Sensory influence of substituted chromanones and furo-chromanones

Abstract

The present invention relates to organoleptic chromanones and furochromanones and their effects on aroma characteristics.

Description

Technical Field

[0001] This invention relates to the sensory effects of chromanone and furochromanone and their use in modulating, for example, enhancing and / or improving the aroma characteristics of flavoring substances, odor substances or fragrances. Background Technology

[0002] Natural cold-pressed lime oil is often described as the "champagne" of citrus oils (Colombo et al., Citrus Oils in Food and Beverages: Uses and Analyses. In Citrus: The Genus Citrus; Dugo, G., Di Giacomo, A., Eds.; Medicinal and Aromatic Plants-Industrial profiles; Taylor & Francis: London; New York, 2002; pp. 539-556). Its unique aroma has been recognized for over 300 years (Flavourings: Production, Composition, Applications, Regulations, 2., compl. rev. ed.; Ziegler, H., Ed.; Wiley-VCH: Weinheim, 2007; Di Giacomo, A. Development of the Citrus Industry: Historical Note. InCitrus: The Genus Citrus; Medicinal and Aromatic Plants-Industrial profiles; Taylor & Francis: London; New York, 2002; pp. 63-70). Currently, the global annual production of limes and lemons exceeds 20 million tons, and approximately 6% of the crop is further processed into juice and essential oils (FAOSTAT: Food and Agriculture Data. Food and Agriculture Organization of the United Nations. Citrus fruit fresh and processed Statistical bulletin 2020; Food and Agriculture Organization of the United Nations Rome, 2021).One important commodity is cold-pressed lime oil, which is widely used in: flavorings for non-alcoholic and alcoholic beverages, confectionery, and flavoring products; and in pharmaceutical preparations, cleaning products, and fragrances (Colombo et al., see the aforementioned literature; Buccellato, F. Citrus Oils in Perfumery and Cosmetic Products. In Citrus: The Genus Citrus; Medicinal and Aromatic Plants-Industrial profiles; Taylor & Francis: London; New York, 2002; pp. 557-567). Recent findings reveal that only two varieties of lime currently have commercial interest: lime (Citrus aurantifolia) and broadleaf lime (Citrus latifolia). Their natural cold-pressed oils can be produced using various methods.

[0003] Cold-pressed lime oil and its volatiles have been extensively analyzed and characterized in the past (Citrus Oils: Composition, Advanced Analytical Techniques, Contaminants, and Biological Activity; Dugo, G., Mondello, L., Eds.; Medicinal and aromatic plants–industrial profiles; CRC Press: Boca Raton, FL, 2011; González-Mas et al. Volatile Compounds in Citrus Essential Oils: A Comprehensive Review. Front. Plant Sci. 2019, 10, 12.). Typical volatile components such as terpenes, sesquiterpenes, aldehydes, oxidized terpenes, and esters contribute significantly to the overall aroma of lime.

[0004] Oxycyclic compounds, including coumarins, are present in the non-volatile fraction of lime oil (Russo et al., Coumarins, Psoralens and Polymethoxyflavones in Cold-Pressed Citrus Essential Oils: A Review. J. Essent. Oil Res. 2021, 33 (3), 221-239). Coumarins and their derivatives are produced during secondary metabolism in various plant species and participate in plant growth, photosynthesis, and defense mechanisms (Önder, A. Anticancer Activity of Natural Coumarins for Biological Targets. In Bioactive Natural Products; Studies in Natural Products Chemistry; Elsevier BV: Amsterdam, 2020; Vol. 64, pp. 85-109).

[0005] Recently, coumarins and psoralens have also attracted attention as markers for the authenticity assessment of lime and lemon juice (Jungen et al., Cousarins, Psoralens, and Quantitative 1H-NMR Spectroscopy for Authentication of Lemon (Citrus Limon [L.] Burm.f.) and Persian Lime (Citrus × Latifolia [Yu.Tanaka] Tanaka) Juices. Food Chem. 2021, 359, 129804.). However, their sensory contribution and influence on the overall lime aroma have not been fully investigated. Although they appear to make a significant contribution to aroma, the coumarin-like flavors in lime, in particular, are not fully understood to this day (Liu et al., Dietary Bioactives and Essential Oils of Lemon and Lime Fruits. Food Sci. Hum. Wellness 2022, 11 (4), 753-764).

[0006] There is a persistent interest in finding substances for generating new fragrances. Natural substances, in particular, that can be produced using green and environmentally friendly methods, are of great interest. Therefore, the object of this invention is to provide compounds with unexplored sensory effects that can be used to modulate the characteristics of aromatic compositions. Detailed Implementation

[0007] By combining distillation, fractionation, olfactory detection, and structural elucidation, key aromatic components influential in cold-pressed lime oil were identified. This distillation ensured the removal of the most volatile compounds, such as monoterpenes, and enriched the classes of compounds of interest. Oxycyclic heterocyclic compounds accumulating in the bottom fraction were further separated by MPLC. Seventeen MPLC fractions were obtained for each oil, which were subsequently studied using GC / MS-O with the assistance of three team members. An unknown peak with a strong coumarin-like odor was detected and characterized based on its MS spectrum. The final identification was completed through the synthesis and analysis of hypothetical compounds. Following this method, the inventors identified a combination of coumarins and their corresponding saturated chromones that significantly contribute to the typical coumarin aroma. This combination comprises three novel components not yet described in the literature as components of lime oil: 7-methoxy-2-chromone (3,4-dihydro-7-methoxy-2H-1-benzopyran-2-one; CAS 20921-02-2), 5,7-dimethoxy-2-chromone (3,4-dihydro-5,7-dimethoxy-2H-1-benzopyran-2-one; CAS 82243-01-4), and 5,6-dihydrobergamot lactone (5,6-dihydro-4-methoxy-7H-furano[3,2-g][1]benzopyran-7-one; CAS 29050-61-1). Sensory evaluation of the effects of these components on the overall lime aroma and their interactions revealed synergistic and moderating effects. The accompanying research activities related to this invention will be submitted for publication (LJ Schulze, U Schäfer, LZygalski, M Verwohlt, S Otte-Hölscher, A Issa, F Hentschel, M Wüst, GKrammer. Sensory Impact of Novel Dihydrocoumarins in Native Lime Oils. JAFC, 2023, in preparation).

[0008] According to the first aspect, the objective is achieved by using at least one compound of formula (I) or (II) for modulating the characteristics of the aromatic composition:

[0009] (I), (II)

[0010] Among them, R1, R2, R3 and R4 are independently selected from the group consisting of H, OH, OCH3 or OCH2CH3, excluding compounds of formula (1) in which R1, R2, R3 and R4 are all H.

[0011] To achieve this invention, the inventors sought to reveal compounds in lime oil that possess unexplored sensory activities. Specifically, distillation and fractionation experiments were conducted using natural cold-pressed broadleaf lime oil and cold-pressed lime oil as starting materials, with a particular focus on: enriching the coumarin-rich fraction of the natural oil; identifying and characterizing unexplored components that contribute sensorily to the typical aroma characteristics of cold-pressed lime oil; and revealing the interactions of these components.

[0012] Three specific compounds have been identified that contribute sensory features to aroma characteristics. These compounds include 3,4-dihydro-7-methoxy-2H-1-benzopyran-2-one (also referred to herein as 7-methoxy-2-chromone), 3,4-dihydro-5,7-dimethoxy-2H-1-benzopyran-2-one (also referred herein as 5,7-dimethoxy-2-chromone), and 5,6-dihydro-4-methoxy-7H-furano[3,2-g][1]benzopyran-7-one (also referred herein as 5,6-dihydrobergamot lactone), which are represented by the following formulas (Ia), (Ib), and (II-a), respectively:

[0013] (Ia)

[0014] (Ib)

[0015] (II-a)

[0016] Other objects, aspects and embodiments of the invention will become apparent from the following description, the appended embodiments, and in particular the appended patent claims.

