The complex aromas of cocktails provide a unique and interesting model system to evaluate the effects of alcohol matrix and aroma-aroma interactions on human aroma perception and partitioning and release of aroma compounds. Here, we study the interactions that occur in an Old-Fashioned cocktail when different types of whiskeys are mixed with different styles of bitters.The interactions are studied in two ways, namely, by sensory descriptive analysis to evaluate changes in human aroma perception, and by headspace solid-phase microextraction and gas chromatography-mass spectrometry to study the volatile profiles individually and upon mixing.Several aroma descriptors showed significant additive and suppressing interaction effects between bitters and whiskeys, and unique sensory characteristics were introduced by both bitters and whiskeys. Volatile compounds also showed suppressing and enhancing effects upon mixing of bitters with whiskeys.Such behaviors point towards chemical mixture effects and the enhancements in two compounds cannot be attributed to just the addition of certain bitters as the effects differ among the four whiskeys.These interactive sensory effects suggest further questions of interest about the inherent sensory complexity of foods and beverages; if sensory qualities in even simple cocktails, such as an Old-Fashioned, only exist upon mixing and for specific combinations of bitters and whiskeys, further unique interactions could be envisioned for more complex mixtures.

Demand for aromatic rice varieties (e.g., Basmati) is increasing in the US. Aromatic varieties typically have elevated levels of the aroma compound 2-acetyl-1-pyrroline (2AP). Due to its very low aroma threshold, analysis of 2AP provides a useful screening tool for rice breeders. Methods for 2AP analysis in rice should quantitate 2AP at or below sensory threshold level, avoid artifactual 2AP generation, and be able to analyze single rice kernels in cases where only small sample quantities are available (e.g., breeding trials). We combined headspace solid phase microextraction with gas chromatography tandem mass spectrometry (HS-SPME-GC-MS/MS) for analysis of 2AP, using an extraction temperature of 40 °C and a stable isotopologue as internal standard. 2AP calibrations were linear between the concentrations of 53 and 5380 pg/g, with detection limits below the sensory threshold of 2AP. Forty-eight aromatic and nonaromatic, milled rice samples from three harvest years were screened with the method for their 2AP content, and overall reproducibility, observed for all samples, ranged from 5% for experimental aromatic lines to 33% for nonaromatic lines.

•A new GC–MS/MS method to quantify four methoxypyrazines in wine was developed•Resolution of IBMP from coeluting peaks achieved•Three extraction techniques (HS-SPME, SBSE, and HSSE) were evaluated and comparedMethoxypyrazines are volatile compounds found in plants, microbes, and insects that have potent vegetal and earthy aromas. With sensory detection thresholds in the low ng L−1 range, modest concentrations of these compounds can profoundly impact the aroma quality of foods and beverages, and high levels can lead to consumer rejection. The wine industry routinely analyzes the most prevalent methoxypyrazine, 2-isobutyl-3-methoxypyrazine (IBMP), to aid in harvest decisions, since concentrations decrease during berry ripening. In addition to IBMP, three other methoxypyrazines IPMP (2-isopropyl-3-methoxypyrazine), SBMP (2-sec-butyl-3-methoxypyrazine), and EMP (2-ethyl-3-methoxypyrazine) have been identified in grapes and/or wine and can impact aroma quality. Despite their routine analysis in the wine industry (mostly IBMP), accurate methoxypyrazine quantitation is hindered by two major challenges: sensitivity and resolution. With extremely low sensory detection thresholds (~8–15 ng L−1 in wine for IBMP), highly sensitive analytical methods to quantify methoxypyrazines at trace levels are necessary. Here we were able to achieve resolution of IBMP as well as IPMP, EMP, and SBMP from co-eluting compounds using one-dimensional chromatography coupled to positive chemical ionization tandem mass spectrometry. Three extraction techniques HS-SPME (headspace-solid phase microextraction), SBSE (stirbar sorptive extraction), and HSSE (headspace sorptive extraction) were validated and compared. A 30 min extraction time was used for HS-SPME and SBSE extraction techniques, while 120 min was necessary to achieve sufficient sensitivity for HSSE extractions. All extraction methods have limits of quantitation (LOQ) at or below 1 ng L−1 for all four methoxypyrazines analyzed, i.e., LOQ’s at or below reported sensory detection limits in wine. The method is high throughput, with resolution of all compounds possible with a relatively rapid 27 min GC oven program.

