ-3-Carene, α-Phellandrene, β-Phellandrene, α-Thuyene, β-Thuyene, Camphene, α-Longipinene, β-Longipinene, α-Humulene, α-Calacorene, α-Bisabolene, β- Bisabolene, β-Caryophyllene, α-Guaiene, β-Guaiene, δ-Amorphene, γ-Cadinene, Terpinolene, Ocimene, Farnesene, Bicyclogermacrene, α-Cedrene, β-Elemene, Valencene, Germacrene, Longifolene, Isocaryophyllene, α-copaene, α-gurjunene, Zingiberene, α-amorphene, α- thujene, β-thujene, α-gurjunene...

[0060] According to one embodiment, the selected plants comprise oxygenated terpenes which may be: Bornyl acetate, Linalool, Menthone, Carvone, Thymol, Menthofuran, Safrole, Anethole, Eugenol, Citronellol, Camphor, Citral, Pulegone, Eucalyptol (1,8-cineole), Terpinene-4-ol, Carvacrol, Myrtenal, Isomenthone, α-Terpineol, Menthol, Isogeraniol, Dihydrocarvone, Isoborneol, Geraniol, β-ionone, Geranyl acetate, α-Bisabolol, Nerol, Lavandulol, Lavandulyl acetate, Thuyan-4-ol, Litseol, Borneol, Citronellyl acetate, Nerolidol, Farnesol, Cinnamal, Estragol, Myristicin, Ascaridol, Linalyl acetate, Guaiol, Sclareol, Santalol...

[0061] In one embodiment of the invention, the vegetable oil is flavored by maceration with plant material, in a mass ratio of 10% (w / w), by hot infusion, cold infusion or any other method known to those skilled in the art.

[0062] The term "plant material" as used herein may refer to any part of the plant, including, but not limited to, flowers or buds, stems, leaves, roots, fruits, bark, or seeds.

[0063] The flavoring process can be carried out using a single plant species or a mixture of several plant species.

[0064] In one embodiment, the plant material is dried in a convection oven equipped with a ventilation system at a temperature not exceeding 50°C. Preferably, the dried plant material has a moisture content of 5% by weight.

[0065] The plant material is preferably fragmented into small pieces of 1 to 2 cm in size in order to increase the contact surface and facilitate the interaction and diffusion of phytochemicals into the vegetable oil.

[0066] In a preferred embodiment, the flavoring process is microwave-assisted infusion.

[0067] The aromatization of vegetable oil is carried out using a domestic microwave oven, which has an irradiation frequency of 2.45 GHz, a power of 400 to 600 Watts and an exposure time of 2 to 5 minutes.

[0068] The direct interaction of microwave irradiation with the mixture of vegetable oil and plant material promotes the release of terpene molecules from the cells of the plant tissue into the oil, in a way that reduces the infusion time.

[0069] Once flavored, the vegetable oil is separated from the residual plant material by a process including, but not limited to, filtration, centrifugation, decantation, and combinations thereof.

[0070] In one embodiment of the invention, the vegetable oil is perfumed by bringing it into contact with an essential oil at a concentration of 1% (w / w) and the mixture is stirred manually or magnetically, homogenized mechanically or treated by sonication or by any other method known to those skilled in the art.

[0071] In a preferred embodiment, the mixture of vegetable oil and essential oil is homogenized while stirring at 200 to 400 rpm using a three-blade propeller stirrer for 10 to 15 minutes at room temperature in order to break up the essential oil droplets and disperse them more evenly in the vegetable oil.

[0072] The present invention considers that the impregnated, flavored and / or perfumed vegetable oil, rich in terpenes, constitutes a green solvent with high solubilizing power for the extraction and / or solubilization of lycopene.

[0073] Although lycopene is a fat-soluble molecule, it is poorly soluble in vegetable oils due to its complex chemical structure, consisting of 40 carbon atoms and 56 hydrogen atoms. Since vegetable oils have a triglyceride structure composed of three fatty acids and one glycerol, their carbon chains are shorter than those of lycopene, making it poorly soluble in these oils.

[0074] According to the invention, the vegetable oil impregnation process is strategically designed to improve the solubility of lycopene in the oil matrix.

[0075] Due to their similar lipophilic properties, terpenes exhibit an affinity for lycopene. Upon contact, hydrophobic interactions promote preferential binding between these compounds. Dispersion forces and van der Waals interactions, as attractive forces between nonpolar molecules, play a central role in this affinity.

