Solid-phase Microextraction Analysis Research Articles

Identification of the Sex Pheromone of the Asparagus Moth, Parahypopta Caestrum (Lepidoptera, Cossidae).

Chemical, electrophysiological, and field trapping experiments were carried out to identify the female-produced sex pheromone of the asparagus moth, Parahypopta caestrum, a very serious pests of asparagus cultivations in southern Europe. Gas chromatography coupled with mass spectrometry and electroantennogram detection (GC-MS-EAD) analysis of hexane and solid-phase microextraction (SPME) extracts of sex pheromone glands of calling females consistently detected four compounds eliciting EAG responses in male moth antennae. According to their GC retention times, mass spectra, and comparative EAG analyses with reference standards, these EAD-active compounds were identified as (Z)-9-tetradecenol (Z9-14:OH), (Z)-5-tetradecenyl acetate (Z5-14:Ac), (Z)-7-tetradecenyl acetate (Z7-14:Ac), and (Z)-9-tetradecenyl acetate (Z9-14:Ac), respectively. In the SPME extracts from the head-space of individual abdominal tips, Z9-14:Ac, Z5-14:Ac, Z7-14:Ac, and Z9:14 OH were detected in the ratio of 82:9:5:4. In EAG dose-response experiments, Z9-14:Ac was the strongest antennal stimulant at different doses tested. In field trapping experiments, Z9-14:Ac, Z7-14:Ac, and Z5-14:Ac proven to be essential for male attraction and a their 85:5:10 blend loaded onto green rubber septum dispensers was significantly more effective than single-, two-, and any other three-component blend of these compounds. The addition of Z9-14:OH to the optimal blend resulted in a significant reduction of male catches. The attractive blend here identified allowed for an effective and accurate monitoring of P. caestrum flight activity in southern Italy.

Chemical bonding of cross-linked glutaraldehyde/chitosan on the surface of a titanium wire to prepare a robust biocompatible SPME fiber for analysis of phytohormones in plants

A robust biocompatible solid-phase microextraction (SPME) fiber, so-called Ti/APTS/GA/CS, was prepared by chemical bonding of cross-linked glutaraldehyde-chitosan to the surface of a titanium wire using APTS. The fiber was applied for sampling of phytohormones in plant tissues, followed by HPLC-UV analysis. The structure and morphology of the fiber coating was investigated by FT-IR, SEM, EDX, XRD, and TGA techniques. A Box-Behnken design was implemented to optimize the experimental variables. The calibration graphs were linear over a wide linear range (0.5–200 μg L−1) with LODs over the range of 0.01–0.06 μg L−1. The intra-day and inter-day precisions were found to be 1.3–6.3% and 4.3–7.3%, respectively. The matrix effect values ranged from 86.5% to 111.7%, indicating that the complex sample matrices had an insignificant effect on the determination of phytohormones. The fiber was successfully employed for the direct-immersion SPME (DI-SPME-HPLC) analysis of the phytohormones in cucumber, tomato, date palm, and calendula samples.

Composition of the Scent in Some Ophrys Orchids Growing in Basilicata (Southern Italy): A Solid-Phase Microextraction Study Coupled with Gas Chromatography and Mass Spectrometry

Several methods have been used to determine the volatile organic compounds emitted by Ophrys orchids. The use of different methods results in incomparable data. Solid-phase microextraction (SPME) has not been used extensively on Ophrys orchids. The main components found in the SPME analysis of the scent in Ophrys orchids were as follows: O. apifera: benzyl benzoate and α-copaene; O. crabronifera subsp. biscutella: pentadecane, heptadecane, and nonadecane; O. bertolonii subsp. bertolonii: pentadecane and heptadecane; O. passionis subsp. garganica: i-propyl palmitate and heptadecane; O. holosericea subsp. apulica: α-copaene, pentadecane, and heptadecane; O. lacaitae: α-copaene, pentadecane, and heptadecane; O. bombyliflora: cyclosativene, pentadecane, and ethyl dodecanoate; O. insectifera: 8-heptadecene and pentadecane; O. lutea: heptadecane and docosane; O. tenthredinifera subsp. neglecta: α-copaene, caryophyllene, and i-propyl palmitate.

Analysis of Volatile Constituent by Hydrodistillation and Solid-Phase Microextraction Techniques and Antimicrobial and Scolicidal Activities of Essential Oil and Soxhlet Extracts of Ulva rigida grown in Turkey.

