The potential of NO+ and O2 +• in switchable reagent ion proton transfer reaction time-of-flight mass spectrometry.

Selected ion flow tube mass spectrometry (SIFT-MS) and proton transfer reaction mass spectrometry with switchable reagent ion capability (PTR+SRI-MS) are analytical techniques for real-time qualification and quantification of compounds in gas samples with trace level concentrations. In the detection process, neutral compounds-mainly volatile organic compounds-are ionized via chemical ionization with ionic reagents or primary ions. The most common reagent ions are H3 O+ , NO+ and O2 +• . While ionization with H3 O+ occurs by means of proton transfer, the ionization via NO+ and O2 +• offers a larger variety on ionization pathways, as charge transfer, hydride abstraction and so on are possible. The distribution of the reactant into various reaction channels depends not only on the usage of either NO+ or O2 +• , but also on the class of analyte compounds. Furthermore, the choice of the reaction conditions as well as the choice of either SIFT-MS or PTR+SRI-MS might have a large impact on the resulting products. Therefore, an overview of both NO+ and O2 +• as reagent ions is given, showing differences between SIFT-MS and PTR+SRI-MS as used analytical methods revealing the potential how the knowledge obtained with H3 O+ for different classes of compounds can be extended with the usage of NO+ and O2 +• .

A study of 9 common breath VOCs in 504 healthy subjects using PTR-TOF-MS.

INTRODUCTION: This study employs Proton-Transfer-Reaction Mass Spectrometry (PTR-MS) to analyze exhaled breath profiles of 504 healthy adults, focusing on nine common volatile organic compounds (VOCs): acetone, acetaldehyde, acetonitrile, ethanol, isoprene, methanol, propanol, phenol, and toluene. PTR-MS offers real-time VOC measurement, crucial for understanding breath biomarkers and their applications in health assessment. OBJECTIVES: The study aims to investigate how demographic factors-gender, age, and smoking history-affect VOC concentrations in exhaled breath. The objective is to enhance our understanding of breath biomarkers and their potential for health monitoring and clinical diagnosis. METHODS: Exhaled breath samples were collected using PTR-MS, measuring concentrations of nine VOCs. The data were analyzed to discern distribution patterns across demographic groups. RESULTS: Males showed higher average VOC levels for certain compounds. Propanol and methanol concentrations significantly increased with age. Smoking history influenced VOC levels, with differences among non-smokers, current smokers, and ex-smokers. CONCLUSION: This research provides valuable insights into demographic influences on exhaled VOC profiles, emphasizing the potential of breath analysis for health assessment. PTR-MS's real-time measurement capabilities are crucial for capturing dynamic VOC changes, offering advantages over conventional methods. These findings lay a foundation for advancements in non-invasive disease detection, highlighting the importance of considering demographics in breath biomarker research.

Influence of Cheese Composition on Aroma Content, Release, and Perception.

The quality of a cheese is determined by the balance of aroma compounds primarily produced by microorganisms during the transformation of milk into ripened cheese. The microorganisms, along with the technological parameters used in cheese production, influence aroma formation. The perception of these compounds is further influenced by the composition and structure of the cheese. This study aimed to characterize how cheese composition affects aroma compound production, release, and perception. Sixteen cheeses were produced under controlled conditions, followed by a quantitative descriptive analysis post ripening. Aroma composition was analyzed using HS-SPME-GC-MS, and a dynamic sensory evaluation (TCATA) was combined with nosespace analysis using PTR-ToF-MS. Image analysis was also conducted to characterize cheese structure. Cheese fat and whey lactose contents were identified as key factors in the variability of sensory attributes. GC-MS analyses identified 27 compounds correlated with sensory attributes. In terms of aroma compound release, 23 ions were monitored, with fat, salt, and lactose levels significantly affecting the release of most compounds. Therefore, cheese fat, salt, and whey lactose levels, as well as the types of microbial strains, play a role in influencing the composition, structure, release of aroma compounds, and sensory perception.