[0017] The compounds of formulas (Ia), (Ib), and (II) form particularly preferred embodiments of the present invention. These compounds have been characterized as making a significant contribution to the typical coumarin aroma of cold-pressed lime oil (broadleaf lime, lemon). These compounds can be used alone or in combination with each other. Refer to the Examples section for this aspect. Specific combinations include the following (the symbols "+" and "-" indicate that the corresponding compounds are included in and / or excluded from the combination, respectively):

[0018]

[0019] In this invention, at least one compound of formula (I) or (II) is used in a sensorily effective amount. This means that the amount used provides a perceptible effect on the aroma characteristics of the composition. To determine the sensorily effective amount, compounds of formula (I) or (II) at naturally occurring concentrations were analyzed individually or in combination in a neutral flavor solution. Furthermore, the sensory effects of compounds of formula (I) or (II) were evaluated by using a lime matrix that reproduces the basic aroma of natural cold-pressed lime oil but lacks coumarin and dihydrocoumarin, and by combining the lime matrix with compounds of formula (I) or (II) individually and in combination with each other. Based on the results, the following amounts are preferred:

[0020] Preferably, the aromatic composition contains at least one compound of formula (I) or (II), said compound being in the following amount:

[0021] Compounds of formula (Ia) with a concentration greater than 10 ppb; and / or

[0022] Compounds of formula (Ib) with a concentration greater than 20 ppb; and / or

[0023] Compounds of formula (II-a) with a concentration greater than 20 ppb.

[0024] Advantageously, the compounds of the present invention possess a wide variety of aroma-modifying capabilities. In many embodiments, the modification is to improve or enhance, or shift towards, an aroma or flavor selected from the group consisting of: coumarin-like, juicy, fruity, floral, honey-like, fresh, lime, lemon, invigorating, herbal, earthy, buttery, anise, caramel, toffee, sweet, sour, or creamy flavors. Based on their capabilities, the compounds of the present invention are particularly suitable (as part of an aromatic composition) for use in food, premium food, food supplements, pharmaceutical preparations, medical preparations, cosmetic preparations, oral hygiene preparations, fragrances, consumer-grade preparations for pleasure, and their semi-finished products.

[0025] Another aspect of the invention is a mixture comprising at least one compound of formula (Ia), (Ib), and (II-a):

[0026] (Ia)

[0027] (Ib)

[0028] (II-a)

[0029] The amount of the compound is:

[0030] (i) Compounds of formula (Ia) with a concentration greater than 10 ppb, preferably greater than 50 ppb, more preferably greater than 500 ppb; and / or

[0031] (ii) Compounds of formula (Ib) with a concentration greater than 20 ppb, preferably greater than 100 ppb, more preferably greater than 1000 ppb; and / or

[0032] (iii) Compounds of formula (II-a) with a content greater than 20 ppb, preferably greater than 100 ppb, and more preferably greater than 1000 ppb.

[0033] The preferred mixture of the present invention contains:

[0034] a. Compounds of formula (Ia) and compounds of formula (Ib); or

[0035] b. Compounds of formula (Ia) and compounds of formula (II-a); or

[0036] c. Compounds of formula (Ib) and compounds of formula (II-a); or

[0037] d. Compounds of formula (Ia), compounds of formula (Ib) and compounds of formula (II-a).

[0038] In the mixture, the amount of the respective compound is independently selected from the amounts given above for the respective compound.

[0039] Preferably, the mixture is not derived using essential oils or extracts. This means, for example, that when the mixture is used in an aromatic composition at a dose that produces a pleasant taste, the amount and / or ratio of compounds of formula (I) and (II) contained in the mixture exceeds the amount of compounds of formula (I) and (II) introduced into the essential oil or extract in a naturally occurring amount.

