Effect of ethanol, temperature, and gas flow rate on volatile release from aqueous solutions under dynamic headspace dilution conditions

On the basis of a mechanistic model, the overall and liquid mass transfer coefficients of aroma compounds were estimated during aroma release when an inert gas diluted the static headspace over simple ethanol/water solutions (ethanol concentration = 120 mL x L(-1)). Studied for a range of 17 compounds, they were both increased in the ethanol/water solution compared to the water solution, showing a better mass transfer due to the presence of ethanol, additively to partition coefficient variation. Thermal imaging results showed differences in convection of the two systems (water and ethanol/water) arguing for ethanol convection enhancement inside the liquid. The effect of ethanol in the solution on mass transfer coefficients at different temperatures was minor. On the contrary, at different headspace dilution rates, the effect of ethanol in the solution helped to maintain the volatile headspace concentration close to equilibrium concentration, when the headspace was replenished 1-3 times per minute.     

Retronasal transport of aroma compounds

A comparison was made between the amounts of volatiles in the headspace above a solution and the breath volatile content (exhaled from the nose or mouth) after consumption of the same solution. The amounts of volatiles in the breath were lower than those in the headspace, with breath exhaled via the mouth containing, on average, 8-fold more volatiles than breath exhaled via the nose. Dilution of the sample by saliva in-mouth did not appear to be a major factor affecting volatile delivery. Instead, the rate of in vivo equilibration (mass transfer) appeared to be the most significant factor, principally affecting volatile delivery from the solution to the gas phase. Thereafter, gas-phase dilution of the volatile as it passed through the upper airway resulted in a further decrease in volatile concentration. The final factor affecting the volatile concentration exhaled from the nose was absorption of volatiles to the nasal epithelia, which was greatest for those compounds with the lowest air/water partition coefficients.     

Solid-phase microextraction method development for headspace analysis of volatile flavor compounds

Solid-phase microextraction (SPME) fibers were evaluated for their ability to adsorb volatile flavor compounds under various conditions with coffee and aqueous flavored solutions. Experiments comparing different fibers showed that poly(dimethylsiloxane)/divinylbenzene had the highest overall sensitivity. Carboxen/poly(dimethylsiloxane) was the most sensitive to small molecules and acids. As the concentrations of compounds increased, the quantitative linear range was exceeded as shown by competition effects with 2-isobutyl-3-methoxypyrazine at concentrations above 1 ppm. A method based on a short-time sampling of the headspace (1 min) was shown to better represent the equilibrium headspace concentration. Analysis of coffee brew with a 1-min headspace adsorption time was verified to be within the linear range for most compounds and thus appropriate for relative headspace quantification. Absolute quantification of volatiles, using isotope dilution assays (IDA), is not subject to biases caused by excess compound concentrations or complex matrices. The degradation of coffee aroma volatiles during storage was followed by relative headspace measurements and absolute quantifications. Both methods gave similar values for 3-methylbutanal, 4-ethylguaiacol, and 2,3-pentanedione. Acetic acid, however, gave higher values during storage upon relative headspace measurements due to concurrent pH decreases that were not seen with IDA.     

Effect of beta-cyclodextrin on aroma release and flavor perception

Binding and release of volatile compounds to and from beta-cyclodextrin were measured in model aqueous systems using static equilibrium headspace and dynamic headspace dilution. Beta-cyclodextrin decreased the static equilibrium headspace for some volatiles (e.g., ethyl octanoate and decanone) due to binding, but dilution studies demonstrated that binding was readily reversible. Dynamic release of hydrophobic volatile compounds was similar to that observed from emulsions. When beta-cyclodextrin was added to fat free yogurt, the release of a commercial lemon flavoring was modified and was similar to release from a regular fat yogurt. Sensory difference testing confirmed the release results. The data demonstrate that beta-cyclodextrin can be used to modify flavor delivery in both model and real systems; the effects in the latter are sensorially significant.     

