USOO59 19991A United States Patent (19) 11 Patent Number: 5,919,991 Subbiah (45) Date of Patent: Jul. 6, 1999

54). SOLID PHASE EXTRACTION OF R.M.E. Richards et al., Preservation of fluorescein Solutions PHENETHYLALCOHOL against contamination with Pseudomonas aeruginosa, J. Pharm. Pharmac. 21, 681–86 (1969). 76 Inventor: Ven Subbiah, 105 Bella Vista Dr., G. Fronza et al., Natural Abundance off H Nuclear Mag Edenton, N.C. 27932 netic Resonance Study of the Origin of 2-Phenylethanol and 2-Phenethyl Acetate, J. Agric. Food Chem. 43, 439-443 21 Appl. No.: 08/674,918 (1995). 22 Filed: Jul. 3, 1996 E. Albertazzi et al., Biogeneration of 2-Phenylethanol and 51) Int. Cl...... C07C 29/74; C12P 7/22 2-Phenylethylacetate Important Aroma Components, Bio 52 U.S. Cl...... 568/810; 435/156 technology Letters 16(5), 491–496 (1994). 58 Field of Search ...... 568/810; 11/702, W.G. Iverson, Ethyl Acetate Extraction of : Quantitative 11/171; 435/156 Determination of Additional Fermentation By-Products, J. Am. Soc. Brew. Chem. 52(3), 91-95 (1994). 56) References Cited F.R. Gabler, Principles of Tangential Flow Filtration: Appli PUBLICATIONS cations to Biological Processing, in Filtration in the Phar maceutical Industry (T.H. Meltzer, ed., Marcel Decker Inc., Chilla et al., Automated Online Solid-Phase Extra New York (1987). tion-High-Performance Liquid Chromatagraphy-Diode array Detection of Phenolic Compounds in Sherry Wine, J. Primary Examiner Michael L. Shippen Chrommatogr., A 4th International Symposium on Hyphen Attorney, Agent, or Firm Myers Bigel Sibley & Sajovec ated Techniques in Chromatography and Hyphen, 1996. Fabre et al., Extraction of 2-phenylethyl by Tech 57 ABSTRACT niques Such as adsorption, inclusion, Supercritical CO2, liquid–liquid and membrane Separations., Perfum. Flavor., A method for isolating phenethyl alcohol in high yield is pp. 27–28, 30–32, 34, 36–40, 1996. provided by fermenting L- within a fermen S. Arctander, Phenylethyl Alcohol, Perfume and Flavor tation bath to provide phenethyl alcohol. The fermented Chemicals vol. 2, monograph 2513 (Montclair, N.J., 1969). phenethyl alcohol is then contacted with an ion-exchange R.M.E. Richards and R.J. McBride, The preservation of resin to extract the phenethyl alcohol. ophthalmic Solutions with antibacterial combinations, J. Pharm. Pharmac. 24, 145-48 (1972). 35 Claims, 1 Drawing Sheet U.S. Patent Jul. 6, 1999 5,919,991

FIG. 1 10 12

L-phenylalanine Fermentation Media 20

Add to fermentation bath 22 30 Inoculation with yeast Fermentation Conditions 40

50 pH to 7.0 with base

Separate yeast cells from Solution 60

Separate large molecular weight proteins from solution

Contact Solution with ion-exchange resin; adsorb phenethyl alcohol 80 Aqueous liquid wash

90

Solvent elution

1 OO / Collect phenethyl alcohol 5,919,991 1 2 SOLID PHASE EXTRACTION OF In particular, a L-phenylalanine Solution is prepared. The PHENETHYLALCOHOL L-phenylalanine Solution is fermented within a fermentation bath, preferably including at least one Strain of yeast, to The present invention relates to methods for isolating provide phenethyl alcohol. The fermented phenethyl alcohol phenethyl alcohol in high yield, and more particularly to Solution is separated to provide a fermented phenethyl methods of extracting phenethyl alcohol from a fermented alcohol Solution essentially devoid of large molecular phenylalanine Solution. weight molecules, and is preferably Separated to provide a fermented phenethyl alcohol Solution essentially free of BACKGROUND OF THE INVENTION yeast cells. The Separated, fermented phenethyl alcohol The production of aroma compounds represents an impor Solution is contacted with an ion-exchange resin to adsorb tant tool in the production of natural flavors for food and phenethyl alcohol thereon. After the phenethyl alcohol is beverage industries. E. AlbertaZZi, et al., Biotech. Lett. 16, adsorbed onto the resin, the resin is eluted with a Solvent, 491-6 (1994). One of the most commonly used aroma and the phenethyl alcohol is collected. compounds is