•A new method for profiling grape monoterpene glycosides using UHPLC-qTOF MS was developed.•Monoterpenol trisaccharide glycosides were tentatively identified here for the first time in any plant.•Monoterpene glycoside profiles varied across different maturity stages.In this work we present a novel approach for the identification of plant metabolites using ultrahigh performance liquid chromatography coupled to accurate mass time-of-flight mass spectrometry. The workflow involves developing an in-house compound database consisting of exact masses of previously identified as well as putative compounds. The database is used to screen accurate mass spectrometry (MS) data to identify possible compound matches. Subsequent tandem MS data is acquired for possible matches and used for structural elucidation. The methodology is applied to profile monoterpene glycosides in Vitis vinifera cv. Muscat of Alexandria grape berries over three developmental stages. Monoterpenes are a subclass of terpenes, the largest class of plant secondary metabolites, and are found in two major forms in the plant, “bound” to one or more sugar moieties or “free” of said sugar moieties. In the free form, monoterpenes are noted for their fragrance and play important roles in plant defense and as attractants for pollinators. However, glycoconjugation renders these compounds odorless, and it is this form that the plant uses for monoterpene storage. In order to gain insight into monoterpene biochemistry and their fate in the plant an analysis of intact glycosides is essential. Eighteen monoterpene glycosides were identified including a monoterpene trisaccharide glycoside, which is tentatively identified here for this first time in any plant. Additionally, while previous studies have identified monoterpene malonylated glucosides in other grapevine tissue, we tentatively identify them for the first time in grape berries. This analytical approach can be readily applied to other plants and the workflow approach can also be used for other classes of compounds. This approach, in general, provides researchers with data to support the identification of putative compounds, which is especially useful when no standard is available.

•16 commercial bitters differed significantly in several sensory attributes.•The bitters samples also differed significantly in their volatile composition.•The volatile profile explained ∼60% of the differences in the sensory responses.•Not all sensory descriptors were explained by the volatile chemical composition.Aromatic cocktail bitters are derived from the alcoholic extraction of a variety of plant materials and are used as additives in mixed drinks to enhance aroma and flavor. In this study sixteen commercial bitters were analyzed using volatile (GC–MS) and sensory profiling and multivariate statistics including Principal Component Analysis (PCA) and Partial Least Squares Regression (PLS). The samples differed significantly in their citrus, celery, and spice characteristics. 148 volatile compounds were tentatively identified and the composition varied significantly with the type of bitters sample evaluated. PLS analysis showed that the volatile data correlated well overall to the sensory data, explaining 60% of the overall variability in the dataset. Primary aldehydes and phenylpropanoids were most closely related to green and spice-related sensory descriptors. However, the sensory impact of terpenoid compounds was difficult to predict in many cases. This may be due to the wide range of aroma qualities associated with terpenes as well as to concentration, synergistic or masking effects.

Catalytic upgrading of bio-oils formed in the pyrolysis of biomass represents a promising approach to the production of renewable fuels and platform chemicals. Development of fundamental information needed for commercialization of bio-oil upgrading requires analytical methods for characterization of complex mixtures. We report data characterizing the conversion of a prototypical bio-oil product, 2-butanone, catalyzed by the zeolite HZSM-5 at 300 °C and a quantitative analysis of the products by a multiple-detector gas chromatographic technique complemented by gas chromatography–mass spectrometry. 2-Butanone was chosen as the reactant because it is converted into a wide range of compounds representing functional groups characteristic of products of bio-oil conversion; 140 products were identified. Among those in the liquid phase for which standards were available, linear (correlation coefficient R > 0.998) calibrations were obtained over concentration ranges typically found in bio-oil products. Results were reproducible with a relative standard deviation (% RSD; n = 3) less than 5% for repeated injections for all products, except n-hexane. Many of the products were recovered in both the gas and liquid phases, and overall recoveries are reported that are dependent upon boiling point and conversion to other products. The reported procedures include an accurate method for measurement of water, a product of almost all bio-oil conversion processes.

Wood samples collected from multiple barrels during the toasting process at a commercial cooperage were analyzed for their content of several volatile compounds related to the aroma of toasted oak wood. Changes in the content of these compounds during the toasting process are presented for two different toasting protocols. Multifactor analysis (MFA) and partial least squares (PLS) regression were done to relate the volatile composition of the wood samples to the stage of the toasting process. These findings may provide objective guidance to coopers for the development of toasting protocols.