Effects of droplet crystallization and melting on the aroma release properties of a model oil-in-water emulsion

Aroma compounds partition between the dispersed and the continuous phases in emulsions, and phase transitions in the lipid droplets profoundly affect the position of the equilibrium. In the present study, the release of ethyl butyrate, ethyl pentanoate, ethyl heptanoate, and ethyl octanoate from a series of sodium caseinate-stabilized, n-eicosane emulsions was investigated as a function of solid and liquid lipid droplet concentration. For all compounds, headspace volatile concentrations above the solid droplet emulsions were higher than those above the liquid droplet emulsions. The interaction with liquid droplets could be modeled in terms of volume-weighted bulk partition coefficients while the more nonpolar volatiles bound to the surface of solid lipid droplets. The amount of volatiles bound to solid surfaces increased with aqueous concentration up to a critical point and then rapidly increased. The critical point corresponds to the dissolution of the solid lipid in a phase of adsorbed volatile. The binding of volatiles to both solid and liquid eicosane droplets is reversible.     

Isolation and quantification of volatiles in fish by dynamic headspace sampling and mass spectrometry.

A dynamic headspace sampling method for isolation of volatiles in fish has been developed. The sample preparation involved freezing of fish tissue in liquid nitrogen, pulverizing the tissue, and sampling of volatiles from an aqueous slurry of the fish powder. Similar volatile patterns were determined by use of this sample preparation method and for samples chewed for 10 s. Effects of sampling time, temperature, and purge flow on level of volatiles were tested. Purging at 340 mL/min for 30 min at 45 degrees C was found to be optimal. Detection limits for a number of aldehydes were 0.2-2.7 microg/kg. Levels of volatiles are given for fresh salmon, cod, saithe, mackerel, and redfish.     

Effect of pH on retention of aroma compounds by beta-lactoglobulin

Interactions between volatile compounds and BLG in aqueous solution were studied using static and dynamic headspace techniques (exponential dilution). The intensity of interactions between methyl ketones (C7-C9), ethyl esters (C6-C9), limonene, myrcene, and beta-lactoglobulin (BLG) were estimated by determination of the relative infinite dilution activity coefficients (gamma(r)). For a constant pH value, the methyl ketones retention by BLG increased significantly with the hydrophobicity of the volatiles, whereas the retention reached a maximum for ethyl octanoate in the ester series, indicating a possible steric hindrance. For limonene and myrcene an unexpected increase in headspace concentration or "salting out" effect was noticed for acid pH. The variations of the retention according to the pH increase of the medium from pH 3 to pH 11 could be related to structural modifications of the BLG. The retention increase observed between pH 3 and pH 9 resulted from the flexibility modification of the protein, allowing better accessibility to the primary or the secondary hydrophobic sites, whereas the dramatic decrease observed at pH 11 was the consequence of the alkaline denaturation of BLG. Electrostatic interactions occurring at pH 7.5 could also explain the observed retention increase.     

Volatile release from an emulsion: headspace and in-mouth studies

The headspace concentrations of three esters above solutions containing emulsified lipids were more resistant to dilution by a stream of gas than those above water alone. The effect was greatest for ethyl octanoate, and least for ethyl butyrate, with ethyl hexanoate showing intermediate behavior. This correlated with their solubility in the lipid fraction of the emulsion. Headspace analysis (comparing the emulsion with water) underestimated the release of the esters during consumption. The ratios observed between water and emulsion systems were different for the maximum breath concentration compared with headspace analysis. The emulsion appears to have acted as a reservoir for volatile release, counteracting the effects of sample dilution by saliva.     

Effect of salivary components on volatile partitioning from solutions

Partition of fourteen volatile compounds, representing the diverse physicochemical properties of aroma compounds, was measured by static equilibrium headspace in solutions containing the components of artificial saliva, either singly or in mixtures. Comparison of a bovine salivary mucin and pig gastric mucin showed no significant difference in partition behavior of the volatiles, so gastric mucin was used. Mucin viscosity changed with pH, but binding of volatile compounds did not show a marked dependence on pH. All combinations of the salivary components were tested for their effect on partition. Three types of behavior were noted. Partition of some compounds was unaffected by mucin, and with other compounds mucin decreased partition, whereas another group showed a decrease with mucin that was affected by the presence of salivary salts and sugar. When volatiles or sugar were added to a mucin solution, the final headspace concentration depended on the order of addition, indicating some competition for binding. These solute-mucin effects are discussed in relation to mucin structure and behavior in solution.     

