Investigation of modulation parameters in multiplexing gas chromatography

Combination of information technology and separation sciences opens a new avenue to achieve high sample throughputs and therefore is of great interest to bypass bottlenecks in catalyst screening of parallelized reactors or using multitier well plates in reaction optimization. Multiplexing gas chromatography utilizes pseudo-random injection sequences derived from Hadamard matrices to perform rapid sample injections which gives a convoluted chromatogram containing the information of a single sample or of several samples with similar analyte composition. The conventional chromatogram is obtained by application of the Hadamard transform using the known injection sequence or in case of several samples an averaged transformed chromatogram is obtained which can be used in a Gauss–Jordan deconvolution procedure to obtain all single chromatograms of the individual samples. The performance of such a system depends on the modulation precision and on the parameters, e.g. the sequence length and modulation interval. Here we demonstrate the effects of the sequence length and modulation interval on the deconvoluted chromatogram, peak shapes and peak integration for sequences between 9-bit (511 elements) and 13-bit (8191 elements) and modulation intervals Δt between 5 s and 500 ms using a mixture of five components. It could be demonstrated that even for high-speed modulation at time intervals of 500 ms the chromatographic information is very well preserved and that the separation efficiency can be improved by very narrow sample injections. Furthermore this study shows that the relative peak areas in multiplexed chromatograms do not deviate from conventionally recorded chromatograms.     

Geographical traceability of Italian white truffle (Tuber magnatum Pico) by the analysis of volatile organic compounds

Results are presented that were obtained on the geographic traceability of the white truffle Tuber magnatum Pico. Solid-phase microextraction coupled to gas chromatography/mass spectrometry (SPME-GC/MS) was employed to characterize the volatile profile of T. magnatum white truffle produced in seven geographical areas of Italy. The main components of the volatile fraction were identified using SPME-GC/MS. Significant differences in the proportion of volatile constituents from truffles of different geographical areas were detected. The results suggest that, besides genetic factors, environmental conditions influence the formation of volatile organic compounds. The mass spectra of the volatile fraction of the samples were used as fingerprints to characterize the geographical origin. Next, stepwise factorial discriminant analysis afforded a limited number of characteristic fragment ions that allowed a geographical classification of the truffles studied.     

Geographical discrimination of Chinese winter wheat using volatile compound analysis by HS‐SPME/GC‐MS coupled with multivariate statistical analysis

This study aimed to develop a potential analytical method to discriminate the Chinese winter wheat according to geographical origin and cultivars. A total of 90 wheat samples of 10 different wheat cultivars among three regions were examined by headspace solid phase microextraction coupled with gas chromatography‐mass spectrometry (GC‐MS). The peak areas of 32 main volatile compounds were selected and subjected to statistical analysis, which revealed significant differences among different regions and cultivars. Multivariate analysis of variance showed a significant influence of regions, wheat genotypes, and their interaction on the volatile composition of wheat. Principal component analysis of the aromatic profile showed better visualization for wheat geographical origins. Finally, a classification model based on the linear discriminant analysis was successfully constructed for the discrimination of regions and cultivars with the correct classification percentages of 90 and 100%, respectively.     

Seasonal Variations of Polycyclic Aromatic Hydrocarbons in Air Particulate Extracts

Aroma-active volatile organic compounds (VOCs) present in brandies produced in the Slovak Republic have been identified by gas chromatography–olfactometry (GC–O) and GC–MS. GC–O data treatment was based on a concept of detection frequency. Direct injection of samples was used due to its simplicity and the fact that determined composition is that of the original samples. This is especially important for correlation of results obtained by sensory evaluation and gas chromatographic measurements. Among 200 organic compounds found by GC only 71 showed olfactive properties at the given concentration levels. In order to identify aroma-active substances, linear retention indices (LRI) have been calculated and compared with available LRI libraries. The most aroma-active compounds have been found in samples of Frucon brandy, while Vinovica showed their minimal content. The results obtained by gas chromatography have been compared with sensory evaluation of brandy odor.     

