Headspace solid-phase microextraction–gas chromatography–mass spectrometry for profiling free volatile compounds in Cabernet Sauvignon grapes and wines

The complex aroma of wine is derived from many sources, with grape-derived components being responsible for the varietal character. The ability to monitor grape aroma compounds would allow for better understanding of how vineyard practices and winemaking processes influence the final volatile composition of the wine. Here, we describe a procedure using GC–MS combined with headspace solid-phase microextraction (HS-SPME) for profiling the free volatile compounds in Cabernet Sauvignon grapes. Different sample preparation (SPME fiber type, extraction time, extraction temperature and dilution solvent) and GC–MS conditions were evaluated to optimize the method. For the final method, grape skins were homogenized with water and 8 ml of sample were placed in a 20 ml headspace vial with addition of NaCl; a polydimethylsiloxane SPME fiber was used for extraction at 40 °C for 30 min with continuous stirring. Using this method, 27 flavor compounds were monitored and used to profile the free volatile components in Cabernet Sauvignon grapes at different maturity levels. Ten compounds from the grapes, including 2-phenylethanol and β-damascenone, were also identified in the corresponding wines. Using this procedure it is possible to follow selected volatiles through the winemaking process.     

Optimization of the Extraction of Proanthocyanidins from Grape Seeds Using Ultrasonication-Assisted Aqueous Ethanol and Evaluation of Anti-Steatosis Activity In Vitro

Conventional extraction methods of proanthocyanidins (PAC) are based on toxic organic solvents, which can raise concerns about the use of extracts in supplemented food and nutraceuticals. Thus, a PAC extraction method was developed for grape seeds (GS) and grape seed powder using food-grade ethanol by optimizing the extraction conditions to generate the maximum yield of PAC. Extraction parameters, % ethanol, solvent: solid (s:s) ratio, sonication time, and temperature were optimized by the central composite design of the response surface method. The yields of PAC under different extraction conditions were quantified by the methylcellulose precipitable tannin assay. The final optimum conditions were 47% ethanol, 10:1 s:s ratio (v:w), 53 min sonication time, and 60 °C extraction temperature. High-performance liquid chromatography analysis revealed the presence of catechin, procyanidin B2, oligomeric and polymeric PAC in the grape seed-proanthocyanidin extracts (GS-PAC). GS-PAC significantly reduced reactive oxygen species and lipid accumulation in the palmitic-acid-induced mouse hepatocytes (AML12) model of steatosis. About 50% of the PAC of the GS was found to be retained in the by-product of wine fermentation. Therefore, the developed ethanol-based extraction method is suitable to produce PAC-rich functional ingredients from grape by-products to be used in supplemented food and nutraceuticals.     

In‐tube extraction for the determination of the main volatile compounds in Physalis peruviana L

An analytical procedure based on in‐tube extraction followed by gas chromatography with mass spectrometry has been developed for the analysis of 24 of the main volatile components in cape gooseberry (Physalis peruviana L.) samples. According to their chemical structure, the compounds were organized into different groups: one hydrocarbon, one aldehyde, four alcohols, four esters, and 14 monoterpenes. By single‐factor experiments, incubation temperature, incubation time, extraction volume, extraction strokes, extraction speed, desorption temperature, and desorption speed were determined as 60°C, 20 min, 1000 μL, 20, 50:50 μL/s, 280°C, 100 μL/s, respectively. Quantitative analysis using authentic standards and external calibration curves was performed. The limit of detection and limit of quantification for the analytical procedure were calculated. Results shown the benzaldehyde, ethyl butanoate, 2‐methyl‐1‐butanol, 1‐hexanol, 1‐butanol, α‐terpineol, and terpinen‐4‐ol were the most abundant volatile compounds in analyzed fruits (68.6–585 μg/kg). The obtained data may contribute to qualify cape gooseberry to the group of superfruits and, therefore, increase its popularity.     