phenethyl alcohol. Phenethyl alcohol (e.g., The present invention provides advantages over prior phenethanol, 2-phenylethanol, or benzeneethanol) is natu 15 methods of isolating phenethyl alcohol. The rate of recovery rally present in many essential oils, and has a rose-like/ is higher when compared to conventional liquid phase honey floral odor. Phenethyl alcohol is chemically synthe extraction techniques, while providing phenethyl alcohol sized or extracted in commercial applications as a flavoring with improved purity. Additionally, the present method or fragrance for consumer goods, Such as perfumes and avoids cumberSome extraction and Separation, particularly food. with , thus also avoiding the common problem of AS a flavorant, phenethyl alcohol is useful in producing a emulsion formation during extraction and reducing the need broad range of desirable flavors, especially imitation butter, for high Volumes of unwieldy and potentially hazardous Strawberry, raspberry, caramel, honey, melon and other fruit organic Solvents. complexes. S. Arctander, Perfume and Flavor Chemicals Vol. 2, 2513 (1969). Phenethyl alcohol is also one of the 25 BRIEF DESCRIPTION OF THE DRAWING most widely used of all perfume chemicals, and is particu FIG. 1 is a Schematic diagram of process Steps represen larly desired as a component in perfumes that are charac tative of one embodiment of the present invention. terized as floral, balsamic, Oriental, mossy, herbaceous, or “modern aldehydic.” Id. In addition to providing its own DETAILED DESCRIPTION OF THE distinctive aroma, phenethyl alcohol may be also blended INVENTION with other aroma compounds (e.g., rose , lily Referring to FIG. 1, L-phenylalanine is provided as a alcohols, and muguet alcohols) to produce floral scents as solution 10, wherein the solution comprises fermentation well as lime and Spice blends. Id. Additionally, phenethyl media 12 containing L-phenylalanine. Alternatively, the alcohol is used as an antimicrobial and as a Solution pre fermentation media is combined with a Solution of servative. See R. M. E. Richards and R. J. McBride, J. 35 L-phenylalanine. The combined fermentation Solution is Pharm. Pharmac. 24, 145-148 (1972); ibid, 21, 681–686 added 20 to a fermentation bath, which fermentation bath is (1969). then inoculated with at least one species of yeast 22. The pH Phenethyl alcohol occurs naturally in alcoholic fermented of the fermentation solution is adjusted 40 to a pH of about foods in amounts of 10-100 ppm. M. E. Kieser, et al., Nature 7.0 with a base (e.g., NaOH) after which the fermentation 204, 887 (1964). This amount, while affecting the quality of 40 Solution is allowed to ferment under fermentation conditions alcoholic beverages, is not Sufficient as a Source for the large-scale production of the natural aroma component. It is 30, providing phenethyl alcohol. therefore of interest to develop methods of producing and The fermented phenethyl alcohol mixture, namely the isolating phenethyl alcohol in large amounts, and of high phenylalanine and fermentation media mixed with yeast purity. 45 cells, is then separated 50 to provide a fermented phenethyl alcohol Solution essentially devoid of yeast cells. The essen Phenethyl alcohol may be produced by fermenting solu tially yeast-free, fermented phenethyl alcohol mixture is tions containing L-phenylalanine under appropriate fermen then further separated 60 to provide a fermented phenethyl tation conditions. See Albertazzi, et al., Supra at 492. Con alcohol Solution essentially devoid of large molecular ventional methods of isolating phenethyl alcohol from Such weight molecules. AS used herein, a large-molecular weight fermented Solutions generally involve liquid-liquid Solvent 50 molecule refers to a molecule that is larger than about 10 extraction utilizing high Volumes of organic Solvents Such as kilodaltons (10 K), and is preferably larger than about 20 butanol. One disadvantage of the Solvent extraction tech kilodaltons. Examples of Such molecules include nique is the formation of emulsions