Aroma Chemicals Research Articles

Antimicrobial and bactericidal activities of esters of 2-endo-hydroxy-1,8-cineole as new aroma chemicals.

Fourteen kinds of alkyl esters of 2-endo-hydroxy-1,8-cineole were synthesized, with yields of 57.8-98.0%. Each ester had a characteristic and unique odor. Especially, the tert-butyl acetate of 2-endo-hydroxy-1,8-cineole had the most interesting odor of all the synthetic esters. The antimicrobial and bactericidal activities of these synthetic esters against test bacteria (Staphylococcus aureus, Escherichia coli, and Pseudomonas fluorescens) were examined using the broth dilution method. As a result, the tert-butyl acetate of 2-endo-hydroxy-1,8-cineole showed the highest antimicrobial and bactericidal activities against all kinds of the test bacteria.

Studies on the autoxidation and sensitizing capacity of the fragrance chemical linalool, identifying a linalool hydroperoxide.

Fragrances are among the most common causes of allergic contact dermatitis. The two monoterpenes linalool and d-limonene are the most frequently incorporated fragrance chemicals in scented products. Previous studies on d-limonene show that this monoterpene oxidizes on air exposure (autoxidation) and that allergenic oxidation products are formed. Due to structural similarities, linalool might also form allergenic oxidation products on air exposure. The aim of the present study was to study the autoxidation of linalool and to investigate the sensitizing potential of linalool before and after air exposure. Linalool was oxidized for 10 weeks and gas chromatographic analyses showed that the content of linalool decreased to about 80%. The chromatograms revealed the formation of other compounds during oxidation. One of the major oxidation products was isolated and identified as 7-hydroperoxy-3,7-dimethyl-octa-1,5-diene-3-ol. This substance is, to the best of our knowledge, described for the first time. In sensitization studies in guinea pigs, linalool of high purity gave no reactions, while linalool that had been oxidized for 10 weeks sensitized the animals. It is concluded that autoxidation of linalool is essential for its sensitizing potential.

Inhibition of malonaldehyde formation from blood plasma oxidation by aroma extracts and aroma components isolated from clove and eucalyptus

The inhibitory effect of aroma extracts isolated from clove buds [ Syzygium aromaticum (L.) Merr. et Perry] and eucalyptus leaves ( Eucalyptus polyanthemos Schauer) on malonaldehyde (MA) formation from horse blood plasma oxidized with Fenton's reagent was determined by gas chromatography. Aroma chemicals such as eugenol, thymol and benzyl alcohol, identified in the aroma extracts, were examined for their inhibitory effect on the same system. Between the two aroma extracts tested, clove exhibited the most potent antioxidant activities. Extracts of eucalyptus and clove inhibited MA formation by 23 and 48%, respectively, at the level of 400 μg/ml, whereas, α-tocopherol and BHT inhibited MA formation by 52 and 70%, respectively, at the same level. Eugenol, thymol and benzyl alcohol inhibited MA formation by 57, 43 and 32%, respectively, at the level of 400 μg/ml.

Fragrance chemicals in domestic and occupational products.

Epidemiological studies have described an increasing prevalence of fragrance allergy and indicated an association with hand eczema. 59 domestic and occupational products intended for hand exposure were subjected to gas chromatography-mass spectrometric (GC-MS) analyses to test the hypothesis that fragrance chemicals known to have the potential to cause contact allergy but not included in fragrance mix (FM) may be common ingredients in these products. A quantitative analysis of 19 selected fragrances was performed by GC-MS. Further analysis of GC-MS data revealed the presence of 43 other fragrance chemicals/groups of fragrance chemicals in the products investigated. Among the 19 target substances the most commonly detected were limonene in 78%, linalool in 61% and citronellol in 47% of the products investigated. The FM ingredients were present in these products with the following frequencies: oak moss (evernic acid methylester) 2%, cinnamic alcohol 2%, cinnamic aldehyde (cinnamal) 3%, isoeugenol 5%, alpha-amylcinnamic aldehyde (amyl cinnamal) 8%, hydroxycitronellal 12%, eugenol 27%, and geraniol 41%. Thus, the chemical analyses of domestic and occupational products indicates that investigation of potential contact allergy related to these products types should consider fragrance allergens additional to those in the FM, since these may occur with high frequency.

