Headspace solid-phase microextraction gas chromatography-mass spectrometry analysis of volatiles in orujo spirits from a defined geographical origin

A headspace solid-phase microextraction (HS-SPME) and gas chromatography-selective ion monitoring/mass spectrometry (GC-SIM/MS) method was optimized for analysis of 22 volatile compounds in orujo spirit samples from the Geographic Denomination "Orujo de Galicia/Augardente de Galicia". HS-SPME experimental conditions, such as fiber coating, extraction temperature, extraction and pre-equilibrium time, sample volume, and the presence of salt, were studied to improve the extraction process. The best results were obtained using a 65 microm Carbowax-divinylbenzene fiber during a headspace extraction at 40 degrees C with constant magnetic stirring for 15 min and after a 5 min period of pre-equilibrium time. The sample volume was 6 mL of orujo containing 25% of NaCl, placed in 12 mL glass vials equipped with a screw cap and PTFE/silicone septum. Desorption was performed directly in the gas chromatograph injector port for 5 min at 250 degrees C using the splitless mode. The proposed method is sensible (with detection limits between 0.0045 and 0.2399 mg/L), precise (with coefficients of variation in the range 0.99-8.18%), and linear over more than 1 order of magnitude. The developed method presented recoveries comprised between 76.0 and 112.4%. The applicability of the new method was demonstrated by determining the considered 22 volatile compounds in nine orujo commercial samples with quality and origin brands.     

Monitoring toasting intensity of barrels by chromatographic analysis of volatile compounds from toasted oak wood.

Toasting changes both the quantity and the quality of the extractable substances in the oak wood of barrels used for the aging of fine wines and spirits. Mastery and repeatability of toasting are vital in the production of quality barrels to be used for the aging of wines and spirits. In this study, we show that it is possible for a given cooperage to differentiate barrel toasting levels by analyzing a certain number of volatile and semivolatile compounds resulting from the thermal degradation of oak. Toasted wood components, which can be extracted by the wines or spirits during the aging process, are normally analyzed after the wood has soaked in standard alcoholic solutions and can be used to control the intensity of the wood toasting. The results of the comparative analysis presented in this work show that headspace analysis with a microextractive method using a stationary polydimethylsiloxane type phase is a promising technique for analyzing toasted oak wood from barrels. It is easier to use than the traditional maceration and extraction method and provides similar information.     

Characterization of volatiles in extracts from oak chips obtained by accelerated solvent extraction (ASE)

Sixteen commercially available oak chips, differing in origin (French or American) and toasting level, were extracted by an accelerated solvent extraction method and characterized by their volatile composition. About 80 compounds were identified and quantified, a great part of them from the thermodegradation of lignin and cellulose. One furanone (solerone) and two C-13 norisoprenoids (3-oxo-alpha-ionol and a 3-oxoretro-alpha-ionol isomer) were also tentatively identified and reported for the first time in toasted wood. Quantitative data demonstrated the oak chips to be not so different from the composition of light- or medium-toasted wood barrels, which was reported by other authors. The same data suggest that toasting level had the strongest influence on the volatile composition of chip samples. Phenyl ketones, volatile phenols, and some furanic compounds were the most influenced. On the other hand, the influence of wood origin was found to be weaker.     

Optimization of multiple headspace solid-phase microextraction for the quantification of volatile compounds in dry fermented sausages

Multiple headspace solid-phase microextraction (HS-SPME) is a stepwise method that eliminates the influence of the matrix sample on the quantitative analysis of solid samples. The process was optimized for the analysis of volatile compounds in dry fermented sausages by gas chromatography and mass spectrometry. Different amounts of fermented sausages and different vial volumes were studied to obtain the theoretical exponential decay of the peak area of the four successive extractions in order to calculate the total area in the sausage. The highest number of volatile compounds analyzed by multiple HS-SPME in dry fermented sausages was obtained in a 10 mL headspace vial with 0.07 g of sample in the presence of water, 0.75 mg butylated hydroxytoluene, and 0.5 g sodium chloride. Finally, the method was characterized in terms of linearity and detection limits and applied to analyze the volatile compounds present in fermented sausages manufactured with either nitrate or nitrite.     

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.     

Using electronic odor sensors to discriminate among oak barrel toasting levels.