Biogenic volatile organic compounds dominated the near-surface ozone generation in Sichuan Basin, China, during fall and wintertime.

The complex air pollution driven by both Ozone (O3) and fine particulate matter (PM2.5) significantly influences the air quality in the Sichuan Basin (SCB). Understanding the O3 formation during autumn and winter is necessary to understand the atmospheric oxidative capacity. Therefore, continuous in-site field observations were carried out during the late summer, early autumn and winter of 2020 in a rural area of Chongqing. The total volatile organic compounds (VOCs) concentration reported by a Proton-Transfer-Reaction Time-of-Flight Mass Spectrometry (PTR-ToF-MS) were 13.66 ± 9.75 ppb, 5.50 ± 2.64 ppb, and 9.41 ± 5.11 ppb in late summer, early autumn and winter, respectively. The anthropogenic VOCs (AVOCs) and biogenic VOCs (BVOCs) were 8.48 ± 7.92 ppb and 5.18 ± 2.99 ppb in late summer, 3.31 ± 1.89 ppb and 2.19 ± 0.93 ppb in autumn, and 6.22 ± 3.99 ppb and 3.20 ± 1.27 ppb in winter. A zero-dimensional atmospheric box model was employed to investigate the sensitivity of O3-precursors by relative incremental reactivity (RIR). The RIR values of AVOCs, BVOCs, carbon monoxide (CO), and nitrogen oxides (NOx) were 0.31, 0.71, 0.09, and -0.36 for late summer, 0.24, 0.59, 0.22, and -0.38 for early autumn, and 0.30, 0.64, 0.33 and -0.70 for winter, and the results showed that the O3 formation of sampling area was in the VOC-limited region, and O3 was most sensitive to BVOCs (with highest RIR values, > 0.6). This study can be helpful in understanding O3 formation and interpreting the secondary formation of aerosols in the winter.

Comparative Analysis of Volatile Organic Compound Purification Techniques in Complex Cooking Emissions: Adsorption, Photocatalysis and Combined Systems.

Volatile organic compounds (VOCs) are molecules present in our everyday life, and they can be positive, such as in the formation of odour and food flavour, or harmful to the environment and humans, and research is focusing on limiting their emissions. Various methods have been used to achieve this purpose. Firstly, we review three main degradation methods: activated carbon, photocatalysis and a synergetic system. We provide a general overview of the operative conditions and report the possibility of VOC abatement during cooking. Within the literature, none of these systems has ever been tested in the presence of complex matrices, such as during cooking processes. The aim of this study is to compare the three methods in order to understand the behaviour of filter systems in the case of realistically complex gas mixtures. Proton transfer reaction-mass spectrometry (PTR-MS) has been used in the real-time monitoring of volatilome. Due to the fact that VOC emissions are highly dependent on the composition of the food cooked, we evaluated the degradation capacity of the three systems for different burger types (meat, greens, and fish). We demonstrate the pros and cons of photocatalysis and adsorption and how a combined approach can mitigate the drawbacks of photocatalysis.

Dynamic Instrumental and Sensory Methods Used to Link Aroma Release and Aroma Perception: A Review.

Perception of flavor is a dynamic process during which the concentration of aroma molecules at the olfactory epithelium varies with time as they are released progressively from the food in the mouth during consumption. The release kinetics depends on the food matrix itself but also on food oral processing, such as mastication behavior and food bolus formation with saliva, for which huge inter-individual variations exist due to physiological differences. Sensory methods such as time intensity (TI) or the more-recent methods temporal dominance of sensations (TDS) and temporal check-all-that-apply (TCATA) are used to account for the dynamic and time-related aspects of flavor perception. Direct injection mass spectrometry (DIMS) techniques that measure in real time aroma compounds directly in the nose (nosespace), aimed at obtaining data that reflect the pattern of aroma release in real time during food consumption and supposed to be representative of perception, have been developed over the last 25 years. Examples obtained with MS operated in chemical ionization mode at atmospheric or sub-atmospheric pressure (atmospheric pressure chemical ionization APCI or proton-transfer reaction PTR) are given, with emphases on studies conducted with simultaneous dynamic sensory evaluation. Inter-individual variations in terms of aroma release and their relevance for understanding flavor perception are discussed as well as the evidenced cross-modal interactions.