The Physicochemical Basis of Perfume Performance in Consumer Products

The literature on physicochemical aspects of perfume performance has been reviewed with emphasis on products for home and body care. Pertinent expressions related to perfume performance have been defined. For the first time a model study is presented to account for the dynamic aspects in applications of perfumed fabric softeners. A new headspace cell, which allows precise control of the atmospheric conditions and modelling of the dynamics of the drying process, has been designed. The determination of the deposition of selected fragrance chemicals and their evaporation, combined with olfactometric dose-response data, produced a reliable measurement of their performance in application.

Vehicle effects on skin sensitizing potency of four chemicals: assessment using the local lymph node assay

The murine local lymph node assay (LLNA) can be used to determine the relative skin sensitizing potency of chemicals via interpolation of the quantitative dose response data generated. Using this approach we have demonstrated previously that the vehicle matrix in which a chemical allergen is encountered on the skin can have a significant influence on sensitizing potency. Estimates of relative potency are calculated from LLNA dose responses as a function of the mathematically derived EC3 value, this being the concentration estimated to induce a stimulation index (SI) of 3. To investigate further the influence of application vehicle on sensitizing potency, the LLNA has been used to examine the activity of four recognized human contact allergens: isoeugenol and cinnamic aldehyde, two fragrance chemicals; 3-dimethylaminopropylamine (a sensitizing impurity of cocamidopropyl betaine, a surfactant used in shower gel) and dibromodicyanobutane (the sensitizing component of Euxyl K 400, a preservative used in cosmetics). The four chemicals were applied in each of seven different vehicles (acetone: olive oil [4 : 1]; dimethylsulphoxide; methylethylketone; dimethyl formamide; propylene glycol; and both 50 : 50 and 90 : 10 mixtures of ethanol and water). It was found that the vehicle in which a chemical is presented to the epidermis can have a marked effect on sensitizing activity. EC3 values ranged from 0.9 to 4.9% for isoeugenol, from 0.5 to 1.7% for cinnamic aldehyde, from 1.7 to > 10% for dimethylaminopropylamine and from 0.4 to 6.4% for dibromodicyanobutane. These data confirm that the vehicle in which a chemical is encountered on the skin has an important influence on the relative skin sensitizing potency of chemicals and may have a significant impact on the acquisition of allergic contact dermatitis. The data also demonstrate the utility of the LLNA as a method for the prediction of these effects and thus for the development of more accurate risk assessments.

Quenching: fact or fiction?

Fragrance chemicals are a frequently reported cause of allergic contact dermatitis (ACD), a matter which has recently come into considerable prominence, to the point that legislation in Europe is under serious consideration. Certain skin-sensitizing fragrance chemicals have been reported by the producing industry to be rendered safe (quenched), at least in terms of ACD, when they are used in the presence of a specific quenching agent. Accordingly, it seemed timely to review this apparent quenching phenomenon, considering the available data and potential mechanistic hypotheses that might be used to explain it. If it is correct, it should be a phenomenon of potentially enormous value in the elimination of the allergenic properties of at least a proportion of common skin sensitizers. Whilst there is some evidence in man for the occurrence of quenching during the induction of skin sensitization, a much more substantial body of work has failed to find supportive evidence in various animals models, at a chemical level or at elicitation in human subjects with existing allergy. On balance, it is concluded that quenching of fragrance allergens is a phenomenon still awaiting positive evidence of existence.

Synthesis and biological activity of alpha-methylene-gamma-lactones as new aroma chemicals.

Seven kinds of alpha-methylene-gamma-lactones with an alkyl group at the C-4 position were synthesized according to a previously described method, with yields of 28-34%. These alpha-methylene-gamma-lactones had characteristic and unique odors. All alpha-methylene-gamma-lactones added a roast-like odor to materials. The antimicrobial effects of alpha-methylene-gamma-lactones were investigated by using a paper disk diffusion method. The results showed the alpha-methylene-gamma-lactones inhibited the growth of three bacteria (Staphylococcus aureus, Escherichia coli, and Pseudomonas fluorescens) and two fungi (Saccharomyces cerevisiae and Aspergillus niger). In particular, alpha-methylene-gamma-undecalactone and alpha-methylene-gamma-dodecalactone exhibited potent inhibition of the growth of these microorganisms compared to butyl p-hydroxybenzoate as standard antibiotic. The umu test revealed that the alpha-methylene-gamma-lactones suppressed the SOS-inducing activity of three mutagens, furylfuramide, UV irradiation, and Trp-P-1, respectively. The antimicrobial effects and the suppressive effects of SOS induction by alpha-methylene-gamma-lactones had a tendency to intensify as the number of carbons in the side chain increased.

Antioxidant properties of aroma compounds isolated from soybeans and mung beans.