Monoterpene and sesquiterpene hydrocarbons of virgin olive oil by headspace solid-phase microextraction coupled to gas chromatography/mass spectrometry

In the present article, a headspace solid-phase microextraction method coupled to GC/MS was developed and applied for the simultaneous determination of mono- and sesquiterpenic hydrocarbons in virgin olive oils of different olive variety and geographical origin. Analysis of various oils resulted in the simultaneous detection of 15 monoterpenes and 30 sesquiterpenes. Some of these hydrocarbons were previously reported to be constituents of virgin olive oil terpenoid fraction, although we also detected some terpenic hydrocarbons that have not previously been documented as present in virgin olive oil. Significant differences were detected in the proportion of terpenic compounds in oils obtained from different olive varieties grown in different geographical areas. The monoterpene, and particularly the sesquiterpene composition of olive oil may be used to distinguish samples from different cultivar and geographical areas.     

Profiling food volatiles by comprehensive two-dimensional ga schromatography coupled with mass spectrometry: Advanced fingerprinting approaches for comparative analysis of the volatile fraction of roasted hazelnuts (Corylus avellana L.) from different origins

This study examined how advanced fingerprinting methods (i.e., non-targeted methods) provide reliable and specific information about groups of samples based on their component distribution on the GC × GC chromatographic plane. The volatile fractions of roasted hazelnuts (Corylus avellana L.) from nine different geographical origins, comparably roasted for desirable flavor and texture, were sampled by headspace-solid phase micro extraction (HS-SPME) and then analyzed by GC × GC-qMS. The resulting patterns were processed by: (a) “chromatographic fingerprinting”, i.e., a pattern recognition procedure based on retention-time criteria, where peaks correspondences were established through a comprehensive peak pattern covering the chromatographic plane; and (b) “comprehensive template matching” with reliable peak matching, where peak correspondences were constrained by retention time and MS fragmentation pattern similarity criteria. Fingerprinting results showed how the discrimination potential of GC × GC can be increased by including in sample comparisons and correlations all the detected components and, in addition, provide reliable results in a comparative analysis by locating compounds with a significant role. Results were completed by a chemical speciation of volatiles and sample profiling was extended to known markers whose distribution can be correlated to sensory properties, geographical origin, or the effect of thermal treatment on different classes of compounds. The comprehensive approach for data interpretation here proposed may be useful to assess product specificity and quality, through measurable parameters strictly and consistently correlated to sensory properties and origin.     

Characterization and classification of Western Greek olive oils according to cultivar and geographical origin based on volatile compounds

The aim of the present study was to characterize and classify olive oils from Western Greece according to cultivar and geographical origin, based on volatile compound composition, by means of Linear Discriminant Analysis. A total of 51 olive oil samples were collected during the harvesting period 2007-2008 from six regions of Western Greece and from six local cultivars. Forty-five of the samples were characterized as extra virgin olive oils. The analysis of volatile compounds was performed by Headspace Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry (HS-SPME-GC/MS). Fifty-three (53) different volatile compounds were tentatively identified and semi-quantified. Using selected volatile compound composition data (selection was based on the application of ANOVA to total volatiles to determine those variables showing substantial differences among samples of different geographical origin/cultivar), the olive oil samples were satisfactorily classified according to geographical origin (87.2%) and cultivar (74%).     

Simultaneous capillary gas chromatographic profiling of medium-and long-chain fatty acid methyl esters with split injection. Correction for injection-related discrimination by the 'bracketing' method

Split injection-related discrimination can be a source of inaccuracy and imprecision in quantitative capillary gas chromatographic profiling methods for compounds with relatively big differences in boiling points, such as the methyl esters of medium- and long-chain fatty acids prepared from biological materials. We systematically investigated a standard containing equal masses of saturated fatty acid methyl esters, with chain lengths from C5 to C26, under different injection conditions, including injection temperature, sample volume and split ratio. Day-to-day performance was studied under one set of conditions. Normalized peak areas, reciprocal response factors, using either C17 or C23 as an internal standard, and 'bracketed' reciprocal response factors (peak area of each analyte divided by half the sum of the peak areas of two adjacent esters were calculated. In all experiments the bracketed reciprocal response factors were found to be closest to unity with the lowest coefficients of variation.     