•UHPLC–QTOF MS was used to differentiate whiskeys based on type, producer and age.•Components used in sample classification were identified by discriminant analysis.•Wood- and fermentation-derived components were identified by MS and MS/MS.•The greatest compositional differences were observed between new (e.g., American) and aged whiskeys.Commercial samples of 63 American whiskeys, including bourbon whiskeys, Tennessee whiskeys, rye whiskeys and other blended whiskeys were analysed using ultra high pressure liquid chromatography (UHPLC) coupled with quadrupole time-of-flight (QTOF) mass spectrometry (MS). The non-volatile composition of the whiskeys was used to model differences among the samples using discriminant analysis. The blended American whiskeys were readily distinguished from the remaining types. Additionally, most Tennessee whiskeys could be differentiated from bourbon and rye whiskeys. Similarly, younger (<4 years old) and older (>8 years old) whiskeys could be separated. The compounds important for differentiating among these whiskeys included wood derived phenolic compounds, lignan derived compounds and several C8 and larger lipids. A number of additional compounds differentiated the whiskeys but could not be identified using MS and MS/MS data alone.

Hydrophobic compounds are important odorants and nutrients in foods and beverages, as well as environmental contaminants and pharmaceuticals. Factors influencing their partitioning within multi-component systems and/or from the bulk liquid phase to the air are critical for understanding aroma quality and nutrient bioavailability. The equilibrium partitioning of hydrophobic analytes between air and water was analyzed using solid phase microextraction (SPME) in the headspace (HS-SPME) and via direct immersion in the liquid (DI-SPME). The compounds studied serve as models for hydrophobic aroma compounds covering a range of air–water partition coefficients that extends over four orders of magnitude. By varying the total amount of analyte as well as the ratio of vapor to liquid in the closed, static system, the partition coefficient, Kv, can be determined without the need for an external calibration, eliminating many potential systematic errors. Kv determination using DI-SPME in this manner has not been demonstrated before. There was good agreement between results determined by DI-SPME and by HS-SPME over the wide range of partitioning behavior studied. This shows that these two methods are capable of providing accurate, complementary measurements. Precision in Kv determination depends strongly on Kv magnitude and the ratio of the air and liquid phases.

2-Methoxy-3-isobutylpyrazine (MIBP) contributes a bell pepper aroma to many grape cultivars and has a reported aroma threshold of ∼2 ng L−1 in water. The purpose of this study was twofold: (1) develop a procedure using headspace solid phase micro-extraction combined with GC–MS in the selected ion monitoring mode (HS-SPME-GC–MS-SIM) for analysis of MIBP in grape berries, and (2) determine the location of MIBP biosynthesis in grapevines by approach grafting clusters of Vitis vinifera L. cvs Cabernet Sauvignon and Muscat blanc onto each other. The soluble solids and pH of the grape juice/homogenate matrix from different grape berry developmental stages influenced the method precision; therefore, quantification via the method of standard addition was used. Using our developed method, the limit of detection (LOD) and limit of quantitation (LOQ) of MIBP were 0.1 ng L−1 and 2 ng L−1, respectively, measured in a model juice and non-MIBP containing Chardonnay juice. Spiked recoveries averaged between 91% and 112% in Cabernet Sauvignon and Pinot noir homogenates and the overall relative standard deviation was less than 10%. The method was used to analyze MIBP in 29 grape cultivars and in fruit from clusters grafted to Cabernet Sauvignon or Muscat vines. Quantifiable levels were found only in Cabernet franc, Cabernet Sauvignon, Merlot, Sauvignon blanc and Semillon, providing information on the genetic connection for the occurrence of MIBP in grapes. No MIBP was detected in the berries of Muscat blanc clusters grafted onto Cabernet Sauvignon vines when sampled at fruit maturity. MIBP was detected in all berries of Cabernet Sauvignon regardless the graft configuration. The data indicate that MIBP or its precursors originate in the berry and its formation depends upon grape genotype.Grape clusters approach grafted to Cabernet Sauvignon or Muscat vines showed that fruit genotype, not shoot genotype, determined the presence or absence of fruit MIBP.

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.