during extraction, polysaccharides, high molecular weight peptides and requiring the use of exceSS Solvent and longer times for proteins, and the like. phase Separation. Additionally, the Solvent extraction tech 55 niques are often cumberSome, and do not provide an optimal The phenethyl alcohol solution devoid of large molecular yield of final product. weight molecules is then contacted with an ion-exchange It would be highly desirable to provide a practical and leSS resin 70 to adsorb the phenethyl alcohol to the resin. The hazardous method of isolating phenethyl alcohol whereby a ion-exchange resin is then washed 80 at least once with an 60 acqueous liquid, and is then further eluted 90 with a solvent. high yield of final product is afforded, and the need for large Finally, the phenethyl alcohol is collected 100 from the Volumes of organic Solvent is obviated. elution volume. SUMMARY OF THE INVENTION Prior to extraction, the ion-exchange resin may be acti The present invention relates to a method of isolating Vated by Washing the resin with an aqueous liquid, and then phenethyl alcohol from an L-phenylalanine fermentation 65 with one or more Solvents. Solution using a Solid-phase extraction involving ion L-phenylalanine is widely commercially available in eXchange resins. powdered form. Solutions of L-phenylalanine may be pro 5,919,991 3 4 Vided by dissolving L-phenylalanine in an appropriate Sol one in the art. Ion-exchange columns are packed with Small vent. Preferably, the L-phenylalanine solution is provided by beads that carry either a positive or a negative charge, Such dissolving the L-phenylalanine using a liquid having an that compounds are fractionated according to the arrange aqueous character. Such a liquid consists primarily of water, ment of charge on their Surface. In an ion-exchange column, normally greater than about 90 percent water, and can be the matrix or resin is insoluble and carries ionic charges that essentially pure water in certain circumstances. For retard molecules of the opposite charge. The Strength of the example, a Solvent having an aqueous character can be asSociation between the dissolved molecules and the ion distilled water, tap water or the like. However, a solvent having an aqueous character can include water having eXchange matrix depends on both the ionic Strength and the Substances Such as pH buffers, pH adjusters, organic and pH of the eluting solution, which may therefore be varied in inorganic Salts, or Surfactants incorporated therein. The a Systematic fashion to achieve effective Separation. Solvent can also be a co-Solvent mixture of water and minor In a preferred embodiment, the resin comprises polymeric amounts of one or more other Solvents (e.g., , beads packed onto a column. The beads may comprise ) which are miscible therewith. polymers of, e.g., Styrene, Styrene divinylbenzene, In one embodiment of the invention, the L-phenylalanine methacrylate, propylene, or derivatives and combinations Solution is provided as a fermentation media Solution con 15 thereof. In one preferred embodiment, the beads comprise taining L-phenylalanine. Alternatively, an L-phenylalanine polymers of Styrene divinylbenzene. The polymeric beads Solution may be prepared Separately from a fermentation may be as large as 600 um in diameter, preferably as large media Solution, and then added to the fermentation media as 400 um in diameter, and more preferably as large as 250 Solution. The fermentation media Solution may contain tim in diameter. In a more preferred embodiment, the resin components essential to the proper fermentation of used is Diaion HP 20 (Mitsubishi Kasei America, Inc., White L-phenylalanine, which components will be apparent to one Plains, N.Y.), a macroporous styrenic-based polymeric bead skilled in the art. In a preferred embodiment of the invention, type resin. the fermentation media contains dextrose monohydrate, After the phenethyl alcohol has been allowed to adsorb KHPO, and KHPO, and is adjusted prior to fermentation onto the ion-exchange resin, the resin may be washed with to a pH of about 6.5 with a base (e.g., NaOH). 