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Multidimensional gas chromatography-olfactometry for the identification and prioritization of malodors from confined animal feeding operations.

Odor profiling efforts were directed at applying to high-density livestock operations some of the lessons learned in resolving past, highly diverse, odor-focused investigations in the consumer product industry. Solid-phase microextraction (SPME) was used for field air sampling of odorous air near and downwind of a beef cattle feedyard and a swine finisher barn in Texas. Multidimensional gas chromatography-olfactometry (MDGC-O) was utilized in an attempt to define and prioritize the basic building blocks of odor character associated with these livestock operations. Although scores of potential odorant volatiles have been previously identified in high-density livestock operations, the odor profile results developed herein suggest that only a very few of these may constitute the preponderance of the odor complaints associated with these environments. This appeared to be especially true for the case of increasing distance from both cattle feedyard and swine barn facilities, with p-cresol consistently taking on the dominant odor impact role with ever increasing distance. In contrast, at- or near-site odor profiles were shown to be much more complex, with many of the well-known lower tier odorant compounds rising in relative significance. For the cattle feedyard at- or near-site odor profiles, trimethylamine was shown to represent a significantly greater individual odor impact relative to the more often cited livestock odorants such as hydrogen sulfide, the organic sulfides, and volatile fatty acids. This study demonstrates that SPME combined with a MDGC-O-mass spectrometry system can be used for the sampling, identification, and prioritization of odors associated with livestock.

Animal Feed↗

Influence of the addition of rosemary essential oil on the volatiles pattern of porcine frankfurters.

The effect of the addition of increasing levels of rosemary essential oil (150, 300, and 600 mg/kg) on the generation of volatile compounds in frankfurters from Iberian and white pigs was analyzed using solid-phase microextraction coupled to gas chromatography and mass spectrometry (SPME-GC-MS). Lipid-derived volatiles such as aldehydes (hexanal, octanal, nonanal) and alcohols (pentan-1-ol, hexan-1-ol, oct-1-en-3-ol) were the most abundant compounds in the headspace (HS) of porcine frankfurters. Frankfurters from different pig breeds presented different volatile profiles due to their different oxidation susceptibilities as a likely result of their fatty acid composition and vitamin E content. Rosemary essential oil showed a different effect on the generation of volatiles depending on the type of frankfurter in which they were added. In frankfurters from Iberian pigs, the antioxidant effect of the essential oil improved with increasing levels, showing the highest activity at 600 mg/kg. In contrast, 150 mg/kg of the essential oil improved the oxidative stability of frankfurters from white pigs, whereas higher levels led to no effect or a prooxidant effect. The activity of the essential oil could have been affected by the different fatty acid compositions and vitamin E contents between types of frankfurters. SPME successfully allowed the isolation and analysis of volatile terpenes from frankfurters with added rosemary essential oil including alpha-pinene, beta-myrcene, l-limonene, (E)-caryophyllene, linalool, camphor, and 1,8-cineole, which might contribute to the aroma characteristics of frankfurters.

Alcohols↗

Composition and Antimicrobial Activity of Anemopsis californica leaf oil.

Isolation and characterization of leaf volatiles in Anemopsis californica (Nutt.) Hook. and Arn. (A. californica) was performed using steam distillation, solid-phase microextraction, and supercritical fluid extraction. Thirty-eight compounds were detected and identified by gas chromatography; elemicin was the major component of the leaf volatiles. While the composition of the leaf volatiles varied with method of extraction, alpha-pinene, sabinene, beta-phellandrene, 1,8-cineole, piperitone, methyl eugenol, (E)-caryophyllene, and elemicin were usually present in readily detectable amounts. Greenhouse-reared clones of a wild population of A. californica had an identical leaf volatile composition with the parent plants. Steam-distilled oil had antimicrobial properties against 3 (Staphylococcus aureus, Streptococcus pneumoniae, and Geotrichim candidum) of 11 microbial species tested. Some of this bioactivity could be accounted for by the alpha-pinene in the oil.

Anti-Infective Agents↗

Identification of metallic-smelling 1-octen-3-one and 1-nonen-3-one from solutions of ferrous sulfate.