[0076] The process for preparing the lycopene-rich impregnated vegetable oil of the present invention mainly consists in directly exposing the lycopene sources or concentrated formulations containing lycopene to the solubilizing action of the different impregnated vegetable oils in varying proportions and at an appropriate temperature.

[0077] In one embodiment of the invention, the impregnated vegetable oil is mixed with tomato peels from industrial tomato processing, for example from production of tomato paste, which are generally considered waste in the food industry.

[0078] In another embodiment, the recovered wet tomato peels are subjected to a preliminary drying step. They can be dried by freeze-drying, drum drying, tray drying or sun drying, among others.

[0079] High moisture content can hinder the direct solubilization process. Preferably, dried tomato peels have a moisture content of less than 7% by weight.

[0080] Dried tomato peels are preferably ground into powder to expose a larger surface area to the impregnated vegetable oil, thus facilitating the lycopene transfer process.

[0081] The present invention reduces the extraction process and improves the solubility of lycopene by extracting lycopene directly from tomato peels using the impregnated vegetable oil, thus providing an environmentally friendly lycopene extraction method.

[0082] Mixing tomato peel powder and impregnated vegetable oil can be done by standard methods such as high pressure homogenization or fluidization, ultrasonic mixing, mixing with colloidal mills, among others.

[0083] In one embodiment, tomato peels less than 0.5 mm in size are mixed with impregnated vegetable oil in a solid-liquid ratio of 2 to 5%, under gentle magnetic stirring for 40 minutes at a temperature of 60°C. The experiments are carried out under reduced light conditions so as to minimize lycopene degradation caused by light exposure.

[0084] In a particular embodiment of the process of the invention, the terpene molecules protect the impregnated vegetable oil and make it less oxidizable, which is why it is not suitable to carry out the maceration under vacuum or in an inert atmosphere.

[0085] Once the mixing process is carried out, the red-colored oily phase constituting the base formulation is separated using conventional separation techniques, which are part of the general knowledge in the field of the invention.

[0086] The recovered red oily phase composed of vegetable oil, phytochemicals, mainly terpenes, and lycopene, constitutes the final product resulting from the process of the present invention.

[0087] According to the invention, the efficiency of the process of direct solubilization of lycopene in vegetable oil depends mainly on the terpene molecules contained in the suspension, which improve the solubility of lycopene in the oil phase.

[0088] In a second method of the invention, the vegetable oil is impregnated and colored by indirect dissolution of the pure and stable extract of lycopene disclosed in our patent application filed under number MA62378.

[0089] Lycopene extract is pure and natural obtained from tomato peels from industrial tomato processing. It is produced by a selective extraction process involving the use of the green ethano-terpene solvent disclosed in our patent application filed under number MA61739. Said method being characterized in that it comprises the following steps: recovery of tomato peels, washing of said peels in order to remove impurities, drying and grinding of the purified peels, contacting of the tomato peel powder with the ethano-terpene solvent at a solid-liquid ratio of 3.5% at a temperature of 70°C for one hour, elimination of the tomato peel residues, concentration of the lycopene extract and recycling of the extraction solvent, drying of said lycopene extract so as to eliminate traces of ethanol and to obtain a pure lycopene extract enriched in terpenes.The said lycopene extract is pure, stable, with biological properties, and characterized by a high extraction yield of around 99%.

[0090] The present invention relates to the application of natural lycopene extract, composed of lycopene molecule and terpenes derived from ethano-terpene solvent, in the process of preparing impregnated vegetable oils rich in lycopene.

[0091] It concerns the application of lycopene extract containing eucalyptol, extracted with ethano-terpene solvent from rosemary, having a lycopene concentration of 4.84 mg / mL.

[0092] It concerns the application of lycopene extract containing thymol, extracted with ethano-terpene solvent from thyme, having a lycopene concentration of 3.9 mg / mL.

[0093] It concerns the application of lycopene extract containing menthol, extracted with ethano-terpene solvent from peppermint, having a lycopene concentration of 4.7 mg / mL.

[0094] It concerns the application of lycopene extract containing citral, extracted with ethano-terpene solvent from verbena, having a lycopene concentration of 4.04 mg / mL.

[0095] It concerns the application of lycopene extract containing linalool, extracted with ethano-terpene solvent from coriander, having a lycopene concentration of 4.73 mg / mL.

[0096] The present invention exploits the fat-soluble characteristics of lycopene, wherein the lycopene extract is processed into an impregnated vegetable oil preparation and can be prepared into a lycopene-rich oil of different contents for different applications.