In the present study, the volatile composition of Ulva rigida (U. rigida) was elucidated by two different methods. As a result of the identification process of volatile components using the GC/MS-FID instrument, 31 compounds were identified by hydrodistillation (HD) method, and 15 compounds were identified by solid-phase microextraction (SPME) method, elucidating the structure of 99.86 % and 92.65 %, respectively. The most abundant compounds in the essential oil of U. rigida were n-hexadecanoic acid and pentadecanal, while the most abundant compound according to the SPME analysis was heptadecyne, a hydrocarbon compound. In the next step, hexane, dichloromethane, chloroform and methanol solvent extracts of U. rigida were prepared and the antimicrobial activities of the extracts and the essential oil obtained by hydro-distillation as well as the scolicidal activities of the solvent extracts were determined. The results of the antimicrobial activity test of the essential oil showed a high level of activity against Bacillus cereus ATCC 10876 and MRSA. The highest activity was found on the microorganism of Pseudomonas aeruginosa ATCC 9027 in chloroform and methanol extracts of U. rigida. Furthermore, viability detection was performed and the scolicidal effects of the extracts on protoscoleces were assessed. The values of lethal concentration doses (LD50 , LD75 and LD90 ) were calculated using probit analysis.

Correlation between physicochemical properties and volatile compound profiles in tilapia muscles subjected to four different thermal processing techniques.

This work studied the physicochemical properties and odor profiles of tilapia muscles after exposure to four types of thermal processing methods: microwaving, roasting, boiling, or steaming. The effect of thermal processing on textural properties followed a pH-water state-water content-tissue microstructure-mass loss-textural properties route, expressed in the following manner: microwaving>roasting>steaming ≈ boiling. After processing, muscle pH increased from 6.59±0.10 to 6.73±0.04-7.01±0.06, and hardness changed from 1468.49±180.77g to 452.76±46.94-10723.66±2898.46g. Gas chromatography-based E-nose analysis confirmed that these methods had significant odor fingerprint effects on the tilapia muscles. Finally, the combined analysis of headspace solid-phase microextraction-gas chromatography-mass spectrometry, statistical MetaboAnalyst, and odor activity value showed that the microwaved, roasted, steamed, and boiled tilapia muscles had, respectively, three (hexanal, nonanal, and decanal), four (2-methyl-butanal, 3-methyl-butanal, decanal, and trimethylamine), one (2-methyl-butanal), and one (decanal) relatively important volatile compounds.

Post-fumigation sub-lethal activities of phosphine and ethyl formate on survivorship, fertility and female sex pheromone production of Callosobruchus chinensis (L.)

Phosphine (PH3) and ethyl formate (EF), the two popular fumigant disinfectants of stored product insect pests, are primarily evaluated for their knock down effects without considering their post-fumigation sub-lethal activities. The sub-lethal activities (adult survivorship, fecundity, sterility and female sex pheromone production) of the fumigants were evaluated on a field-to-storage insect pest adzuki bean beetle, Callosobruchus chinensis (L.). The adults’ survivorship and female fecundity, both were dose-dependently affected by sub-lethal PH3 and EF fumigation exposures. Hatchability of the eggs laid by fumigated female adults were also significantly affected. Gas-chromatography mass-spectrometry analysis of solid-phase micro-extraction from virgin fumigated C. cinensis females revealed that the PH3 LC25 (the lethal concentration required to kill the 25% of the population) fumigated female C. chinensis released significantly less amount of the pheromone components. In contrast, EF LC25 exposure did not affect the pheromone release. This study unveils the facts that the EF and PH3 fumigation have detrimental bioactivities against C. chinensis. Notably, this suggests to consider the sub-lethal EF and PH3 fumigation rather than the dose required to instantly kill all the C. chinensis individuals for disinfestation of stored adzuki bean.

An electrochemically fabricated ZIF-67/[HOEMIM]BF4 coating for the solid-phase microextraction and detection of polycyclic aromatic hydrocarbons.