25 deionized water. The phenethyl alcohol may then be eluted The L-phenylalanine Solution, preferably an aqueous from the resin by repeated washing with solvents. Preferred L-phenylalanine Solution, is fermented in a fermentation Solvents include acetone, methanol, ethanol, with ethanol bath under fermentation conditions. The basic reaction and methanol being particularly preferred. involves the decomposition of the amino acid into an alcohol The phenethyl alcohol isolated by any embodiment of the and carbon dioxide. The fermentation reaction is induced by present invention is useful as a fragrance or as a flavoring in a living organism or enzyme, Specifically microorganisms foods and beverages, and primarily in non-alcoholic bever occurring in one-celled plants (i.e. yeast, mold or fungi). ageS. Preferably, the L-phenylalanine solution is fermented in a The following examples are provided in order to further fermentation bath including at least one Strain of yeast. 35 illustrate various embodiments of the invention and are not Preferred yeasts include Strains of the genera Hansenula to be construed as limiting the Scope thereof. In the follow (e.g., H. anomala), Pichia (e.g., P. pastoris, P. etchelsii), ing examples, lbS. means pounds, um means micrometers, Kloeckera (e.g., K. Saturnus), Klyveromyces (e.g., K. cm means centimeters, it means microliters, mL means fragilis), and Saccharomyces (e.g., S. distaticus, S. millilters, L means liters, ppm means parts per million, C. delbruecki, and S. cerevisiae), Sometimes referred to as 40 means temperature in degrees Centigrade, and dH2O means “brewer's yeast.” Most commercially available yeast such as deionized water. Fleischmann's Yeast, Red Star yeast, Ale Beer Yeast, Geor die yeast, Montrachet yeast, Laaglander Irish ale yeast, etc. EXAMPLE 1. are mixtures of various yeast Strains. Fermentation and Filtration of Phenylalanine Typically the L-phenylalanine Solution is fermented at 45 ambient or room temperature over a period of at least one Six hundred and forty-two (642) lbs. of dextrose mono day, and up to about fifteen days or more. In the resulting hydrate is added to a fermentation bath containing 700 fermented L-phenylalanine Solution, phenethyl alcohol typi gallons of water heated to a temperature of 30° C. This cally comprises at least about 60% of the fermented solution mixture is stirred until the dextrose monohydrate is dis by weight, and preferably at least about 80% of the fer 50 solved. Forty-four (44) lbs. of L-phenylalanine (Fisher mented Solution by weight, as measured by a commercially Scientific, Raleigh, N.C.) is added to the solution, along with available vinometer. 9.9 lbs. of KHPO and 5.4 lbs of KHPO. The solution is Yeast cells are removed from the fermented solution by stirred until all Solutes have dissolved, after which NaOH any appropriate filtration or Separation techniques which (25% solution) is added until the pH of the fermentation will be readily apparent to one skilled in the art of fermen 55 Solution is 4.5. The fermentation Solution is then inoculated tation. In a preferred embodiment, yeast cells are removed with 58.4 lbs. (approximately 0.08 pounds/gallon) of Red by crossflow filtration, utilizing a filter with a pore size of Star Dry Yeast and allowed to ferment under aeration and not more than 0.2 um and preferably not more than 0.15um. agitation conditions for 24 hours. Large molecular-weight molecules are removed from the After fermentation, the pH of the fermented solution is fermented Solution by any appropriate filtration or Separa 60 adjusted to pH 7.0 with NaOH (25% solution). The solution tion techniques which will be readily apparent to one skilled is then transferred to a crossflow filtration tank and run, with in the art of protein Separation. In a preferred embodiment, pressure, through a crossflow Maxcell(R) filter of pore size large molecular-weight molecules are also removed by 0.2 um (A/G Technology Corporation, Boston, Mass.) to crossflow filtration, utilizing a filter with a pore size of not remove yeast cells. After this first filtration, the resulting more than about 10 kilodaltons (K). 