Using Design of Experiments to Optimize a Screening Analytical Methodology Based on Solid-Phase Microextraction/Gas Chromatography for the Determination of Volatile Methylsiloxanes in Water

Volatile methylsiloxanes (VMSs) constitute a group of compounds used in a great variety of products, particularly personal care products. Due to their massive use, they are continually discharged into wastewater treatment plants and are increasingly being detected in wastewater and in the environment at low concentrations. The aim of this work was to develop and validate a fast and reliable methodology to screen seven VMSs in water samples, by headspace solid-phase microextraction (HS-SPME) followed by gas chromatography with flame ionization detection (GC-FID). The influence of several factors affecting the extraction efficiency was investigated using a design of experiments approach. The main factors were selected (fiber type, sample volume, ionic strength, extraction and desorption time, extraction and desorption temperature) and optimized, employing a central composite design. The optimal conditions were: 65 µm PDMS/Divinylbenzene fiber, 10 mL sample, 19.5% NaCl, 39 min extraction time, 10 min desorption time, and 33 °C and 240 °C as extraction and desorption temperature, respectively. The methodology was successfully validated, showing low detection limits (up to 24 ng/L), good precision (relative standard deviations below 15%), and accuracy ranging from 62% to 104% in wastewater, tap, and river water samples.     

Optimization of HS-SPME for GC-MS Analysis and Its Application in Characterization of Volatile Compounds in Sweet Potato

Volatile compounds are the main chemical species determining the characteristic aroma of food. A procedure based on headspace solid-phase microextraction (HP-SPME) coupled to gas chromatography-mass spectrometry (GC-MS) was developed to investigate the volatile compounds of sweet potato. The experimental conditions (fiber coating, incubation temperature and time, extraction time) were optimized for the extraction of volatile compounds from sweet potato. The samples incubated at 80 °C for 30 min and extracted at 80 °C by the fiber with a divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) coating for 30 min gave the most effective extraction of the analytes. The optimized method was applied to study the volatile profile of four sweet potato cultivars (Anna, Jieshu95-16, Ayamursaki, and Shuangzai) with different aroma. In total, 68 compounds were identified and the dominants were aldehydes, followed by alcohols, ketones, and terpenes. Significant differences were observed among the volatile profile of four cultivars. Furthermore, each cultivar was characterized by different compounds with typical flavor. The results substantiated that the optimized HS-SPME GC-MS method could provide an efficient and convenient approach to study the flavor characteristics of sweet potato. This is the basis for studying the key aroma-active compounds and selecting odor-rich accessions, which will help in the targeted improvement of sweet potato flavor in breeding.     

固相微萃取-顶空-气相色谱-质谱法测定美甲贴中6种挥发性有机物的含量

称取美甲贴样品0.0010g置于20mL顶空瓶中,在选定的条件下[孵化温度为36℃,孵化时间为15min,振动器转速为300r·min^-1,固相微萃取(SPME)温度为常温和样品的萃取次数为10次]进行顶空采样和固相微萃取,并将所得挥发性有机物(VOCs,包括所需测定的丙酮、正己烷、乙酸乙酯、乙酸异丙酯、甲苯和乙酸丁酯等6种化合物)引入气相色谱-质谱仪中进行分析。用HP-VOC色谱柱(60m×0.25mm,0.25μm)在40~250℃区间以程序升温模式进行色谱分离,并在电子轰击离子源和选择离子扫描模式等条件下进行质谱测定。采用外标法定量。测得上述6种VOCs的质量在相同的范围(2~250ng)内与其相应的峰面积呈线性关系,其检出限(3.14s)也相同(0.5ng)。以实际样品为基体,在5个浓度水平上加入标准溶液进行回收试验,测得其回收率为80.0%~110%,测定值的相对标准偏差(n=7)为0.030%~5.0%。     

Effect of Extraction Solvent and Temperature on Polyphenol Profiles,Antioxidant and Anti-Inflammatory Effects of Red Grape Skin By-Product