Aroma compounds contained in the extracts of soybean and mung bean that possess antioxidant activity were identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major aroma constituents of soybeans were 1-octen-3-ol (13.699 ppm), maltol (1.662 ppm), phenylethyl alcohol (1.474 ppm), hexanol (1.430 ppm), and gamma-butyrolactone (1.370 ppm). The major aroma constituents of mung beans were hexanol (3.234 ppm), benzyl alcohol (2.060 ppm), gamma-butyrolactone (1.857 ppm), 2-methyl-2-propenal (1. 633 ppm), and pentanol (1.363 ppm). The major aroma chemicals of soybeans and mung beans were examined for antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol showed potent antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited the oxidation of hexanal by 100%, 93%, 84%, and 32%, respectively, for a period of 40 days at the 500 microg/mL level. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited malonaldehyde (MA) formation from cod liver oil by 91%, 78%, 78%, and 78%, respectively, at the 160 microg/mL level. The antioxidative activity of eugenol was comparable to that of the natural antioxidant alpha-tocopherol (vitamin E).

Chemical Characterization of basil (Ocimum Spp.) found in the markets and used in traditional medicine in Brazil

Ocimum species are used both in Brazilian traditional medicine against bronchitis, coughs, and sorethroat, and in foods and flavorings. The traditional uses of basil in Brazil are reviewed and fourteen accessions of basil, O. americanum (1), O. basilicum (3), O. campechianum (3), O. gratissimum (5), and O. selloi (2) were collected and brought from Brazil to the USA and grown at Purdue University. Upon flowering, the volatile oils were extracted by hydrodistillation and chemically analyzed by GC/ MS. Accessions ofO. gratissimum showed high eugenol (40-66%) and high thymol (31%), as a relative percentage of total volatile oil. Ocimum campechianum accessions showed high 1,8-cineole (62%) and s-caryophyllene (78.7%). Accessions of O. basilicum were found to be rich in 1,8-cineole (22%), linalool (49.7%), methyl chavicol (47%) or methyl (E)-cinnamate (65.5%). An O. americanum var. americanum accession showed high methyl (E)-cinnamate (>90%). Volatile oils of O. basilicum and O. gratissimum naturalized in Brazil reflected the range of chemotypes found in their country of origin. Unusual basils from other Ocimum species were identified that can serve as genetic sources of aroma chemicals for crop improvement.

Effect of Increased Yeast Alcohol Acetyltransferase Activity on Flavor Profiles of Wine and Distillates

The distinctive flavor of wine, brandy, and other grape-derived alcoholic beverages is affected by many compounds, including esters produced during alcoholic fermentation. The characteristic fruity odors of the fermentation bouquet are primarily due to a mixture of hexyl acetate, ethyl caproate (apple-like aroma), iso-amyl acetate (banana-like aroma), ethyl caprylate (apple-like aroma), and 2-phenylethyl acetate (fruity, flowery flavor with a honey note). The objective of this study was to investigate the feasibility of improving the aroma of wine and distillates by overexpressing one of the endogenous yeast genes that controls acetate ester production during fermentation. The synthesis of acetate esters by the wine yeast Saccharomyces cerevisiae during fermentation is ascribed to at least three acetyltransferase activities, namely, alcohol acetyltransferase (AAT), ethanol acetyltransferase, and iso-amyl AAT. To investigate the effect of increased AAT activity on the sensory quality of Chenin blanc wines and distillates from Colombar base wines, we have overexpressed the alcohol acetyltransferase gene (ATF1) of S. cerevisiae. The ATF1 gene, located on chromosome XV, was cloned from a widely used commercial wine yeast strain of S. cerevisiae, VIN13, and placed under the control of the constitutive yeast phosphoglycerate kinase gene (PGK1) promoter and terminator. Chromoblot analysis confirmed the integration of the modified copy of ATF1 into the genome of three commercial wine yeast strains (VIN7, VIN13, and WE228). Northern blot analysis indicated constitutive expression of ATF1 at high levels in these yeast transformants. The levels of ethyl acetate, iso-amyl acetate, and 2-phenylethyl acetate increased 3- to 10-fold, 3.8- to 12-fold, and 2- to 10-fold, respectively, depending on the fermentation temperature, cultivar, and yeast strain used. The concentrations of ethyl caprate, ethyl caprylate, and hexyl acetate only showed minor changes, whereas the acetic acid concentration decreased by more than half. These changes in the wine and distillate composition had a pronounced effect on the solvent or chemical aroma (associated with ethyl acetate and iso-amyl acetate) and the herbaceous and heads-associated aromas of the final distillate and the solvent or chemical and fruity or flowery characters of the Chenin blanc wines. This study establishes the concept that the overexpression of acetyltransferase genes such as ATF1 could profoundly affect the flavor profiles of wines and distillates deficient in aroma, thereby paving the way for the production of products maintaining a fruitier character for longer periods after bottling.