Toasting changes both the quantity and the quality of the extractable substances in oak wood of barrels used for the aging of fine wines and spirits. Mastery and repeatability of toasting are vital in the production of quality barrels to be used for aging wines and spirits. Toasted wood components, which can be extracted by the wines or spirits during the aging process, are normally analyzed by maceration in standard alcohol solutions at concentrations adapted to the various products and can be used to control the intensity of the wood toasting. These kinds of analyses are accurate but time-consuming and need specialized laboratories. In this work, the feasibility of monitoring barrel toasting levels using an electronic nose with a metal oxide odor sensor array (MOS) was studied. The results of oak toasting level differentiation obtained via the MOS network were identical to those obtained by analyzing extractable compounds in liquid or gas phase as described in a previous paper. The results presented in this work at the laboratory scale could be used to implement a nondestructive monitoring system based on the analysis of headspace of barrels under industrial conditions.     

Characterization of aroma compounds in apple cider using solvent-assisted flavor evaporation and headspace solid-phase microextraction

The aroma-active compounds in two apple ciders were identified using gas chromatography-olfactometry (GC-O) and GC-mass spectrometry (MS) techniques. The volatile compounds were extracted using solvent-assisted flavor evaporation (SAFE) and headspace solid-phase microextraction (HS-SPME). On the basis of odor intensity, the most important aroma compounds in the two apple cider samples were 2-phenylethanol, butanoic acid, octanoic acid, 2-methylbutanoic acid, 2-phenylethyl acetate, ethyl 2-methylbutanoate, ethyl butanoate, ethyl hexanoate, 4-ethylguaiacol, eugenol, and 4-vinylphenol. Sulfur-containing compounds, terpene derivatives, and lactones were also detected in ciders. Although most of the aroma compounds were common in both ciders, the aroma intensities were different. Comparison of extraction techniques showed that the SAFE technique had a higher recovery for acids and hydroxy-containing compounds, whereas the HS-SPME technique had a higher recovery for esters and highly volatile compounds.     

Determination of 2-methylisoborneol and geosmin produced by Streptomyces sp. and Anabaena PCC7120

A new sample preparation and enrichment technique, headspace liquid-phase microextraction (HS-LPME) linked to gas chromatography-mass spectrometry (GC-MS), was developed for the determination of the off-flavor odorants, 2-methylisoborneol and geosmin, produced by Streptomyces sp. and Anabaena PCC7120. Some of the factors that influence the extraction efficiency of HS-LPME, such as the type of extraction solvent, ionic strength of sample solution, and sample agitation rate, were studied and optimized by a single factor test. Other factors, including extraction temperature, extraction time, microdrop volume, and headspace volume were optimized by orthogonal array design. Extraction of 2-methylisoborneol and geosmin was conducted by exposing 2.5 microL of 1-hexanol for 9 min at 50 degrees C in the headspace of a 20 mL vial with a 10 mL of sample solution saturated by NaCl and stirred at 800 rpm. The developed protocol demonstrated good repeatability (relative standard deviations (RSDs) < 5%), wide linear ranges (10-5000 ng/L, r2 > 0.999), and low limits of detection (LODs) for 2-methylisoborneol and geosmin (0.05 ng/L for both analytes). Subsequently, the method was successfully applied to extract the analytes in bacterial cultures with high recoveries (from 94% to 98%). Compared with headspace solid-phase microextraction (HS-SPME), HS-LPME demonstrates better linearity, precision, and recovery. Importantly, the sensitivity is about 1 order of magnitude higher than that of most HS-SPME. The results showed that HS-LPME coupled with GC-MS is a simple, convenient, rapid, sensitive, and effective method for the qualitative and quantitative analysis of 2-methylisoborneol and geosmin.     

Immunochemical analysis of 2,4,6-tribromophenol for assessment of wood contamination