Taste threshold tests of ferrous sulfate (FeSO4) solutions have been confounded by the presence of putative odorants. To detect the presence of odorants released from these solutions solid-phase microextraction (SPME) was used to collect volatiles in the headspace above FeSO4 solutions. Gas chromatography-olfactometry of samples collected over three time periods (1, 5, and 16 h) and at two temperatures (22 and 37 degrees C) revealed the presence of several metallic-smelling odorants in the headspace. Using authentic standards, two of the odorants were conclusively identified as 1-octen-3-one and 1-nonen-3-one. Trace levels of other odorants were also detected, but dilution experiments indicated that 1-nonen-3-one was at least 10 times more potent than anything else released from the solutions. 1-Octen-3-one and 1-nonen-3-one are excellent candidates for the metallic odor responses often observed in threshold testing of solutions of FeSO4.

Chromatography, Gas↗

Identification of components responsible for the odor of cigar smoker's breath.

Following smoking 1/2 of a cigar, the most odorous cigar tobacco smoke components extracted from the surface of the tongue by nylon-meshed swabs and then extracted from the swab headspace by solid phase microextraction were ethyl pyrrole, 2,3-dimethyl pyrazine, and 2-ethyl pyridine. Similar classes of compounds were identified from the headspace of an aqueous simulated saliva solution treated with cigar smoke. The most odorous compounds were 2,3,5-trimethyl pyridine, 2,5-dimethyl pyrazine, and 2-ethyl-3,5-dimethyl pyridine. Pyridines and pyrazines, the most prominent classes of odorous compounds identified in this experiment, may be generated during cigar pyrrolysis by cleavage of nicotine or by Maillard reaction.

Breath Tests↗

Role of the solvent glycerol in the Maillard reaction of d-fructose and l-alanine.

The volatiles in the headspace above a solution of [(13)C(6)]fructose and alanine in glycerol/water, heated in a closed vial at 130 degrees C for 2 h, were analyzed by solid-phase microextraction in tandem with GC-MS. Carbonyl compounds and pyrazines were among the detected components. The examination of their mass spectra showed that most of the 1-hydroxy-2-propanone and 2,3-pentanedione were (13)C(3)-labeled, the majority of the 2-methylpyrazine and 2-ethyl-3-methylpyrazine were (13)C(5)-labeled, and 2,5-dimethylpyrazine and 3-ethyl-2,5-dimethylpyrazine were mainly (13)C(6)-labeled. This is in agreement with the literature, and corresponds to the incorporation of fructose carbons, and in the case of 2,3-pentanedione, 2-ethyl-3-methylpyrazine, and 3-ethyl-2,5-dimethylpyrazine alanine carbons, into the molecules. However, minority fractions of 1-hydroxy-2-propanone (10%) and 2,3-pentanedione (14%) were found unlabeled, 2-methylpyrazine (10%) and 2-ethyl-3-methylpyrazine (11%) only doubly labeled, and 2,5-dimethylpyrazine (20%) and 3-ethyl-2,5-dimethylpyrazine (27%) only triply labeled, suggesting they contain carbons originating from the solvent glycerol. This could be confirmed by reaction of fructose and alanine in [(13)C(3)]glycerol/water, which produced the same volatiles, with 11-27% existent in their (13)C(3)-labeled form. Hence, glycerol participated not only as a solvent but also as a precursor in the reaction.

Alanine↗

Comparison of aroma volatiles in commercial Merlot and Cabernet Sauvignon wines using gas chromatography-olfactometry and gas chromatography-mass spectrometry.

Seventy-four aroma active compounds were observed in Merlot and Cabernet Sauvignon wines produced in California and Australia. Volatiles were sampled using solid phase microextraction and analyzed using time-intensity gas chromatography-olfactometry and gas chromatography-mass spectrometry (GC-MS). The most intense odorants were 3-methyl-1-butanol, 3-hydroxy-2-butanone, octanal, ethyl hexanoate, ethyl 2-methylbutanoate, beta-damascenone, 2-methoxyphenol, 4-ethenyl-2-methoxy-phenol, ethyl 3-methylbutanoate, acetic acid, and 2-phenylethanol. Aroma compounds were classified according to their aroma descriptor similarity and summed into nine distinct categories consisting of fruity, sulfury, caramel/cooked, spicy/peppery, floral, earthy, pungent/chemical, woody, and green/vegetative/fatty. Both Merlot and Cabernet Sauvignon wines were characterized by high fruity, caramel, green, and earthy aroma totals. Although there were distinct quantitative differences between Merlot and Cabernet wines, the relative aroma category profiles of the four wines were similar. Of the 66 volatiles identified by GC-MS, 28 were esters and 19 were minor alcohols. Between 81 and 88% of the total MS total ion chromatogram peak areas from each wine type were produced from only eight compounds: ethanol, ethyl octanoate, ethyl decanoate, ethyl acetate, 3-methyl-1-butanol, ethyl hexanoate, diethyl succinate, and 2-phenylethanol. Merlot wines from both Australia and California contained 4-5 times more ethyl octanoate than Cabernet Sauvignon wines from the same sources.