[0040] In another preferred embodiment, the mixture of the present invention comprises (iv) a flavoring agent other than the compounds of formulas (I) and (II). The other flavoring agent may be selected from the group consisting of: aliphatic flavoring substances, especially saturated fatty alcohols (such as ethanol, isopropanol, butanol, isopentanol, hexanol, 2-heptanol, octanol (1 / 2 / 3), decanol); unsaturated fatty alcohols (such as cis-2-pentenol, cis-3-hexenol, trans-2-hexenol, trans-3-hexenol, cis-2-octenol, 1-octen-3-ol, cis-6-nonen-1-ol, trans-2, cis-6-nonadienol); fatty aldehydes, such as saturated fatty aldehydes (e.g., acetaldehyde, propionaldehyde, butyraldehyde, isobutyraldehyde, pentanol, isopentanol, succinyl acetaldehyde, hexanal, 3-methylhexanal, octanol, nonanol) or monounsaturated fatty aldehydes or polyunsaturated fatty aldehydes (such as 2-methylbut-2-enal). Trans-2-hexenal, cis-3-hexenal, cis-4-hexenal, trans-2-octenal, trans-2-nonenal, cis-6-nonenal, trans-2,cis-6-nonadienal, trans-2-decenal, trans-2,trans-decadienal; aliphatic ketones, such as saturated ketones (e.g., 2-butanone, 2-pentanone, 2-heptanone, 2-octanone, 2-methylheptan-3-one, 2-decanone, 2-undecanone), unsaturated ketones (e.g., 1-penten-3-one, 1-hexen-3-one, 5-methyl-3-hexenone, 3-hepten-2-one, 1-octen-3-one, 2-octen-4-one, 3-octen-2-one, 3-nonen-2-one); aliphatic diketones and aliphatic diketols. Diketoles (e.g., diacetyl, acetylmethylethanol, 2,3-hexanedione); fatty acids, such as straight-chain saturated acids (e.g., acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, heptanoic acid, octanoic acid, decanoic acid), branched-chain saturated acids (e.g., 2-methylheptanoic acid, 4-ethyloctanoic acid), and unsaturated acids (e.g., 2-butenoic acid, 2-pentenoic acid, 4-pentenoic acid, 2-methylpentenoic acid, trans-3-hexenoic acid, cis-3-hexenoic acid, 3-octenic acid, linoleic acid); fatty acid esters, such as saturated esters (e.g., methyl acetate, methyl butyrate, methyl 2-methylbutyrate, methyl hexanoate, ethyl acetate, ethyl butyrate, ethyl 2-methylbutyrate, ethyl 3-methylbutyrate, ethyl hexanoate, ethyl decanoate). Isopropyl acetate, isobutyl acetate, isobutyl valerate, isoamyl acetate, isoamyl butyrate, isoamyl isovalerate, hexyl acetate, hexyl hexanoate, 3-octyl acetate), and unsaturated esters (such as methyl 2-hexenoate, allyl hexanoate, cis-3-hexenol acetate, cis-3-hexenol butyrate); aliphatic thiols and dithiols (e.g., propanethiol, allylthiol, 1-methoxy-3-methylbutane-3-thiol, dimethyl sulfide, dimethyl trisulfide, dipropyl sulfide, diallyl trisulfide); other aliphatic sulfur compounds (such as 2-mercapto-3-butanol, methylthiopropionaldehyde, 3-mercapto-pentanone, 4-methoxy-2-methyl-2-mercaptobutanone, methyl thiobutyrate, methyl thiobutyrate).3-Methylthiopropionate; aliphatic nitrogen compounds (such as butylamine, trimethylamine, allyl isothiocyanate, isopropyl isothiocyanate); alicyclic compounds, such as alicyclic ketones (e.g., cis-jasmone, isophorone, 4-ketoisophorone), alicyclic esters (e.g., methyl jasmone, dihydrojasmone); terpenes, such as terpenols (e.g., linalool, citronellol, geraniol, nerol, α-terpineol, menthol, 8-menthene-1,2-diol, fentanyl, borneol, nerolidol, dehydrolinalool), terpenalaldehydes (e.g., geraniol, nerol, citronellol, β-sweet orange aldehyde), terpenones (e.g., α-ionone, (D)-carvone, (L)-carvone, nocaconone, piperonone, menthone, α-turfaconone, β-turfaconene, turfaconene). Ketones), terpene esters (such as linalyl acetate, geraniol acetate, citronellol acetate, carvyl acetate, fennel acetate), terpene sulfur compounds (4-menth-8-thiol-3-one, thiogeraniol, p-menth-1-en-8-thiol, mercapto-p-menth-3-one), terpene alkenes (such as D-limonene, L-limonene, α-pinene, β-pinene, ocimene, α-terpinene, γ-terpinene, β-bisabolene, valerene), terpene oxides (such as 1,8-cineole, rose ether, mentholactone, menthofuran); aromatic compounds, such as aromatic alcohols (such as benzyl alcohol, cinnamyl alcohol, 2-phenyl alcohol), aromatic aldehydes (such as benzaldehyde, cinnamaldehyde, 5-methyl-2-phenylhexenal, salicylaldehyde, 4-hydroxybenzaldehyde, tartrazine aldehyde). Aromatic acids (such as 2-phenyl-2-butenal), aromatic esters (such as benzyl acetate, benzyl salicylate, anisyl acetate, methyl phenyl acetate, methyl benzoate, methyl salicylate, methyl cinnamate), aromatic phenols (such as phenol, o-cresol, p-cresol, 2,3-dimethylphenyl, 2-ethylphenol, 2,3,5-trimethylphenol, 4-vinylphenol, guaiacol, 4-vinylguaiacol, eugenol, thymol, carvacrol), aromatic sulfur compounds (such as thiophenol, diphenyl disulfide), aromatic nitrogen compounds (such as methyl anthranilate, N-methyl anthranilate), aromatic ethers (such as vanillin, ethyl vanillin, anethole), aromatic oxides (such as heliotropein). (e.g., diphenyl ethers), aromatic lactones (such as coumarins, dihydrocoumarins); heterocyclic compounds, such as heterocyclic lactones (e.g., γ-butyrolactone, γ-nonanolactone, γ-decanolactone, δ-decanolactone, jasmonicol, δ-dodecanolactone, asterolactone), heterocyclic furans (such as furfuryl alcohol, furfural, 2-acetylfuran, theanosterane, 2-methyltetrahydrofuran-3-one, furfuryl thiol, 2-methyl3-furanthiol, 2-methyl3-tetrahydrofuranthiol, difurfuryl sulfide, difurfuryl disulfide), heterocyclic pyrans (such as maltol, ethyl maltol, rose ether, maltol isobutyrate), heterocyclic pyrroles (such as indole, 2-acetylpyrrole, pyrrolidine), heterocyclic pyrazines (such as 2-methylpyrazine, 2,3-dimethylpyrazine, 2-methyl3-ethylpyrazine).Trimethylpyrazine, 2-acetylpyrazine, 2-methoxy-3-methylpyrazine, 2-methoxy-3-ethylpyrazine, 2-methoxy-3-isobutylpyrazine, 2-ethyl-3-methylthiopyrazine); heterocyclic thiazoles (such as thiazole, 2-methylthiazole, 4-methyl-5-vinylthiazole, 2-isobutylthiazole, 2-acetylthiazole); flavoring ingredients and flavoring preparations, such as essential oils, extracts, absolutes, extracts or tinctures derived from citrus fruits, etc. Fruits (e.g., lemon, lime, mandarin orange, bergamot, grapefruit, bitter orange, peel or essential oil), herbs (dill, parsley, cumin, rosemary, sage, clary sage, basil, tarragon, thyme, oregano, savory, marjoram, allspice powder, nutmeg seed coat, nutmeg, clove leaves, clove buds, caraway seeds, cinnamon leaves, cinnamon bark, low-grade cinnamon, cardamom seeds, ginger, galangal, turmeric, coriander seeds, coriander leaves, etc.) Fenugreek, juniper berries, wormwood, bay leaves, eucalyptus, white pepper, green bell pepper, white pepper, carrot seeds, celery seeds, angelica leaves, asafoetida, onions, leeks, garlic, mustard, horseradish, chili peppers, red chili powder, seaweed, valerian oil, fir needles, spearmint, mint, holly, broom leaves, blackcurrant buds, fennel, star anise, wax apple (jumbu), long pepper, artemisia, iris, mimosa, acacia, violet leaves, camphor leaves, jasmine, ylang-ylang, canan Osmanthus, Angelica dahurica, Clary sage, Okra seeds, Hops, Chamomile, Lavender, Rose, Geranium, Lemongrass, Palmarosa, Litsea cubeba, Lemongrass, Marigold, Orange blossom, Petitgrain, Yerba mate, Congee oil, Coffee, Kola nuts, Cocoa, Green tea, Black tea, White tea, Gentian, Turmeric balm, Benzoin gum, Peruvian balm, Aquilaria sinensis, White pine resin, Vetiver, Labdanum, Patchouli, Sandalwood, Cedarwood, Guaiac wood, Oak, Massoi bark Bark, vanilla pods, tonka bean and their enriched fractions; juice concentrates, such as orange juice, lemon juice, strawberry, cherry juice or passion fruit juice concentrates; aqueous phases and recoveries from the following raw materials, such as citrus fruits (lemon, lime, orange, tangerine, grapefruit), apple, pear, quince, mispel, red fruits (raspberry, strawberry, blueberry, blackberry, trefoil (June plum), rosehip, cranberry, plum, prune, red currant and blackcurrant, etc.), yellow fruits (peach, apricot, nectarine, banana, etc.), tropical fruits ( Mangoes, passion fruit, pineapples, lychees, etc.), vegetables (such as cucumbers and tomatoes), and spices (such as ginger); acetophenone, allyl hexanoate, α-ionone, β-ionone, anisaldehyde, anisyl acetate, anisyl formate, benzaldehyde, benzothiazole, benzyl acetate, benzyl alcohol, benzyl benzoate, β-ionone, butyl butyrate, butyl hexanoate, 3,3-butenylphthalide, carvone, camphene, caryophyllene, eucalyptol, cinnamyl acetate, citral, citronellol, citronellol, citronellol acetate, cyclohexyl acetate, cymene, damascone, decanolactone, dihydrocoumarin.Dimethyl anthranilate, dodecyl lactone, ethoxyethyl acetate, ethyl butyric acid, ethyl butyrate, ethyl decanoate, ethyl hexanoate, ethyl crotonate, ethyl furanone, ethyl guaiacol, ethyl isobutyrate, ethyl isovalerate, ethyl lactate, ethyl methyl butyrate, ethyl propionate, eucalyptol, eugenol, ethyl heptanate, 4-(p-hydroxyphenyl)-2-butanone, γ-decyl lactone, geraniol, geraniol acetate, geraniol acetate, grapefruitaldehyde, methyl dihydrojasmone (e.g., Hedion®), jasmine aldehyde, 2-heptanone, 3-heptanone, 4-heptanone, trans-2-heptenal, cis-4-heptenal, trans-2-hexenal, cis-3-hexenol, trans-2-hexenoic acid, trans-3-hexenoic acid, cis-2-hexene Acrylates, cis-3-hexenyl acetate, cis-3-hexenyl hexanoate, trans-2-hexenyl hexanoate, cis-3-hexenyl carboxylate, cis-2-hexenyl acetate, cis-3-hexenyl acetate, trans-2-hexenyl acetate, cis-3-hexenyl carboxylate, p-hydroxybenzylacetone, isoamyl alcohol, isoamyl isovalerate, isobutyl butyrate, isobutyraldehyde, isoeugenol methyl ether, isopropyl methylthiazole, lauric acid, levulinic acid, linalool, linalool oxide, linalyl acetate, menthol, menthol furan, methyl anthranilate, methyl butanol, methyl butyric acid, 2-methylbutyl acetate, methyl hexanoate, methyl cinnamate, 5-methylfurfural, 3,2,2-methylcyclopentenolone, 6,5,2-methylheptenone Methyl dihydrojasmonic acid, methyl jasmonic acid, 2-methyl methyl butyrate, 2-methyl-2-pentenolic acid, methyl thiobutyrate, 3,1-methylthiohexanol, 3-methylthiohexanol acetate, nerol, nerol acetate, trans, trans-2,4-nonadienal, 2,4-nonadienol, 2,6-nonadienol, 2,4-nonadienol, nocaketone, δ-octanolide, γ-octanolide, 2-octanol, 3-octanol, 1,3-octenol, 1-octyl acetate, 3-octyl acetate, palmitic acid, paraldehyde, phellandrene, pentylene, phenethyl acetate, phenethyl alcohol, phenethyl isovalerate, piperaldehyde, propionaldehyde, propyl butyrate, menthol, menthol, sweet orange aldehyde, thiothiazole, terpinene, terpineol, terpineyl oleylene, 8,3-s-thio Menthol, 4,4,2-thiomethylpentanone, thymol, δ-undecyl lactone, γ-undecyl lactone, valerene, valeric acid, vanillin, acetoin, ethyl vanillin, ethyl vanillin isobutyrate (=3-ethoxy-4-isobutyryloxybenzaldehyde), 2,5-dimethyl-4-hydroxy-3(2H)-furanone and its derivatives (preferably cyclohomanthone (=2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone), soy sauce ketone (=2-ethyl-5-methyl-4-hydroxy-3(2H)-furanone and 5-ethyl-2-methyl-4-hydroxy-3(2H)-furanone)), maltol and maltol derivatives (preferably ethyl maltol), coumarin and coumarin derivatives,γ-lactone (preferably γ-undecylactone, γ-nonylactone, γ-decylactone), δ-lactone (preferably 4-3,4-methylδdecylactone, masoyanalactone, δdecylactone, tuberolactone), methyl sorbate, divanillin, 4-hydroxy-2 (or 5)-ethyl-5 (or 2)-methyl-3(2H)furanone, 2-hydroxy-3-methyl-2-cyclopentenone, 3-hydroxy-4,5-dimethyl-2(5H)-furanone, isoamyl acetate, ethyl butyrate, n-butyl butyrate, isoamyl butyrate, 3-methylethyl butyrate, ethyl hexanoate, allyl hexanoate, n-butyl hexanoate, ethyl octanoate, 3 ethyl methyl-3-phenyl glycidyl ester, ethyl 2-trans-4-cis-decadienoate, 4-(p-hydroxyphenyl)-2-butanone, 1,1-dimethoxy-2,2,5-trimethyl-4-hexane, 2,6-dimethyl-5-hepten-1-aldehyde and phenylacetaldehyde, 2-methyl-3-(methylthio)furan, 2-methyl-3-furanthiol, bis(2-methyl-3-furanyl)disulfide, furfuryl thiol, methylthiopropionaldehyde, 2-acetyl-2-thiazoline, 3-mercapto-2-pentanone, 2,5-dimethyl-3-furanthiol, 2 4,5-Trimethylthiazole, 2-acetylthiazole, 2,4-dimethyl-5-ethylthiazole, 2-acetyl-1-pyrrolline, 2-methyl-3-ethylpyrazine, 2-ethyl-3,5-dimethylpyrazine, 2-ethyl-3,6-dimethylpyrazine, 2,3-diethyl-5-methylpyrazine, 3-isopropyl-2-methoxypyrazine, 3-isobutyl-2-methoxypyrazine, 2-acetylpyrazine, 2-pentylpyridine, (E,E)-2,4-decadienal, (E,E)-2,4-nonadienal, (E)-2-octenal (E)-2-nonenal, 2-undecenal, 12-methyltridecaldehyde, 1-penten-3-one, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, guaiacol, 3-hydroxy-4,5-dimethyl-2(5H)-furanone, 3-hydroxy-4-methyl-5-ethyl-2(5H)-furanone, cinnamaldehyde, cinnamyl alcohol, methyl salicylate, isopreneol, and (not explicitly stated here) stereoisomers, enantiomers, positional isomers, diastereomers, cis / trans isomers, or epimers of these substances.