[0097] According to the conventional approach to obtaining lycopene-enriched oils, the oleoresins of a lycopene-enriched product, being obtained by extraction using an organic solvent whose presence is undesirable in the final formulation, require the use of advanced homogenization, emulsification or encapsulation techniques in order to obtain a uniform dispersion in the oil matrix.

[0098] According to the invention, the pure and stable lycopene extract is characterized by the presence of terpenes making lycopene more soluble in vegetable oil, thus avoiding the problem of residues caused by the dissolution of an organic solvent.

[0099] Furthermore, the mixture of vegetable oil and lycopene extract of the present invention is produced by magnetic or mechanical maceration allowing to obtain a complete dilution of the extract in the oil matrix, thus ensuring the dissolution of lycopene in the vegetable oil at a maximum concentration.

[0100] In one embodiment of the invention, the pure and stable lycopene extract is added to the vegetable oil at a concentration of 2 to 40 mg / 100g, and the mixture is magnetically stirred for 40 minutes at a temperature of 60°C.

[0101] The lycopene-rich impregnated vegetable oil of the present invention is in the form of a red-colored liquid suitable for filling softgel capsules for direct consumption by the consumer, and is packaged in opaque containers that protect them from the effects of exposure to light.

[0102] The present invention relates to a kind of new lycopene-based vegetable oil, studied based on the synergistic effect of lycopene and suspended terpenes.

[0103] As natural antioxidants, the terpenes present in Lycopene-Rich Infused Vegetable Oil react synergistically with lycopene to provide increased protection of the vegetable oil against oxidation, thereby improving the stability of the oil and preserving and enhancing the bioactivity of lycopene.

[0104] According to the invention, the impregnated vegetable oil is characterized by a synergy between the oil and the suspended terpenes, which makes it possible to improve the absorption and bioavailability of lycopene through lipophilic interactions.

[0105] The simultaneous implementation of both synergies in the lycopene-rich impregnated vegetable oil of the present invention makes it possible to overcome the negative effect of oil oxidation on health and to strengthen the body's immunity.

[0106] The preparation method of the present invention validates the stability, uniformity and potential of solubility, bioavailability and absorption of lycopene in the enriched vegetable oil in a safe manner without the use of chemical intermediates.

[0107] The prepared lycopene-rich infused vegetable oil comprises between 5 and 95% lycopene, and between 0.5 and 20% terpenes.

[0108] The lycopene-rich impregnated vegetable oil of the present invention is a functional oil with improved nutritional value and high stability, with natural antioxidant capacity, various biological properties, special odor and flavor, and long shelf life.

[0109] In one aspect of the invention, the lycopene-rich impregnated vegetable oil is used to manufacture nutraceutical, cosmetic, pharmaceutical or food products.

[0110] The lycopene-rich impregnated vegetable oil of the invention can be used for the manufacture of various food products such as mayonnaise, salad dressings, sauces, margarines, among others, which become new sources of lycopene in food products.

[0111] The lycopene-rich infused vegetable oil of the invention constitutes a nutritional formulation with greater added value. In addition to lycopene, it provides an additional content of other phytochemical compounds, terpenes, with antioxidant potential and synergistic action.

[0112] The method of the present invention is simple to produce on a large scale and is widely applicable to all raw materials rich in lycopene.

[0113] METHOD OF IMPLEMENTATION

[0114] According to a general concept of the invention, the impregnated vegetable oil rich in lycopene is prepared according to two methods: By direct solubilization respecting the following steps: Drying the plant material to a moisture content of 5% and grinding said plant material to a particle size of less than 1 to 2 cm; Maceration of vegetable oil with plant material in a mass ratio of 10% (w / w); Separation of flavored vegetable oil and plant residues; Maceration of vegetable oil with essential oil at a concentration of 1% (w / w) and recovery of the fragrant vegetable oil; Drying tomato peels to a moisture content of less than 7% and grinding said peels to a powder size of 0.5 mm; Maceration of tomato peel powder with impregnated, flavored and / or perfumed vegetable oil, in a solid-liquid ratio of 2.5%, under reduced light conditions and recovery of the impregnated oil enriched with lycopene. By indirect dissolution by respecting the following steps: Preparation of lycopene extract by maceration of tomato peel powder, with a moisture content of less than 7% and a size of 0.5 mm, with the ethano-terpene solvent in a solid-liquid ratio of 3.5%; Recovery and concentration of the lycopene extract by vacuum evaporation until it contains only 5 to 10% of the ethano-terpene solvent; Drying the recovered concentrated lycopene extract to remove any lingering traces of ethanol and obtain a pure and stable condensed lycopene extract containing terpenes; Maceration of vegetable oil by direct dissolution of pure lycopene extract, containing terpenes, and recovery of impregnated vegetable oil enriched with lycopene.