Automated screening procedure using gas chromatography-mass spectrometry for identification of drugs after their extraction from biological samples

A novel analytical screening procedure has been developed, using computer-controlled gas chromatography-mass spectrometry (GC-MS), to detect 120 drugs of interest to road safety. This paper describes GC-MS methodology suitable for use on extracts of biological origin, while extraction procedures will be the subject of a future communication. The method was devised to identify drugs in extracts of blood samples, as part of an investigation into the involvement of drugs, other than alcohol, in road accidents. The method could be adapted to screen for other substances. The method depends on a "macro" program which was written to automate the search of GC-MS data for target drugs. The strategy used was to initially search for each drug in the database by monitoring for a single characteristic ion at the expected retention time. If a peak is found in this first mass chromatogram, a peak for a second characteristic ion is sought within 0.02 min of the first and, if found, the ratio of peak areas calculated. Probable drug identification is based on the simultaneous appearance of peaks for both characteristic ions at the expected retention time and in the correct ratio. If the ratio is outside acceptable limits, a suspected drug (requiring further investigation) is reported. The search macro can use either full mass spectra or, for enhanced sensitivity, data from selected ion monitoring (which requires switching between groups of ions during data acquisition). Quantitative data can be obtained in the usual way by the addition of internal standards.     

Microwave-Assisted Extraction Coupled to HPLC-UV Combined with Chemometrics for the Determination of Bioactive Compounds in Pistachio Nuts and the Guarantee of Quality and Authenticity

Two novel microwave-assisted extraction (MAE) methods were developed for the isolation of phenols and tocopherols from pistachio nuts. The extracts were analyzed by reversed-phase high-pressure liquid chromatography coupled with a UV detector (RP-HPLC-UV). In total, eighteen pistachio samples, originating from Greece and Turkey, were analyzed and thirteen phenolic compounds, as well as α-tocopherol, (β + γ)-tocopherol, and δ-tocopherol, were identified. The analytical methods were validated and presented good linearity (r² > 0.990) and a high recovery rate over the range of 82.4 to 95.3% for phenols, and 93.1 to 96.4% for tocopherols. Repeatablility was calculated over the range 1.8–5.8%RSD for intra-day experiments, and reproducibility over the range 3.2–9.4%RSD for inter-day experiments, respectively. Principal component analysis (PCA) was employed to analyze the differences between the concentrations of the bioactive compounds with respect to geographical origin, while agglomerative hierarchical clustering (AHC) was used to cluster the samples based on their similarity and according to the geographical origin.     

Development of a voltammetric electronic tongue for discrimination of edible oils

In this paper, we propose a novel strategy to perform cyclic voltammetric measurements with a platinum microelectrode directly in edible oil samples. The microelectrode was employed as an electronic tongue that, along with the application of chemometrics to the current–potential responses, proved useful for discriminating oils on the basis of their quality and geographical origin. The method proposed here is based on the use of suitable room temperature ionic liquids, added to oils as supporting electrolytes to provide conductivity to the low-polarity samples. The entire voltammograms, recorded directly on the oil/RTIL mixtures, were processed via principal component analysis and a classification technique (K nearest neighbors), to extract information on samples characteristics. Data processing showed that oils having different nature (i.e. maize and olive) or geographical origin (i.e. olive oils coming from different regions) can be distinguished.     

Chemical differentiation of volatile compounds in crude and processed Atractylodis Macrocephalae Rhizoma by using comprehensive two‐dimensional gas chromatography with time‐of‐flight mass spectrometry combined with multivariate data analysis

In the present study, comprehensive 2D GC—TOF‐MS combined with multivariate data analysis was applied to analyze the differences of the volatile components in crude and processed Atractylodis Macrocephalae Rhizoma (AMR) samples. As a result, 26 compounds that were found in crude AMR samples disappeared in processed AMR samples, and 19 compounds were newly generated and identified in AMR after processing with wheat bran. Meanwhile, principal component analysis demonstrated that there were significant chemical differences between crude and processed AMR samples, and processing procedure caused obvious quantitative and qualitative changes of volatile components in AMR. The established method could be used to explain the chemical differentiation between crude and processed AMR, and to further understand the processing mechanism of herbal medicines.     