Effect of copper on the volatility of aroma compounds in a model mouth system

Copper is thought to influence aroma perception by affecting volatility of aroma compounds in the mouth through interaction with salivary components, especially proteins. Our objective was to identify the effect of copper on the volatility of aroma compounds and the role of copper-protein interaction in volatile chemistry in the mouth. Copper (2.5 mg/L) and four aroma compounds (hexanal, butyl acetate, 2-heptanone, and ethyl hexanoate, 0.5 microL/L each) were added to model systems containing water, electrolytes, and artificial saliva at different pH levels. Headspace concentration of each volatile was measured using SPME-GC analysis. Copper in the model systems increased headspace concentration of volatiles at pH 6.5, but no change in volatility was observed at pH 7.0. At pH 7.5, the presence of copper in the artificial saliva system containing mucin and alpha-amylase decreased headspace volatile concentration, whereas histatin did not cause any changes in volatility. Effect of copper on volatiles at pH 6.5 may be due to increased solubility of copper at lower pH. Salivary proteins seem to interact with copper at pH 7.5. The interaction may change configuration of binding sites for aroma compounds in mucin.     

Interactions between methyl ketones and beta-lactoglobulin: sensory analysis, headspace analysis, and mathematical modeling

Interaction of flavor compounds with proteins is known to have an influence on the release of flavor from food. Hydrophobic interactions were found between beta-lactoglobulin and methyl ketones; the affinity constant increases by increasing the hydrophobic chain. Addition of beta-lactoglobulin (0.5 and 1%) to aroma solutions (12.5, 50, and 100 microL L(-)(1)) of three methyl ketones induces a significant decrease in odor intensity. The chosen methyl ketones were 2-heptanone (K(b) = 330), 2-octanone (K(b) = 950), and 2-nonanone (K(b) = 2440). The release of these flavor compounds (50 microL L(-)(1)) was studied by static headspace in water solution (50 mM NaCl, pH 3) with different concentrations of beta-lactoglobulin (0, 0.5, 1, 2, 3, and 4%). Increasing the concentration of protein increases the retention of volatiles, and this effect is greatest for 2-nonanone, the compound with the highest affinity constant, and lowest for 2-heptanone. A mathematical model previously developed to describe flavor release from aqueous solutions containing flavor-binding polymers (Harrison, M.; Hills, B. P. J. Agric. Food Chem. 1997, 45, 1883-1890) was used to interpret the data. The model assumes that the polymer-flavor interaction is reversible and the rate-limiting step for release is the transfer of volatiles across the macroscopic gas-liquid interface. This model was used to predict the equilibrium partitioning properties and the rate of release of the three methyl ketones. Increasing the affinity constant leads to decreased release rates and a lower final headspace aroma concentration.     

Dynamic flavor release from sucrose solutions

The initial dynamic flavor release from sucrose solutions was modeled. Modeling was based on the theoretical hydration behavior of sucrose, theoretical physicochemical data of flavor volatiles, and process parameters of a headspace apparatus used for model validation. The rate-limiting factor determining the initial flavor release was the hydration of sucrose, which in turn depends on the molarity of sucrose in the solution and, therefore, on the actual amount of nonbound water. Improved solubility of the more hydrophilic compounds due to their orientation toward the hydration shells of the sugar molecules was considered. The viscosity of nonassociated water forming the microregion for mass transfer of volatiles was considered instead of the bulk solution viscosity. Experimental validation of the model by real-time measurements of dynamic flavor release using foodlike flavor concentrations confirmed the above theory. Increasing sucrose concentrations resulted predominantly in increased flavor release, and bulk solution viscosity showed no effect.     

