ESSENTIAL OIL

Abstract: A process for the preparation of hypoallergenic moss oils, comprising reacting the starting moss oil, a concrete or preferably an absolute thereof, with an aldehyde reducing agent, in an organic solvent medium.

Description: Essential Oil The present invention relates to the preparation of moss oils which are characterized by hypoallergenicity, i.e. by a strongly reduced allergenic potential and excellent olfactive performance in perfume compositions. Moss oils are highly appreciated by perfumers for their typical woody notes and play an important role in the creation of perfumes, e.g. of the so-called "Chypre" or "Fougέre" type. They are obtained by solvent extraction of lichens including, in particular Evernia prunastri L. for the Oakmoss oil and Evernia furfuracea L. for Treemoss oil. The total amount of lichens treated worldwide for perfumery use may be estimated to 6000 tons/year (P. Vigne, Parfums, Cosmetiques, Arόmes, (78), p 97-105, 1987) and represents an estimated annual turnover close to $ 35 millions. Moss extracts, e.g. moss absolutes or concretes, which are the most frequently used moss oil products have been reported to cause contact sensitization on human skin, and several groups of researchers have shown that some components of moss oils, particularly ethyl hematommate I, ethyl chlorohematommate II, atranorin III and chloroatranorin IV as depicted on page 3 are involved in these allergenic reactions. European patent publication No. 202,647 (Shiseido Company Ltd.) describes a process for the preparation of hypoallergenic moss oils by removing allergenic substances from moss oils by chromatography, solvent extraction, countercurrent partition and membrane separation followed by a catalytic hydrogenation and/or an alkaline treatment. The allergens removed in this way are aldehydes which include the above mentioned ethyl hematommate I, ethyl chlorohematommate II, atranorin III and chloroatranorin IV. The more recent European patent publication No. 468,189 (Roure S.A.) describes a process for the preparation of hypoallergenic moss oils by reacting, e.g. moss absolutes or concretes in alcoholic solution with a ino acid(s) under mono-phasic conditions followed by removal of the insolubilized allergenic substances I-VL II III IV VI The concentrations of the allergens I-VI achieved in this way are clearly below the critical levels (0.05 - 1%) established experimentally via allergenicity tests. The goal of the present invention was to eliminate now substantially further the above-mentioned aldehydes I-VI by using an economical process without affecting significantly the original color and olfactive performance of the starting moss oil, thereby providing moss absolutes or concretes with a strongly reduced allergenic potential. It was achieved by reacting moss extracts e.g. concretes or absolutes with appropriate reducing agents, e.g. alkali metal borohydrides, which specifically and exclusively reduce the allergenic aldehydes I-VI to the corresponding primary alcohols. A further advantage of the novel process consists in the avoidance of concomitant formation of trace amounts of colorants, which can cause inconveniences for some perfumery applications. The present invention describes thus the reaction between moss extracts, e.g. oils, concretes or absolutes, with appropriate aldehyde reducing agents, e.g. complex metal hydrides, and preferably with alkali metal borohydrides, e.g. lithium or sodium borohydride, in organic solvent systems. The starting moss extracts subjected to this treatment are suitably obtained by solvent extraction of lichens and include in particular the Oakmoss concrete (Evernia prunastri L.) and the Treemoss concrete (Evernia furfuracea L.) and, preferably, the absolutes thereof. It is known from the literature [e.g. Reagents for Organic Synthesis, L.F. Fieser and . Fieser, p 599-603 and 1049-1055, Editor John Wiley and Sons, Inc., 1967] that complex alkali metal hydrides, e.g. borohydrides or aluminium hydrides are able to reduce aldehydes, ketones and even esters. The aldehydic allergens I-VI contribute insignificantly to the total odour of moss extracts, but numerous esters, other aldehydes and ketones are known to be olfactively important minor or major constituents of these extracts [R. Ter Heide et al., Qualitative Analysis of the Odoriferous Fraction of Oakmoss (Evernia prunastn (L.) Ach.), J. Agric. Food Chem., 23 (5) p 950-957 (1975)]. It was therefore surprising to find, that the novel process allowed the selective reduction of allergens I-VI without organoleptically deteriorating the moss oil, or in other words, none of the above- mentioned organoleptically active constituents, e.g. no organoleptically relevant esters appear to have been removed from the original moss extract, as was demonstrated by GC data. In addition, the concentration levels found for I-VI are far below the required limits (cf. Table 1) and those achieved in earlier publications [EP publication No. 468,189 and C. Ehret, P. Maupetit, M. Petrzilka, G. Klecak, Int. J. of Cosm. Science, 2A, 121-130 (1992)]. Finally, the colors of the resulting non-allergenic moss oils are very close to the original ones and are therefore suitable for most