A full-factorial design was used to assess the matrix effects of ethanol, glucose, glycerol, catechin, and proline on the volatile partitioning of 20 volatile compounds considered to play a role in wine aroma. Analysis of variance showed that the two-way interactions of ethanol and glucose, ethanol and glycerol, and glycerol and catechin significantly influenced headspace partitioning of volatiles. Experiments were conducted to observe the effect of varied ethanol and glucose concentrations on headspace partitioning of a constant concentration of volatiles. Analysis of variance and linear regression analysis showed that the presence of glucose increased the concentration of volatiles in the headspace, whereas increasing ethanol concentration was negatively correlated with headspace partitioning of volatiles. A subsequent study assessed the effect of diluting white and red wines with water and ethanol. It was again observed that increased ethanol concentration significantly reduced the relative abundance of volatile compounds in the sample headspace. This study investigates some of the complex matrix interactions of the major components of grape and wine that influence volatile compound headspace partitioning. The magnitude of each matrix−volatile interaction was ethanol > glucose > glycerol > catechin, whereas proline showed no apparent interaction. The results clearly identify that increasing ethanol concentrations significantly reduce the headspace concentration of volatile aroma compounds, which may contribute to explaining recent sensory research observations that indicate ethanol can suppress the fruit aroma attributes in wine.

Over the past century, advances in analytical chemistry have played a significant role in understanding wine chemistry and flavor. Whereas the focus in the 19th and early 20th centuries was on determining major components (ethanol, organic acids, sugars) and detecting fraud, more recently the emphasis has been on quantifying trace compounds including those that may be related to varietal flavors. In addition, over the past 15 years, applications of combined analytical and sensory techniques (e.g., gas chromatography−olfactometry) have improved the ability to relate chemical composition to sensory properties, whether identifying impact compounds or elucidating matrix effects. Many challenges remain, however. This paper discusses some of the recent research aimed at understanding how viticultural and enological practices influence grape and wine volatiles. In addition, the challenges in linking composition to sensory properties will also be reviewed. Finally, future advances in linking grape, yeast, and human genomics to wine chemistry and flavor will be briefly discussed.

Although hundreds of chemical compounds have been identified in grapes and wines, only a few compounds actually contribute to sensory perception of wine flavor. This critical review focuses on volatile compounds that contribute to wine aroma and provides an overview of recent developments in analytical techniques for volatiles analysis, including methods used to identify the compounds that make the greatest contributions to the overall aroma. Knowledge of volatile composition alone is not enough to completely understand the overall wine aroma, however, due to complex interactions of odorants with each other and with other nonvolatile matrix components. These interactions and their impact on aroma volatility are the focus of much current research and are also reviewed here. Finally, the sequencing of the grapevine and yeast genomes in the past ∼10 years provides the opportunity for exciting multidisciplinary studies aimed at understanding the influences of multiple genetic and environmental factors on grape and wine flavor biochemistry and metabolism (147 references).

The DNA maintenance enzyme, topoisomerase I, is thought to play crucial roles in all living cells and for this reason inhibitors of this enzyme have been much studied. In this paper we describe a gel electrophoresis method capable of characterizing and quantifying inhibition of topoisomerase I by selected compounds. Inhibitors of topoisomerase I are often associated with intercalative binding to DNA and the method can simultaneously determine intercalative binding (as DNA unwinding) except in the cases where inhibition is prohibitively strong. The method uses closed circular (plasmid) DNA and can separate single-strand nicked, linearized (double-strand nicked), fully relaxed, partially relaxed (topoisomers), and supercoiled forms of the plasmid so that topoisomerase-dependent DNA cleavage (poisoning) can also be determined. By quantifying poisoning, inhibition, and intercalation simultaneously and separately in relation to reference compounds it is possible to make quantitative determinations of these phenomena for comparative purposes. Data for the topoisomerase I inhibitor, luteolin, are presented.

Although hundreds of chemical compounds have been identified in grapes and wines, only a few compounds actually contribute to sensory perception of wine flavor. This critical review focuses on volatile compounds that contribute to wine aroma and provides an overview of recent developments in analytical techniques for volatiles analysis, including methods used to identify the compounds that make the greatest contributions to the overall aroma. Knowledge of volatile composition alone is not enough to completely understand the overall wine aroma, however, due to complex interactions of odorants with each other and with other nonvolatile matrix components. These interactions and their impact on aroma volatility are the focus of much current research and are also reviewed here. Finally, the sequencing of the grapevine and yeast genomes in the past ∼10 years provides the opportunity for exciting multidisciplinary studies aimed at understanding the influences of multiple genetic and environmental factors on grape and wine flavor biochemistry and metabolism (147 references).