Chromatography, Gas↗

Effect of soybean lipoxygenase on volatile generation and inhibition of Aspergillus flavus mycelial growth.

Volatiles generated from lipoxygenase (LOX) normal and LOX deficient soybean (Glycine max) varieties with and without added lipase inhibited Aspergillus flavus mycelial growth and aflatoxin production. Soybean volatiles were analyzed using a solid phase microextraction (SPME) method combined with gas chromatography-mass spectrometry (GC-MS). Twenty-one compounds, including 11 aldehydes, three alcohols, four ketones, one furan, one alkane, and one alkene were detected in the LOX normal soybean line. However, only nine volatile compounds were observed in the LOX deficient soybean variety. The antifungal aldehydes hexanal and (E)-2-hexenal were observed in both LOX normal and LOX deficient lines and were detected at significantly higher amounts in soybean homogenate with added lipase. These aldehydes may be formed through alternate pathways, other than the LOX pathway, and may account for the inhibition of A. flavus growth observed. Other volatiles detected, particularly the ketones and alcohols, may contribute to the antifungal activity observed in both LOX normal and LOX deficient soybean lines. These results suggest that other factors, other than LOX activity, may better explain why soybeans are generally not as severely affected by A. flavus and aflatoxin contamination as other oilseed crops.

Aflatoxins↗

Analysis of volatile compounds and triglycerides of seed oils extracted from different poppy varieties (Papaver somniferum L.).

Poppy seed oil (Oleum Papaveris Seminis) is used for culinary and pharmaceutical purposes, as well as for making soaps, paints, and varnishes. Astonishingly, hardly anything was yet known about the volatile compounds of this promising comestible. Likewise, there are no current published data about the triglyceride (TAG) composition of poppy seed oils available. In this investigation solid-phase microextraction (SPME) with DVB/Carboxen/PDMS Stable-Flex fiber was applied to the study of volatile compounds of several seed oil samples from Papaver somniferum L. (Papaveraceae). 1-Pentanol (3.3-4.9%), 1-hexanal (10.9-30.9%), 1-hexanol (5.3-33.7%), 2-pentylfuran (7.2-10.0%), and caproic acid (2.9-11.5%) could be identified as the main volatile compounds in all examined poppy seed oil samples. Furthermore, the TAG composition of these oils was analyzed by MALDI-ReTOF- and ESI-IT-MS/MS. The predominant TAG components were found to be composed of linoleic, oleic, and palmitic acid, comprising approximately 70% of the oils. TAG patterns of the different poppy varieties were found to be very homogeneous, showing also no significant differences in terms of the applied pressing method of the plant seeds.

Chromatography, Gas↗

Model studies on the efficacy of protein homogenates from raw pork muscle and dry-cured ham in binding selected flavor compounds.

The binding of sarcoplasmic and myofibrillar proteins extracted from postrigor pork muscle and from 7 and 12 months dry-cured hams with volatile compounds such as 3-methyl-butanal, 2-methyl-butanal, 2-pentanone, hexanal, methional, and octanal was studied using solid phase microextraction and gas chromatography analysis. The binding ability of sarcoplasmic proteins from pork muscle was higher than the ability shown by 7 and 12 months dry-cured ham sarcoplasmic homogenates and also higher than the binding ability of myofibrillar homogenates. The effect of the ionic strength on the binding was also studied. This effect was more important on myofibrillar proteins due to its ability to produce changes on the protein conformation that affect their binding ability. However, the sarcoplasmic protein binding ability was more related to the small compounds present in this homogenate than with the aqueous phase ionic strength.

Aldehydes↗

Quantification of flavor-related compounds in the unburned contents of bidi and clove cigarettes.

