COMPOUNDS FOR A CONTROLLED RELEASE OF ACTIVE MOLECULES

Abstract

The present invention relates to the field of perfumery. More particularly, it concerns compounds comprising at least one -glucuronide moiety capable of liberating a perfuming alcohol. The present invention concerns also the use of said compounds in perfumery as well as the perfuming compositions or perfumed articles, in particular deodorants or antiperspirants comprising the invention's compounds.

Claims

1. A compound of formula (I): ##STR00002## wherein: R represents an perfuming alcohol selected from the group consisting of 4-allyl-2-methoxyphenol, 3-benzyl-3-pentanol, 4-cyclohexyl-2-methylbutan-2-ol, 2-cyclohexylpropanol, decanol, 9-decenol, (2,4-dimethylcyclohex-3-enyl)methanol, (2,4-dimethylcyclohexyl)methanol, 2-(1,1-dimethylethyl)-4-methylcyclohexanol, 2,6-dimethylheptan-2-ol, 3,7-dimethyl-7-hydroxyoctanal, 2,5-dimethyl-2-indanmethanol, 3,7-dimethyl-1,6-nonadien-3-ol, 6,8-dimethylnonan-2-ol, 4,8-dimethyl-7-nonen-2-ol, (E)-3,7-dimethyl-2,6-octadienol, (Z)-3,7-dimethyl-2,6-octadienol, 3,7-dimethyl-3,6-octadienol, 3,7-dimethyl-1,6-octadien-3-ol, 3,7-dimethyloctane-1,7-diol, 3,7-dimethyloctanol, 2,6-dimethyloctan-2-ol, 3,7-dimethyloctan-3-ol,3,7-dimethylocten-3-ol, 3,7-dimethyloct-6-enol, 3,7-dimethyloct-7-enol, 2,6-dimethyloct-7-en-2-ol, (E)-3,3-dimethyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-4-penten-2-ol, dodecanol, 1,8-epoxy-p-menthane, 2-ethoxy-5-(1-propenyl)phenol, 2-ethyl-1-hexanol, ethyl 3-hydroxy hexanoate, 4-ethyl-2-methoxyphenol, 6-ethyl-3-methyl-5-octenol, 5-ethylnonan-2-ol, 2-ethyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enol, 1-heptanol, hexanol, hexan-2-ol, 3-hexenol, 4-hexenol, 3-hydroxybutan-2-one, 4-hydroxy-3-ethoxybenzaldehyde, 4-hydroxy-3-methoxybenzaldehyde, 4-(4-hydroxy-3-methoxyphenyl)butan-2-one, 2-(hydroxymethyl)nonan-2-one, 4-(4-hydroxy-1-phenyl)butan-2-one, 1-(N-indolyl)-3,7-dimethyloctane-1,7-diol, 4-isopropyl-1-benzenemethanol, 4-isopropylcyclohexanol, 1-(4-isopropyl-1-cyclohexyl)ethanol, (4-isopropyl-1-cyclohexyl)methanol, 2-isopropyl-5-methylphenol, 5-isopropyl-2-methylphenol, (4-i sopropylphenyl)methanol, 7-p-menthanol, p-menthan-3-ol, p-menthan-8-ol, p-menthen-4-ol, p-menthen-8-ol, p-menth-8-enol, p-menth-8-en-2-ol, p-menth-8-en-3-ol, 4-methoxy-1-benzenmethanol, 7-methoxy-3,7-dimethyloctan-2-ol, 2-methoxy-4-methylphenol, 2-methoxyphenol, 2-methoxy-2-phenylethanol, (4-methoxyphenyl)methanol, 2-methoxy-4-(1-propenyl)phenol, 2-methoxy-4-propyl-1-cyclohexanol, 2-methoxy-4-propylphenol, 2-methoxy-4-vinylphenol, 3-(4-methylcyclohex-3-enyl)butanol, 4-methyl-3-decenol, 4-methyl-3-decen-5-ol, 4-(1-methylethyl)cyclohexylmethanol, 4-methylphenol, 2-methyl-4-phenylbutan-2-ol, 