[0115] According to a first aspect of the invention, the plant material is gently dried at 40°C so as not to degrade the phytochemical compounds of interest.

[0116] Preferably, the plant material is dried until its moisture content is less than 5%, to prevent high water content from spoiling the oil.

[0117] According to a particular aspect of the invention, the infusion of the vegetable oil with the plant material is treated by a method promoting the release of the terpene molecules from the cells of the plant tissue into the oil, thereby reducing the infusion time.

[0118] According to a second aspect of the invention, the maceration of the vegetable oil with the essential oil is carried out using a technique which makes it possible to break up the droplets of essential oil and disperse them more uniformly in the vegetable oil.

[0119] Preferably, the impregnated vegetable oil is macerated with tomato peels whose moisture content is less than 7%, so as not to hinder the process of direct solubilization of lycopene in the oil.

[0120] According to a third aspect of the invention, the impregnation of the vegetable oil with the lycopene extract, containing terpenes, is carried out by adding this extract to the vegetable oil at a concentration of 2 to 40 mg / 100g, so as to recover an impregnated vegetable oil having a lycopene content of 10 to 150 mg / 100g.

[0121] EXAMPLES

[0122] Example 1: Preparation of lycopene-rich impregnated vegetable oil by direct solubilization

[0123] The fresh plant is washed, gently dried at 40°C to a moisture content of 5% and broken into small 2 cm pieces. The plant material is mixed with vegetable oil in a mass ratio of 10% (w / w). The mixture is processed in a domestic microwave oven at a power of 600 watts for 4 minutes. The two phases are then separated by centrifugation at 3000 rpm for 10 minutes. The liquid phase is a formulation of flavored vegetable oil with dissolved terpenes.

[0124] The essential oil (EO) is added to the vegetable oil at a concentration of 1% (w / w) and the mixture is homogenized under stirring at 400 rpm using a three-blade propeller stirrer for 10 minutes at room temperature. A vegetable oil scented by the diffused terpenes is thus obtained.

[0125] Dried tomato peels (7% moisture) are ground to a powder of less than 0.5 mm, then mixed with impregnated vegetable oil in a solid-liquid ratio of 2.5%. A magnetic stirrer is used to macerate the mixture in a thermostatically controlled bath for 40 minutes at a temperature of 60°C. The experiments are carried out under reduced light conditions. After maceration, the tomato peel residue is removed by filtration through a 2 μm cellulose filter to obtain a very strongly red-colored flavored and / or perfumed vegetable oil. Example 2: Lycopene recovery rate in the solubilization process

[0126] The recovery rate (RR%) of lycopene in the different samples of impregnated vegetable oil was determined by an HPLC-DAD system using a C18 column (4.6 x 250 mm, 3 μm). The different samples were diluted in ethyl acetate at a dilution factor of 0.1. The aliquots were filtered through 0.2 μm cellulose syringe filters and the filtrates were stored in HPLC vials as the final solution. The chromatographic analysis was performed in isocratic elution mode with a mixture of acetonitrile and dichloromethane (75:25; v / v) applied at a constant flow rate of 1.5 mL / min and measured at a wavelength of 470 nm. The retention times and spectra were compared with those of the lycopene standard analyzed under the same conditions. Figures 1 to 5 illustrate the chromatographic profile of lycopene recovered by the different impregnated vegetable oils. The suspended terpene molecules improve the solubility of lycopene in vegetable oil, which allows an effective recovery of between 93% and more than 99%.

[0127] Example 3: Preparation of lycopene-rich impregnated vegetable oil by indirect dissolution

[0128] A weighed quantity of 50 g of tomato peel powder is subjected to the lycopene extraction process with the ethano-terpene solvent (SET) in a solid-liquid ratio of 3.5%, under magnetic stirring at 400 rpm for 1 h at 70°C. After stirring, the residue is removed to obtain a very strongly red colored solution, from which 90% of the ethano-terpene solvent is then removed by vacuum evaporation, while the lycopene extract, with 10% of the ethano-terpene solvent, is recovered and dried overnight at room temperature to remove traces of ethanol and recover a pure and stable lycopene extract containing terpenes.