[0041] Preferably, the flavoring agent contains a flavor-modifying component selected from the group consisting of: hesperidin, hesperidin dihydrochalcone, phloretin, naringin, fosetyl-aluminum, acetylated trifolin, sennaol, hypersennaol, podophyllin, asiaticoside, asiaticoside B, 3',7-dihydroxy-4'-methoxyflavan, 1-(2,4-dihydroxyphenyl)-3-(3-hydroxy-4-methoxyphenyl)prop-1-one, physalisol, and mixtures thereof. More preferably, the flavoring agent is selected from flavoring agents of the lime, lemon, vanilla, caramel, green tea, tomato, cherry, and raspberry types.

[0042] Further components of the preferred mixtures of the present invention include (v) sweeteners, acidifiers and / or (vi) water or milk.

[0043] In the context of this invention, sweeteners can be one or more substances selected from the group consisting of:

[0044] - Natural sweeteners, preferably naturally occurring sweeteners, including plant extracts, sweet proteins (such as miracle fruit protein, petidine, indigofera protein, sematriol, curculigofera protein, carrageenan, areca nut protein), D-amino acids (such as D-phenylalanine, D-tryptophan) or extracts or fractions obtained from natural sources containing these amino acids and / or proteins and physiologically acceptable salts of these amino acids and / or proteins, particularly their sodium, potassium, calcium or ammonium salts; neohesperidin dihydrochalcone, naringin dihydrochalcone, steviol glycosides, steviol glycosides, steviol disaccharides, rebaudioside, rebaudioside A, Rebaudiside B, Rebaudiside C, Rebaudiside D, Rebaudiside E, Rebaudiside F, Rebaudiside G, Rebaudiside H, Rebaudiside M, Rebaudiside N, Rebaudiside X, Duke glycoside, Stevia glycoside, Mogroside (such as Mogroside V), Succinylcholine, Monatose, Glycyrrhetinic acid and its derivatives (especially glycyrrhizic acid glycosides, preferably glycyrrhizic acid ammonium salts) or other natural sweeteners as described in WO2022148540; extracts or enriched fractions of such extracts, such as subspecies of the genera *Thaumatococcus* or *Stemisia*, especially *Stemisia*. Extracts of *Rebaudiana*, *Siraitia grosvenorii*, especially *Momordica / Siraitia grosvenorii / Luo-Han-Guo*, *Glycyrrhiza* subspecies, especially *Glycyrrhiza uralensis*, *Rubus ssp.*, especially *Rubus suavissimus*, *Lippia dulcis*, *Mycetia balansae*, preferably containing balansin A and / or balansin B, or other extracts as described in EP 11168468.

[0045] - Synthetic sweeteners, preferably synthetic sweet substances, preferably selected from the group consisting of: magap, sodium cyclolatate or other physiologically acceptable salts of cyclolatate, acesulfame K, neohesperidin dihydrochalcone, naringin dihydrochalcone, saccharin, sodium saccharin, aspartame, super aspartame, neotame, alitame, advans, perilla stigma, sucralose, lugduname, carrelame, sucrononate or sucrooctate;

[0046] - Sweet carbohydrates, preferably selected from the group consisting of: sucrose, trehalose, lactose, maltose, melitriose, melibiose, raffinose, paraginose, lactulose, D-fructose, D-glucose, D-galactose, L-rhamnose, D-sorbose, D-mannose, D-tagatose, D-alulose, D-arabinose, L-arabinose, D-ribose, D-glyceraldehyde, maltodextrin; and plant preparations containing one or more of these carbohydrates in amounts preferably at least 5 wt.-% and preferably at least 15 wt.-% of each, wherein in these preparations the carbohydrates may be present as naturally occurring or synthetic carbohydrates or mixtures thereof, such as preferably honey, invert sugar syrup from corn starch or highly enriched fructose syrup and / or physiologically acceptable salts of these carbohydrates, particularly their sodium, potassium, calcium or ammonium salts;

[0047] - Sugar alcohols, preferably naturally occurring sugar alcohols, preferably selected from the group consisting of: glycerol, erythritol, threitol, arabinitol, ribitol, xylitol, sorbitol, mannitol, maltitol, isomaltitol, euonymol, lactitol, and their physiologically acceptable salts, especially their sodium, potassium, calcium, or ammonium salts.

[0048] - or a combination of the above two or more.