3-methyl-4-phenylbutan-2-ol, 1-(4-methylphenyl)ethanol, 2-(2-methylphenyl)ethanol, 2-methyl-4-phenylpentanol, 2-methyl-5-phenylpentanol, 3-methyl-5-phenylpentanol, 4-methyl-1-phenylpentan-2-ol, 2-methyl-1-phenylpropan-2-ol, 2-(4-methylphenyl)propan-2-ol, 3-methyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-4-penten-2-ol, 2-(2-methylpropyl)-4-hydroxy-4-methyl-tetrahydropyrane, 2-methyl-4-(2,3,3-trimethyl-2-cyclopenten-1-yl)-2-butenol, 3-methyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)pentan-2-ol, 2-methyl-4-(2,2,3-trimethyl-3-cyclopenten-1-yl)pent-4-enol, 3-methyl-5-(2,2,3-trimethylcyclopentyl-3-enyl)pent-4-en-2-ol, 2,6-nonadienol, 1-nonanol, 6-nonenol, 1,2,3,4,4a,5,8,8a-octahydro-2,2,6,8-tetramethyl-1-naphthalenol, octahydro-2,5,5-trimethyl-2-naphthalenol, octanol, octan-2-ol, octan-3-ol, 1-octen-3-ol, 3,4,5,6,6-pentamethylheptan-2-ol, 2-pentyl-1-cyclopentanol, perhydro-4,8a-dimethyl-4a-naphthalenol, 2-phenoxyethanol, 4-phenylbutan-2-ol, 4-phenyl-3-buten-2-ol, 1-phenylethanol, 2-phenylethanol, 1-phenylhexan-2-ol, phenylmethanol, 1-phenylpentan-2-ol, 2-phenylpropanol, 2-phenylpropanol, 3-phenylpropanol, 1-phenylpropan-2-ol, 3-phenyl-2-propenol, 2-tert-butylcyclohexanol, 4-tert-butylcyclohexanol, 1-(2-tert-butyl-cyclohexyloxy)butan-2-ol, 2-tert-butyl-4-methyl-1-cyclohexanol, tetrahydro-2-isobutyl-4-methyl(2H)pyran-4-ol, 2-(tetrahydro-5-methyl-5-vinyl-2-furyl)propan-2-ol, 1-(2,2,3,6-tetramethylcyclohex-1-yl)hexan-3-ol, 2,4,6,8-tetramethylnonan-1-ol, 3,6,7-tetramethylnonan-1-ol, 2,6,10,10-tetramethyl-1-oxaspiro[4.5]decan-6-ol, 2,6,6,8-tetramethyltricyclo[5.3.1.0(1,5)]undecan-8-ol, (+)-(1R,2R)-1,3,3-trimethylbicyclo[2.2.1]heptan-2endo-ol, (+)-(1R,2S)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-01, 2,6,6-trimethylbicyclo[3.1.1]heptan-3-ol, 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexanol, 4-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexanol, 3,3,5-trimethylcyclohexanol, 4-(2,6,6-trimethyl-2-cyclohexen-1-yl)butan-2-ol, 4-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3-buten-2-ol, (E)-4-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3-buten-2-ol, (2,4,6-trimethylcyclohex-3-enyl)methanol, 1-(2,2,6-trimethyl-1-cyclohexyl)hexan-3-ol, 5-(2,2,3-trimethyl-3-cyclopentenyl)-3-methylpentan-2-ol, 4,7,9-trimethyldecan-2-ol, 4,6,8-trimethyldecan-2-ol, 3,8,9-trimethyldecan-2-ol, 3,7,11-trimethyl-1,6,10-dodecatrien-3-ol, 3,3,5-trimethylhexanol, undecanol, undecan-2-ol and 10-undecenol, provided that phenethyl -D-glucopyranosidiuronic acid; -D-glucopyranosiduronic acid 4-(3-oxobutyl)phenyl; glucopyranosiduronic acid 1-menthyl; -D-glucopyranosiduronic acid 4-formyl-2-methoxyphenyl; and -D-glucopyranosiduronic acid 2-methoxy-4-(2-propen-1-yl)phenyl are excluded.