Coatings are considered to play a crucial role in solid-phase microextraction (SPME). In this work, a novel coating named ZIF-67/[HOEMIM]BF4 was fabricated through in situ potentiostatic electrodeposition in methanol solutions containing ZIF-67 precursors and 1-(2'-hydroxyethyl)-3-methylimidazolium tetrafluoroborate ([HOEMIM]BF4). Compared with the traditional solvothermal method, this method reduced the synthesis time and enabled ZIF-67 to grow directly on the surface of stainless-steel wire, effectively simplifying the preparation process and improving the coating reproducibility. Owing to the inherent characteristics such as high porosity and high thermal and mechanical stability, and the impressive morphological regulation and extraction function of [HOEMIM]BF4, the developed coating exhibited a prolonged service life and a better extraction capacity for trace polycyclic aromatic hydrocarbons (PAHs) compared to single ZIF-67 and commercial fibers. Under the optimal conditions, the linear range of the ZIF-67/[HOEMIM]BF4-based SPME-GC method was 0.01-500 μg L-1, and the detection limit was 0.27-5.2 ng L-1. When applied in the determination of PAHs in a real water sample, recoveries between 85.6-117.3% were obtained, indicating the potential of ZIF-67/[HOEMIM]BF4 in the high efficiency SPME and GC analysis of PAHs.

Analysis of Volatile Compounds in Jinhua ham Using Three Extraction Methods Combined with Gas Chromatography-Time-of-Flight Mass Spectrometry.

The volatile compounds in Jinhua ham samples after different aging times were characterized using solvent-assisted flavor evaporation (SAFE), solid-phase microextraction (SPME), and needle trap (NT) extraction methods combined with gas chromatography-time-of-flight mass spectrometry (GC-TOF/MS). Hundreds of aroma compounds were identified, including aldehydes, alcohols, ketones, furans, esters, acids, pyrazines, and sulfides. The results showed that NT extracted the greatest number of volatile compounds, whereas the extraction efficiency of SPME headspace adsorption was highest among the three sample preparation methods. Principal component analysis of SPME effectively distinguished the variation in the aroma of the Jinhua hams specific to aging time. Butyrolactone, 2,6-dimethylpyrazine, 2,3,5-trimethylpyrazine, phenylacetaldehyde, and acetic acid were considered as the main volatile compounds in the Jinhua ham samples at three years of aging. The results showed that SPME-GC-TOF/MS effectively discriminated among samples by age. By comparing the three extraction methods, this study provides a theoretical basis for the selection of extraction methods of volatile aroma compounds in Jinhua ham.

Applications of headspace solid-phase microextraction in human biological matrix analysis

Abstract The inherent complexity of the human biological matrix and its importance in modern medical diagnosis and medical research promote the development of modern analytical technology. Solid-phase microextraction technology has been widely used in the treatment and analysis of different complex biological matrices due to its smaller sample size, simpler sample preparation and setting, and lower consumption of harmful chemicals. This review provides updated information on headspace solid-phase microextraction combined with gas chromatography technique applications, focusing on the analysis of volatile and semivolatile compounds in human biological matrices. The application of headspace solid-phase microextraction combined with gas chromatography techniques in human biological matrix analysis is mainly summarized into three aspects, namely, to discover biomarkers, to investigate volatile metabolomics, and to explore the effect of the external environment on volatile metabolomics of the human biological matrix. In addition, the frequently used statistical analytical methods are summarized, and the application prospect of solid-phase microextraction in the analysis of human biological matrices is proposed.

The Scent of Himantoglossum Species Found in Basilicata (Southern Italy)

The SPME (Solid Phase Microextraction) analysis of the scent of H. hircinum showed the presence of elemicin in the presence of a relevant amount of eugenol. The scent of the sample of H. adriaticum collected in Abruzzo showed the presence 4-amino-5-(4-morpholinylmethyl)-2-oxazolidinone, β-ocimene, decyl decanoate, and 9-tricosene as main components. The sample of H. adriaticum collected at Marsico Nuovo has an aroma where the main component was pentadecyl hexanoate, 9-tricosene, methyleugenol, tetradecane, pentadecane, and elemicin. The samples of H. adriaticum collected at Viggianello showed some similarities in the scent: the main components were 9-tricosene and methyleugenol.