[0049] In the context of this invention, the acidifier may contain at least one compound that imparts a sour taste, preferably one or more food acids, preferably selected from the group consisting of: acetic acid, adipic acid, aspartic acid, benzoic acid, sorbic acid, caffeic acid, citric acid, isocitric acid, fumaric acid, galacturonic acid, gluconic acid, glucuronic acid, glyceric acid, glycolic acid, ketoglutaric acid, [α]-ketoglutaric acid, lactic acid, lactic acid isocitric acid, malic acid, maleic acid, oxaloacetic acid, oxalic acid, phosphoric acid, pyruvic acid, quinic acid, shikimic acid, succinic acid, tannic acid, and tartaric acid, particularly preferably one or more food acids selected from adipic acid, citric acid, fumaric acid, gluconic acid, glucuronic acid, lactic acid, malic acid, phosphoric acid, succinic acid, and tartaric acid.

[0050] Another aspect of the invention relates to a product selected from the group consisting of food, premium food, food supplements, pharmaceutical preparations, medicinal preparations, cosmetic preparations, oral hygiene preparations, fragrances, pleasure-enhancing consumer preparations, and semi-finished preparations thereof, the product comprising mixtures as defined herein.

[0051] Preferred products are selected from baked goods (e.g., bread, biscuits, cakes, other pastry products), sweets (e.g., chocolate, chocolate bars, other bar products, fruit gummies, hard and soft caramel, chewing gum), alcoholic or non-alcoholic beverages (e.g., coffee, tea, iced tea, wine, wine-containing beverages, beer, beer-containing beverages, liqueurs, distilled spirits, brandy, (carbonated) fruit-containing beverages, (carbonated) isotonic beverages, (carbonated) soft drinks, fruit juices, sparkling wines, fruit and vegetable juices, fruit or vegetable juice preparations), instant beverages (e.g., instant cocoa beverages, instant tea beverages, instant coffee beverages, instant fruit beverages), meat products (e.g., ham, fresh or smoked sausage products, fresh or smoked meat products with added spices or cured), eggs or egg products (dried eggs, egg whites, egg yolks), cereal products (e.g., breakfast cereals, oat bars, pre-cooked instant rice products), and dairy products (e.g., milk beverages, skim milk beverages, milk ice cream, yogurt, kefir, curd cheese). Soft cheese, hard cheese, milk powder, whey, whey beverages, butter, skim milk, partially or fully hydrolyzed milk protein products), products made from soy protein or other soy fractions (e.g., soy milk and products made therefrom, soy protein-containing fruit beverages, soy lecithin-containing preparations, fermented products such as tofu or tempeh or products made therefrom), protein products from other plant sources (e.g., oat protein beverages), fruit preparations (e.g., jams, fruit ice cream, fruit sauces, fruit fillings, frozen fruit products or fruit smoothies), vegetable preparations (e.g., tomato sauce, sauces, dried vegetables, deep-frozen vegetables, pre-cooked vegetables, pickled vegetables), snack products (e.g., baked or fried potato chips or potato cake products, corn-based or peanut-based extrudates), fat-based or oil-based products or their emulsions (e.g., mayonnaise, remola sauce, seasonings), other ready-to-eat meals and soups (e.g., dry soups, instant soups, pre-cooked soups), spices, seasoning mixtures, especially powdered seasonings for snack applications, for example. The formulations / products used for the purposes of this invention can also be used as semi-finished formulations for the production of other nutritional or pleasurable formulations / products. The formulations / products used for the purposes of this invention can also be nutritional supplements in the form of capsules, tablets (uncoated and coated tablets, e.g., anti-gastric acid coated), sugar-coated tablets, granules, pills, solid mixtures, liquid dispersions, emulsions, powders, solutions, pastes, or other swallowable or chewable formulations. Beverages and dairy products are most preferred.

[0052] According to a preferred embodiment, the product of the present invention comprises (in addition to compounds of formula (I) and / or (II) as specified above):

[0053] (iv) The flavoring agent or at least one component or mixture thereof, other than the compounds of formulas (I) and (II), in an amount ranging from 0.001 wt.% to 5 wt.%, preferably from 0.005 wt.% to 2 wt.%, more preferably from 0.01 wt.% to 1 wt.% relative to the total weight of the product; and / or

[0054] (v) the sweetener in an amount ranging from 0.001 wt.% to 15 wt.% relative to the total weight of the product, preferably from 0.2 wt.% to 10 wt.%, more preferably from 0.5 wt.% to 8 wt.%, and / or the acidifier in an amount ranging from 0.001 wt.% to 5 wt.%, preferably from 0.01 wt.% to 2 wt.%, more preferably from 0.1 wt.% to 1 wt.%, relative to the total weight of the product; and / or

[0055] (vi) The amount of the water or the milk relative to the total weight of the product is in the range of 60 wt.% to 99.5 wt.%, preferably 70 wt.% to 99.5 wt.%, more preferably 80 wt.% to 99.5 wt.%, and most preferably 90 wt.% to 99.5 wt.%.

[0056] According to a final aspect of the invention, a method for producing compounds of formula (I) or (II) as defined herein comprises the following steps:

[0057] Provide olefin reductase or microorganisms containing olefin reductase; and

[0058] The double bond of the corresponding coumarin or furanocoumarin used as a precursor is reduced to produce the compound of formula (I) or (II).

[0059] Enyl reductase can be provided as an isolated enzyme, a cell lysate containing the enzyme, a partially purified fraction thereof, a microorganism naturally expressing the enzyme, or a genetically modified microorganism expressing the enzyme. For example, microorganisms containing enyl reductase can be recombinant Escherichia coli strains (such as strain K12 W3110) or yeasts (such as Kluyveromyces marxianus, Torulaspora delbrueckii, or Saccharomyces cerevisiae).

[0060] Preferably, the compound of formula (I) or (II) is selected from the group consisting of compounds of formulas (Ia), (Ib), and (II-a) as defined herein. The precursor is correspondingly selected from the group consisting of compounds of formulas (III-a), (III-b), and (III-c):

[0061] (III-a)(7-methoxycoumarin);

[0062] (III-b)(5,7-dimethoxycoumarin); and

[0063] (III-c)(5-methoxypsoralen)(bergamot lactone).

[0064] The precursor can be obtained synthetically or naturally.

[0065] The invention will be explained in more detail below with reference to representative embodiments.

[0066] Example

[0067] 1. Chemical Synthesis of Chondroitin

[0068] 1.1 Chromone [Ia] (CAS 20921-02-2)

[0069] 3.80 g of 7-methoxycoumarin (21.0 mmol, 1.00 equivalent) was dissolved in 100 mL of acetic acid, and 0.76 g of 5% palladium / carbon (20 wt%) was added. The suspension was stirred at room temperature under a hydrogen atmosphere for 4.5 h. The mixture was filtered through a diatomaceous earth pad, and the solvent in the filtrate was concentrated under reduced pressure. The residue was added to 150 mL of water, and the aqueous layer was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate solution (3 × 50 mL) and saturated aqueous NaCl solution (50 mL), dried over sodium sulfate, and the solvent was removed under reduced pressure to give 3.8 g of 7-methoxy-2-cryocystone [Ia] (21.3 mmol, 99%) as a white solid.

[0070] NMR: 1 H-NMR (400 MHz, CDCl3): δ = 7.07 (dtd, 1, J = 8.3 Hz, 0.9 Hz, 0.3Hz, C Ar HC Ar HC Ar ), 6.64 (dd, 1, J = 8.3 Hz, 2.6 Hz, C Ar OCH3C ArH C Ar H), 6.60 (dq, 1,J = 2.5 Hz, 0.3 Hz, C Ar C Ar H C Ar OCH3), 3.78 (s, 3, C Ar HC Ar O CH3 C Ar H), 2.96 - 2.91 (m, 2, C) Ar CH2 CH2), 2.79-2.74 (m, 2, CH2 CH2 CO2). 13 C-NMR (100 MHz, CDCl3): δ =168.55, 159.65, 152.70, 128.44, 114.38, 110.40, 102.53, 55.53, 29.48, 23.00.