A fully-detailed LC-MS qualitative profiling of red grape skin, extracted with a mixture of ethanol and water (70:30 v:v) has permitted the identification of 65 compounds which can be classified into the following chemical classes: organic and phenolic acids (14 compounds), stilbenoids (1 compound), flavanols (21 compounds), flavonols (15 compounds) and anthocyanins (14 compounds). The extraction yield obtained with water at different temperatures (100 °C, 70 °C, room temperature) was then evaluated and the overall polyphenol content indicates that EtOH:H₂O solvent is the most efficient and selective for polyphenol extraction. However, by analyzing the recovery yield of each single polyphenol, we found that water extraction under heating conditions is effective (extraction yield similar or even better in respect to the binary solvent) for some polyphenolic classes, such as hydrophilic procyanidins, phenolic acids, flavonol glucosides and stilbenoids. However, according to their lipophilic character, a poor yield was found for the most lipophilic components, such as flavonol aglycones, and in general for anthocyanins. The radical scavenging activity was in accordance with the polyphenol content, and hence, much higher for the extract obtained with the binary solvent in respect to water extraction. All the tested extracts were found to have an anti-inflammatory activity in the R3/1 cell line with NF-kb reporter challenged with 0.01 µg/mL of IL-1α, in a 1 to 250 µg/mL concentration range. An intriguing result was that the EtOH:H₂O extract was found to be superimposable with that obtained using water at 100 °C despite the lower polyphenol content. Taken together, the results show the bioactive potentialities of grape skin extracts and the possibility to exploit this rich industrial waste. Water extraction carried out by heating is an easy, low-cost and environmentally friendly extraction method for some polyphenol classes and may have great potential for extracts with anti-inflammatory activities.     

Novel sample preparation technique with needle-type micro-extraction device for volatile organic compounds in indoor air samples

A novel needle-type sample preparation device was developed for the effective preconcentration of volatile organic compounds (VOCs) in indoor air before gas chromatography–mass spectrometry (GC–MS) analysis. To develop a device for extracting a wide range of VOCs typically found in indoor air, several types of particulate sorbents were tested as the extraction medium in the needle-type extraction device. To determine the content of these VOCs, air samples were collected for 30 min with the packed sorbent(s) in the extraction needle, and the extracted VOCs were thermally desorbed in a GC injection port by the direct insertion of the needle. A double-bed sorbent consisting of a needle packed with divinylbenzene and activated carbon particles exhibited excellent extraction and desorption performance and adequate extraction capacity for all the investigated VOCs. The results also clearly demonstrated that the proposed sample preparation method is a more rapid, simpler extraction/desorption technique than traditional sample preparation methods.     

Development of solid-phase microextraction followed by gas chromatography-mass spectrometry for rapid analysis of volatile organic chemicals in mainstream cigarette smoke

In this work, a novel, simple and efficient method based on solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS) was developed to the analysis of volatile organic chemicals (VOCs) in mainstream cigarette smoke (MCS). Using a simple home-made smoking machine device, extraction and concentration of VOCs in MCS were performed by SPME fiber, and the VOCs adsorbed on fiber were desorbed, and analyzed by GC-MS. The extraction fiber types and the desorption conditions were studied, and the method precision was also investigated. After the investigation, the optimal fiber was divinylbenzene/carboxen/polydemethylsiloxane (DVB/CAR/PDMS), and the optimal desorption condition was 250 degrees C for 3 min. The method precision was from 2% to 11%. Finally, the proposed method was tested by its application of the analysis of VOCs in MCS from 10 brands of cigarettes and one reference cigarette. A total of 70 volatile compounds were identified by the proposed method. The experimental results showed that the proposed method was a simple, rapid, reliable, and solvent-free technique for the determination of VOCs in MCS.     

An Extraction Method for Obtaining the Maximum Non-Anthocyanin Phenolics from Grape Berry Skins

To obtain maximum non-anthocyanin phenolics from grape skins, various extraction conditions were evaluated according to the amounts of 8 non-anthocyanin phenolics. The result showed that the optimal extraction was performed in ethyl acetate solution at 25°C, with liquid/solid ratio of 10:1 (ml/g) and for 30 min. On this basis, the non-anthocyanin extracts from two red and two white grape cultivars were analyzed using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC- MS/MS), and twenty-four compounds were qualified and simultaneously quantified, which indicated that the present extraction method was suitable for the study of non-anthocyanin in grape skins.     