65 essentially yeast-free fermented Solution is then run through Preferably, the ion-exchange resin is immobilized onto a another crossflow Maxcell filter of pore size 10 kilodaltons column, the use and Selection of which will be apparent to to remove large molecular weight molecules. 5,919,991 S 6 COMPARATIVE EXAMPLE A gel by passing the aqueous fermented Solution through the gel. Subsequently, the gel was eluted with 1 L of deionized Solvent Extraction of Phenethyl Alcohol water, followed by 1 L each of methanol and acetone. Phenethyl alcohol was isolated using conventional liquid An aliquot of each collected fraction was directly injected phase Solvent extraction techniques. Additionally, different into a gas chromatograph for quantitative phenethyl alcohol Solvents that are used in the conventional method were determination. compared, as measured by the yield of phenethyl alcohol provided when each Solvent was used to extract phenethyl alcohol. Elution fraction Quantity Phenethyl alcohol (ppm) Two liters of fermented L-phenylalanine was collected HP 20 wash-direct 500 mL. 3.0 DI water 1000 mL. O.O from a large fermenter. Two hundred mL aliquots of the Methanol 1000 mL. 3O14 fermented Solution were extracted three times with an equal Methanolfacetone 1000 mL. 60 Volume of one of the following Solvents: hexane, ethyl ether, ethyl acetate, or butanol. The organic phase of each Solvent 15 was pooled and concentrated by evaporating the Solvent to These results indicate that after adsorbing onto the ion near dryneSS. Additionally, Solvent from the trap, as well as eXchange resin, phenethyl alcohol is eluted from the resin left-over aqueous fractions were collected for quantitative with methanol in a concentration of approximately 3000 analysis. parts per million, which is a concentration of approximately Individual collected Samples were analyzed by gas chro 20 percent greater than that achieved with conventional matography (injection volume, 1 till of respective Solvent or butanol eXtraction. aqueous fractions) for quantitative estimation of concentra EXAMPLE 2 tion of phenethyl alcohol present in each fraction. The results are presented below: Activation of HP-20 Gel 25 One particular advantage of using the ion exchange resin is that the resin may be reactivated after use in order to be Sample Phenethyl alcohol (ppm) utilized repeatedly for the purpose of isolating phenethyl Original fermented broth 2376 alcohol. (no extraction) 200 ml Hexane extraction 1995 In order to activate the HP-20 gel, approximately 1.5 L of trap fraction 692 HP-20 gel was stirred in 3 L of acetone, and after settling, hexanefaqueous 833 the acetone was decanted to remove the fines. The Support 200 mL Ethyl ether extraction 2582 was transferred to a 3 L coarse fitted filter and was washed ethyl etherlaqueous 462 200 mL Ethyl acetate extraction 3715 successively with methanol (2 L), water (2 L), 30% w/v ethyl acetate trap 323 potassium hydroxide, water (4 L), 10% hydrochloric acid (2 ethyl acetate/aqueous 8.O 35 L), water (until effluent was neutral), acetone (4 L), metha 200 mL. Butanol extraction 2441 butanol trap 222 nol (4 L) and water (4 L). Using this process, the HP-20 gel butanol?water 2.5 is available for reuse repeatedly. EXAMPLE 3 These results indicate that when conventional liquid 40 phase Solvent extraction techniques are used to isolate Comparative Ethanol Washes of Resin phenethyl alcohol, ethyl acetate appears to provide the Phenethyl alcohol was isolated onto an ion eXchange resin highest concentration of phenethyl alcohol. However, the using the method described above in Comparative Example undesired formation of phenethyl acetate is a disadvantage B. However, instead of methanol, ethanol in varying con in terms of the practical use of ethyl acetate in phenethyl 45 centrations was used to elute the column in order to evaluate alcohol