Isolation and functional characteristics of the fungus Hypoxylon spp. Sj18 with biocontrol potential

A fungus with biocontrol potential was isolated from the roots of hickory trees. The strain named sj18 was classified as a member of the genus Hypoxylon (Hypoxylaceae) after multigene phylogenetic analysis (beta-tubulin gene, internal transcribed spacer, 28S large subunit ribosomal RNA gene, and RNA polymerase II subunit gene). The strain grew well on a PDA with an optimum temperature range between 32 and 34 °C. The fungus had obvious inhibitory effects on Botryosphaeria dothidea, Colletotrichum gloeosporioides, and Gibberella moniliformis in fumigation experiments on solid agar plates. In an inoculation experiment of Chinese cabbage, the fungus was also found to have an obvious repellent effect on cabbage caterpillars. In vitro experiments on Petri dishes showed that the fermentation broth of the sj18 strain could kill 100% of Bursaphelenchus xylophilus within 8 h even if the fermentation broth was diluted 8 times. The inoculation test of Arabidopsis thaliana showed that the fungus could promote the lateral root formation of plants and significantly increase their aboveground biomass. Through the analysis of solid phase microextraction (SPME), it was found that the main volatile components of the fermentation products were azulene 65.39% (61.77% + 3.62%), caryophyllene 7.41%, and eucalyptol 6.83% according to the peak area ratio. Therefore, sj18 can be used as a candidate for the further research and development of biocontrol agents.

Antinociceptive Activity of Compounds from the Aqueous Extract of Melampodium divaricatum.

A decoction prepared from the aerial parts of Melampodium divaricatum showed antinociceptive and antihyperalgesic responses when tested in the formalin model in mice. From the CH2 Cl2 fraction of the decoction, two non-previously reported secondary metabolites, 3-O-β-D-glucopyranosyl-16α-hydroxy-ent-kaurane (1) and melampodiamide (2) [(2'R*,4'Z)-2'-hydroxy-N-[(2S*,3S*,4R*)-1,3,4-trihydroxyoctadec-2-yl]tetracos-4-enamide] were separated and characterized by spectroscopic, spectrometric, and computational techniques. The flavonoids isoquercitrin and hyperoside, which possessed noted antinociceptive properties, were obtained from the active AcOEt fraction of the decoction. The chemical composition of the essential oil of the plant was also analyzed by gas chromatography-mass spectrometry. The major constituents were (E)-caryophyllene, germacrene D, β-elemene, δ-elemene, γ-patchoulene, and 7-epi-α-selinene. Headspace solid-phase microextraction analysis detected (E)-caryophyllene as the main volatile compound of the plant.

Identification of Volatile and Semi-Volatile Compounds in Polymeric Coatings Used in Metal Cans by GC-MS and SPME.

Polymeric coatings are used as a protective layer to preserve food or beverage quality and protect it from corrosion and avoid a metallic taste. These types of materials can contain some chemicals that are susceptible to migrate to food and constitute a risk for consumers’ health. This study is focused on the identification of volatile and semi-volatile low molecular weight compounds present in polymeric coatings used for metal food and beverage cans. A method based on solid–liquid extraction followed by gas chromatography–mass spectrometry (GC-MS) was optimized for the semi-volatile compounds. Different solvents were tried with the aim of extracting compounds with different polarities. Furthermore, a method based on solid-phase microextraction (SPME) in headspace (HS) mode and gas chromatography coupled with mass spectrometry (HSSPME-GC-MS) was developed for the identification of potential volatile migrants in polymeric coatings. Some parameters such as extraction time, equilibrium temperature, or the type of fiber were optimized. Different compounds, including aldehydes such as octanal or nonanal, alcohols such as α-terpineol or 2-butoxyethanol, ethers, alkenes, or phthalic compounds, among others, were identified and confirmed with analytical standards both via SPME analysis as well after solvent extraction.

Inhibitory effect of cinnamon ( Cinnamomum tamala (Buch.-Ham.) T.Nees & Eberm.) essential oil and its aldehyde constituents on growth and spore germination of phytopathogenic fungi

Plant pathogens lead to severe damage of the agricultural crops, worldwide. The disease management is currently being done by the synthetic chemicals. In the present report, Cinnamomum tamala (Buch.-Ham.) T. Nees & Eberm. oil was analyzed by gas chromatography-FID and GC/MS, which revealed (E)-cinnamaldehyde (40.6%), (E)-cinnamyl acetate (19.6%), linalool (10.2%) and benzaldehyde (3.6%) as marker constituents. Contact, volatile and spore germination inhibition assays were performed to determine the antifungal activities of Cinnamomum tamala (Buch.-Ham.) T.Nees & Eberm. against Rhizoctonia solani AG 4, Choanephora cucurbitarum and Bipolaris australiensis. C. tamala oil 500-560 ppm showed complete fungal growth inhibition in poison food technique over concentration range of. Solid-phase microextraction analysis of drop diffusion assay revealed the presence of benzaldehyde in headspace of Petri-plate, and found to be one of the strongest antifungal compounds against tested phytopathogens. Similarly, an oil concentration of 500 ppm inhibits spore germination. Therefore, products containing such compositions can be used for the plant diseases management.