Solid phase microextraction with gas chromatography-mass spectrometry: a very rapid method for identification of volatile organic compounds emitted by Carum copticum

The investigation of flower scent represents an important field of modern biological research which is directed towards special theories of biological recognition. The headspace solid phase microextraction coupled with gas chromatography-mass spectrometry was used to identify the volatile components of Carum copticum (C. copticum) cultivated in Iran. The compounds were identified according to their retention indices and mass spectra (EI, 70 eV). The effects of different parameters, such as the desorption time, the extraction temperature, the sample mass, the addition of salt, the pre-equilibration time and the extraction time, on the extraction efficiency were investigated. The optimized conditions were: the desorption time, 2 min; the extraction temperature, 58 degrees Celsius; the sample mass, 1.000 g in 4.0 mL 2.0 M NaCl solution; the pre-equilibration time, 25 min; the extraction time, 20 min. Finally, ten components were identified in the volatile components of C. copticum. The major components of C. copticum were thymol (68.2%), gamma-terpinene (13.9%), p-cymene (11.6%), myrcene (1.0%) and beta-pinene (0.6%). Precision of the proposed method is good and %RSD less than 14 was obtained.     

A Green Method of Extracting and Recovering Flavonoids from Acanthopanax senticosus Using Deep Eutectic Solvents

In recent years, green extraction of bioactive compounds from herbal medicines has generated widespread interest. Deep eutectic solvents (DES) have widely replaced traditional organic solvents in the extraction process. In this study, the efficiencies of eight DESs in extracting flavonoids from Acanthopanax senticosus (AS) were compared. Response surface methodology (RSM) was employed to optimize the independent variable including ultrasonic power, water content, solid-liquid ratio, extraction temperature, and extraction time. DES composed of glycerol and levulinic acid (1:1) was chosen as the most suitable extraction medium. Optimal conditions were ultrasonic power of 500 W, water content of 28%, solid-liquid ratio of 1:18 g·mL⁻¹, extraction temperature of 55 °C, and extraction time of 73 min. The extraction yield of total flavonoids reached 23.928 ± 0.071 mg·g⁻¹, which was 40.7% higher compared with ultrasonic-assisted ethanol extraction. Macroporous resin (D-101, HPD-600, S-8 and AB-8) was used to recover flavonoids from extracts. The AB-8 resin showed higher adsorption/desorption performance, with a recovery rate of total flavonoids of up to 71.56 ± 0.256%. In addition, DES solvent could efficiently be reused twice. In summary, ultrasonic-assisted DES combined with the macroporous resin enrichment method is exceptionally effective in recovering flavonoids from AS, and provides a promising environmentally friendly and recyclable strategy for flavonoid extraction from natural plant sources.     

Optimisation of solid-phase microextraction combined with gas chromatography-mass spectrometry based methodology to establish the global volatile signature in pulp and skin of Vitis vinifera L. grape varieties

The volatiles (VOCs) and semi-volatile organic compounds (SVOCs) responsible for aroma are mainly present in skin of grape varieties. Thus, the present investigation is directed towards the optimisation of a solvent free methodology based on headspace-solid-phase microextraction (HS-SPME) combined with gas chromatography-quadrupole mass spectrometry (GC-qMS) in order to establish the global volatile composition in pulp and skin of Bual and Bastardo Vitis vinifera L. varieties. A deep study on the extraction-influencing parameters was performed, and the best results, expressed as GC peak area, number of identified compounds and reproducibility, were obtained using 4 g of sample homogenised in 5 mL of ultra-pure Milli-Q water in a 20 mL glass vial with addition of 2 g of sodium chloride (NaCl). A divinylbenzene/carboxen/polydimethylsiloxane fibre was selected for extraction at 60 °C for 45 min under continuous stirring at 800 rpm. More than 100 VOCs and SVOCs, including 27 monoterpenoids, 27 sesquiterpenoids, 21 carbonyl compounds, 17 alcohols (from which 2 aromatics), 10 C₁₃ norisoprenoids and 5 acids were identified. The results showed that, for both grape varieties, the levels and number of volatiles in skin were considerably higher than those observed in pulp. According to the data obtained by principal component analysis (PCA), the establishment of the global volatile signature of grape and the relationship between different part of grapes—pulp and skin, may be an useful tool to winemaker decision to define the vinification procedures that improves the organoleptic characteristics of the corresponding wines and consequently contributed to an economic valorization and consumer acceptance.     