Volatile sulfur production by pig cecal bacteria in batch culture and screening inhibitors of sulfate reducing bacteria.

We studied the effects of specific inhibitors of methanogenesis (2-bromoethane sulfonate, BES) and sulfate reduction (sodium molybdate) on volatile sulfur production in batch cultures of pig cecal bacteria. The volatile sulfur concentration in headspace gas was determined by flame-photometric detector gas chromatography. BES stimulated production of hydrogen sulfide (H2S) and methanethiol, and sodium molybdate completely inhibited the production of these volatile sulfur compounds. The results indicated that dissimilate sulfate reduction is mainly responsible for volatile sulfur production in the hindgut. Therefore the extracts of herbs, food colors, and aroma chemicals were tested for their inhibitory effects on H2S production by a dissimilatory sulfate-reducing bacteria. Desulfovibrio desulfuricans DSM642. H2S was measured by the chromatography of the headspace gas, using a flame photometric detector. Of 306 herbal extracts tested, 69 extracts from 38 herbs inhibited H2S production at 1.0 mg/mL. Sisymbrium officinale (hedge mustard) was the most potent inhibitor. Six pigments inhibited H2S release. Erythrosine and rose bengal showed inhibitory effects at 0.01 mg/mL. Peppermint oil and 96 aroma chemicals were assayed for their effects on H2S release. Thirty-two aroma chemicals suppressed H2S production at 0.1 mg/mL, and camphene, 1-decanol, and 2-nonanone were effective at 0.01 mg/mL.

Antioxidative activities of aroma extracts isolated from natural plants

Natural leaves and flowers containing numerous aroma chemicals are widely used in aromatherapy since ancient times. In addition to their pleasant smells, aroma chemicals might have some beneficial health effects. Aroma extracts, isolated from coffee beans, soybeans, and mung beans by steam distillation under mild conditions (55 degrees C and 85 mm Hg) were examined for their antioxidative activities. The inhibitory effect of these extracts toward hexanal/hexanoic acid conversion was measured in the testing solution over prolonged time periods. The inhibitory effects of these extracts toward malonaldehyde formation from lipids oxidized by Fenton's reagent were also measured. The antioxidative activity of these extracts, in particular coffee bean extract, was consistent with that of BHT or alpha-tocopherol (vitamin E). Soybeans and mung beans extract contained maltol, which inhibits hexanal oxidation significantly. Eugenol, which is one of the major constituents of mung bean extract, exhibited potent antioxidative activity in an aldehyde/carboxylic acid assay. Antioxidants such as eugenol and maltol may play an important role in the pharmaceutical activities of natural plant extracts used for aromatherapy.

Aroma chemicals isolated and identified from leaves of Aloe arborescens Mill. Var. Natalensis Berger.

Extracts from leaves of aloe (Aloe arborescens Mill. var. natalensis Berger) were obtained using two methods: steam distillation under reduced pressure followed by dichloromethane extraction (DRP) and simultaneous purging and extraction (SPE). A total of 123 aroma chemicals were identified in the extracts obtained by both methods using gas chromatography and gas chromatography/mass spectrometry. There were 42 alcohols, 23 terpenoids, 21 aldehydes, 9 esters, 8 ketones, 6 acids, 5 phenols, and 9 miscellaneous compounds. The major aroma constituents of this extract by DRP were (Z)-3-hexenol (29.89%), (Z)-3-hexenal (18.86%), (E)-hexenal (7.31%), 4-methyl-3-pentenol (5.66%), and butanol (4.29%). The major aroma constituents of this extract by SPE were (E)-2-hexenal (45.46%), (Z)-3-hexenal (32.12%), hexanal (9.14%), (Z)-3-hexenol (1.60%), and 3-pentanone (1.41%). Terpenoids were also found as one of the major constituents. The fresh green note of aloe leaves is due to the presence of these C(6) alcohols and aldehydes as well as terpenoids.

Biotechnological production of flavours and fragrances.

The biotechnological generation of natural aroma compounds is rapidly expanding. Aroma chemicals, such as vanillin, benzaldehyde (bitter almond, cherry) and 4-(R)-decanolide (fruity-fatty) are marketed on a scale of several thousand tons per year. Their possible production by single-step biotransformations, bioconversions and de novo synthesis using microorganisms, plant cells or isolated enzymes is shown. The perspectives of bioprocesses for the oxifunctionalisation of lower terpenes by genetically modified organisms and economic aspects are discussed.