Composition of Eucalyptus camaldulensis volatiles using direct thermal desorption coupled with comprehensive two-dimensional gas chromatography-time-of-flight-mass spectrometry

The direct qualification and quantitation of the volatile organic components of four Eucalyptus camaldulensis fruit samples, obtained from different geographical areas in Turkey, is studied using a direct thermal desorption (DTD) technique coupled with comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry. It is found that the E. camaldulensis sample from Adrasan gave a slightly higher oil yield (1.18%) than the others. The number of components quantitatively identified from Adrasan, Belek, Kuyucak, and Cesme were 46, 54, 55, and 59, respectively. The main compounds found in the volatile oils were: aromadendrene (6.45-15.02%), eucalyptol (0.17-12.61%), gamma-gurjunene (8.40-10.08%), terpinolen (1.98-8.39%), spathulenol (1.42-8.34%), alpha-pinene (0.85-6.81%), ledene (0.94-6.72%), and longifonene (0.07-6.22%). The composition of the volatiles desorbed from samples from all four different areas varied qualitatively and quantitatively. All identified compounds were quantitated using total ion chromatogram peak areas. DTD is a good method for qualitative and/or quantitative analysis of complex mixtures, and in particular for quantitative analysis of plant samples, which can yield data without the traditional obligation for costly and time consuming extraction techniques.     

Untargeted metabolomics liquid chromatography-high resolution mass spectrometry approach for the geographical origin assessment of Italian dehydrated apples

Geographical provenience is nowadays a relevant aspect of the authenticity and the quality of many food commodities. Dehydrated apple cubes/slices represent an ingredient commonly used by food companies for bakery products. However, this apple-based matrix is not so known and studied from an analytical point of view. In the present work, seven compounds were identified as key molecules to distinguish between Italian and non-Italian samples, through an untargeted ultrahigh-pressure liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) approach. This methodology was merged with multivariate statistical analysis, and the principal features were studied and identified considering several identification steps. Samples from 2020 and 2021 harvesting campaigns, with partial and total dehydration rates, with or without peel, and from different apple varieties were considered for the study, for a total of 91 samples. Afterward, the same analysis protocol was applied to an external set (n = 12 samples), included in the statistical models, searching for the key compounds identified in the training set. Interesting and significant results underlined the potentiality of the UHPLC-HRMS technology as a confirmatory strategy for the geographical origin assessment of dehydrated apple commodities.     

Headspace solid-phase microextraction-gas chromatographic-time-of-flight mass spectrometric methodology for geographical origin verification of coffee

Increasing consumer awareness of food safety issues requires the development of highly sophisticated techniques for the authentication of food commodities. The food products targeted for falsification are either products of high commercial value or those produced in large quantities. For this reason, the present investigation is directed towards the characterization of coffee samples according to the geographical origin. The conducted research involves the development of a rapid headspace solid-phase microextraction (HS-SPME)-gas chromatography-time-of-flight mass spectrometry (GC-TOFMS) method that is utilized for the verification of geographical origin traceability of coffee samples. As opposed to the utilization of traditional univariate optimization methods, the current study employs the application of multivariate experimental designs to the optimization of extraction-influencing parameters. Hence, the two-level full factorial first-order design aided in the identification of two influential variables: extraction time and sample temperature. The optimum set of conditions for the two variables was 12 min and 55 degrees C, respectively, as directed by utilization of Doehlert matrix and response surface methodology. The high-throughput automated SPME procedure was completed by implementing a single divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) 50/30 microm metal fiber with excellent durability properties ensuring the completion of overall sequence of coffee samples. The utilization of high-speed TOFMS instrument ensured the completion of one GC-MS run of a complex coffee sample in 7.9 min and the complete list of benefits provided by ChromaTOF software including fully automated background subtraction, baseline correction, peak find and mass spectral deconvolution algorithms was exploited during the data evaluation procedure. The combination of the retention index (RI) system using C(8)-C(40) alkanes and the mass spectral library search was utilized for the confirmation of analyte identity in the reference authentic Brazilian coffee sample. The semi-quantitative results were then submitted to statistical evaluation, namely principal component analysis (PCA) for the establishment of geographical origin discriminations.     

Effects of electron beam irradiation on cork volatile compounds by gas chromatography-mass spectrometry

Summary The effects of electron beam irradiation on cork volatile compounds was studied at different doses (25, 100, 1000 kGy). Volatiles were isolated from cork using the dynamic headspace-sampling technique, then identified by gas chromatography-mass spectrometry (GC-MS). Similar gas chromatographic profiles were obtained for non-irradiated and irradiated corks. Quantitative differences induced by the three doses were evaluated by calculating peak areas for each compound identified. The quantitative differences between non-irradiated corks and those irradiated at 25 kGy were significant for only a few substances, whereas significant quantitative differences were found in samples irradiated at 100 and 1000 kGy. For these doses, the content of volatile compounds generally increased, especially that of aliphatic hydrocarbons and carbonyl compounds. The behaviour of radiolytic hydrocarbons indicates that the mechanisms proposed for their formation in irradiated foods could take place even in cork.     