Headspace Solid-Phase Microextraction Analysis of Volatile Components in Peanut Oil.

Peanut oil is favored by consumers due to its rich nutritional value and unique flavor. This study used headspace solid-phase microextraction (HS-SPME) combined with gas chromatography (GC) and gas chromatography–mass spectrometry (GC-MS) to examine the differences in the peanut oil aroma on the basis of variety, roasting temperatures, and pressing components. The results revealed that the optimal conditions for extracting peanut oil were achieved through the use of 50/30 μm DVB/CAR/PDMS fibers at 60 °C for 50 min. The primary compounds present in peanut oil were pyrazines. When peanuts were roasted, the temperature raised from 120 °C to 140 °C and the content of aldehydes in peanut oil increased; however, the content of aldehydes in No. 9 oil at 160 °C decreased. The components of peanut shell oil varied depending on the peanut variety. The most marked difference was observed in terms of the main compound at the two roasting temperatures. This compound was a pyrazine, and the content increased with the roasting temperature in hekei oils. When the roasting temperature was lower, No. 9 oil contained more fatty acid oxidation products such as hexanal, heptanal, and nonanal. When the roasting temperature increased, No. 9 oil contained more furfural and 5-methylfurfural. Heren oil was easier to oxidize and produced nonanal that possessed a fatty aroma.

Chemical composition and in vitro antihyperglycemic potential of Kainth fruit (Pyrus pashia Buch.-Ham ex D. Don)

The Kainth (Pyrus pashia Buch.-Ham. ex D. Don) fruit is an underutilized, variety of the Pyrus family. The fruit has a typical wild aroma, astringent taste and has not been explored for its flavor and chemical components, which are important for consumer acceptance and commercialization. Therefore, the composition of the fresh Kainth fruit pulp in terms of sugars, organic acids and volatile aroma compounds was investigated using the UPLC-ELSD, HPLC-PDA and GC-MS/MS, respectively. Fructose (7.29 g/100 g) and glucose (6.69 g/100 g) were the dominant sugars while glucuronic acid (85.4 mg/100 g) was the major organic acid in the fresh fruit. The bound phenolics showed a high in vitro antihyperglycemic potential when compared to acarbose and free phenolics. The major volatile compounds identified were esters (79.1% of total volatiles area) whereas in the dried fruit, volatile fatty acids (FA, 53.4%) were significant. The solid-phase microextraction analysis of the fresh fruit showed a dominance of lower molecular weight compounds such as esters (60.7%) and alkanes (12.1%), while the dried fruit had a predominance of aldehydes and alcohols. The odor activity value (OAV) and relative odor contribution (ROC) value were also calculated for the volatiles and the study suggested that Kainth has an aroma similar to citrus fruits: sweet and rose-like smell. This study showed the major volatile compounds, sugars and organic acids present in Kainth fruit. Further, the fruit or its dried powder could be used to develop food products with a unique flavor and potential health benefits.

Multi-class pesticide analysis in cannabis oil using coated blade spray and solid-phase microextraction with liquid chromatography coupled to mass spectrometry

The application of solid-phase microextraction (SPME) coupled to mass spectrometry to provide a high-throughput and cost-effective solution to multi-residue analysis of pesticides in cannabis oil samples has not been extensively explored. In this work, the method development steps for the extraction of an initial target list of 74 pesticides from cannabis oil via SPME for analysis with both LC-MS/MS and coated blade spray (CBS) are presented. The exploration of a washing step to remove adhered oil whilst minimally desorbing extracted analytes along with the implementation of central composited design investigation to examine compound extraction kinetics in the non-polar matrix yielded a workflow that was validated via both instrumental techniques. Of the initial target list, 37 pesticides were found to be suitable for screening or quantitation via CBS with performance validated via LC-MS/MS. The majority of compounds were found to meet the EU SANTE guidelines for analysis (i.e. linearity, precision, accuracy) whilst reaching limits of quantitation below or at Health Canada minimum regulatory limits (majority at 10 ng/mL). Examination of factors contributing to poor quantitation of pesticides via CBS are shared and explored, such as contributing isobaric interference sourced from plant byproducts and carrier oil, and comparison of signal-to-noise values achieved in cannabis oil when compared to the cannabis-free medium-chain triglyceride oil used as a carrier oil to stress the importance of matrix-match method development.