[0071] GC / MS (EI, 70 eV, two of the strongest ions out of every 14 ions above m / z 20): 27 (2), 29 (1), 39 (9), 42 (3), 50 (5), 51 (11), 53 (5), 63 (9), 65 (15), 77 (22), 78 (19), 91 (11), 92 (4), 107 (10), 108 (29), 121 (15), 124 (6), 136 (32), 137 (16), 149 (26), 150 (41), 161 (7), 162 (1), 178 (100), 179 (11).

[0072] Purity (GC / MS): 99%

[0073] 1.2 Chromone [Ib] (CAS 82243-01-4)

[0074] 1.00 g of 5,7-dimethoxycoumarin (4.75 mmol, 1.00 equivalent) was dissolved in 15 mL of tetrahydrofuran, and 250 mg of 5% palladium / carbon (25 wt%) was added. The suspension was stirred in an autoclave at 80 °C for 2 hours under 10 bar of hydrogen. The mixture was cooled to room temperature, filtered through an injection filter, and the solvent in the filtrate was removed under reduced pressure. The crude product was purified by silica column chromatography (n-hexane / ethyl acetate 100:0 -> 80:20) to give 0.95 g of 5,7-dimethoxy-2-cryomenone [Ib] (4.56 mmol, 96%) as a white solid.

[0075] NMR: 1 H NMR (400 MHz, CDCl3): δ = 6.25 (d, 1, J = 2.3 Hz, C Ar OCH3C Ar H C Ar OCH3), 6.23 (d, 1, J = 2.3 Hz, C Ar OCH3C Ar H C Ar ), 3.82 (s, 3,C Ar HC Ar O CH3 C Ar ), 3.78 (s, 3, C Ar HC Ar O CH3 C Ar H), 2.92 - 2.84 (m, 2, C Ar CH2 CH2), 2.77–2.68 (m, 2, CH2) CH2 CO2). 13 C NMR (101 MHz, CDCl3): δ = 168.73, 160.12, 157.33, 153.23, 103.42, 94.73, 93.88, 77.35, 77.04, 76.72, 55.71, 55.57, 28.86,17.12.

[0076] GC / MS (EI, 70 eV, two of the strongest ions out of every 14 ions above m / z 20): 27 (1), 29 (1), 39 (5), 42 (3), 51 (6), 53 (5), 65 (7), 69 (13), 77 (11), 78 (7), 91 (4), 95 (11), 108 (4), 109 (14), 121 (5), 123 (8), 137 (18), 138 (37), 149 (5), 151 (5), 165 (19), 166 (71), 179 (6), 180 (5), 191 (3), 193 (1), 208 (100), 209 (12).

[0077] Purity (GC / MS): 99%

[0078] 1.3 Chromone [II-a] (CAS 29050-61-1)

[0079] Under a nitrogen atmosphere, 1.00 g of bergamot lactone (4.62 mmol, 1.00 equivalent) was dissolved in 58 mL of toluene. 5.80 mL of triethylsilane (36.3 mmol, 8.00 equivalent) and 230 mg of triphenylphosphine rhodium chloride (0.25 mmol, 0.015 equivalent) were added, and the suspension was stirred at 65 °C for 6 hours. The mixture was cooled to room temperature, filtered through an injection filter, and the solvent in the filtrate was removed under reduced pressure. The crude product was purified by silica column chromatography (cyclohexane / ethyl acetate 100:0 -> 70:30) to give 155 mg of 5,6-dihydrobergamot lactone [II-a] (0.71 mmol, 16%) as a white solid.

[0080] NMR: 1 H NMR (400 MHz, CDCl3): δ = 7.54 (d, 1, J = 2.3 Hz, C Ar HC Ar HO), 6.96(s, 1, C Ar C Ar H C Ar ), 6.91 (dd, 1, J = 2.3 Hz, 1.0 Hz, C Ar C Ar H C Ar H), 4.12 (s, 3,CO CH3), 3.05 (dd, 2, J = 8.2 Hz, 6.3 Hz, CH2 CH2 C Ar ), 2.78 - 2.72 (m, 2,CO2 CH2 CH2). 13 C NMR (101 MHz, CDCl3): δ = 168.85, 155.44, 150.32, 149.53, 144.20, 114.11, 108.07, 104.57, 94.86, 59.91, 28.96, 17.94.

[0081] GC / MS (EI, 70 eV, two of the strongest ions out of every 14 ions above m / z 20): 27 (1), 29 (1), 38 (2), 39 (6), 50 (8), 51 (19), 63 (11), 65 (8), 77 (15), 89 (11), 90 (6), 91 (14), 104 (2), 105 (5), 118 (5), 131 (6), 133 (37), 134 (4), 146 (40), 147 (26), 161 (37), 162 (5), 176 (100), 177 (12), 189 (2), 190 (12), 202 (1), 203 (0.5), 218 (96), 219 (13).

[0082] Purity (GC / MS): 99%

[0083] 2. Biotechnical Synthesis of Chromone

[0084] 2.1 Transformation of plasmid DNA into E. coli cells

[0085] Plasmid DNA was converted into chemically competent *E. coli* DH5α cells (New England Biolabs, Frankfurt am Main, Germany) for plasmid proliferation. To produce the expression strain, the plasmid DNA was converted into chemically competent *E. coli* BL21(DE3) cells.

[0086] 50 µL of the corresponding *E. coli* strain was incubated on ice for 5 minutes. After adding 1 µL of plasmid DNA, the suspension was mixed and incubated on ice for 30 minutes. Transformation was performed by incubating the cell suspension in a heating block at 42°C for 45 seconds, followed by incubation on ice for 2 minutes. After adding 350 µL of SOC Outgrowth medium (New England Biolabs, Frankfurt am Main, Germany), the cells were incubated at 37°C and 200 rpm for 1 hour. Subsequently, the cell suspension was spread on LB agar plates (Car Roth GmbH, Karlsruhe, Germany) containing the corresponding antibiotics and incubated at 37°C for 16 hours.

[0087] 2.2 Generation of expression plasmids

[0088] The sequences encoding sequences according to SEQ ID NO 1 to SEQ ID NO 16, respectively, were synthesized and cloned into the NcoI and XhoI restriction sites in the pET28a vector (Twist BioScience, San Francisco, USA) to obtain the plasmids listed in Table 1. These expression vectors were then transformed into E. coli BL21(DE3) cells according to Example 1 as described in Example 1.

[0089] Table 1: Source organisms of the obtained plasmids and enzyme sequences.

[0090]

[0091]

[0092] 2.3 Culture Escherichia coli cells and perform biotransformation using olefin reductase

[0093] E. coli BL21(DE3) cells containing one of the pET28a plasmids listed in Table 1 were seeded into 5 mL of LB medium (Carl Roth GmbH, Karlsruhe, Germany) containing the necessary antibiotics. After incubation for 16 hours (37°C, 200 rpm), the cells were inoculated at 0.1 OD280. 600 Cells were inoculated into 50 mL of TB medium (CarlRoth GmbH, Karlsruhe, Germany) containing the necessary antibiotics. Cells were allowed to grow (37°C, 200 rpm) to an OD of 0.5–0.8. 6001 mM isopropyl-β-D-thiogalactoside was added to the culture. The cell culture was incubated for 16 h (22 °C, 200 rpm), centrifuged (10 min, 10,000 rpm), and the supernatant was discarded. Cell clumps were lysed using B-PER protein extraction reagent (Thermo Fisher Scientific, Bonn, Germany) according to the manufacturer's instructions. After a subsequent centrifugation (10 min, 20,000 rpm), biotransformation was performed using the supernatant by adding 1.5 mM nicotinamide adenine dinucleotide phosphate, 1.5 mM nicotinamide adenine dinucleotide, 1 M glucose, 1 U glucose dehydrogenase, and 50 ppm substrate. Bergamot lactone, 7-methoxycoumarin, or 5,7-dimethoxycoumarin were used as substrates. The reaction mixture was incubated at 30 °C for 16 h. After terminating the reaction with acetonitrile (1 volume of reaction mixture + 1 volume of acetonitrile and 0.2% formic acid), the sample was centrifuged (20 min, 20,000 rpm) and the supernatant was used for LC and LC-MS analysis.