Recognition of volatile compounds as markers in geographical discrimination of Spanish extra virgin olive oils by chemometric analysis of non-specific chromatography volatile profiles

Chromatographic profiles obtained by headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography (GC) were processed as continuous and non-specific signals through multivariate analysis techniques in order to select and identify the most discriminate volatile marker compounds related to the geographical origin of extra virgin olive oils. The blind analysis of the chromatographic profiles was carried out on several steps including preliminary mathematical treatments, explorative analysis, feature selection and classification. The results obtained through the application of stepwise linear discriminant analysis (SLDA) method revealed a perfect discrimination between the different Spanish geographical regions considered (La Rioja, Andalusia and Catalonia). The assignment success rate was 100% in both classification and prediction by using cross validation procedure. In addition, it must be noted that the proposed strategy was able to verify the geographical origin of the samples involving only a reduced number of discriminate retention times selected by the stepwise procedure. This fact emphasizes the quality of the accurate results obtained and encourages the feasibility of similar procedures in olive oil quality and traceability studies. Finally, volatile compounds corresponding to the predictors retained were identified by gas chromatography-mass spectrometry (GC-MS) for a chemical interpretation of their importance in quality virgin olive oils.     

基于化学模式识别的大蒜傅里叶变换红外光谱指纹图谱多样性

以不同地理居群的大蒜为研究对象,采用傅里叶变换红外光谱法获取各样品的红外谱图及不同波数下各样品的透过率数据,运用主成分分析及聚类分析等统计学方法对谱图数据进行了比较分析。结果发现,不同地理居群大蒜的红外指纹图谱存在差异,统计学分析结果显示25个品种大致可分为4大类(其中2个样品,4号及25号与这4类间有较大差异),此现象一定程度上反映了大蒜种植地居群环境对大蒜理化性质的影响。     

pH-mediated field-amplified sample stacking of pharmaceutical cations in high-ionic strength samples

Capillary electrophoretic separation of samples of physiological origin typically have both poor resolution and efficiency due to destacking. We have previously reported a stacking method for concentration of catecholamines in artificial dialysate, or Ringer's solution. However, pH-mediated sample stacking of other cations has not been investigated. In this report, pH-mediated stacking has been extended to eletripan, dofetilide, doxazosin, sildenafil, UK-103,320, UK-202,581, and CP-122,288. These compounds were chosen without prior structural screening except that they were cationic at the pH of our background electrolyte (BGE). Capillary electrophoretic behavior of samples in BGE is compared with those of samples in Ringer's solution with and without pH-mediated acid stacking. Results indicate that the peak heights and efficiencies for acid-stacked samples are increased compared to the unstacked samples in Ringer's solution or BGE. For example, the peak efficiencies for 5 s injections of eletriptan in BGE and Ringer's solution are 138,000 and 72,000 plates, respectively. In contrast, a 10 s injection of eletriptan followed by acid injection for 16 s produces a peak with 246,000 plates. Evaluation of the stacking effect was performed by comparison of the peak height at similar peak efficiencies for samples in Ringer's solution with and without stacking. Using this method, pH-mediated acid stacking provides a 10- to 27-fold sensitivity enhancement for the seven cations.     

Combination of cationic surfactant-assisted solid-phase extraction with field-amplified sample stacking for highly sensitive analysis of chlorinated acid herbicides by capillary zone electrophoresis

This report describes a novel online field-amplified sample stacking (FASS) procedure to analyze 16 chlorinated acid herbicides. By using a poly(vinyl alcohol) (PVA)-coated capillary to reduce electroosmotic flow and introducing a methanol-water plug before sample loading, the sample injection time could be very long without loss of sample and separation efficiency. Under the optimized condition, the FASS procedure could provide great sensitivity enhancement (5000-10 000-fold) and satisfactory reproducibility (relative standard deviations of migration times less than 2.4%, relative standard deviations of peak areas less than 8.0%). Combined with cationic surfactant-assisted solid-phase extraction (CSA-SPE), the limit of detection of the herbicides ranged from 0.269 to 20.3 ppt, which are two orders lower than those of the US Environmental Protection Agency standard method 515.1. The CSA-SPE-FASS-CE method was successfully applied to the analysis of local pond water.