Trends and applications in plant volatile sampling and analysis.

Volatile organic compounds (VOCs) released by plants serve as information and defense chemicals in mutualistic and antagonistic interactions and mitigate effects of abiotic stress. Passive and dynamic sampling techniques combined with gas chromatography-mass spectrometry analysis have become routine tools to measure emissions of VOCs and determine their various functions. More recently, knowledge of the roles of plant VOCs in the aboveground environment has led to the exploration of similar functions in the soil and rhizosphere. Moreover, VOC patterns have been recognized as sensitive and time-dependent markers of biotic and abiotic stress. This focused review addresses these developments by presenting recent progress in VOC sampling and analysis. We show advances in the use of small, inexpensive sampling devices and describe methods to monitor plant VOC emissions in the belowground environment. We further address latest trends in real-time measurements of volatilomes in plant phenotyping and most recent developments of small portable devices and VOC sensors for non-invasive VOC fingerprinting of plant disease. These technologies allow for innovative approaches to study plant VOC biology and application in agriculture.

Watching Paint Dry: Organic Vapor Emissions from Architectural Coatings and their Impact on Secondary Organic Aerosol Formation.

Emissions from volatile chemical products (VCPs) are emerging as a major source of anthropogenic secondary organic aerosol (SOA) precursors. Paints and coatings are an important class of VCPs that emit both volatile and intermediate volatility organic compounds (VOCs and IVOCs). In this study, we directly measured I/VOC emissions from representative water- (latex) and oil-based paints used in the U.S. Paint I/VOC emissions vary by several orders of magnitude by both the solvent and gloss level. Oil-based paints had the highest emissions (>105 µg/g-paint), whereas low-gloss interior paints (Flat, Satin, and Semigloss) all emitted ∼102 µg/g-paint. Emissions from interior paints are dominated by VOCs, whereas exterior-use paints emitted a larger fraction of IVOCs. Extended emission tests showed that most I/VOC emissions occur within 12-24 h after paint application, though some paints continue to emit IVOCs for 48 h or more. We used our data to estimate paint I/VOC emissions and the subsequent SOA production in the U.S. Total annual paint I/VOC emissions are 48-155 Gg (0.15-0.48 kg/person). These emissions contribute to the formation of 2.2-7.5 Gg of SOA annually. Oil-based paints contribute 70-98% of I/VOC emissions and 61-99% of SOA formation, even though they only account for a minority of paint usage.

Per-Person and Whole-Building VOC Emission Factors in an Occupied School with Gas-Phase Air Cleaning.

Using real-time measurements of CO2 and volatile organic compounds (VOCs) in the air handler of an occupied middle school, we quantified source strengths for 249 VOCs and apportioned the source to the building, occupants and their activities, outdoor air, or recirculation air. For VOCs quantified in this study, there is a source to the outdoors of 8.6 ± 1.8 g/h in building exhaust air, of which 5.9 ± 1.7 g/h can be attributed to indoor sources (the building and occupants and their activities). The corresponding whole-building area emission factor from indoor sources is 1020 ± 300 µg/(m2 h), including reactive VOCs like isoprene and monoterpenes (33 ± 5.1 and 29 ± 5.7 µg/(m2 h), respectively). Per-person emission factors are calculated for compounds associated with occupants and their activities, e.g., monoterpenes are emitted at a rate of 280 ± 80 µg/(person h). The air handler included carbon scrubbing, reducing supply air concentrations of 125 compounds by 38 ± 19% (mean ± std. dev.) with a net removal of 2.4 ± 0.4 g/h of organic compounds from the building. This carbon scrubber reduces steady-state indoor concentrations of organics by 65 µg/m3 and the contribution of indoor sources of VOCs to the outdoor environment by ∼40%. These data inform the design and operation of buildings to reduce human exposure to VOCs inside buildings. These data indicate the potential for gas-phase air cleaning to improve both indoor air quality and reduce VOC emissions from buildings to the outdoor environment.