2. The compound of claim 1, wherein R is selected from the group consisting of (E)-3,7-dimethyl-2,6-octadienol, 3,7-dimethyl-1,6-octadien-3-ol, 3,7-dimethyloct-6-enol, 2,6-dimethyloct-7-en-2-ol, 3-hexenol, 2-phenylethanol, (E)-3,3-dimethyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-4-penten-2-ol, 7-p-menthanol, 3-methyl-5-phenylpentanol, 2,6-nonadienol, 1-phenylhexan-2-ol, tetrahydro-2-isobutyl-4-methyl (2H)pyran-4-ol, 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexanol, 3-(4-methylcyclohex-3-enyl)butanol, 4-cyclohexyl-2-methylbutan-2-ol, 4-methyl-3-decen-5-ol, 4-allyl-2-methoxyphenol, 4-(4-hydroxy-1-phenyl)butan-2-one, p-menthan-3-ol, 4-hydroxy-3-methoxybenzaldehyde, 3,7-dimethyl-1,6-nonadien-3-ol, 3,7-dimethyloctanol, 1-(2,2,6-trimethyl-1-cyclohexyl)hexan-3-ol, 2-ethyl-4-[2,2,3-trimethylcyclopent-3-enyl)but-2-enol, (E)-3,3-dimethyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-4-penten-2-ol and 2-(1,1-dimethylethyl)-4-methylcyclohexanol.

3. The compound of claim 2, wherein R is selected from the group consisting of 4-allyl-2-methoxyphenol, 4-(4-hydroxy-1-phenyl)butan-2-one, p-menthan-3-ol, 4-hydroxy-3-methoxybenzaldehyde, 2-phenylethanol, 3,7-dimethyloct-6-enol and 2,6-dimethyloct-7-en-2-ol.

4. A perfuming composition comprising: i) as perfuming ingredient, at least one compound of formula (I) as defined in claim 1; and ii) at least one ingredient selected from the group consisting of a perfumery carrier and at least a perfuming co-ingredient.

5. A perfuming composition according to claim 4, further comprising at least one perfumery adjuvant.

6. A perfuming consumer product which comprises i) as perfuming ingredient, at least one compound of formula (I) as defined in claim 1; and ii) a perfumery consumer base.

7. A perfuming consumer product according to claim 6, wherein the perfumery consumer base is a liquid or solid detergent, a fabric softener, a fabric refresher, a bleach, a deodorant or antiperspirant, a perfumed soap, shower or bath mousse, a hygiene product, an air freshener, a ready to use powdered air freshener, a dish detergent or hard-surface detergent.

8. A perfuming consumer product according to claim 6, wherein the perfumery consumer base is a deodorant or antiperspirant.

Description

EXAMPLE 1

[0048] Synthesis of Compounds of Formula (I)

[0049] Experimental:

[0050] UPLC-MS. Analyses were performed on a Waters (Baden-Dttwil, Switzerland) Acquity system coupled to a mass spectrometer. The separations were performed on an Acquity BEH-C18 column (2.1 mm i.d.100 mm, 1.7 m). The elution solvents were CH.sub.3CN containing 0.1% of formic acid (solvent B) and water containing 0.1% of formic acid (solvent A). The gradient profile started at 10% of B, which was held for 0.5 min, increased to 90% of B in 7.5 min. The flow rate was 0.3 mL/min. The mass spectrometer was a Thermo Finnigan LXQ with an ESI source (HESI-II) operated in negative mode. The spray voltage was 4.0 kV, the vaporizer temperature was 250 C., the capillary temperature was 350 C. The sheath gas was nitrogen at a flow rate of 50 (Finnigan arbitrary units). The auxiliary gas was also nitrogen at a flow rate of 20 (Finnigan arbitrary units). The events were full scan [80-800] which was linked with a data dependent MS/MS of the 1.sup.st most intense ion from scan event 1, collision energy 35 V.