An Optimized SPME-GC-MS Method for Volatile Metabolite Profiling of Different Alfalfa (Medicago sativa L.) Tissues

Solid-phase microextraction (SPME) was coupled to gas chromatography mass spectrometry (GC-MS) and a method optimized to quantitatively and qualitatively measure a large array of volatile metabolites in alfalfa glandular trichomes isolated from stems, trichome-free stems, and leaves as part of a non-targeted metabolomics approach. Major SPME extraction parameters optimized included SPME fiber composition, extraction temperature, and extraction time. The optimized SPME method provided the most chemically diverse coverage of alfalfa volatile and semi-volatile metabolites using a DVB/CAR/PDMS fiber, extraction temperature of 60 °C, and an extraction time of 20 min. Alfalfa SPME-GC-MS profiles were processed using automated peak deconvolution and identification (AMDIS) and quantitative data extraction software (MET-IDEA). A total of 87 trichome, 59 stem, and 99 leaf volatile metabolites were detected after background subtraction which removed contaminants present in ambient air and associated with the fibers and NaOH/EDTA buffer solution containing CaCl₂. Thirty-seven volatile metabolites were detected in all samples, while 15 volatile metabolites were uniquely detected only in glandular trichomes, 9 only in stems, and 33 specifically in leaves as tissue specific volatile metabolites. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) of glandular trichomes, stems, and leaves showed that the volatile metabolic profiles obtained from the optimized SPME-GC-MS method clearly differentiated the three tissues (glandular trichomes, stems, and leaves), and the biochemical basis for this differentiation is discussed. Although optimized using plant tissues, the method can be applied to other types of samples including fruits and other foods.     

Coupling of stir bar sorptive extraction with single photon ionization mass spectrometry for determination of volatile organic compounds in water

A home-made stir bar sorptive extraction (SBSE) apparatus was combined to a single photon ionization time-of-flight mass spectrometer (SPI-TOFMS) for rapid and sensitive determination of trace volatile organic compounds (VOCs) in water. The home-made SBSE bar, low-cost and disposable, was used for VOCs extraction. A thermal desorption (TD) device was designed to desorb the analytes from the SBSE bar, and a high throughput interface was developed to transfer the analytes into the ionization chamber of the SPI-TOFMS. The combination of large extraction volume of SBSE bar, and the direct measurement power of SPI-TOFMS enable a short analysis time for VOCs in water with high sensitivity, for example the limits of detection (LODs) were in the range of 7.4-11.1 ng L(-1) for benzene, toluene, and p-xylene (BTX) within 15 min. BTX aqueous solutions were chosen to demonstrate the quantitative capability, the linear range was from 0.05 to 100 μg L(-1) and the correlation coefficients were better than 0.996. The proposed method was successfully applied for the analysis of VOCs in urban river water.     

Ultrasound-Assisted Extraction of Carotenoids from Carrot Pomace and Their Optimization through Response Surface Methodology

Ultrasound-assisted extraction (UAE) was used to extract carotenoids from the carrot pomace. To investigate the effect of independent variables on the UAE, the response surface methodology (RSM) with central-composite design (CCD) was employed. The study was conducted with three independent variables including extraction time (min), temperature (°C), and ethanol concentration (%). The results showed that the optimal conditions for UAE were achieved with an extraction time of 17 min, temperature of 32 °C, and ethanol concentration of 51% of total carotenoids (31.82 ± 0.55); extraction time of 16 min, temperature of 29 °C, and ethanol concentration of 59% for a combination of β-carotene (14.89 ± 0.40), lutein (5.77 ± 0.19), and lycopene (2.65 ± 0.12). The non-significant (p > 0.05) correlation under optimal extraction conditions between predicted and experimental values suggested that UAE is the more productive process than conventional techniques for the extraction of carotenoids from the carrot pomace.     