extraction. Furthermore, under neutral pH (7.0) the effectiveness of ethanol as a Solvent in the practice of the ethyl acetate is known to be non-reactive. present invention. These results are presented below. Butanol is an often-used Solvent in the conventional practice of phenethyl alcohol eXtraction, and these results indicate that it extracts phenethyl alcohol in a concentration 50 Elution Volume (mL) Phenethyl alcohol (ppm) of approximately 2400 parts per million (ppm). fermented broth 1OOO 2377 HP-20 wash 1OOO 1O COMPARATIVE EXAMPLEB dEH2O wash 1OOO 3 10% ethanol 500 O 55 25% ethanol 500 O Solid Phase Extraction of Phenethyl Alcohol 50% ethanol 500 O Phenethyl alcohol was isolated using one method of the 100% ethanol 500 3457 present invention in order to compare the efficacy of the method with conventional methods in terms of final yield of These results indicate that ethanol is useful in the practice phenethyl alcohol. 60 of the present invention as a Solvent for eluting phenethyl 500 mL of fermented L-phenylalanine was collected from alcohol that has been adsorbed onto an ion-exchange resin. a large fermentor. The HP-20 ion-exchange resin In the Specification and examples, there have been dis (Mitsubishi Kasei Corp., White Plains, N.Y.) was packed on closed preferred embodiments of the invention. Although a column (2.5x30 cm) and activated by passing 2 bed Specific terms are employed in these examples, they are used Volume each of acetone and methanol over the resin. The gel 65 in a generic and descriptive Sense only and not for the was rinsed with deionized water and remained wet prior to purpose of limitation, the Scope of the invention being extraction. The organic molecules were adsorbed onto the defined by the following claims. 5,919,991 7 8 That which is claimed is: 19. The process according to claim 18, whereby said 1. A proceSS for preparing phenethyl alcohol comprising: Solvent is Selected from the group consisting of ethanol, (a) providing an L-phenylalanine Solution; methanol, and acetone. (b) fermenting the L-phenylalanine Solution within a 20. A process for preparing phenethyl alcohol having a 5 high yield, the proceSS comprising: fermentation bath to provide a fermented phenethyl (a) preparing a L-phenylalanine Solution; alcohol Solution; (b) fermenting the L-phenylalanine Solution within a (c) separating the phenethyl alcohol Solution to provide a fermentation bath containing at least one Strain of yeast fermented phenethyl alcohol Solution essentially to provide a fermented phenethyl alcohol Solution; devoid of large molecular-weight molecules, (c) separating the phenethyl alcohol Solution to provide a (d) isolating the fermented phenethyl alcohol Solution fermented phenethyl alcohol Solution essentially essentially devoid of large molecular-weight mol devoid of yeast cells, ecules, (d) Separating the phenethyl alcohol Solution essentially (e) contacting the fermented phenethyl alcohol Solution of devoid of yeast cells to provide a fermented phenethyl Step (d) with an ion-exchange resin under conditions 15 alcohol Solution essentially devoid of large molecular sufficient to adsorb phenethyl alcohol thereon; and weight molecules, (f) collecting the phenethyl alcohol. 2. The process according to claim 1, whereby the (e) isolating the fermented phenethyl alcohol Solution L-phenylalanine Solution is a liquid having an aqueous essentially devoid of large molecular-weight mol character. ecules, 3. The process according to claim 1, whereby the (f) contacting the fermented phenethyl alcohol Solution of L-phenylalanine Solution comprises a fermentation media Step (e) with an ion-exchange resin to adsorb phenethyl Solution containing L-phenylalanine. alcohol thereon, and 4. The process according to claim 1, whereby the (g) collecting the phenethyl alcohol. L-phenylalanine Solution is added to a fermentation media 25 21. The process according to claim 20, whereby the Solution to provide a fermentation media Solution containing L-phenylalanine Solution is a liquid having an aqueous L-phenylalanine. character. 