2,4,6-Tribromophenol (2,4,6-TBP) has been used as a wood preservative and flame retardant and is a synthetic intermediate of the most important brominated flame retardants (BFR) produced. The use of TBP-contaminated wood materials in the food industry poses a risk of significant economical losses due to food contamination. In this work an efficient and reliable immunochemical method for analysis of TBP in wood samples has been established consisting of alkaline wood extraction followed by analysis on a microplate ELISA (enzyme-linked immunosorbent assay). TBP is efficiently extracted from wood samples in 10 min and directly measured after 10-fold buffer dilution to avoid matrix interferences. The analytical procedure has a limit of detection of 45 ng g (-1) of TBP in wood (1.5 microg L (-1) in extracts). The method has been applied to the analysis of contaminated real wood samples, showing that the levels of contamination can reach high TBP concentrations (up to 2000 microg L (-1)). An excellent correlation was observed between TBP levels in wood extracts determined by ELISA and gas chromatography-mass spectrometry (GC-MS) analysis ( R (2) = 0.990, N = 19). The precision found is below 22% CV. The immunoanalytical method developed is fast, reliable, and cost-effective, shows good high-throughput screening capabilities, and can be an excellent tool for assessment of wood contamination at lumber mills or related industries. The TBP ELISA has the potential to be used for environmental, food, and biological monitoring of brominated phenolic compounds considered nowadays as emerging pollutants.     

Improved malonaldehyde assay using headspace solid-phase microextraction and its application to the measurement of the antioxidant activity of phytochemicals

A modified malonaldehyde (MA) assay for antioxidant activity, which involves derivatization and headspace solid-phase microextraction (HS-SPME) was developed and validated. The recovery of MA as 1-methylpyrazole (product of MA and N-methylhydrazine) from a headspace of an aqueous solution containing MA, buffer, surfactant, and cod liver oil using HS-SPME with a PDMS/DVB fiber was 91.3 +/- 3.38%. MA was analyzed by a gas chromatograph with a nitrogen-phosphorus detector, and its detection limit was 0.0103 nmol/mL. The antioxidant activities of natural compounds were determined as the percentage inhibition of MA formed from cod liver oil oxidized by Fenton's reagents in the above aqueous solution. Sesamol inhibited MA formation most (86.1%), followed by eugenol (84.4%), capsaicin (80.7%), ethylvanillin (45.3%), and vanillin (31.6%) at a level of 50 microg/mL. This method did not require any organic solvents and is a simple, fast, and a highly sensitive method for MA determination.     

Influence of oak wood on the aromatic composition and quality of wines with different tannin contents

The aromatic compounds associated with aging in wood have been studied in Monastrell wines containing different quantities of tannin. Analyses were carried out before and after a six month period in wooden barrels (225 L) of American and French (Allier) medium-toasted oak. Three classes of wine were made: one from a free-run must; one to which enological tannin was added; and one to which was added the wine obtained by pressing the grape pomaces. The aromas were determined by GC-MS, and the quality was evaluated by a group of expert tasters. The aromatic compounds, some of which came from the wines themselves and others which came from the wood, all increased in concentration by the end of the aging process. The ratio between cis- and trans-whiskylactone contents in wines reflected the different types of oak wood used. The note "vanilla" used by the tasters is probably not totally due to the presence of vanillin. The wines of best quality were those with a natural tannin content which had been aged in American oak, whereas the wines to which enological tannin had been added but aged in the same wood were considered the worst. An equalizing effect on the sensorial wines qualities was seen to result from the use of French oak. The furfural, eugenol, and cis-, and in some cases trans-, whiskylactone contents identified the wines from the different classes of wood. The wood from which the barrels were made had a greater effect on wines differentiation than the tannin content.     

Essential oil composition of sachalinmint from Norway detected by solid-phase microextraction and gas chromatography-mass spectrometry analysis

The essential oil of leaves and flowers of sachalinmint [Mentha sachalinensis (Briq.) Kud?] grown in Norway (Trondheim) has been studied by headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry analysis (GC-MS). The essential oil content increased linearly in acropetal direction from 1.08% (0-20 cm plant height) to 1.75% (60-80 cm; young leaves and flowers). The steam-distilled samples showed a minor complex matrix with a very high menthol and a much lower menthone content (87.89 and 4.05%, respectively). From testing of HS-SPME unequilibrated exposure times ranging from 10 s to 5 min, an extraction time of 30 s was found to be sufficient to detect both low- and high-eluting compounds. Comparison of HS-SPME and steam-distilled samples established that the same tendencies of increasing menthol/menthone content in the basipetal/acropetal direction could be detected by both analysis methods. With regard to the extraction efficiency, HS-SPME gave additional detailed information about less important terpenic compounds.     