Analysis of volatile organic compounds from deep airway in the lung through intubation sampling.

Exhaled volatile organic compounds (VOCs) have been widely applied for the study of disease biomarkers. Oral exhalation and nasal exhalation are two of the most common sampling methods. However, VOCs released from food residues and bacteria in the mouth or upper respiratory tract were also sampled and usually mistaken as that produced from body metabolism. In this study, exhalation from deep airway was first directly collected through intubation sampling and analyzed. The exhalation samples of 35 subjects were collected through a catheter, which was inserted into the trachea or bronchus through the mouth and upper respiratory tract. Then, the VOCs in these samples were detected by proton transfer reaction mass spectrometry (PTR-MS). In addition, fast gas chromatography proton transfer reaction mass spectrometry (FGC-PTR-MS) was used to further determine the VOCs with the same mass-to-charge ratios. The results showed that there was methanol, acetonitrile, ethanol, methyl mercaptan, acetone, isoprene, and phenol in the deep airway. Compared with that in oral exhalation, ethanol, methyl mercaptan, and phenol had lower concentrations. In detail, the median concentrations of ethanol, methyl mercaptan, and phenol were 7.3, 0.6, and 23.9 ppbv, while those in the oral exhalation were 80.0, 5.1, and 71.3 ppbv, respectively, which meant the three VOCs mainly originated from the food residues and bacteria in the mouth or upper respiratory tract, rather than body metabolism. The research results in our study can provide references for expiratory VOC research based on oral and nasal exhalation samplings, which are more feasible in clinical practice.

Nanoplastics measurements in Northern and Southern polar ice.

It has been established that various anthropogenic contaminants have already reached all the world's pristine locations, including the polar regions. While some of those contaminants, such as lead and soot, are decreasing in the environment, thanks to international regulations, other novel contaminants emerge. Plastic pollution has been shown as a durable novel pollutant, and, since recently, smaller and smaller plastics particles have been identified in various environments (air, water and soil). Considerable research already exists measuring the plastics in the 5 mm to micrometre size range (microplastics). However, far less is known about the plastics debris that fragmented to the sub-micrometre size (nanoplastics). As these small particles are light, it is expected that they have already reached the most remote places on Earth, e.g. transported across the globe by air movement. In this work, we used a novel method based on Thermal Desorption - Proton Transfer Reaction - Mass Spectrometry (TD-PTR-MS) to detect and measure nanoplastics of different types in the water sampled from a Greenland firn core (T2015-A5) and a sea ice core from Antarctica. We identify polyethylene (PE), polypropylene (PP), polyethylene terephthalate (PET), polystyrene (PS), polyvinyl chloride (PVC), and Tire wear nanoparticles in the 14 m deep Greenland firn core and PE, PP and PET in sea ice from Antarctica. Nanoplastics mass concentrations were on average 13.2 ng/mL for Greenland firn samples and 52.3 ng/mL for Antarctic sea ice. We further discuss the possible sources of nanoplastics that we found at these remote locations, which likely involve complex processes of plastic circulation (emission from both land and sea surface, atmospheric and marine circulation).

Tracer-based characterization of source variations of ambient isoprene mixing ratios in a hillocky megacity, India, influenced by the local meteorology.