Bidi cigarettes, small hand-rolled cigarettes produced primarily in India, are sold in the United States in a wide variety of candy-like flavors (e.g. dewberry, chocolate, clove) and are popular with adolescents. Many flavored bidis contain high concentrations of compounds such as eugenol, anethole, methyleugenol, pulegone, and estragole; several of these compounds have known toxic or carcinogenic properties. Clove cigarettes, or kreteks, are another highly flavored tobacco product with high levels of eugenol due to clove buds present in the tobacco filler. In this study, compounds in the burnable portion-the filler and wrapper material actually consumed during the smoking of bidis, kreteks, and U.S. cigarettes-were analyzed. Flavor-related compounds were solvent extracted from the burnable portion of each cigarette with methanol. An aliquot of the methanol extract was heated, and the sample headspace was sampled with a solid-phase microextraction fiber and introduced into a gas chromatograph-mass spectrometer for analysis in selected-ion monitoring mode. High levels of eugenol were detected in five clove-flavored bidi brands ranging from 78.6 to 7130 microg/cigarette (microg/cig), whereas diphenyl ether (128-3550 microg/cig) and methyl anthranilate (154-2360 microg/cig) were found in one grape-flavored bidi brand. A nontobacco herbal bidi brand contained the greatest variety of compounds, including anethole (489-665 microg/cig), eugenol (1670-2470 microg/cig), methyleugenol (27.7-36.6 microg/cig), safrole (32.4-34.4 microg/cig), myristicin (170-247 microg/cig), and elemicin (101-109 microg/cig). Filler from kreteks was found to contain high levels of eugenol, anethole, and coumarin. Flavored bidis and clove cigarettes contain a number of compounds that are present at levels far exceeding those reported in U.S. cigarette tobacco. Research is underway to determine the levels of these compounds delivered in smoke. It is not known what effect inhalation of these compounds has on smokers.

Filtration↗

Changes in headspace volatile concentrations of coffee brews caused by the roasting process and the brewing procedure.

Headspace-solid-phase microextraction technique (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-olfactometry (GC-O) were used to characterize the aroma compounds of coffee brews from commercial conventional and torrefacto roasted coffee prepared by filter coffeemaker and espresso machine. A total of 47 volatile compounds were identified and quantified. Principal component analysis (PCA) was applied to differentiate coffee brew samples by volatile compounds. Conventional and torrefacto roasted coffee brews were separated successfully by principal component 1 (68.5% of variance), and filter and espresso ones were separated by principal component 2 (19.5% of variance). By GC olfactometry, a total of 34 aroma compounds have been perceived at least in half of the coffee extracts and among them 28 were identified, among which octanal was identified for the first time as a contributor to coffee brew aroma.

Coffee↗

Analysis of volatile compounds as spoilage indicators in fresh king salmon (Oncorhynchus tshawytscha) during storage using SPME-GC-MS.

A method was developed for the analysis of salmon volatiles using solid-phase microextraction and gas chromatography-mass spectrometry. This method was used to monitor the volatiles of fresh king salmon (Oncorhynchus tshawytscha) stored in ambient air or in a 40:60 (v/v) mixture of CO2:N2 over time. The levels of several of the volatile compounds were found to change during storage, with some showing a clear difference between storage in air and storage in CO2:N2. Of these, several alcohols (cyclopentanol, Z-2-penten-1-ol, 1-penten-3-ol, and 1-octen-3-ol) and aldehydes (hexanal, octanal, E-2-pentenal, and E-2-hexenal) were identified as potential markers for salmon freshness. Several other volatiles (acetoin, ethyl benzene, propyl benzene, styrene, 3-methyl butanoic acid, and acetic acid) were identified as potential markers for salmon spoilage. A comparison of salmon harvested with and without the "rested harvesting" technique showed that E- and Z-isoeugenol levels were increased by the use of the isoeugenol based anesthetic. The use of the anesthetic did not affect the levels of any of the other compounds identified.

Air↗

Sample extraction method combining micellar extraction-SPME and HPLC for the determination of organochlorine pesticides in agricultural soils.

A method for the determination of organochlorine pesticides in soil samples combining microwave assisted micellar extraction (MAME) with solid-phase microextraction (SPME) and high-performance liquid chromatography-UV has been developed. A mixture of two nonionic surfactants (polyoxyethylene 10 lauryl ether and polyoxyethylene 10 stearyl ether) was used for the extraction of pesticides from agricultural soils, and different types of SPME fibers were compared. The different parameters which affect extraction efficiency in the SPME procedure were optimized such as extraction time and temperature. The method developed involves extraction and preconcentration for the target analytes in soil samples. The analytical parameters were also studied and good recoveries obtained, RSD being lower than 10% and detection limits ranging between 36 and 164 ng g(-1) for the pesticides studied. The proposed method was successfully applied to the determination of some organochlorine pesticides in several kinds of agricultural soil samples with different characteristics.