Validation of a mass spectrometry method to quantify oak ellagitannins in wine samples

Detection and individual quantification of oak wood ellagitannins in oak barrel aged red wine samples are difficult mainly due to their low levels and the similarity between their structures. In this work, a quantification method using mass spectrometry has been developed and validated to quantify wine ellagitannins after sample fractionation with a previously reported method. The use of an internal standard is a requirement to correct mass signal variability. (-)-Gallocatechin, among the different tested compounds, was the only one that proved to be a suitable internal standard making possible the accurate and individual quantification of the main oak wood ellagitannins. The developed methodology has been used to detect and quantify these ellagitannins in different Spanish commercial wines, proving its usefulness.     

Volatile compounds in a spanish red wine aged in barrels made of Spanish, French, and American oak wood

A red Rioja wine was aged in barrels made of Spanish oak wood (Quercus robur, Quercus petraea,Quercus pyrenaica, and Quercus faginea) during 21 months. The concentrations of some volatile compounds [syringaldehyde, vanillin, eugenol, maltol, guaiacol, 4-ethylphenol, cis and trans isomers of beta-methyl-gamma-octalactone, 2-furfuraldehyde, 5-methyl-2-furfuraldehyde, 5-(hydroxymethyl)-2-furfuraldehyde, and furfuryl alcohol] were studied in these wines and compared with those of the same wine aged in barrels made from French oak of Q. robur (Limousin, France) and Q. petraea (Allier, France) and American oak of Quercus alba (Missouri). Similar concentrations of these compounds were found in wines aged in Spanish and French oak wood barrels, and significantly different concentrations were found with respect to wines aged in barrels made of American oak wood, indicating a different behavior. Thus, wines with different characteristics were obtained, depending on the kind of wood. Also, the kind of wood had an important influence on sensory characteristics of wine during the aging process. Spanish oak wood from Q. robur, Q. petraea, and Q. pyrenaica can be considered to be suitable for barrel production for quality wines, because a wine aged in barrels made of these Spanish oak woods showed similar and intermediate characteristics to those of the same wine aged in French and American oak woods usually used in cooperage.     

Rapid headspace gas chromatographic method for assessment of oxidative stability of cooked chicken meat

Experiments were conducted to determine the volatile compounds in cooked chicken meat using a static headspace gas chromatographic (GC) technique. Preheating conditions for samples in vials were tested at 70, 80, and 90 degrees C for 20-120 min at each temperature. The majority of the peaks increased in size as the temperature and time increased. Optimum conditions were established as preheating at 80 degrees C for 30 min followed by analysis on a packed column of 8% Poly MPE on Tenax GC with the temperature programmed from 50 to 200 degrees C at 10 degrees/min. Coefficients of variation for major peaks ranged from 8.3 to 14.7%. These results were compared with those obtained with a capillary column analysis of samples preheated at the same conditions. Cooked and stored chicken patties, pretreated with different levels of sodium tripolyphosphate, were analyzed by the thiobarbituric acid (TBA) method and the headspace GC technique. Significant positive correlations were obtained between TBA numbers and the areas of 3 major peaks of the headspace profiles, indicating the applicability of the rapid headspace GC method for the determination of oxidative changes in chicken meat.     

Assessment of strawberry aroma through SPME/GC and ANN methods. Classification and discrimination of varieties

To provide an efficient and running analytical tool to strawberry plant breeders who have to characterize and compare the aromatic properties of new cultivars to those already known, a HS-SPME/GC-MS analysis method has been coupled with a statistical treatment method issued from the current development of artificial neuron networks (ANN), and more specifically, the unsupervised learning systems called Kohonen self-organizing maps (SOMs). So, 70 strawberry samples harvested at CIREF from 17 known varieties have been extracted by using a DVB/Carboxen/PDMS SPME fiber according to the headspace procedure, and then chromatographed. A panel of 23 characteristic aromatic constituents has been selected according to published results relative to strawberry aroma. The complex resulting matrix, collecting the relative abundance of the 23 selected constituents for each sample, has been input into the SOM software adapted and optimized from the Kohonen approach described by one of the authors. After a period of training, the self-organized system affords a map of virtual strawberries to which real samples are compared and plotted in the best matching unit (BMU) of the map. The efficiency for discriminating the real samples according to their variety is dependent on the number of units selected to define the map. In this case, a 24-unit map allowed the complete discrimination of the 17 selected varieties. Moreover, to test the validity of this approach, two additional samples were blind-analyzed and the results were computed according to the same procedure. At the end of this treatment, both samples were plotted into the same unit as those of the same variety used for training the map.     