Volatile composition of coffee berries at different stages of ripeness and their possible attraction to the coffee berry borer Hypothenemus hampei (Coleoptera: Curculionidae)

The analysis of volatile emissions of coffee berries in different physiological states of ripeness was performed using dynamic headspace and gas chromatography/mass spectrometry analysis for Coffea arabica, var. Colombia. The composition of the volatiles emitted by coffee berries is dominated by very high levels of alcohols, mainly ethanol, in all stages of ripeness in comparison with other compounds. Overripe coffee berries have high volatile emissions and show a composition dominated mainly by esters followed by alcohols, ketones, and aldehydes. The lowest level compounds were monoterpenes. 2-Methyl furan was detected in various ripening stages; this compound has not been previously reported as a coffee berry volatile. The presence of ethanol and other alcohols in the volatile composition might explain the effectiveness of using traps with mixed alcohols for detection and capture of coffee berry borers.     

Optimization of solid phase microextraction analysis for the headspace volatile compounds of parmesan cheese

Optimum conditions of solid phase microextraction (SPME) analysis of the headspace volatile compounds of Parmesan cheese in airtightly sealed 100-mL bottles were developed. The coefficient of variation of SPME analysis on the headspace volatile compounds of Parmesan cheese was 2%. The reproducibility of SPME was improved by a combination of sampling at -10 degrees C, controlling the sample temperature, and uniform magnetic stirring of samples during equilibrium and isolation steps. The sensitivity of SPME increased by 125% in total peak areas by a combination of 40 min of sonication and 25% (w/v) sodium phosphate solution, compared with that of samples containing deionized water only (P < 0.05). The addition of salt solution or sonication treatment in samples increased the headspace volatile compounds of cheese quantitatively without producing any new volatile compounds.     

Volatile release from aqueous solutions under dynamic headspace dilution conditions.

Static equilibrium was established between the gas phase (headspace) and an unstirred aqueous phase in a sealed vessel. The headspace was then diluted with air to mimic the situation when a container of food is opened and the volatiles are diluted by the surrounding air. Because this first volatile signal can influence overall flavor perception, the parameters controlling volatile release under these conditions are of interest. A mechanistic model was developed and validated experimentally. Release of compounds depended on the air-water partition coefficient (K(aw)) and the mass transport in both phases. For compounds with K(aw) values <10(-)(3), K(aw) was the factor determining release rate. When K(aw) was >10(-)(3), mass transport in the gas phase became significant and the Reynolds number played a role. Because release from packaged foods occurs at low Reynolds numbers, whereas most experiments are conducted at medium to high Reynolds numbers, the experimentally defined profile may not reflect the real situation.     

Modeling the partition of volatile aroma compounds from a cloud emulsion

Parameters determining the partitioning behavior of volatile compounds between a cloud emulsion and the gas phase were measured under static equilibrium headspace conditions, using volatiles (e.g., ethyl hexanoate, cymene, and octanol) representing different volatilities and different degrees of hydrophobicity. The significant factors were the molecular characteristics of the volatile and the concentration of the oil phase. The nature of the lipid (C8 and C12 triglycerides), particle size, and emulsifier type (modified starch and gum arabic) did not significantly alter volatile partitioning. An empirical model based on the partition behavior and physicochemical parameters of 67 volatile compounds was produced. This predicted the partition of volatiles (R(2) = 0.83) in cloud emulsions as a function of lipid content. The significant terms (P < 0.05) in the empirical model were Log P, Log solubility, the dipole vector, and the oil fraction.     