Estimating temporal and spatial levels of PAHs in air using rain samples and SPME analysis: Feasibility evaluation in an urban scenario

There is a growing interest in evaluating the role of concentration changes of contaminants in temporal and spatial gradients. This is often relevant for fast moving environmental phases such as air and water. In this paper, small volumes of rainwater were sampled as proxy for air concentrations of Polycyclic Aromatic Hydrocarbons (PAHs): rain was collected in three sampling sites (high traffic, restricted traffic and a low traffic zone) in Como. Solid phase micro extraction (SPME) was used for the extraction to reduce required sample volumes, allowing the acquisition of more samples in time. Rain samples highlighted a spatial and temporal variability along a traffic gradient in the Como city, especially for the most abundant PAH, e.g. phenanthrene. Air concentrations were then estimated from rain concentrations. The results show that this is a cheap and promising method, although requiring rainfall/snowfall conditions, that can be used to perform monitoring campaign of air concentrations at a higher temporal and spatial resolution than the adopted standard methods (e.g. high-volume air samplers). The results could be employed for evaluation of the exposure, emission profiles and calibration of fate models.

Metabolome-Based Analysis of Herbal Cough Preparations Via Headspace Solid-Phase Microextraction GC/MS and Multivariate Data Analyses: A Prospect for Its Essential Oil Equivalency.

Liquid cough preparations containing essential oils pose a challenge for isolating and quantifying their volatile components from such a complex matrix enriched with nonvolatile constituents and excipients. This study aims to develop a strategy integrating QC analysis of seven natural cough preparations in the Egyptian market and to assess volatile variation among the preparations using multivariate data analyses. Cough preparations were subjected to headspace solid-phase microextraction (HS-SPME) for determination of their essential oil composition mediating for their actions and to assess volatile differences among them. HS-SPME is a suitable technique for sample preparation that allows for extraction and enrichment of volatiles from complex nonvolatile matrices and their direct desorption into the gas chromatography analytical system. A total of 88 volatile components were identified belonging to seven classes, viz. aromatics, aliphatic hydrocarbons, mono/sesquiterpene hydrocarbons, and oxygenated mono/sesquiterpenes. Oxygenated monoterpenes, viz., menthol, cineole, thymol, and (E)-anethole, were the major volatiles identified in five cough preparations (79.5–98.6%), whereas aromatics, chiefly cinnamate derivatives, constituted the second class amounting for 50.5 and 27.4% in the other two cough preparations. Meaningful results regarding the products’ efficacy and safety were extrapolated from this analytical procedure, where artificial preservatives (parabens) were detected in five cough preparations. This study established an efficient strategy for exploring volatile profiling and defining different markers among the different cough preparations. Additionally, authenticity of listed herbal ingredients in the cough preparations was also confirmed in certain preparations, while other formulations failed to show representative volatile components. Volatile variation among preparations was assessed using multivariate data analyses in an attempt to prioritize cough preparations for usage, suggesting the preference of Bronchicum and Babetone among examined cough products.

Application of vacuum solid-phase microextraction for the analysis of semi-hard cheese volatiles

Solid-phase microextraction (SPME) is a well-established technique in the analysis of food volatile compounds, usually performed for qualitative analysis. This paper presents an elaboration of conditions for SPME analysis of main volatile compounds in hard cheese and an evaluation of suitability of vacuum to improve the transfer of volatile compounds towards SPME fiber. Compounds representing the main groups of hard cheese volatiles were investigated: 1-pentanol, butanoic acid, 2,3-butanedione and 2-heptanone. Parameters for SPME extraction (temperature, water, sodium carbonate addition, time, vacuum) were evaluated. Application of vacuum had a positive effect on all analytes when extraction was performed from water, but in the cheese matrix the effect was significant only for butanoic acid. Extraction time was the most significant factor for extraction efficiency in examined cheeses, while temperature had a minor effect on the amount of extracted volatiles. The method was applied on Edam, Emmentaler, Gouda and Maasdam cheeses obtained from the market.