Vinegar classification based on feature extraction and selection from headspace solid-phase microextraction/gas chromatography volatile analyses: a feasibility study

Headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography (GC) and multivariate data analysis were applied to classify different vinegar types (white and red, balsamic, sherry and cider vinegars) on the basis of their volatile composition. The collected chromatographic signals were analysed using the stepwise linear discriminant analysis (SLDA) method, thus simultaneously performing feature selection and classification. Several options, more or less restrictive according to the final number of considered categories, were explored in order to identify the one that afforded highest discrimination ability. The simplicity and effectiveness of the classification methodology proposed in the present study (all the samples were correctly classified and predicted by cross-validation) are promising and encourage the feasibility of using a similar strategy to evaluate the quality and origin of vinegar samples in a reliable, fast, reproducible and cost-efficient way in routine applications. The high quality results obtained were even more remarkable considering the reduced number of discriminant variables finally selected by the stepwise procedure. The use of only 14 peaks enabled differentiation between cider, balsamic, sherry and wine vinegars, whereas only 3 variables were selected to discriminate between red (RW) and white wine (WW) vinegars. The subsequent identification by gas chromatography-mass spectrometry (GC-MS) of the volatile compounds associated with the discriminant peaks selected in the classification process served to interpret their chemical significance.     

Varietal flavour compounds of four grape varieties producing Madeira wines

Boal, Malvasia, Sercial and Verdelho are the main white grape varieties used in Madeira wine production. To estimate the free fraction of varietal aroma compounds of these varieties, 39 samples of musts were analysed to determine their content of monoterpenols and C₁₃ norisoprenoids (terpenoids), using dynamic headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The r-values for linearity studies of the analytical method used, varied between 0.977 (nerolidol) and 0.999 (linalool). The repeatability for each compound varied between 2.5% (citronellol) and 11.8% (β-ionone).The mean values from three vintages (1998, 1999 and 2000) confirmed that these musts have differentiated contents of terpenoids. In opposition to Verdelho musts, Malvasia showed the highest free terpenoids content. In order to establish relations between the compounds and the varieties under investigation, principal component analysis and linear discriminant analysis were applied to the data, revealing a good separation and classification power between the four groups as a function of varietal origin.     

Headspace solid-phase microextraction for the determination of volatile organic sulphur and selenium compounds in beers,wines and spirits using gas chromatography and atomic emission detection

A rapid and solvent-free method for the determination of eight volatile organic sulphur and two selenium compounds in different beverage samples using headspace solid-phase microextraction and gas chromatography with atomic emission detection has been developed. The bonded carboxen/polydimethylsiloxane fiber was the most suitable for preconcentrating the analytes from the headspace of the sample solution. Volumes of 20 mL of undiluted beer were used while, in the case of wines and spirits, sample:water ratios of 5:15 and 2:18, respectively, were used, in order to obtain the maximum sensitivity. Quantitation was carried out by using synthetic matrices of beer and wine, and a spiked sample for spirits, and using ethyl methyl sulphide and isopropyl disulphide as internal standards. Detection limits ranged from 8 ng L⁻¹ to 40 ng mL⁻¹, depending on the compound and the beverage sample analyzed, with a fiber time exposure of 20 min at ambient temperature. The optimized method was successfully applied to different samples, some of the studied compounds being detected at concentration levels in the 0.04–152 ng mL⁻¹ range.     

Ultrasound-assisted extraction of volatile compounds from wine samples: optimisation of the method

This study describes a liquid–liquid extraction technique for extracting volatile compounds from wine using dichloromethane and ultrasounds. This technique permits the simultaneous extraction of different samples with high reproducibility. After the preliminary tests, several parameters (sample volume, solvent volume and extraction time) were optimised using a factorial design to obtain the most relevant variables. The analytical characteristics were obtained such as calibration graphs, detection limits ranging from 0.0238 mg L⁻¹ for 1-pentanol to 0.261 mg L⁻¹ for octanoic acid, quantification limits and relative standard deviation from 2.1 to 6.2%. Extraction yields were calculated giving a range 9.16–1.2%. The optimised conditions were applied to the extraction of samples of young wines from the Denominación de Origen Calificada Rioja category using gas chromatography and a flame ionisation detector.     