The ambient biogenic volatile organic compounds (BVOCs), mainly isoprene, are potentially involved in the formation of secondary pollutants, hence, they are significant in terms of air quality and climate. Although the largest sources of BVOCs are tropical regions, the measurements of isoprene in the Indian subcontinent are limited. We conducted the measurements of isoprene, benzene, and toluene at an urban site in a hillocky megacity of India using a high-sensitivity proton transfer reaction quadrupole mass spectrometer (PTR-QMS). The mixing ratios of isoprene were compared with those of aromatic compounds like benzene and toluene, which represent typical anthropogenic VOCs. Isoprene and isoprene/benzene (>5 ppbv ppbv-1) showed higher levels in the pre-monsoon months, most likely due to large emissions by urban vegetation during physiological activities in plants which was enhanced by the high ambient temperatures and solar radiation. While Benzene and toluene showed higher mixing ratios during winter, which were due to shallower boundary layer depths and transport of air masses from polluted Indo-Gangetic Plain during this season. The mixing ratios of VOCs show significant diurnal variation as a result of their different origins and the role of different meteorological parameters. The robust emission ratios of isoprene/benzene obtained from nighttime data were used to separate the non-anthropogenic and anthropogenic isoprene emissions. ∼30% enhancement observed in non-anthropogenic emissions to isoprene from winter to pre-monsoon season when temperatures and solar radiation were stronger, although traffic in the city. Isoprene/benzene ratio at lower temperatures (<25 °C) and solar radiation (<100 W m-2) was predominantly controlled by anthropogenic sources. Overall, toluene and isoprene are the most frequent species in terms of having the highest ozone-forming potential (OFP) values but biogenic isoprene became more important to ozone formation during the afternoon hours in the pre-monsoon months with high air temperatures (>25 °C).

Proton-transfer-reaction mass spectrometry (PTR-MS) for online monitoring of glucose depletion and cell concentrations in HEK 293 gene therapy processes.

OBJECTIVES: The applicability of proton-transfer-reaction mass spectrometry (PTR-MS) as a versatile online monitoring tool to increase consistency and robustness for recombinant adeno-associated virus (rAAV) producing HEK 293 bioprocesses was evaluated. We present a structured workflow to extract process relevant information from PTR-MS data. RESULTS: Reproducibility of volatile organic compound (VOC) measurements was demonstrated with spiking experiments and the process data sets used for applicability evaluation consisted of HEK 293 cell culture triplicates with and without transfection. The developed data workflow enabled the identification of six VOCs, of which two were used to develop a soft sensor providing better real-time estimates than the conventional capacitance sensor. Acetaldehyde, another VOC, provides online process information about glucose depletion that can directly be used for process control purposes. CONCLUSIONS: The potential of PTR-MS for HEK 293 cell culture monitoring has been shown. VOC data derived information can be used to develop soft sensors and to directly set up new process control strategies.

The Effect of Corn Dextrin on the Rheological, Tribological, and Aroma Release Properties of a Reduced-Fat Model of Processed Cheese Spread.

Low-calorie and low-fat foods have been introduced to the market to fight the increasing incidence of overweightness and obesity. New approaches and high-quality fat replacers may overcome the poor organoleptic properties of such products. A model of processed cheese spread (PCS) was produced as a full-fat version and with three levels of fat reduction (30%, 50%, and 70%). Fat was replaced by water or by corn dextrin (CD), a dietary fiber. Additionally, in the 50% reduced-fat spreads, fat was replaced by various ratios of CD and lactose (100:0, 75:25, 50:50, 25:75, and 0:100). The effect of each formulation was determined by measuring the textural (firmness, stickiness, and spreadability), rheological (flow behavior and oscillating rheology), tribological, and microstructural (cryo-SEM) properties of the samples, as well as the dynamic aroma release of six aroma compounds typically found in cheese. Winter's critical gel theory was a good approach to characterizing PCS with less instrumental effort and costs: the gel strength and interaction factors correlated very well with the spreadability and lubrication properties of the spreads. CD and fat exhibited similar interaction capacities with the aroma compounds, resulting in a similar release pattern. Overall, the properties of the sample with 50% fat replaced by CD were most similar to those of the full-fat sample. Thus, CD is a promising fat replacer in PCS and, most likely, in other dairy-based emulsions.