Headspace solid-phase microextraction method for the study of the volatility of selected flavor compounds

Changes in the volatility of selected flavor compounds in the presence of nonvolatile food matrix components were studied using headspace solid-phase microextraction (HS-SPME) combined with GC-MS quantification. Time-dependent adsorption profiles to the SPME fiber and the partition coefficients between different phases were obtained for several individual volatiles, showing that HS-SPME analysis with a short sampling time can be used to determine the "true" headspace concentration at equilibrium between the headspace and a sample matrix. Equilibrium dialysis followed by HS-SPME/GC-MS was carried out to confirm the ability of HS-SPME extraction for monitoring the free volatile compounds in the presence of proteins. In particular, a short sampling time (1 min) avoided additional extraction of volatiles bound to the protein. Interactions between several selected flavor compounds and nonvolatile food matrix components [beta-lactoglobulin or (+)-catechin] were also studied by means of HS-SPME/GC-MS analysis. The volatility of ethyl hexanoate, heptanone, and hexanal was significantly decreased by the addition of beta-lactoglobulin compared to that of isoamyl acetate. Catechin decreased the volatility of ethyl hexanoate and hexanal by 10-20% and increased that of 2-heptanone by approximately 15%. This study indicates that HS-SPME can be a useful tool for the study of the interactions between volatile compounds and nonvolatile matrix components provided the kinetic and thermodynamic behavior of the volatiles in relation to the fiber chosen for the studies is carefully considered.     

Measurement of free zinc concentration in wine with AGNES

AGNES (absence of gradients and Nernstian equilibrium stripping), a voltammetric technique recently introduced to measure free metal concentration in solution and checked with different natural and synthetic aqueous media, has been applied here to determine free Zn concentration in wine. The content of ethanol in a solution increases its viscosity, and, so, the diffusion coefficient decreases. Another added effect in ethanolic solutions is the increase of the activity of the metal ions, due to the decrease of the permittivity in the alcoholic medium with respect to the aqueous one. With this taken into account, a specific methodology has been developed to apply AGNES in ethanolic media. A relevant point in this methodology has been the introduction of a new kind of blank, the EDTA blank, able to be applied in the same natural sample and with the same potential program. The free Zn concentrations of the two wines analyzed, a red and a white Raimat wine, were 4.5(2) x 10 (-7) and 7.2(4) x 10 (-7) M, respectively. These represent around 5% of the total Zn content. In the wine samples analyzed, it was checked that intermetallic formation of Zn-Cu does not affect the measurement of free Zn in a significant way.     

Quantitative analysis of aroma compounds in carrot (Daucus carota L.) cultivars by capillary gas chromatography using large-volume injection technique

Dynamic headspace sampling was used to collect aroma compounds from raw samples of four carrot (Daucus carota L.) cultivars (Brasilia, Duke, Fancy, and Cortez). The collected volatiles were analyzed by capillary GC-FID and GC-MS using large-volume cool on-column injection (LVI-COC). Of the 36 compounds identified, 6 had not been previously detected in carrots. Significant differences between the carrot cultivars were found for 31 of the identified volatiles as well as for total monoterpenes, sesquiterpenes, and total volatile content. Mono- and sesquiterpenes accounted for about 98% of the total volatile mass in all cultivars. LVI-COC injection was used to determine the loss of carrot volatiles during concentration of headspace samples under a stream of nitrogen. The loss among major monoterpenes in the concentrated samples varied from 16% for p-cymene to >40% for alpha-pinene as compared to nonconcentrated samples. The loss among high-boiling sesquiterpenes varied from not detectable (beta-caryophyllene, alpha-humulene, and caryophyllene oxide) to approximately 7% for (E)- and (Z)-gamma-bisabolene.     

Optimization of extraction of apple aroma by dynamic headspace and influence of saliva on extraction of volatiles

The dynamic headspace procedure of aroma extraction was optimized on Gala apples (Malus domestica). Two parameters affecting the extractability of compounds were studied: temperature and purge time. The influence of artificial saliva was also included. An increase in purge time and temperature caused an increase in the extraction of volatiles from the apple matrix. The optimum point of extraction was 40 degrees C and 70 min of purge. The study also showed that the addition of saliva influenced the extraction of volatile compounds, but this effect was different from one compound to another. To verify that the headspace extracts presented a global odor representativeness of fresh apple under these conditions of extraction, eight assessors compared the odor of extracts with fresh fruit odor for three different cultivars. With regard to the sensory profiles of extracts, the optimal conditions of extraction were suitable for extraction of volatile compounds, even if cooked apple odor appeared in some extracts. The similarity marks of extracts were low but acceptable.