Multivariate analysis for the classification and differentiation of Madeira wines according to main grape varieties

In order to differentiate and characterize Madeira wines according to main grape varieties, the volatile composition (higher alcohols, fatty acids, ethyl esters and carbonyl compounds) was determined for 36 monovarietal Madeira wine samples elaborated from Boal, Malvazia, Sercial and Verdelho white grape varieties. The study was carried out by headspace solid-phase microextraction technique (HS-SPME), in dynamic mode, coupled with gas chromatography–mass spectrometry (GC–MS). Corrected peak area data for 42 analytes from the above mentioned chemical groups was used for statistical purposes. Principal component analysis (PCA) was applied in order to determine the main sources of variability present in the data sets and to establish the relation between samples (objects) and volatile compounds (variables). The data obtained by GC–MS shows that the most important contributions to the differentiation of Boal wines are benzyl alcohol and (E)-hex-3-en-1-ol. Ethyl octadecanoate, (Z)-hex-3-en-1-ol and benzoic acid are the major contributions in Malvazia wines and 2-methylpropan-1-ol is associated to Sercial wines. Verdelho wines are most correlated with 5-(ethoxymethyl)-furfural, nonanone and cis-9-ethyldecenoate. A 96.4% of prediction ability was obtained by the application of stepwise linear discriminant analysis (SLDA) using the 19 variables that maximise the variance of the initial data set.     

Gas chromatography-mass spectrometry analysis of volatile compounds from Houttuynia cordata Thunb after extraction by solid-phase microextraction, flash evaporation and steam distillation

Various sampling techniques including flash evaporation (FE), headspace solid-phase microextraction (HS-SPME) and steam distillation (SD) were compared for the gas chromatography-mass spectrometry of volatile constituents present in Houttuynia cordata Thunb (HCT). 2-Undecanone (22.21%) and houttuynum (7.23%) were predominant components of HCT samples obtained by HS-SPME whereas those levels were 3.95 and 3.60% in the same samples by FE and 25.93 and 6.60% in those by SD, respectively. SPME with polydimethylsiloxane (PDMS) fibre was more selective and particularly efficient for the isolation of biologically active compounds and afforded a higher yield of total compounds than FE and SD. A total of 60 compounds were detected in SPME extracts. While in FE and SD extracts, the detected compounds were 41 and 51, respectively. The total amount of compounds isolated by SPME was much larger than that isolated by FE or SD. Some minor constituents were isolated by SPME, but not by SD and FE. This carries great significance because of the importance of the oil volatiles to clinical therapy. HS-SPME is a powerful tool for determining the volatile constitutes present in the TCMs.     

Optimisation of stir bar sorptive extraction for the analysis of volatile phenols in wines

An easy, fast and reliable analytical method is proposed for the determination of the concentration of volatile phenols (ethyl- and vinylphenols) in wines. The novel stir bar sorptive extraction (SBSE) technique is employed, following a simple and fast procedure that allows 15 samples to be extracted simultaneously using very small sample volume. Extracts are desorbed in a thermodesorption system (TDS) coupled on-line to a gas chromatograph-mass spectrometry system. The SBSE offers better recovery and linear regression coefficient (r2) for the four volatile phenols than solid-phase extraction (SPE). The mass spectrometric detection in selected ion monitoring mode contributes to the lower detection limit and good sensibility obtained with this method.     

Headspace solid-phase micro-extraction gas chromatography-mass detection method for the determination of butyltin compounds in wines

Butyltin compounds are widespread contaminants which have also been found in some wines, determined by liquid-liquid extraction followed by alkylation with a Grignard reagent and gas chromatography-mass spectrometric (GC-MS) analysis. A promising alternative to this extraction/derivatization method is the one-step tetraethylborate in situ ethylation/solid-phase micro-extraction (SPME) method. In this work, a SPME-GC-MS method for the determination of butyltin compounds in wine was optimised. The optimised parameters concerned the pre-treatment with tetramethylammonium hydroxide, matrix modification with sodium chloride, tetraethylborate concentration, extraction time and temperature, and the GC separation program. The analytical figures of merit of the optimised method (range, limit of detection (LOD) and reproducibility) were evaluated. The sensitivity (range 20-1421 kcounts μg⁻¹ l⁻¹ as Sn) and LOD (range, 0.01-0.2 μg l⁻¹ as Sn) depended greatly on the butyltin species to be measured and on the type of wine. For the tested species (monobutyltin, dibutyltin and tributyltin) the highest sensitivities were achieved for Port wine samples, followed by red wine>white wine>white Verde wine. The method allowed acceptable repeatability (relative standard deviation (R.S.D.), 6-8%; n=4) and reproducibility (R.S.D., 8-9%; n=3).     