荧光衍生化-磁分散固相萃取/高效液相色谱检测池塘水与蔬菜中3种挥发性异味物质

以6-碳酰氯左氧氟沙星(LFC-Cl)为衍生试剂,磁性氧化石墨烯(MGO)为吸附剂,建立了2-甲基异莰醇(2-MIB)、土臭素(GSM)和3-甲基-1-丁醇(3-MB)的高效液相色谱荧光检测(HPLC-FLD)分析方法。在浓度为0.05 mol/L的4-二甲氨基吡啶(DMAP)乙腈溶液中,衍生温度60℃,反应时间90 min,超声波辅助条件下,可完成上述3种醇类挥发性异味物质(VOCs)的衍生化。在吸附剂用量20 mg、萃取时间20 min、解吸剂为乙腈(含1%甲酸)、解吸时间3 min条件下,能实现3种异味物质衍生物的富集和净化。在优化的HPLC条件下,15 min内可实现3种衍生物的基线分离和高灵敏检测。方法检出限(LOD)为0.020~0.95 ng/L,定量下限(LOQ)为0.10~3.3 ng/L,线性、精密度和回收率良好。与已报道的方法相比,该方法具有灵敏度高、样品前处理简单、仪器普适性好等优势,可用于池塘水、蔬菜中VOCs的快速、定量测定,为食品和环境水样监测提供了一种新方法。     

Rapid removal of selected volatile organic compounds from gaseous mixtures using a new dispersive vapor extraction technique: A feasibility study

A new dispersive vapor extraction (DVE) technique for rapid removal of selected volatile organic compounds (VOCs) from gaseous mixtures was investigated. In this technique, less than 1.0 mL of a volatile solvent was vaporized for 8 min in a 250-mL flask containing a gaseous mixture. The flask was then cooled under running tap water for 2-3 min to induce condensation of the vapor and co-extraction of the VOCs from the headspace. The technique was tested over a concentration range of 4-23 ppb, and resulted in extraction efficiencies ranging from 80 to 97% for the VOCs tested. Because of its simplicity and the relatively short sampling time, DVE could potentially lead to high sample throughput and rapid air analysis.     

Electrochemical immunosensor for prostate-specific antigen using a glassy carbon electrode modified with a nanocomposite containing gold nanoparticles supported with starch-functionalized multi-walled carbon nanotubes

We report on a simple, rapid, and efficient method for the extraction of volatile organic compounds (VOCs; including methanol, tetrahydrofuran, 2-hexanone and benzene) from air and solid samples. The system is based on the use of a laboratory-made syringe as the extractor. The needle of the syringe is placed in a chamber cooled by liquid nitrogen. The tip of the needle is placed in the headspace of a vial containing the sample. The headspace components then are circulated with a pump to pass the needle, and this results in freeze-trapping of the VOCs on the inner surface of the needle. The circulation of the headspace components is continued for 15 min, and the syringe is then removed and placed in a GC injector. The effects of volume of the sample vial, headspace flow rate, temperature and time of extraction and desorption were optimized. The overall time for sampling and analysis is <30 min. The method displays an extraction efficiency of >80%) and a good sample transfer efficiency into the GC column due to the absence of a sorbent inside the needle. No carry-over was observed after 30?s desorption at 260?°C. An external standard method was used for quantitative analysis. The relative standard deviation values are below 10% and the limits of detection range from 1.3 to 4.6?ng?g^?1.

Extraction Optimization of Astragaloside IV by Response Surface Methodology and Evaluation of Its Stability during Sterilization and Storage

Radix Astragali is referred to as a variety of food-medicine herb, and it is commonly applied as Traditional Chinese Medicine (TCM). However, it is extremely difficult to extract its bio-active compounds (astragaloside IV) and apply it in food processing efficiently, which restricts its practical applications. In this study, the conditions required for the extraction of astragaloside IV were optimized by following the response surface methodology. More specifically, ammonia with a concentration of 24% was used as an extracting solvent, the solid–liquid ratio was 1:10 (w:v); the Radix Astragali was soaked at 25 °C for 120 min in advance and then stirred at 25 °C for 52 min (150 rpm) to extract astragaloside IV. This method promoted the transformation of other astragalosides into astragaloside IV and replaced the traditional approach for extraction, the solvent reflux extraction method. The yield of astragaloside IV reached the range of 2.621 ± 0.019 mg/g. In addition, the stability of astragaloside IV was evaluated by detecting its retention rate during sterilization and 60-day storage. As suggested by the results, the astragaloside IV in acidic, low-acidic, and neutral solutions was maintained above 90% after sterilization (95 °C and 60 min) but below 60% in an alkaline solution. High temperature and short-term sterilization approach is more appropriate for astragaloside IV in an alkaline solution. It was also found out that the astragaloside IV obtained using our method was maintained over 90% when stored at room temperature (25 °C), and there was no significant difference observed to low temperature (4 °C) in solutions regardless of acidity.