Fragrance allergens: Classification and ranking by QSAR

Quantitative structure-activity relationship (QSAR) models which predict both skin penetration and cell mediated immunity for small molecular weight non-electrolytes developed earlier were validated on 74 known allergens and non-allergens chosen among fragrance chemicals in common use to test discriminating and grading power. While the test set used for classification was based on experience in humans exclusively, the rank model was tested for sensitization potency including guinea pig data also. In the classification test, 12 of 74 compounds fell in the indeterminate range and were non-classifiable by the present QSAR model. On the remaining 62 compounds the model performs with 90% sensitivity and 100% specificity at 92% concordance. The rank model correctly grades 65 of 74 compounds (88% concordance), with 60% specificity based on exact prediction of non-allergens (NON), and 95% sensitivity on allergens (ACD) allowing for a variance of ± one level among weak, moderate and severe ratings. In combination, the two models perform with 93% overall concordance on the test set of 74 compounds.

Production of methoxyphenol-type natural aroma chemicals by biotransformation of eugenol with a new Pseudomonas sp.

In our screening program for microorganisms that are able to metabolize eugenol, the main component of the essential oil of the clove tree Syzigium aromaticum (sy. Eugenia cariophyllus), we found a new Pseudomonas sp. that produces several substituted methoxyphenols when eugenol is fed to the culture. A taxonomic characterization of this new organism has been performed. Examples of the biotransformation products, produced in high amounts, were vanillic acid with 3.25 g/l within 99 h, ferulic acid with 5.8 g/l within 75 h and coniferyl alcohol with 3.22 g/l within 47.5 h. By changing the culture conditions the ratio of the different metabolites could be varied. Based on these results a scheme for the degradation of eugenol by this strain has been established.

Enhancing the productivity of secondary metabolites via induced polyploidy: a review

Artificial polyploidy generally enhances the vigour of determinate plant parts and may be favourable where vegetative organs and biomass constitute the economic product. Giving suitable examples, it is shown that genomic multiplication can confer enhanced production and/or qualitative improvement in the biochemical profile of secondary metabolites. Allopolyploidisation can bring about natural complementation of biosynthetic pathways to harness the useful metabolites. A plea is made to utilise the induced polyploidy approach as a rapid means to attain enhanced production of secondary metabolites: pharmaceuticals, aroma chemicals, etc. The necessary prerequisites that may be needed for achieving genetic stability and reproductive success of the induced polyploids are outlined.

Volatile constituents of Dactylanthus taylorii flower nectar in relation to flower pollination and browsing by animals

The volatile fraction of Dactylanthus taylorii flower nectar from male and female plants has been shown to contain squalene as a major constituent. The inflorescence from male plants contained approximately twice the amount of squalene as that detected in the inflorescence of female plants. In addition, the nectar contained C 21 to C 31 hydrocarbons and in one of the male plants samples, ethyl and benzyl esters of hexadecanoic and C 18 to C 23 polyunsaturated fatty acids as other significant lipid components. The fragrance chemicals in the nectar comprised a relatively simple mixture of butenols, butanols and pentanols, benzyl and phenylethyl alcohols, nerol, geraniol and ethyl esters of benzoic, salicylic and cinnamic acids. The inflorescence from female plants contained approximately five times the amount of benzyl alcohol compared with the amount from the inflorescences of male plants. The importance of the nectar volatile compounds in bat pollination and browsing of Dactylanthus inflorescences by possums and ship rats is briefly discussed.

Microbial biocatalysis in the generation of flavor and fragrance chemicals.

Microbial biocatalysis is used in the commercial production of many flavor and fragrance chemicals. Bulk flavoring chemicals such as citric acid, high fructose corn syrup, and glutamic acid are produced in millions of pounds annually using microbial processes. In the past few years, biocatalysis has also begun to play an increasingly important role in the production of many flavor and fragrance aroma chemicals. Microbial processes have traditionally played an integral role in the development of complex mixtures of flavor and aroma chemicals since the discovery of beer, wine, cheese, and soy sauce thousands of years ago. Today, contemporary microbiological techniques are being increasingly applied to enhance the efficiency of many microbial biocatalysts for the production of specific flavor and fragrance chemicals. However, to ensure commercial implementation of these new microbial processes, much more needs to be learned about the basic biochemistry and genetics of these novel biocatalysts.