Discrimination of French wine brandy origin by PTR-MS headspace analysis using ethanol ionization and sensory assessment.

The headspace volatile organic compound (VOC) fingerprints (volatilome) of French wine brandies were investigated by proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS). Protonated ethanol chemical ionization was used with dedicated experimental conditions that were previously validated for model wines. These included a reference vial containing a hydro-alcoholic solution with the same ethanol content (20% v/v) as the diluted sample spirits, which was used to establish steady-state ionization conditions. A low electric field strength to number density ratio E/N (85 Td) was used in the drift tube in order to limit the fragmentation of the protonated analytes. The obtained headspace fingerprints were used to investigate the origin of French brandies produced within a limited geographic production area. Brandies of two different vintages (one freshly distilled and one aged for 14 years in French oak barrels) were successfully classified according to their growth areas using unsupervised (principal component analysis, PCA) and supervised (partial least squares regression discriminant analysis, PLS-DA) multivariate analyses. The models obtained by PLS-DA allowed the identification of discriminant volatile compounds that were mainly characterised as key aroma compounds of wine brandies. The discrimination was supported by sensory evaluation conducted with free sorting tasks. The results showed that this ethanol ionization method was suitable for direct headspace analysis of brandies. They also demonstrated its ability to distinguish French brandies according to their growth areas, and this effect on brandy VOC composition was confirmed at a perceptive level.

Estimating person-to-person variability in VOC emissions from personal care products used during showering.

An increasing fraction of volatile organic compounds (VOC) emissions come from the domestic use of solvents, contained within myriad commonplace consumer products. Emission rates are often poorly characterized and depend significantly on individual behavior and specific product formulation and usage. Time-concentration profiles of volatile organic compounds (VOCs) arising from the use of a representative selection of personal care products (PCPs) during showering are generated, and person-to-person variability in emissions calculated. A panel of 18 participants used a standardized set of products, dosages, and application times during showering in a controlled indoor bathroom setting. Proton transfer mass spectrometry was used to measure the in-room VOC evolution of limonene (representing the sum of monoterpenes), benzyl alcohol, and ethanol. The release of VOCs had reproducible patterns between users, but noticeable variations in absolute peak concentrations, despite identical amounts of material being used. The amounts of VOC emitted to air for one showering activity were as follows: limonene (1.77 mg ± 42%), benzyl alcohol (1.07 mg ± 41%), and ethanol (0.33 mg ± 78%). Real-world emissions to air were between 1.3 and 11 times lower than bottom-up estimates based on dynamic headspace measurements of product emissions rates, likely a result of PCPs being washed away before VOC evaporation could occur.

Hyphenated Mass Spectrometry versus Real-Time Mass Spectrometry Techniques for the Detection of Volatile Compounds from the Human Body.

Mass spectrometry (MS) is an analytical technique that can be used for various applications in a number of scientific areas including environmental, security, forensic science, space exploration, agri-food, and numerous others. MS is also continuing to offer new insights into the proteomic and metabolomic fields. MS techniques are frequently used for the analysis of volatile compounds (VCs). The detection of VCs from human samples has the potential to aid in the diagnosis of diseases, in monitoring drug metabolites, and in providing insight into metabolic processes. The broad usage of MS has resulted in numerous variations of the technique being developed over the years, which can be divided into hyphenated and real-time MS techniques. Hyphenated chromatographic techniques coupled with MS offer unparalleled qualitative analysis and high accuracy and sensitivity, even when analysing complex matrices (breath, urine, stool, etc.). However, these benefits are traded for a significantly longer analysis time and a greater need for sample preparation and method development. On the other hand, real-time MS techniques offer highly sensitive quantitative data. Additionally, real-time techniques can provide results in a matter of minutes or even seconds, without altering the sample in any way. However, real-time MS can only offer tentative qualitative data and suffers from molecular weight overlap in complex matrices. This review compares hyphenated and real-time MS methods and provides examples of applications for each technique for the detection of VCs from humans.