[0051] Nuclear Magnetic Resonance (NMR) Spectra. 1H- and 13C-NMR spectra were recorded on a Bruker AV-500 (Zurich, Switzerland) spectrometer at 500.13 and 125.76 MHz or AV-600 spectrometer at 600.34 and 150.96 MHz respectively. The chemical shifts are referenced to TMS as internal standard. The assignments have been confirmed by 2D NMR (HSQC, HMBC and COSY).

(i) Synthesis of 2-phenylethyl -D-glucopyranosiduronic acid (phenethylol glucuronide)

[0052] -Glucopyranosides were prepared according to standard procedures, starting from commercially available pentaacetylbromo-D-glucose as the glycosyl donor and the free alcohols as the respective glycosyl acceptors.

[0053] Phenethylol glucoside (973 mg, 3.42 mmol), PIPO (12 mg), KBr (41 mg, 0.34 mmol) and tetrabutylammonium chloride (57 mg, 0.2 mmol) were mixed in methylene chloride (11 mL) and NaHCO.sub.3 sat. (6.6 mL). After cooling at 0 C., a mixture of NaOCl 10% (8.6 mL) and NaCl sat (15 mL) was added dropwise and the reaction was stirred for 1.5 h at room temperature. Methylene chloride was distilled off and the mixture was acidified with concentrated HCl to pH 2-2.5. The product was purified by flash chromatography over RP-18 (water 9:ethanol 1) afforded phenethylol glucuronide (581 mg) in 57% yield. MW: 298.3 (C.sub.14H.sub.18O.sub.7).

[0054] UPLC-MS H-ESI.sup. (r.t. 3.14 min)

[0055] MS-MS: M-H=296.9, dependant scan: MS.sup.n: 112.6 (100% relative intensity), 156.7 (42), 174.7 (30%), 86.6 (27%), 236.9 (26%), 192.8 (25%).

[0056] .sup.13C NMR (125 MHz, MeOD): 37.2 (t), 72.0 (t), 73.7 (d), 74.9 (d), 76.2 (d), 77.8 (d), 104.4 (d), 127.2 (d), 129.4 (d), 130.1 (d), 140.0 (s), 177.4 (s).

(ii) Synthesis of (4-hydroxyphenyl)-2-butanone--D-glucopyranosiduronic acid (raspberry ketone glucuronide)

[0057] Raspberry ketone glucoside (1000 mg, 3.06 mmol), PIPO (12 mg), KBr (36.5 mg, 0.306 mmol) and tetrabutylammonium chloride (51.1 mg, 0.184 mmol) were mixed in methylene chloride (11 mL) and NaHCO3 sat. (6.6 mL). After cooling at 0 C., a mixture of NaOCl 10% (7.72 mL) and NaCl sat (15 mL) was added dropwise and the reaction was stirred for 2.5 h at R.T. Methylene chloride was distilled off and the mixture was acidified with concentrated HCl to pH 2-2.5. The product was purified by flash chromatography on SiO2 (ethyl acetate 6:methanol 4) afforded raspberry ketone glucuronide 10 (228 mg) in 22% yield.

[0058] MW: 340.3 (C.sub.16H.sub.20O.sub.8).

[0059] UPLC-MS H-ESI.sup. (r.t. 2.67 min)

[0060] MS-MS: M-H=339, dependant scan: MS.sup.n: 112.6 (100% relative intensity), 174.7 (66%), 86.6 (27%), 162.8 (7%).