[0129] 2 mg, 10 mg, 15 mg, 30 mg and 40 mg of lycopene extract are added to each 100 g of vegetable oil, and magnetically stirred for 20 minutes at room temperature until a uniform red color is obtained. 10-150 mg / 100 g of vegetable oil containing lycopene thus obtained. Example 4: Determination of the oxidative stability of impregnated vegetable oil rich in lycopene

[0130] The various impregnated vegetable oils rich in lycopene are placed for 30 weeks in a closed environment at a temperature of 40°C and a humidity of 80%.

[0131] The degree of rancidity of lycopene-rich impregnated vegetable oil is determined by measuring the acidity value and the peroxide value. 5 g of the oil sample is dissolved in 30 mL of peroxide solution (glacial acetic acid and chloroform (3:2 v / v)). Saturated potassium iodide (0.5 mL) and distilled water (30 mL) are then added with stirring for 1 minute. The mixture is titrated with sodium thiosulfate (0.01%) with stirring, 0.5 mL of 1% starch solution is added, and the titration is continued with vigorous stirring until colorless. The peroxide value is calculated as the milliequiv of oxygen induced per kilogram of oil (meq O2 / kg), which could oxidize potassium iodide with subsequent release of iodine. 2 g of oil sample are dissolved in 50 mL of ethanol (96%). After heating, the solution is titrated with potassium hydroxide solution (0.01%) in the presence of the indicator phenolphthalein until it is colorless. The acidity value is calculated as the amount of free fatty acids present in the oil and expressed in milligrams of potassium hydroxide required to neutralize the free acidity contained in one gram of oil (mg KOH / g).

[0132] The peroxide and acidity values of the different lycopene-enriched impregnated vegetable oils are respectively less than 12 meq O2 / kg and 3 mg KOH / g, which clearly confirms that the degree of rancidity of the oil of the present invention is remarkable, and that the synergy of lycopene and terpenes therefore protects the vegetable oil from rancidity. References cited in the description Aluyor, E.O., & Ori-Jesu, M. (2008). The use of antioxidants in vegetable oils-A review. African Journal of Biotechnology, 7(25). Henry, L. N. (2016). Effect of light and air on the quality and stability of selected, vegetable oils. International Journal of Innovative Research in Science, Engineering and Technology. Vol. 5, 6609-16. Karpinski, T. M. (2020). Essential oils of Lamiaceae family plants as antifungals. Biomolecules, 10(1), 103. Kubola, J., Meeso, N., & Siriamornpun, S. (2013). Lycopene and beta carotene concentration in aril oil of gac (Momordica cochinchinensis Spreng) as influenced by aril-drying process and solvents extraction. Food Research International, 50(2), 664-669. Liu, R., Lu, M., Zhang, T., Zhang, Z., Jin, Q., Chang, M., & Wang, X. (2020). Evaluation of the antioxidant properties of micronutrients in different vegetable oils. European Journal of Lipid Science and Technology, 122(2), 1900079. Moghaddam, M., & Mehdizadeh, L. (2017). Chemistry of essential oils and factors influencing their constituents. In Soft chemistry and food fermentation (pp. 379-419). Academic Press. Zuorro, A., Lavecchia, R., Gonzalez, E., & Kafarov, V. (2018). Kinetics of lycopene degradation in sunflower and grape seed oils. Oriental Journal of Chemistry, 34(5), 2229.

Claims

Claims 1. Process for the preparation of impregnated vegetable oil rich in lycopene, comprising lycopene and terpenes of natural origin, characterized in that it consists of a direct solubilization method or an indirect dissolution method.

2. Process for preparing the impregnated vegetable oil rich in lycopene according to claim 1, by a direct solubilization method, characterized in that it comprises the following steps: Drying and grinding of plant material; Maceration of vegetable oil with plant material in a mass ratio of 10% (w / w); Separation of flavored vegetable oil and plant residues; Maceration of vegetable oil with essential oil at a concentration of 1% (w / w) and recovery of the fragrant vegetable oil; Drying and grinding tomato peels; Maceration of tomato peel powder with impregnated, flavored and / or perfumed vegetable oil, in a solid-liquid ratio of 2.5%, under reduced light conditions and recovery of the impregnated oil enriched with lycopene.

3. Process for preparing the impregnated vegetable oil rich in lycopene according to claims 1 and 2, characterized in that the vegetable oil and the impregnating agent, the plant and / or the essential oil are chosen so as to obtain a fragrant and tasty harmony.

4. Process for preparing the impregnated vegetable oil rich in lycopene according to claims 1 to 3, characterized in that the oil is impregnated with a food plant, a pharmaceutical plant, an aromatic plant, a perfume plant, and / or an essential oil derived from one of these plants.