Volatile flavour constituent patterns of Terras Madeirenses red wines extracted by dynamic headspace solid-phase microextraction

A suitable analytical procedure based on static headspace solid-phase microextraction (SPME) followed by thermal desorption gas chromatography-ion trap mass spectrometry detection (GC-(ITD)MS), was developed and applied for the qualitative and semi-quantitative analysis of volatile components of Portuguese Terras Madeirenses red wines. The headspace SPME method was optimised in terms of fibre coating, extraction time, and extraction temperature. The performance of three commercially available SPME fibres, viz. 100 mum polydimethylsiloxane; 85 mum polyacrylate, PA; and 50/30 mum divinylbenzene/carboxen on polydimethylsiloxane, was evaluated and compared. The highest amounts extracted, in terms of the maximum signal recorded for the total volatile composition, were obtained with a PA coating fibre at 30 degrees C during an extraction time of 60 min with a constant stirring at 750 rpm, after saturation of the sample with NaCl (30%, w/v). More than sixty volatile compounds, belonging to different biosynthetic pathways, have been identified, including fatty acid ethyl esters, higher alcohols, fatty acids, higher alcohol acetates, isoamyl esters, carbonyl compounds, and monoterpenols/C(13)-norisoprenoids.     

Analysis of volatile secondary metabolites from Colombian Xylopia aromatica (Lamarck) by different extraction and headspace methods and gas chromatography

Hydrodistillation (HD), simultaneous distillation-solvent extraction (SDE), microwave-assisted hydrodistillation (MWHD), and supercritical fluid (CO2) extraction (SFE), were employed to isolate volatile secondary metabolites from Colombian Xylopia aromatica (Lamarck) fruits. Static headspace (S-HS), simultaneous purge and trap (P&T) in solvent (CH2Cl2), and headspace (HS) solid-phase microextraction (SPME) were utilised to obtain volatile fractions from fruits of X. aromatica trees, which grow wild in Central and South America, and are abundant in Colombia. Kováts indices, mass spectra or standard compounds, were used to identify more than 50 individual components in the various volatile fractions. beta-Phellandrene was the main component found in the HD and MWHD essential oils, SDE and SFE extracts (61, 65, 57, and ca. 40%, respectively), followed by beta-myrcene (9.1, 9.3, 8.2 and 5.1%), and alpha-pinene (8.1, 7.3, 8.1 and 5.9%). The main components present in the volatile fractions of the X. aromatica fruits, isolated by S-HS, P&T and HS-SPME were beta-phellandrene (53.8, 35.7 and 39%), beta-myrcene (13.3, 12.3 and 10.1%), p-mentha-1(7),8-diene (7.1, 10.6 and 10.4%), alpha-phellandrene (2.2, 5.0 and 6.4%), and p-cymene (2.2,4.7 and 4.4%), respectively.     

The use of headspace solid phase microextraction for the characterization of volatile compounds in olive oil matrices

Two different fibre coatings, for solid phase microextraction (SPME) sampling, poly(dimethylsiloxane) (PDMS) and poly(acrylate) (PA), were studied in order to test, for olive oil matrixes, two mathematical models that relate the directly proportional relationship between the amount of analyte absorbed by a SPME fibre and its initial concentration in the sample matrices. Although the PA fibre was able to absorb higher amounts of compounds from the olive oil sample, the equilibrium was reached later then with the PDMS fibre. In both cases, the amount of analyte present affected the time profile or the equilibrium time in two of the concentrations studied, 0.256 μL/kg, 2.56 μL/kg and for 2-ethylfuran, pentan-3-one, pent-1-en-3-one, hexanal, trans,trans-non-2,4-dienal and in the four concentrations studied, 0.256 μL/kg, 2.56 μL/kg, 6.25 μL/kg and 400 μL/kg, for 4-methyl-pent-3-en-2-one, 2-methylbutan-1-ol, methoxybenzene, hexan-1-ol, cis-hex-3-en-1-ol, trans-hex-2-en-1-ol, 2-ethyl-hexan-1-ol and trans,trans-dec-2,4-dienal. Comparing the mathematical models of both fibres, the PA-coated fibre showed direct proportionality between the initial concentration and amount extracted, that allows the possibility of relative quantification in a non-equilibrium state in non-aqueous media. The same was not observed for the PDMS fibre.     