[0094] 2.4 Yeast culture and substrate biotransformation

[0095] The microorganisms listed in Table 2 were inoculated into 5 mL of YPD medium (Carl Roth GmbH, Karlsruhe, Germany). After incubation for 16 hours (30°C, 200 rpm), the cells were cultured at an OD of 0.2. 600 Cells were seeded in 50 mL of YPD medium (Carl Roth GmbH, Karlsruhe, Germany). Cells were allowed to grow (30°C, 200 rpm) to an OD value of 1. 600 50 ppm of the corresponding substrate (bergamot lactone, 7-methoxycoumarin, or 5,7-dimethoxycoumarin) was added to the culture. The cell culture was incubated for 16 h (30 °C, 200 rpm) and terminated with acetonitrile (1 volume of culture + 1 volume of acetonitrile and 0.2% formic acid). The samples were centrifuged (20 min, 20,000 rpm) and the supernatant was used for LC and LC-MS analysis.

[0096] Table 2: Microorganisms used for substrate biotransformation.

[0097]

[0098] According to the literature, the biotransformation of simple coumarin to dihydrocoumarin and its corresponding acid has been studied (Häser et al. Biocatalytic Production of Dihydrocoumarin from Coumarin by Saccharomycescerevisiae. J. Agric. Chem. 2006, 54, 6236-6240; Serra et al. Biocatalytic Synthesis of Natural Dihydrocoumarin by Microbial Reduction of Coumarin. Catalysts 2019, 9, 665; Serra et al. Two Biotechnological Approaches to the Preparative Synthesis of Natural Dihydrocoumarin. Catalysts 2022, 12, 28).

[0099] Table 3 shows the results of the above-mentioned biotransformation of substituted coumarins (7-methoxycoumarin, 5,7-dimethoxycoumarin and bergamot lactone) as precursors to 7-methoxy-2-chromone, 5,7-dimethoxy-2-chromone, 5,6-dihydrobergamot lactone and their corresponding acids.

[0100] Table 3: Results of biotransformation screening experiments (+ = detected, trace = detected trace, - = not detected, * = preliminarily identified).

[0101]

[0102]

[0103] 3. Application Examples

[0104] 3.1 Example 1

[0105] In lemonade made from 91.84% water, 8% sucrose, 0.15% citric acid, and 0.01% lime-type flavoring agents, a distinctly perceptible coumarin flavor is produced by adding 0.1 ppm (parts per million) of 7-methoxy-2-chromone. This coumarin flavor is described as typical of virgin cold-pressed lime oil. Furthermore, the juiciness and authenticity of the product are significantly improved. The coumarin-like flavor is further enhanced by adding a combination of 0.1 ppm of 7-methoxy-2-chromone and 1 ppm of 5,7-dimethoxy-2-chromone, while another combination of 0.1 ppm of 7-methoxy-2-chromone, 1 ppm of 5,7-dimethoxy-2-chromone, and 1 ppm of 5,6-dihydrobergamot lactone produces a very typical and well-rounded character reminiscent of cold-pressed lime juice.

[0106] List of ingredients for lime-based flavoring agents (based on a 100-unit scale):

[0107] A lime oil formulation (38) consisting of an extractable fraction of a mixture of 16% distilled lime peel oil, 16% cold-pressed lime peel oil and 68% 1,2-propanediol; a lemon oil formulation (18) consisting of an extractable fraction of 14% cold-pressed lemon peel oil and 86% 1,2-propanediol; a grapefruit oil formulation (41) consisting of an extractable fraction of 5% grapefruit peel oil and 95% 1,2-propanediol; and citral (3).

[0108] 3.2 Example 2

[0109] In lemonade containing 91.8% water, 8% sucrose, 0.15% citric acid, and 0.05% lemon-based flavorings, the addition of 0.8 ppm of 5,7-dimethoxy-2-chromone improved juiciness and sweetness, as well as the perception of herbaceous, earthy notes reminiscent of distilled lime. This flavor is also found in clear lemon and lime soda. The lime-like flavor was further enhanced by adding a combination of 0.8 ppm of 5,7-dimethoxy-2-chromone and 0.08 ppm of 7-methoxy-2-chromone. A third combination of 0.8 ppm of 5,7-dimethoxy-chromone, 0.08 ppm of 7-methoxy-2-chromone, and 0.8 ppm of 5,6-dihydrobergamot lactone significantly modulated the lemongrass aroma characteristics into a cold-pressed lime with pronounced floral, juicy, and candy-like notes.

[0110] List of ingredients for lemon-based flavoring agents (based on a 100-unit scale):

[0111] Enriched orange peel oil (0.4), enriched lemon peel oil (4), lemon peel oil terpenes (2.5), citral (1), 96% vol ethanol (77.1) and water (15).

[0112] 3.3 Example 3

[0113] In a dairy beverage composed of 93.9% whole milk containing 3.8% fat, 6% sucrose, 0.1% natural vanilla extract, and 10 ppm hesperidin, the addition of 1 ppm 7-methoxy-2-chromone significantly mimicked flavor perception. Subjects described a significant enhancement in buttery, anise-like, and caramel-like flavors, as well as overall aroma intensity.

[0114] The addition of 1 ppm of 5,7-dimethoxy-2-chromone produced an additional effect: the complexity and intensity of the flavor were significantly enhanced, and the added creaminess completed the overall character.

[0115] Components of vanilla extract (list based on 100):

[0116] Vanilla bean equivalent (27), 96% by volume ethanol and water.

[0117] 3.4 Example 4

[0118] In a milk beverage composed of 93.95% whole milk containing 3.8% fat, 6% sucrose, and 0.05% caramel-type aromatic compounds, the addition of 1 ppm 7-methoxy-2-cryomenone resulted in a noticeable shift in aroma characteristics towards toffee, butter, and cream, while also rounding out previously present baked flavors. Simultaneously, the addition of 1 ppm 5,7-dimethoxy-2-cryomenone to this milk beverage significantly enhanced the typical characteristics of the milk and the creamy flavor. The combination of 1 ppm each of 7-methoxy-2-cryomenone and 5,7-dimethoxy-2-cryomenone in this beverage example showed the most pronounced effect: it highlighted the toffee, butter, and cream flavors in a very positive manner, and further enhanced the overall aroma intensity.

[0119] Components of caramel-based flavoring (list based on 100):

[0120] 1,2-Propanediol (83.3), furanone (3), cocoa powder extract (8.5) consisting of 15% cocoa powder extract, 50% 1,2-propanediol and 50% water, acetylmethylethanol (1), diacetyl (1), methylcyclopentenolone-3,2,2 (0.7), maltol (0.5), vanilla extract consisting of 27% vanilla bean extract, 96% vol% ethanol and water.

[0121] 3.5 Example 5

[0122] In a green tea beverage composed of 99.2% water and 0.8% green tea extract, the addition of 0.8 ppm of 5,7-dimethoxy-2-chromone produced a very typical fermented flavor of green tea. However, overall, the tea-like flavor was also significantly enhanced. Therefore, this substance can optimize the authentic taste of green tea.

[0123] Ingredients of green tea preparations (detailed list based on 100):

[0124] 96% vol% ethanol (34.47), green tea CO2 extract composed of 30% extractable green tea components and 70% 96% vol% ethanol (2), black tea CO2 extract composed of 35% extractable black tea components and 65% 96% vol% ethanol (0.5), tea distillate composed of 10% distillable black tea components, 10% 96% vol% ethanol and 90% water (51), yerba mate leaf distillate composed of 20% distillable yerba mate components, 45% 96% vol% ethanol and 55% water (12), linalool (0.027).

[0125] 3.6 Example 6

[0126] A tomato-based flavoring agent (0.06%) was incorporated into a flavored matrix consisting of 98.91% water, 0.2% salt, 0.8% sucrose, and 0.03% citric acid. The addition of 1 ppm of 5-7-dimethoxy-2-chromone significantly enhanced key tomato-type flavor characteristics, such as fruitiness, spiciness, and tomato paste flavor.