Influence of Prebiotic Fructans on Retronasal Aroma from Elderly Individuals.

This study investigates for the first time the role of fructans with prebiotic effects (oligofructose and inulin) on retronasal aroma among elderly individuals. The impact of oligofructose (20% w/w) on retronasal aroma release was investigated using proton transfer reaction-mass spectrometry (PTR-MS) after 73 elderly individuals consumed aqueous solutions aromatized with five aroma compounds (pentan-2-one, nonan-2-one, hexan-2,3-dione, octanal and linalool). The influence of oligofructose and inulin (10% w/w) on the perceived intensity (n = 26) of two aroma descriptors (butter and floral) was also studied together with the possibility of a dumping effect on aroma evaluation due to the sweetness provided by the fructans. The results showed that the presence of oligofructose produced a significant reduction in retronasal aroma release, which could be generally explained by the physicochemical properties of aroma compounds. The presence of prebiotic fructans did not significantly affect the perceived intensity of butter and floral notes, although a dumping effect for the butter descriptor in the presence of oligofructose was observed. To conclude, these findings suggest that although fructans can exert an impact on retronasal aroma, they can be used at precise concentrations to increase the prebiotic fibre content of food products without affecting the aroma profile of foods.

Identification of the volatile profiles of 22 traditional and newly bred maize varieties and their porridges by PTR-QiTOF-MS and HS-SPME GC-MS.

BACKGROUND: Low adoption of maize varieties bred to address the nutritional needs of the growing African population limits their impact. Aroma is essential in consumer preference, but has hitherto hardly been studied. We analysed the volatile organic compounds of flours and porridges of 22 maize varieties belonging to four nutritionally distinct groups, namely provitamin A maize, quality protein maize, yellow and white maize. RESULTS: Proton-transfer-reaction quadrupole ion time-of-flight mass spectrometry (PTR-QiTOF-MS) analysis generated 524 mass peaks ranging from 16.007 to 448.089 m/z. Principal component analysis separated the varieties belonging to the four groups. With headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME GC-MS), 48 volatile compounds were identified in maize flour and 21 in maize porridge, including hexane, nonane, pentanoic acid, 1-octen-3-ol, 1-hexanol, hexanal, nonanal, 2-pentylfuran and 2-heptanone. Volatile compounds such as 1,2,4-trimethyl benzene, associated with thermal degradation of carotenoids, increased in the porridge of yellow and provitamin A maize. CONCLUSION: The results indicate that PTR-QiTOF-MS and HS-SPME GC-MS combined with multivariate analysis are instrumental to study the volatile aroma compounds of different maize varieties.

High-throughput screening for in planta characterization of VOC biosynthetic genes by PTR-ToF-MS.

Functional characterization of plant volatile organic compound (VOC) biosynthetic genes and elucidation of the biological function of their products often involve the screening of large numbers of plants from either independent transformation events or mapping populations. The low time resolution of standard gas chromatographic methods, however, represents a major bottleneck for in planta genetic characterization of VOC biosynthetic genes. Here we present a fast and highly-sensitive method for the high-throughput characterization of VOC emission levels/patterns by coupling a Proton Transfer Reaction Time-of-Flight Mass Spectrometer to an autosampler for automation of sample measurement. With this system more than 700 samples per day can be screened, detecting for each sample hundreds of spectrometric peaks in the m/z 15-300 range. As a case study, we report the characterization of VOC emissions from 116 independent Arabidopsis thaliana lines transformed with a putative isoprene synthase gene, confirming its function also when fused to a C-terminal 3×FLAG tag. We demonstrate that the method is more reliable than conventional characterization of transgene expression for the identification of the most highly isoprene-emitting lines. The throughput of this VOC screening method exceeds that of existing alternatives, potentially allowing its application to reverse and forward genetic screenings of genes contributing to VOC emission, constituting a powerful tool for the functional characterization of VOC biosynthetic genes and elucidation of the biological functions of their products directly in planta.