Cooled internal reflection element for infrared chemical sensing of volatile to semi-volatile organic compounds in the headspace of aqueous solutions

In this study, the cooling effect was applied to an evanescent wave type infrared (IR) chemical sensing method to effectively trap volatile organic compounds (VOCs), which have been absorbed in the hydrophobic film coated around the internal reflection element (IRE). The detection of VOCs in aqueous solutions was taken in the headspace of the aqueous solution. This method eliminates the long-term instability of hydrophobic film soaked in an aqueous solution and the potential spectral interference caused by the matrix of the aqueous solution. Thermal energy has been applied to the aqueous solution to assist in the evaporation of VOCs out of the aqueous matrix. By applying a cooling system to the IRE, the excess thermal energy can be removed leading to more stable IR signals. After examination of organic compounds with vapour pressure (P_v) ranging from 0.017 to 150 Torr, significant differences were found between IR signals from cooled and un-cooled systems. Because the thermal conductivity of the IRE used in IR detection is typically low; the efficiency in removing the thermal energy is limited. By heating the aqueous solutions to different temperatures, the IR signals showed that the sample temperature was limited to around 80 °C. The IR signal determination results for five different volatility organic compounds indicated that the optimal heating temperature was not necessary to match with the volatilities of organic compounds in cooling system. The linear regression coefficient (R²) of the standard curve for sample concentrations in the range 5-200 μg ml⁻¹ was generally higher than 0.991 and the detection limit was around a few hundred ng ml⁻¹, which was two to three times lower than that of un-cooled system.     

Elimination of matrix effects for headspace solid-phase microextraction of important volatile compounds in red wine using a novel coating

In this paper, hydroxy-terminated silicone oil-butyl methacrylate-divinylbenzene (OH-TSO-BMA-DVB) copolymer was first synthesized and used as stationary phase with the aid of γ-methacryloxypropyltrimethoxylsilane (KH-570) as bridge in solid-phase microextraction (SPME) using sol–gel method and cross-linking technique. It has high extraction efficiency for both polar alcohols and fatty acids and nonpolar esters in comparison with commercial PDMS, PDMS-DVB and PA fibers. A simple and sensitive headspace SPME-gas chromatography (HS-SPME-GC) method using the novel fiber was presented for the simultaneous analysis of both polar alcohols and fatty acids and nonpolar esters in wine. To check the matrix effects, various model wine matrices, including distilled water; 11.5% ethanol/water (v/v) solution; a concentrated synthetic wine; a ‘volatile-free’ wine and a real wine were investigated in detail. Matrix effects were compensated for by using internal standard method and selecting the ‘volatile-free’ wine as working standard. The method presented in this study showed satisfactory linearity, precision, detection limits and accuracy. The recoveries obtained ranged from 85.87 to 104.2%, and the relative standard deviation values were below 9%. The results obtained indicated that the present method is a validated and accurate procedure for the simultaneous determination of both polar and nonpolar aroma compounds in wine.     

Direct thermal extraction and gas chromatographic-mass spectrometric determination of volatile compounds of extra-virgin olive oils

The instrumental performances of a Thermo Desorption-Cooled Injection System coupled with a gas chromatography-mass spectrometer (GC-MS) were improved by a Plackett-Burman experimental design for the direct thermal extraction of volatile compounds from extra-virgin olive oils. The obtained experimental conditions were applied to the analysis of samples from West Liguria (cv. Taggiasca > or = 90%) and Spain (cv. Arbequina), which shared such similar sensorial features that Taste Panel did not distinguish them. Principal component analysis (PCA) was then applied to the experimental data. Three linear combinations of the amounts of the lipoxygenase oxidation products proved to be decisive and sufficient in the differentiation of the two groups of samples.     