perfumery applications. Table 1 Concentration levels required Aldehydes for moss absolute with reduced allergenic potential (%) Ethyl hematommate I ≤l Ethyl chlorohematommate II ≤0.05 Atranorins III + IV ≤0.15 Atranol V ≤0.2 Chloratranol VI ≤0.2 In the broadest context of the present invention, the allergenic moss oil is dissolved in an appropriate organic solvent and treated with preferably an excess of an appropriate aldehyde reducing reagent. The suitable reducing agents are those which are able to reduce exclusively, or at least preferentially the aldehydes over the esters and belong to various types (cf. R.C. Larock, Comprehensive Organic Transformations, A Guide to Functional Group Preparations, published by VCH PubHshers, Inc., New- York, 527-535 [1989],) e.g. : complex metal or ammonium hydrides, such as sodium, lithium, potassium, zinc, tetraethylammonium borohydrides, etc., substituted complex metal or ammonium hydrides, such as sodium triacetoxyborohydride, potassium triacetoxyborohydride, sodium cyanoborohydride or tetra-n-butylammonium triacetoxyborohydride, metal hydrides, such as diborane or an alkali or aluminium hydride, etc. The preferred reducing agents are lithium borohydride and sodium borohydride. The reduction can be carried out according to know methods. It is usually carried out in an organic medium, e.g. in solution using optionally halogenated, aliphatic or aromatic hydrocarbon solvents, such as hexane, cyclohexane or toluene, etc., ester solvents such as ethyl acetate, isopropyl acetate etc., or alcoholic solvents, such as methanol, ethanol etc. Alternatively ether solvents such as t-butyl methyl ether, tetrahydrofuran etc., or halogenated solvents such as methylene chloride may also be used. Another possibility consists in using mixtures of the above-mentioned solvents. The concentrations of moss extracts applied in the reaction may vary between ca. 5-50%, preferably between ca. 5-15% (w/w). Convenient amounts of reducing agents, e.g. alkali metal borohydrides, are ca. 0.02-0.1g, preferably ca. 0.03-0.07g per g of moss extract. This amount represents a ca. 2 to 5 fold molar excess, i.e. a ca. 8 to 20 fold reducing equivalent excess. The reaction temperature is ca. 20°-80°C, preferably ca. 20°-30°C, if, e.g. lithium borohydride is used, and preferably reflux temperature, e.g. that of an alcanol, e.g. ethanol, if sodium borohydride is used. - 7 - The reaction is usually quenched after ca. 30 minutes to 3 hours, preferably after ca. 30 to 60 minutes, if sodium borohydride is used, and after ca. 1-2 hours, if lithium borohydride is used. If water insoluble solvents are used for the reaction, such as hydrocarbons, esters, halogenated and aliphatic ether solvents, work up consists in extensively washing the reaction mixture with water or aqueous acids (e.g. 1-10%, preferably 1-3% aq. HC1 solution) followed by water until neutral. Finally the organic solvent is distilled off at reduced pressure without exceeding a temperature of ca. 85°C. Alternatively, if water soluble solvents are used, e.g. an alcohol or a cyclic ether, such as tetrahydrofuran, the solvent is first removed by distillation at reduced pressure. The remaining residue is then redissolved in a water insoluble solvent, e.g. the solvents mentioned above, and worked up as in the previous case. Examples 1) Allergenicitv The strongly reduced allergenic potential in the product was in each case determined by conventional, fully established skin sensitization and skin response methods, i.e. in concreto the so-called * Modified BUEHLER method using guinea pigs, and the * RIPT (Repeated Insult Patch Test) using human subjects. 2) Analysis Content of aldehydes I. II. V and VI The contents of products I, II, V and VI are suitably determined by GC analysis, using an internal standard and working under the following conditions : * Column : 50m x 0.32mm inner diameter, fused silica * Stationary phase : CP Sil 5CB (a silicone) * Detector : FID (flame ionisation detector) * Vector gas : Helium, 2ml/mn * Temperature program : 100-240°C, 2°C/min. * Internal standard : methyl 2,4-dihydroxy-3,6-dimethyl-benzoate - 8 - Content of aldehydes III and W The contents of aldehydes HI and IV are suitably determined by HPLC, using an external standard and working under the following conditions : 5 * Column : 250mm length, 4.6mm i.d. * Stationary phase : RP 18 (reverse phase, particle size : 7μm) * Detector : UV at 260nm * Mobile phase : H 2 O acidified to pH 2.8 with cone. H3PO 4 B : acetronitrile 10 * Gradient : Timβ fmin.) %A %B Flow fail/min 0 80 20 1 30 5 95 1 40 5 95 1 This gradient allows the effective separation of the above non- 15 volative aldehydes HI and IV. .