[0127] The ingredients of the tomato-based flavoring agent (data based on 100): 1,2-propanediol (88.98), eugenol acetate (3), furanone (0.6), dimethyl sulfide (2), maltol (4), 3-methylthiopropional (0.2), geraniol butyrate (1), β-ionone (0.2), isobutylthiazole (0.02).

[0128] 3.7 Example 7

[0129] 1 ppm of 5,7-dimethoxy-2-chromone was added to a flavored matrix consisting of 93.97% water, 6% sucrose, and 0.03% cherry-type flavoring agents. The moderating properties of 5,7-dimethoxy-2-chromone enhanced the floral and fruity aroma characteristics, particularly highlighting the juiciness and typical flavor of Black Morelo cherry.

[0130] Ingredients of cherry-flavored seasoning (data based on 100)

[0131] Ethyl acetate (0.4%), ethyl butyrate (0.2%), cinnamaldehyde (0.1%), 3Z-hexenol (0.05%), ethyl benzoate (0.02%), butyric acid (0.04%), benzyl acetate (0.02%), β-ionone (0.02%), isoamyl isovalerate (0.02%), anisaldehyde (0.01%), and 96% vol ethanol (99.12%).

[0132] 3.8 Example 8

[0133] 0.4 ppm of 7-dimethoxy-2-chromone was added to a flavored matrix consisting of 93.99% water, 6% sucrose, and 0.01% raspberry-type flavoring agents. This significantly mimicked the previous slightly fruity, juicy, ester-like, and grassy characteristics. The characteristics produced by adding 7-methoxy-2-chromone were described as sweeter, more balanced, more mature, and reminiscent of candy. Conversely, a combination of 0.4 ppm of 7-methoxy-2-chromone and 1 ppm of 5,7-dimethoxy-2-chromone mimicked warm woody and balsam notes.

[0134] Raspberry-based flavoring ingredients (in units of 100).

[0135] Ethyl acetate (2), hydroxybenzylacetone (2.5), benzyl acetate (1.5), maltol (1), furanone (0.3), α-ionone (0.5), isoamyl acetate (0.5), 3Z-hexenol (0.5), 3Z-hexenyl acetate (0.5), acetaldehyde (0.25), β-dukethenone (0.05), 96% vol ethanol (90.9).

Claims

1. Use of at least one compound of formula (I) or (II) for modifying the characteristics of an aromatic composition, （I） (II) in, R1, R2, R3, and R4 are independently selected from the group consisting of H, OH, OCH3, or OCH2CH3, excluding compounds of formula (I) where R1, R2, R3, and R4 are all H.

2. The use according to claim 1, wherein, The compound of at least one formula (I) or (II) is selected from the group consisting of compounds of formulas (Ia), (Ib) and (II-a) and mixtures of the compounds. (I-a) (I-b) (II-a)。 3. The use according to claim 2, wherein, The aromatic composition contains: Compounds of formula (Ia) with a concentration greater than 10 ppb; and / or Compounds of formula (Ib) with a concentration greater than 20 ppb; and / or Compounds of formula (II-a) with a concentration greater than 20 ppb.

4. The use according to any one of the preceding claims, wherein, The adjustment is to improve or enhance the aroma of a group selected from the following groups or to shift the aroma towards the following groups: coumarin-like, juicy, fruity, floral, honey-like, fresh, lime, lemon, invigorating, herbal, earthy, buttery, anise, caramel, toffee, milky, spicy, earthy, sweet, sour, or creamy.

5. The use according to any one of the preceding claims, wherein, The aromatic composition is a product selected from the group consisting of: food, premium food, food supplement, pharmaceutical preparation, medicinal preparation, cosmetic preparation, oral hygiene preparation, fragrance, pleasure-oriented consumer preparation, and its semi-finished preparation.

6. A mixture comprising at least one compound of formula (I) or (II) according to claim 1, preferably at least one compound of formula (Ia), (Ib) and (II-a) according to claim 2. The amount of the compound is: (i) Compounds of formula (I), preferably formula (Ia), with a concentration greater than 10 ppb, preferably greater than 50 ppb, more preferably greater than 500 ppb; and / or (ii) Compounds of formula (I), preferably formula (Ib), with a concentration greater than 20 ppb, preferably greater than 100 ppb, more preferably greater than 1000 ppb; and / or (iii) Compounds of formula (II), preferably formula (II-a), with a content greater than 20 ppb, preferably greater than 100 ppb, more preferably greater than 1000 ppb.

7. The mixture according to claim 6, wherein, The mixture contains: a. Compounds of formula (Ia) and compounds of formula (Ib); or b. Compounds of formula (Ia) and compounds of formula (II-a); or c. Compounds of formula (Ib) and compounds of formula (II-a); or d. Compounds of formula (Ia), compounds of formula (Ib) and compounds of formula (II-a).

8. The mixture according to claim 6 or 7, wherein, The mixture is not derived from essential oils or extracts.

9. The mixture according to any one of claims 6 to 8, wherein the mixture further comprises: (iv) Flavoring agents other than the compounds of formulas (I) and (II), wherein, The flavoring agent is preferably selected from flavoring agents of the lime, vanilla, caramel, green tea, tomato, cherry and raspberry types.

10. The mixture according to any one of claims 6 to 9, wherein the mixture further comprises one or more of the following: (v) sweeteners and / or acidifiers; and (vi) Water or milk.

11. A product selected from the group consisting of food, premium food, food supplements, pharmaceutical preparations, medical preparations, cosmetic preparations, oral hygiene preparations, fragrances, pleasure-enhancing consumer preparations, and semi-finished preparations thereof, the product comprising a mixture according to any one of claims 6 to 10.

12. The product of claim 11, wherein the product is selected from the group consisting of: baked goods; confectionery; alcoholic or non-alcoholic beverages; meat products; eggs or egg products; cereal products; dairy products; products made from soy protein or other soy fractions; protein products from other plant sources, such as oat protein beverages; fruit preparations; vegetable preparations; snack products; fat-based or oil-based products or emulsions thereof; other ready-to-eat meals and soups; spices; seasoning mixtures; nutritional supplements in the form of capsules, tablets, sugar-coated tablets, granules, pellets, solid mixtures, liquid dispersions as emulsions, powders, solutions, pastes or other swallowable or chewable preparations, preferably beverages and dairy products.

13. The product according to claim 11 or 12, wherein the product comprises: (iv) The flavoring agent or at least one component or mixture thereof, other than the compounds of formulas (I) and (II), in an amount ranging from 0.001 wt.% to 5 wt.%, preferably from 0.005 wt.% to 2 wt.%, more preferably from 0.01 wt.% to 1 wt.% relative to the total weight of the product; and / or (v) the sweetener in an amount ranging from 0.001 wt.-% to 15 wt.-%, preferably from 0.2 wt.-% to 10 wt.-%, more preferably from 0.5 wt.-% to 8 wt.-%, relative to the total weight of the product; and / or the acidifier in an amount ranging from 0.001 wt.-% to 5 wt.-%, preferably from 0.01 wt.-% to 2 wt.-%, more preferably from 0.1 wt.-% to 1 wt.-%, relative to the total weight of the product; and / or (vi) The amount of the water or the milk relative to the total weight of the product is in the range of 60 wt.% to 99.5 wt.%, preferably 70 wt.% to 99.5 wt.%, more preferably 80 wt.% to 99.5 wt.%, and most preferably 90 wt.% to 99.5 wt.%.

14. A method for producing a compound of formula (I) or (II) according to claim 1, the method comprising: Provide olefin reductase or microorganisms containing olefin reductase; and The double bond of the corresponding coumarin or furanocoumarin used as a precursor is reduced to produce the compound of formula (I) or (II).

15. The method according to claim 14, wherein, The compound of formula (I) or (II) is selected from the group consisting of compounds of formula (Ia), (Ib) and (II-a) according to claim 2; and / or the precursor is selected from the group consisting of compounds of formula (III-a), (III-b) and (III-c): (III-a) (III-b), and (III-c)。

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