Different sorption behaviors for wine polyphenols in contact with oak wood

The evolution of polyphenols of enological interest- monomeric anthocyanins, (+)-catechin, (-)-epicatechin, gallic acid, and trans-resveratrol-in the presence of oak wood was investigated in aging-model conditions. Disappearance kinetics showed that, except for gallic acid, all of the wine polyphenols tend to disappear from the model wine in presence of oak wood, to reach an equilibrium after 20 days of contact. At equilibrium, the higher disappearance rates were obtained for monomeric anthocyanins and trans-resveratrol with values of 20 and 50%, respectively. For monomeric anthocyanins, the rate of disappearance seemed to be independent of their nature. In order to evaluate the contribution of sorption to oak wood in the disappearance phenomena, sorption kinetics were determined for trans-resveratrol and malvidin-3-glucoside through the extraction and the quantification of the fraction sorbed to wood. These curves showed that the wood intake of trans-resveratrol and malvidin-3-glucoside followed a two-step behavior, with a higher rate during the first 2 days, likely due to a surface sorption mechanism, and then a slower rate to reach the equilibrium, which could be related to a diffusion mechanism. The comparison of disappeared and sorbed amounts at equilibrium showed that a minor part of the disappeared monomeric anthocyanins were sorbed by wood. In contrast, half of the concentration decrease of trans-resveratrol in wine finds its origin in a sorption mechanism by oak wood. Results in real wine show similar sorption kinetics.     

The effects of sample preparation and gas chromatograph injection techniques on the accuracy of measuring guaiacol, 4-methylguaiacol and other volatile oak compounds in oak extracts by stable isotope dilution analyses

The deuterium-labeled standards [(2)H(3)]-guaiacol and [(2)H(3)]-4-methylguaiacol were synthesized and utilized in a method employing gas chromatography-mass spectrometry to determine the concentration of guaiacol and 4-methylguaiacol in wine or extracts of oak shavings. The method was combined with previously published methods for 4-ethylphenol, 4-ethylguaiacol, cis- and trans-oak lactone and vanillin, so that all these compounds could be quantified in a single analysis. The method can employ either liquid-liquid extraction or headspace solid-phase microextraction (SPME) and is rapid, robust, precise, and accurate. Under certain conditions, there was artifactual generation, to varying degrees, of guaiacol, 4-methylguaiacol, cis-oak lactone, and vanillin during the analysis of oak extracts, especially when diethyl ether extraction and injector block temperatures at or above 225 degrees C were employed. The most substantial effects were observed for guaiacol, in which results could be exaggerated by over 10 times. These artifacts could be avoided by using headspace SPME or by preparing liquid-liquid extracts with pentane or pentane/diethyl ether (2:1) injected at 200 degrees C providing spot checks using headspace SPME were performed. Data obtained for previously published quantitative determination of guaiacol in oak extracts should be reexamined carefully, with special attention paid to their respective methods of sample preparation and analysis.     

Concurrent phenomena contributing to the formation of the aroma of wine during aging in oak wood: an analytical study

Red wine was stored in different oak barrels or in stainless steel, and samples were taken for two years to determine 79 aroma compounds. Aging in oak affects 41 compounds. The type of wood affects 11 compounds. At least seven different processes seem to take place concurrently in aroma evolution, and five such processes, affecting 37 compounds, are linked to the oak cask. These are extraction from the wood, oxidation of wine alcohols and amino acids, microbiological formation of ethyl phenols, sorption processes, and condensation of acetaldehyde with polyphenols. The wood can release linear gamma- and delta-lactones, beta-damascenone, and ionones. Some compounds are released very fast from wood, which suggests they lie in the external part of the wood. Some extraction profiles are too complex to be explained by physical processes. Finally, the levels of 2,5-dimethyl-4-hydroxy-3(2H)-furanone and 2-ethyl-5-methyl-4-hydroxy-3(2H)-furanone increase even in the reference wine, which suggests the presence of a precursor.     

Preliminary study of oaklins, a new class of brick-red catechinpyrylium pigments resulting from the reaction between catechin and wood aldehydes

Several structurally related pigments were found to result from the reaction between catechin and coniferaldehyde/sinapaldehyde extracted from oak wood. Their structures were tentatively identified by mass spectrometry, and their formation was studied in different pH and temperature conditions for several days. They were all found to have a characteristic catechinpyrylium core, thereby constituting a new class of compounds named as oaklins. One of the main oaklins was also detected in a commercial table red wine aged in oak barrels.