τnτnp1p 1 Productio of Oakmoss absolute with strongly reduced alleryem>. potential usin-f Kthinm horohvdride A 500ml three-necked flask equipped with a mechanical stirrer, 20 a condenser and a dropping funnel was charged with 15g of a commercially available melted Oakmoss absolute (Givaudan-Roure, mp about 70°C), which then was dissolved in 200ml of cyclo- hexane/isopropyl acetate 3:1 at room temperature and under N 2 . To this homogeneous solution was then added dropwise a suspension of 25 480mg (22 mmol) of lithium borohydride in 100ml of cyclo- hexane/isopropyl acetate 3:1 during ca. 30 minutes. Immediately after addition a precipitation occurred and a slight increase of the tem¬ perature of the reaction mixture (ca. 6°C) was observed. After stirring the reaction mixture for an additional 2 hours at room temperature, it 30 was carefully quenched with 150ml of 0.5% (w/w) aqueous HC1 and extracted with cyclohexane/isopropylacetate 3:1 (3 x 300ml). The organic layers were washed with water (1 x 150ml), combined and concentrated at reduced pressure (20 mbars) on a water bath without exceeding a temperature of ca. 85°C. An Oakmoss absolute (12.78g, 85.2% yield) was obtained in this way, which according to GC- and HPLC- analysis contained extremely small amounts of aldehydes I-VI (cf. Table 2). Aldehyde Ethyl hematommate I Ethyl chlorohematommate H Atranorins in + TV Atranol V Chloratranol VI Production of Oakmoss absolute with strongly reduced allergenic potential υsιτψ ~-Λ..-τr. borohydride A 250ml three-necked flask equipped with a mechanical stirrer, a condenser and a dropping funnel was charged with 14.9g of a com¬ mercially available melted Oakmoss absolute (Givaudan-Roure, mp about 70°C), which then was dissolved in 90ml of ethanol 96% at room temperature and under N2. To this solution was then added dropwise a suspension of lg (26.4 mmol) of sodium borohydride in 60ml of ethanol 96% during ca. 5 minutes. During the addition a slight increase of the temperature (ca. 12°C) was observed. After stirring the reaction mixture at reflux temperature during 45 minutes, the ethanol was distilled off at reduced pressure (20 mbars) and the residue was taken up in 300ml of t-butyl methyl ether (TBME). The reaction mixture was then carefully quenched with 150ml of water and extracted with TBME (3 x 300ml). The organic layers were washed with water (1 x 150ml), combined and concentrated at reduced pressure (20 mbars) on a water bath without exceeding a temperature of ca. 65°C. An Oakmoss absolute (12.78g, 85.2% yield) was obtained in this way, which according to GC- and HPLC- analysis contained extremely small amounts of aldehydes I-VI (cf. Table 3). A 250ml three-necked flask equipped with a mechanical starrer, a condenser and a dropping funnel was charged with 15g of a com¬ mercially available (Givaudan-Roure), melted Oakmoss absolute (mp about 70°C), which then was dissolved in 100ml of ethanol 96% at room temperature and under N 2 . To this solution was then added dropwise a suspension of lg (26.4 mmol) of sodium borohydride in 60ml of EtOH 96% during ca. 5 minutes. During the addition a slight increase of the temperature (ca. 15°C) was observed. After stirring the reaction mixture at reflux temperature during 45 minutes ethanol was distilled off at reduced pressure (20 mbars) and the residue was taken up in 300ml of t-butyl methyl ether. The reaction mixture was then carefully quenched with 100ml of water and acidified to pH = 1.5 with ca. 15ml of 6% aqueous HC1. The organic layer was washed with water (1 x 150ml) and concentrated at reduced pressure (20mbars) on a water bath without exceeding a temperature of ca. 65°C. An Oakmoss absolute (14.42g, 96.1% yield) was obtained in this way, which according to GC- and HPLC- analysis contained extremely small amounts of aldehydes I-VI (cf. Table 4).

Claims: Claims 1. A process for the preparation of hypoallergenic moss oils, comprising reacting the starting moss oil, a concrete or preferably an absolute thereof, with an aldehyde reducing agent, in an organic solvent medium. 2. A process according to claim 1, wherein the reagent is an alkali metal hydride, an optionally substituted complex metal or ammonium hydride, preferably an alkali metal borohydride, e.g. preferably lithium or sodium borohydride. 3. A process according to claim lor 2, wherein the solvent used is an optionally halogenated, aliphatic or aromatic hydrocarbon, an ester, an alcohol, an ether or a mixture of such solvents. 4. A process according to any one of the claims 1 to 3, wherein the solvent used is substantially water insoluble, e.g. a mixture of a aliphatic hydrocarbon, preferably hexane or cyclohexane, with a alkane carboxylic acid ester, preferably ethyl acetate or isopropyl acetate or an aliphatic ether, preferably t-butyl methyl ether. 5. A process according to any one of the claims 1 to 3, wherein the solvent used is water soluble, preferably an alkanol, e.g. ethanol or a cyclic ether, e.g. tetrahydrofuran. 6. A process according to claim 5, wherein the work up is carried out in a water insoluble solvent, preferably a solvent of claim 4. 7. A process according to any one of claims 1 to 6, wherein the reaction is carried out within a temperature range of ca. 20°-80°C, preferably at ca. 20°-30°C if lithium borohydride is used and preferably at ca. 60°-80°C if sodium borohydride is used. 8. Hypoallergenic moss oils, whenever prepared according to a process as claimed in any one of claims 1 to 7 or by an obvious chemical equivalent thereof. ***

Inventors: EHRET CHARLES, PETRZILKA MARTIN, EHRET, CHARLES, PETRZILKA, MARTIN
Applicants: EHRET CHARLES, GIVAUDAN ROURE INT, PETRZILKA MARTIN, GIVAUDAN-ROURE (INTERNATIONAL) SA
Application year: 1993
Main classification: C11B9
IPC codes: A61K8, A61Q13, C11B9