A useful approach for the differentiation of wines according to geographical origin based on global volatile patterns

In this study, the feasibility of solid‐phase extraction combined with gas chromatography and mass spectrometry in tandem with partial least squares discriminant analysis was evaluated as a useful strategy to differentiate wines according to geographical origin (Azores, Canary and Madeira Islands) and types (white, red and fortified wine) based on their global volatile patterns. For this purpose, 34 monovarietal wines from these three wine grape‐growing regions were investigated, combining the high throughput extraction efficiency of the solid‐phase extraction procedure with the separation and identification ability. The partial least squares discriminant analysis results suggested that Madeira wines could be clearly discriminated from Azores and Canary wines. Madeira wines are mainly characterized by 2‐ethylhexan‐1‐ol, 3,5,5‐trimethylhexan‐1‐ol, ethyl 2‐methylbutanoate, ethyl dl ‐2‐hydroxycaproate, decanoic acid, 3‐methylbutanoic acid, and (E)‐whiskey lactone, whereas 3‐ethoxypropan‐1‐ol, 1‐octen‐3‐ol, (Z)‐3‐hexenyl butanoate, 4‐(methylthio)‐1‐butanol, ethyl 3‐hydroxybutanoate, isoamyl lactate, 4‐methylphenol, γ‐octalactone and 4‐(methylthio)‐1‐butanol, are mainly associated with Azores and Canary wines. The data obtained in this study revealed that solid‐phase extraction combined with gas chromatography and quadrupole mass spectrometry data and partial least squares discriminant analysis provides a suitable tool to discriminate wines, both in terms of geographical origin as well as wine type and vintage.     

Volatile compounds as markers of ageing in Tempranillo red wines from La Mancha D.O. stored in oak wood barrels

Solid-phase extraction cartridges (SPE)-GC/MS method was used to analyse red wines aromas. The matrix effect was studied with chemicals standard prepared in synthetic wines with water/alcohol solutions (12% ethanol, v/v) following the procedure proposed. The method offers good reproducibility since the relative standard deviations (RSD%) for the volatile compounds levels were less than 9%. This method was used to differentiate the aroma of one hundred mono-varietal young, crianza, reserva and gran reserva La Mancha D.O. wines (cv. Tempranillo) on the basis of oak barrel contact period. Samples were checked at ten time points over 36 months. Sixty important wine odorants, such as volatile phenols, vanillin derivatives, lactones, norisoprenoids, benzene compounds, esters and terpenols, can be quantitatively determined in a single run. Results showed significant quantitative differences in the volatile profiles of wines depending on the length of time in contact with wood, even in wines belonging to the same commercial category. Stepwise multiple linear regression (SMLR) was used to obtain a model that predicted the time of barrel ageing to which wines were submitted in relation with the wine volatile composition. A successful function based on eight compounds with a mean deviation of 0.37 months in the predictions, was obtained.     

Study of the effect of different fiber coatings and extraction conditions on dry cured ham volatile compounds extracted by solid-phase microextraction (SPME)

Extraction of dry cured ham volatile compounds by solid-phase microextraction (SPME) was optimized. Different fiber coatings (carboxen/polydimethylsiloxane (CAR/PDMS), divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS), polydimethylsiloxane (PDMS), polydimethylsiloxane/divinylbenzene (PDMS/DVB)), times of extraction (15, 30, 60 min) and sample preparation (ground samples and homogenates with NaCl saturated solution) were assayed. CAR/PDMS and DVB/CAR/PDMS fiber coatings extracted more than 100 volatile compounds and showed the highest area counts for most volatile compounds. CAR/PDMS coating extracted better those compounds whose Kovats index (KI) was lower than 980 (on average) and DVB/CAR/PDMS those with higher KI. Fifteen minutes of extraction provided a volatile compound profile with lower area counts for most compounds and qualitatively different to that obtained with 30 and 60 min of extraction. Homogenates gave a different profile compared to ground samples, with lower total counts for most compounds but higher proportion of aldehydes, and presence of several compounds not found in ground samples.     