Agriculture↗

Effect of high-pressure-moderate-temperature processing on the volatile profile of milk.

The effects of high hydrostatic pressure on volatile generation in milk were investigated in this study. Raw milk samples were treated under different pressures (482, 586, and 620 MPa), temperatures (25 and 60 degrees C), and holding times (1, 3, and 5 min). Samples submitted to heat treatments alone (25, 60, and 80 degrees C for 1, 3, and 5 min) were used for comparison. Trace volatile sulfur compounds were analyzed using solid-phase microextraction (SPME) and gas chromatography (GC) with pulsed-flame photometric detection (PFPD), whereas the rest of the volatile compounds were analyzed using SPME-GC with flame ionization detection (FID). Multivariate analysis of variance (MANOVA) and principal component analysis (PCA) were used to study the effect of pressure, temperature, and time on volatile generation. Relative concentration increases of 27 selected volatile compounds were compared to an untreated sample. It was found that pressure, temperature, and time, as well as their interactions, all had significant effects (P < 0.001) on volatile generation in milk. Pressure and time effects were significant at 60 degrees C, whereas their effects were almost negligible at 25 degrees C. The PCA plot indicated that the volatile generation of pressure-heated samples at 60 degrees C was different from that of heated-alone samples. Heat treatment tended to promote the formation of methanethiol, hydrogen sulfide, methyl ketones, and aldehydes, whereas high-pressure treatment favored the formation of hydrogen sulfide and aldehydes.

Animals↗

Analytical discrimination of poisonous and nonpoisonous chemotypes of giant fennel (Ferula communis L.) through their biologically active and volatile fractions.

Giant fennel (Ferula communis L.) from Sardinia is characterized by two chemotypes with different biological activities. One chemotype is poisonous, due to prenylcoumarins, and responsible for ferulosis, which mainly affects sheep and goats, cattle, and horses; the other chemotype is nonpoisonous and contains daucane esters. The two chemotypes cannot be distinguished botanically. High-performance liquid chromatography-diode array-ultraviolet detection-mass spectrometry (HPLC-DAD-UV-MS) analysis of the composition of the fractions containing the biologically active metabolites and of the volatile fractions, by gas chromatography-mass spectrometry (GC-MS), of both essential oil and headspace sampled by headspace solid-phase microextraction (HS-SPME) are here shown to be effective in discriminating the poisonous and nonpoisonous chemotypes. HS-SPME with CAR/PDMS/DVB in combination with GC-MS has also been found to be a successful, fully automated one-step method for rapid and unequivocal discrimination of the two chemotypes, using aristolene and allohedycaryol as markers of the poisonous and nonpoisonous chemotypes, respectively.

Chromatography, High Pressure Liquid↗

Comparison of the amounts of volatile compounds in French protected designation of origin virgin olive oils.

Headspace solid-phase microextraction (HS-SPME) -gas chromatography using flame ionization detection and multivariate analysis were applied to the study of the specificity of protected designation of origin (PDO) virgin olive oils produced in a southern French region (Alpes-Maritimes) based on their volatile compounds. A total of 35 PDO olive oils from Nice, 6 commercial oils, and 12 other French PDO olive oils were analyzed. Recorded data were subjected to principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA). The method developed here was able to perfectly distinguish different qualities of olive oils. Representative samples from each class obtained by chemometric treatment were analyzed by HS-SPME and GC-MS. PCA and SIMCA of chromatographic data were related to sensory analysis and led to a better understanding of the chemical features and observed sensory effects of olive oils.

Chromatography, Gas↗

Quantification of jasmonic acid by SPME in tomato plants stressed by ozone.

Jasmonates are signalling molecules induced in plants as a response to various biotic and/or abiotic stresses. As ozone is known to activate defense responses in plants, we have monitored the concentration of jasmonic acid in tomato leaves during and after an acute exposure to this abiotic elicitor. In this experiment, we observed that the maximum induction of jasmonic acid in O3-fumigated plants occurred 9 h after the end of treatment and the concentration of jasmonic acid in stressed plants increased 13-fold. However, the level of endogenous methyl-jasmonate was constant during the observed period. The extraction and quantification of jasmonic acid as its methyl ester was performed by headspace-solid-phase microextraction (or HS-SPME) in combination with GC-FID and GC-MS. The sensitivity (LOD = 2 ng/g) of this method permitted the detection and quantification of jasmonic acid present in plant tissues at very low concentrations.

Chromatography, Gas↗