[0061] .sup.13C NMR (125 MHz, MeOD): 30.0 (t), 30.0 (q), 46.0 (t), 73.6 (d), 74.7 (d), 76.4 (d), 77.7 (d), 102.6 (d), 118.1 (d), 130.3 (d), 136.6 (s), 157.6 (s), 177.0 (s), 211.1 (s).

(iii) Synthesis of methyl 2,3,4-tri-O-acetyl-1-O-(2,2,2-trichloroethanimidoyl)-D-glucopyranuronate

[0062] Methyl 2,3,4-tri-O-acetyl-D-glucopyranuronate (1.5 g, 4.49 mmol) was dissolved in CH.sub.2Cl.sub.2 (18 mL) followed by trichloroacetonitrile (4.34 g, 30.1 mmol) and K.sub.2CO.sub.3 (4.15 g, 30.1 mmol). The mixture was stirred 15 hours at 21 C. The crude mixture was loaded on a SiO.sub.2 column containing (4 cm high, about 6 g SiO.sub.2), eluted with Et.sub.2O (100 mL) and concentrated on rotary evaporator to give 1.79 g ,100% -isomer, yield 83%.

[0063] .sup.13C NMR (125 MHz, CDCl.sub.3): 20.4 (q), 20.5 (q), 20.7 (q), 53.0 (q), 69.0 (d), 69.1 (d), 69.5 (d), 70.5 (d), 90.5 (s), 92.6 (d), 160.6 (s), 167.2 (s), 169.5 (s), 169.7 (s), 169.8 (s).

(iv) Synthesis of 3,7-dimethyloct-6-en-1-yl -D-glucopyranosiduronic acid (citronellol glucuronide)

[0064] Methyl 2,3,4-tri-O-acetyl-1-O-(2,2,2-trichloroethanimidoyl)-D-glucopyranuronate (190 mg, 0.397 mmol) and citronellol (62 mg, 0.397 mmol) were stirred 1 hour in CH.sub.2Cl.sub.2 (7 mL), in presence of molecular sieves 4A. The reaction was cooled down at 15 C., 1 hour. Then BF.sub.3Et.sub.2O (25.4 mg, 0.179 mmol) was added. The reaction was monitored by TLC (thin layer chromatography, SiO.sub.2). After 30 min. the whole imidate was consumed. EtOAc was added (30 mL) and the organic phase was washed with NaHCO.sub.3 aqueous saturated solution, then with brine. The solvent was removed on rotary evaporator under vacuum. The crude product was purified by flash chromatography on SiO.sub.2 (13 g), elution with pentane/Et.sub.2O 2/3 and we obtained 85 mg (yield 45%) of the protected sugar, 100% isomer.

[0065] The purified product (74 mg, 0.157 mmol) was diluted in MeOH (2 mL). At 0 C. aqueous NaOH 5 M (0.16 mL) was added. The product was stirred one hour while coming back at room temperature. The MeOH was removed under vacuum, the residue was then loaded in solution in water on a chromatography column (Si=2-RP18, 2.5 g). The column was eluted with a gradient starting at 100% water and the product was in fractions eluted with a mixture of water/EtOH 4/1. We obtained the title compounds as a mixture of two diastereoisomers, pure with a stereochemistry: 28 mg (yield 54%).

[0066] MW: 332.4 (C.sub.16H.sub.28O.sub.7).

[0067] UPLC-MS H-ESI.sup. (r.t. 4.93 min)

[0068] MS-MS: M-H=331.2 dependant scan: MS.sup.n: 112.6 (100% relative intensity), 156.7 (58%), 128.7 (39%), 271.0 (36%).

[0069] .sup.13C NMR (151 MHz, MeOD) (* stereoisomer): 17.6 (q); 20.0 (q); 25.9 (q); 26.5 (t); 30.7 (t); 30.7 (t); 73.6 (d); 69.3, 69.4* (t); 73.7, 75.0* (d); 75.0, 76.1* (d); 77.9 (d); 104.3, 104.4* (d); 125.9, 126.0 (s); 177.0 (s).