Optimisation of stir bar sorptive extraction and liquid desorption combined with large volume injection-gas chromatography-quadrupole mass spectrometry for the determination of volatile compounds in wines

Stir bar sorptive extraction and liquid desorption followed by large volume injection coupled to gas chromatography-quadrupole mass spectrometry (SBSE-LD/LVI-GC-qMS) had been applied for the determination of volatiles in wines. The methodology was optimised in terms of extraction time and influence of ethanol in the matrix; LD conditions, and instrumental settings. The optimisation was carried out by using 10 standards representative of the main chemical families of wine, i.e. guaiazulene, E,E-farnesol, β-ionone, geranylacetone, ethyl decanoate, β-citronellol, 2-phenylethanol, linalool, hexyl acetate and hexanol. The methodology shows good linearity over the concentration range tested, with correlation coefficients higher than 0.9821, a good reproducibility was attained (8.9-17.8%), and low detection limits were achieved for nine volatile compounds (0.05-9.09 μg L⁻¹), with the exception of 2-phenylethanol due to low recovery by SBSE. The analytical ability of the SBSE-LD/LVI-GC-qMS methodology was tested in real matrices, such as sparkling and table wines using analytical curves prepared by using the 10 standards where each one was applied to quantify the structurally related compounds. This methodology allowed, in a single run, the quantification of 67 wine volatiles at levels lower than their respective olfactory thresholds. The proposed methodology demonstrated to be easy to work-up, reliable, sensitive and with low sample requirement to monitor the volatile fraction of wine.     

Ultrasonic nebulization extraction coupled with headspace single-drop microextraction of volatile and semivolatile compounds from the seed of Cuminum cyminum L

Ultrasonic nebulization extraction (UNE) coupled with headspace single-drop microextraction (HS-SDME) was developed. In the UNE process, the analytes were transferred from the aqueous phase to the gas phase. Then the analytes were transferred from the gas phase to the solvent phase by the carrier gas and extracted and enriched with suspended microdrop solvent. Finally, the microdrop solvent injected into GC-MS system. The parameters affecting extraction performance, such as type of suspended solvent, microdrop volume, flow rate of carrier gas, temperature of extraction vessel and extraction time were investigated and optimized. The proposed method can be applied for the extraction and enrichment of the volatile and semivolatile compounds simultaneously. The extraction efficiency of the proposed method was compared with that of ultrasonic extraction (UE) and UE-HS-SDME. Compared with UE-HS-SDME, the contents of constituents in the extract obtained by the proposed method were closer to those obtained by hydrodistillation (HD), which is a standard extraction method.     

Comparison of different extraction methods for the analysis of volatile secondary metabolites of Lippia alba (Mill.) N.E. Brown, grown in Colombia, and evaluation of its in vitro antioxidant activity

Hydrodistillation (HD), simultaneous distillation solvent extraction (SDE), microwave-assisted hydrodistillation (MWHD), and supercritical fluid (CO2) extraction (SFE) were employed to isolate volatile secondary metabolites from fresh leaves and stems of Colombian Lippia alba (Mill.) N.E. Brown. Kovàts indices, mass spectra or standard compounds were used to identify around 40 components in the various volatile fractions. Carvone (40-57%) was the most abundant component, followed by limonene (24-37%), bicyclosesquiphellandrene (5-22%), piperitenone (1-2%), piperitone (ca. 1.0%), and beta-bourbonene (0.6-1.5%), in the HD, SDE, MWHD, and SFE volatile fractions. Static headspace (S-HS), simultaneous purge and trap in solvent (CH2Cl2) (P&T), and headspace solid-phase microextraction (HS-SPME) were used to sample volatiles from fresh L. alba stems and leaves. The main components isolated from the headspace of the fresh plant material were limonene (27-77%), carvone (14-30%), piperitone (0.3-0.5%), piperitenone (ca. 0.4%), and beta-bourbonene (0.5-6.5%). The in vitro antioxidant activity of L. alba essential oil, obtained by hydrodistillation was evaluated by determination of hexanal, the main carbonyl compound released by linoleic acid subjected to peroxidation (1 mm Fe2+, 37 degrees C, 12 h), and by quantification of this acid as its methyl ester. Under the same conditions, L. alba HD-essential oil and Vitamin E exhibited similar antioxidant effects.