5. Process for the preparation of the impregnated vegetable oil rich in lycopene according to claims 1 to 4, characterized in that the plants are chosen from Lamiaceae, Asteraceae, Rosaceae, Ericaceae, Apiaceae, Fabaceae, Schisandraceae, Myrtaceae, Rubiaceae, Rutaceae, Betulaceae, Brassicaceae, Monimiaceae, Malvaceae, Poaceae, Cupressaceae, Lauraceae, Pinaceae, Aquifoliaceae, Hamamelidaceae, Myristicaceae, Mimosaceae, Balsaminaceae, Myricaceae, Dipterocarpaceae, Cardiopteridaceae, Costaceae, Cistaceae, Winteraceae, Styracaceae, Papaveraceae, Acoraceae, Annonaceae, Caryophyllaceae, Amaranthaceae, Burseraceae, Amaryllidaceae, Geraniaceae, Zingiberaceae, Caprifoliaceae, Cannabaceae, Oleaceae, Liliaceae, Parmeliaceae, Iridaceae, Santalaceae, Asparagaceae, Anacardiaceae, Verbenaceae, Violaceae, Chenopodiaceae, Piperaceae, Theaceae and Tiliaceae.

6. Process for the preparation of impregnated vegetable oil rich in lycopene according to claims 1 to 5, characterized in that the impregnated vegetable oil constitutes a green solvent with high solubilizing power for the extraction and / or solubilization of lycopene.

7. Process for the preparation of the impregnated vegetable oil rich in lycopene according to claims 1 to 6, characterized in that the suspended terpenes improve the solubility, absorption and bioavailability of lycopene in the impregnated vegetable oil.

8. Process for preparing the impregnated vegetable oil rich in lycopene according to claims 1 to 7, characterized in that the lycopene recovery rate is between 93% and more than 99%.

9. Process for preparing the impregnated vegetable oil rich in lycopene according to claim 1, by an indirect dissolution method, characterized in that it comprises the following steps: Preparation of lycopene extract by selective extraction of tomato peels with ethano-terpene solvent; Recovery and concentration of the lycopene extract until it contains only 5 to 10% of the ethano-terpene solvent; Drying the recovered concentrated lycopene extract to remove any lingering traces of ethanol and obtain a pure and stable condensed lycopene extract containing terpenes; Maceration of vegetable oil by direct dissolution of the pure and stable extract of lycopene, containing terpenes, and recovery of the impregnated vegetable oil enriched with lycopene.

10. Process for the preparation of the impregnated vegetable oil rich in lycopene according to claims 1 and 9, characterized in that the lycopene is extracted by the ethano-terpene solvent rich in terpenes having an affinity and a selectivity for lycopene.

11. Process for the preparation of the impregnated vegetable oil rich in lycopene according to claims 1 to 10, characterized in that the vegetable oil is of food and / or pharmaceutical quality, or a combination thereof.

12. Process for preparing the impregnated vegetable oil rich in lycopene according to claims 1 to 11, characterized in that the vegetable oil is preferably low in natural antioxidants.

13. Impregnated vegetable oil rich in lycopene prepared by one of the two processes according to the preceding claims, characterized in that it comprises between 5 to 95% lycopene, and between 0.5 to 20% terpenes.

14. Impregnated vegetable oil rich in lycopene prepared by one of the two processes according to claims 1 to 13, characterized by a synergy of lycopene and terpenes in suspension, which generates a stability of the vegetable oil with a peroxide index of the order of 11 meq O2 / kg and an acidity index of the order of 2 mg KOH / g.

15. Lycopene-rich impregnated vegetable oil prepared by one of the two processes according to claims 1 to 14, characterized by a synergy of lycopene and terpenes in suspension, which reinforces the stability and bioactivity of lycopene.

16. Lycopene-rich impregnated vegetable oil prepared by one of the two processes according to claims 1 to 15, characterized by a synergy of lycopene and suspended terpenes, which gives the vegetable oil a natural antioxidant capacity, various biological properties, a special odor and flavor and a long shelf life.

17. Impregnated vegetable oil rich in lycopene prepared by one of the two processes according to claims 1 to 16, characterized in that it is in the form of a red-colored liquid suitable for filling soft capsules intended for direct consumption.

18. Use of the impregnated vegetable oil rich in lycopene prepared by one of the two processes according to claims 1 to 17, in the manufacture of nutraceutical, cosmetic, pharmaceutical or food products.

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