(v) Synthesis of 2,6-dimethyloct-7-en-2-yl -D-glucopyranosiduronic acid (dihydromyrcenol glucuronide)

[0070] Methyl 2,3,4-tri-O-acetyl-1-O-(2,2,2-trichloroethanimidoyl)-D-glucopyranuronate (190 mg, 0.397 mmol) and dihydromyrcenol (62 mg, 0.397 mmol) were stirred 1 hour in presence of molecular sieves 4A. The reaction was cooled down at 15 C. (1 hour). Then BF.sub.3Et.sub.2O (25.4 mg, 0.179 mmol) was added. The reaction was monitored by TLC (thin layer chromatography, SiO.sub.2). After 30 min. the whole imidate was consumed. EtOAc was added (30 mL) and the organic phase was washed with NaHCO.sub.3 aqueous saturated solution, then with brine. The solvent was removed on rotary evaporator under vacuum. The crude product was purified by flash chromatography on SiO.sub.2 (13 g), elution with pentane/Et.sub.2O 2/3 and we obtained 73 mg (yield 39%) of the protected sugar, 100% isomer.

[0071] The purified product (60 mg, 0.127 mmol) was diluted in MeOH (2 mL). At 0 C. aqueous NaOH 5 M (0.13 mL) was added. The product was stirred one hour while coming back at room temperature. The MeOH was removed under vacuum, the residue was then loaded in solution in water on a chromatography column (Si=2-RP18, 2.5 g). The column was eluted with a gradient starting at 100% water and the product was in fractions eluted with a mixture of water/EtOH 4/1. We obtained the title compounds as a mixture of two diastereoisomers, pure with a stereochemistry: 21 mg (yield 50%).

[0072] MW: 332.4 (C.sub.16H.sub.28O.sub.7).

[0073] UPLC-MS H-ESI.sup. (r.t. 4.64 min)

[0074] MS-MS: M-H=331.2 dependant scan: MS.sup.n: 112.6 (100% relative intensity), 156.7 (47%), 128.7 (29%), 174.8 (7%).

[0075] .sup.13C NMR (151 MHz, MeOD) (* stereoisomer): 20.1 (q); 23.0 (t); 26.8 (q), 38.5 (t); 39.1 (d); 48.6, 48.7* (t); 73.8 (t); 75.0 (d); 76.5 (t); 78.1 (d); 79.2 (s); 98.5 (d); 112.9 (t); 146.1 (d); 177.0 (s).

EXAMPLE 2

[0076] Performance of Compounds of Formula (I) as Pro-Perfume

[0077] The liberation of perfuming alcohols from the present invention's compounds of formula (I) was tested in vitro during incubation with defined skin bacteria strains.

[0078] Incubations

[0079] 1. Preparation of Bacterial Solution

[0080] a) Media: Trypticase Soja Agar (TSA) solid medium was purchased from Beckton Dickinson, Pont de Claix, France. Schaedler agar containing 5% sheep blood (SCH) solid medium was purchased from Biomerieux, Lyon, France.

[0081] b) Reagent: sterile Sodium Chloride solution at 0.9% (NaCl 0.9%) was prepared in the laboratory. Sodium Chloride powder was purchased from Carlo Erba, Val de Reuil, France.

[0082] c) Bacterial solutions: Two underarm bacterial isolates, Propionibacterium acnes ATCC 6919 and Staphylococcus warneri DSM 20316 (identified by 16S rDNA sequencing by Deutsche Sammlung von Mikroorganismen and ZellKulturen DSMZ, Germany), were cultivated on SCH agar at 37 C. during 72 h in anaerobic conditions and on TSA at 37 C. during 24 h, respectively.

[0083] One isolated colony of P. acnes from the pure primary culture on SCH agar was inoculated on a new SCH agar and incubated at 37 C., 72 h in anaerobic conditions. After incubation, the whole culture was dissociated by shaking in 20 ml of NaCl 0.9% then centrifuged 10 min at 5000 rpm at room temperature. The pellet was washed in NaCl 0.9% and concentrated two times in NaCl 0.9%.