5. The process according to claim 1, whereby said fer 22. The process according to claim 20, whereby the menting Step comprises the addition of at least one Strain of L-phenylalanine Solution comprises a fermentation media yeast. Solution containing L-phenylalanine. 6. The process according to claim 5, whereby the yeast is 23. The process according to claim 20, whereby the Selected from the group consisting of Strains of kluyvero L-phenylalanine Solution is added to a fermentation media myces and Saccharomyces. Solution to provide a fermentation media Solution containing 7. The process according to claim 5, further comprising L-phenylalanine. the additional Step of Separating the fermented phenethyl 24. The proceSS according to claim 20, whereby the yeast alcohol Solution to provide a fermented phenethyl alcohol 35 is Selected from the group consisting of Strains of Kluyvero Solution essentially free of yeast cells. myces and Saccharomyces. 8. The process according to claim 7, whereby said addi 25. The process according to claim 20, whereby said first tional Separation Step comprises crossflow filtration. Separation Step comprises crossflow filtration. 9. The process according to claim 1, whereby the fermen 26. The process according to claim 20, whereby said tation bath is a liquid having an aqueous character. 40 Second Separation Step comprises crossflow filtration. 10. The process according to claim 1, whereby the 27. The process according to claim 20, whereby the L-phenylalanine Solution is fermented over a period of at fermentation bath is a liquid having an aqueous character. least 24 hours. 28. The process according to claim 20, whereby the 11. The process according to claim 1, whereby the L-phenylalanine Solution is fermented over a period of at L-phenylalanine Solution is fermented to provide a fer 45 least 24 hours. mented phenethyl alcohol Solution having a phenethyl alco 29. The process according to claim 20, whereby the hol content of at least 60% by weight. L-phenylalanine Solution is fermented to provide a fer 12. The process according to claim 1, whereby Said mented phenethyl alcohol Solution having a phenethyl alco Separation Step comprises crossflow filtration. hol content of at least 60% by weight. 13. The process according to claim 1, wherein Said 30. The process according to claim 20, whereby the ion contacting Step comprises contacting the fermented phen 50 eXchange resin is activated prior to Said contacting Step. ethyl alcohol Solution with an ion-exchange resin compris 31. The process according to claim 30, whereby said ing polymeric beads of average particle size not greater than activation comprises washing the ion exchange resin first 250 um in diameter. with deionized water and then with a solvent. 14. The process according to claim 1, whereby the ion 32. The process according to claim 31, whereby said eXchange resin is activated prior to Said contacting Step. 55 Solvent is Selected from the group consisting of ethanol, 15. The process according to claim 14, whereby said methanol, and acetone. activation comprises washing the ion exchange resin first 33. The process according to claim 20, further comprising with deionized water and then with a solvent. Washing the resin adsorbed to phenethyl alcohol with deion 16. The process according to claim 15, whereby said ized water prior to the collection Step. Solvent is Selected from the group consisting of ethanol, 60 34. The process according to claim 20, whereby said methanol, and acetone. collecting Step comprises repeated washings of the ion 17. The proceSS according to claim 1, further comprising eXchange resin with a Solvent. Washing the resin adsorbed to phenethyl alcohol with deion 35. The process according to claim 34, whereby said ized water prior to the collection Step. Solvent is Selected from the group consisting of ethanol, 18. The process according to claim 1, whereby Said methanol, and acetone. collecting Step comprises repeated washings of the ion 65 eXchange resin with a Solvent. k k k k k