[0084] Similarly, one colony of S. warneri from the pure primary culture on TSA was inoculated on a new TSA and incubated for 24 h at 37 C. After incubation, the whole culture was dissociated by shaking in 20 ml of NaCl 0.9% then centrifuged 15 min at 5000 rpm, 4 C. The pellet was washed in NaCl 0.9% and concentrated two times in NaCl 0.9%.

[0085] 2. General Method for Incubation

[0086] A pro-perfume glucuronides mixture containing menthol glucuronide (6 mg/kg), phenethylol -glucuronide (10.75 mg/kg) and raspberry ketone -glucuronide (13 mg/kg) in water was incubated with either: [0087] 500 L of the previously described bacterial solution [0088] 10 L of commercial enzymes -glucuronidase/arylsulfatase from Helix pomatia (RocheDiagnostic, Indianapolis, USA) and 490 L of NaCl 0.9% as a positive control [0089] 500 l of NaCl 0.9% as a negative controlfor the incubation with bacteria.

[0090] Each solution was prepared in duplicate (in glass vials).

[0091] The vials were placed in a heating block at 37 C. under gentle stirring from 1 to 7 days depending on the bacteria solutions. At the end of the incubation time, vials were cooled down to room temperature, 150 L was mixed with 50 L of LC-MS solvent B, filtered through acrodisc and 1 L was injected on LC-MS (peak areas considered were measured in SIM mode according to their respective molecular weights). Comparisons were made between glucuronides (HESI) or glucosides (HESI+) areas in incubated solutions and glucuronides (HESI) or glucosides (HESI+) areas in the blanks (fixed as 100%).

[0092] 3. Analysis of the Volatiles After Incubation

[0093] General Method

[0094] In a 2 mL glass vial, a solution of Internal Standard (IS) (Eicosane) in CH.sub.2Cl.sub.2 (200 L, 0.2 g) and 200 L of CH.sub.2Cl.sub.2 were added to the solution of incubation, well mixed then centrifuged at 3260 g for 3 min. The lower layer was removed with a pipette, dried on Na.sub.2SO.sub.4 filtered on cotton and concentrated under on argon flow to about 20 L. 1 L was injected onto the GC-MS. The concentration was determined from the GC-MS peak areas of a-androstenol (m/z 274) compare to the a-androstanol (m/z 276) and corrected by the response factor value (0.38450.03).

TABLE-US-00001 TABLE 1 Glucuronide-conjugates remaining after incubation with Propionibacterium acnes (in percentage) Released raspberry corresponding alcohol ketone 2-phenylethanol p-menthan-3-ol After 1 day No transformation After 5 days incubation 1 61 94 5 incubation 2 58 85 9 incubation 3 57 84 7 Average 58 88 7

TABLE-US-00002 TABLE 2 Glucuronide-conjugates remaining after incubation with S. warneri (in percentage). Released raspberry corresponding alcohol ketone 2-phenylethanol p-menthan-3-ol After 1 day incubation 1 0 0 1 incubation 2 0 0 0 Average 0 0 0

[0095] The data show that in presence of Propionibacterium acnes, while no transformation is observed after 1 day, alcohols are released after 5 days incubation. Turning to Staphylococcus warneri, there is a complete transformation and no remaining starting material after only 1 day, which demonstrates a quicker release of the corresponding alcohols.

TABLE-US-00003 TABLE 3 Glucuronide-conjugates remaining after incubation with S. warneri (in percentage). Released corresponding alcohol vanilline eugenol menthol After 1 day incubation 1 84 130 72 incubation 2 96 120 77 Average 100 100 74 After 5 day incubation 1 0 0 36 incubation 2 0 0 31 Average 0 0 34

[0096] The results show an efficient release of alcohols from glucoside conjugates after 5 day-incubation with S. warneri.