Determination of citrus juice coumarins, furanocoumarins and methoxylated flavones using solid phase extraction and HPLC with photodiode array and fluorescence detection.

A synergistic combination of analytical techniques was developed for the simultaneous determination of the three most biologically active chemical families in citrus juices: methoxylated flavones, coumarins, and furanocoumarins. No rapid methodology has been available to determine them together. A solid phase extraction concentrated these groups and a ternary reverse phase HPLC gradient completely resolved them from other juice components. Two coumarins, isomeranzin and osthole, were identified in a sweet orange (C. sinensis) cultivar, Changyecheng, for the first time. Pummelo juice was characterized by coumarin and furanocoumarin epoxides such as meranzin and epoxybergamottin. No epoxides were observed in the more acidic juices. Added furanocoumarin epoxides hydrolyzed rapidly in the most acidic juices. The ratios of the UV peak areas at 320 nm to the fluorescence emission peaks as well as the ratio of fluorescence emission peaks at 450-400 nm could be used to identify chromatographic peaks.

Comparison of two extraction techniques, solid-phase microextraction versus continuous liquid-liquid extraction/solvent-assisted flavor evaporation, for the analysis of flavor compounds in gueuze lambic beer.

Lambic is a beer style that undergoes spontaneous fermentation and is traditionally produced in the Payottenland region of Belgium, a valley on the Senne River west of Brussels. This region appears to have the perfect combination of airborne microorganisms required for lambic's spontaneous fermentation. Gueuze lambic is a substyle of lambic that is made by mixing young (approximately 1 year) and old (approximately 2 to 3 years) lambics with subsequent bottle conditioning. We compared 2 extraction techniques, solid-phase microextraction (SPME) and continuous liquid-liquid extraction/solvent-assisted flavor evaporation (CCLE/SAFE), for the isolation of volatile compounds in commercially produced gueuze lambic beer. Fifty-four volatile compounds were identified and could be divided into acids (14), alcohols (12), aldehydes (3), esters (20), phenols (3), and miscellaneous (2). SPME extracted a total of 40 volatile compounds, whereas CLLE/SAFE extracted 36 volatile compounds. CLLE/SAFE extracted a greater number of acids than SPME, whereas SPME was able to isolate a greater number of esters. Neither extraction technique proved to be clearly superior and both extraction methods can be utilized for the isolation of volatile compounds found in gueuze lambic beer.

Comparison of aroma active and sulfur volatiles in three fragrant rice cultivars using GC-olfactometry and GC-PFPD.

Aroma volatiles from three cooked fragrant rice types (Jasmine, Basmati and Jasmati) were characterised and identified using SPME GC-O, GC-PFPD and confirmed using GC-MS. A total of 26, 23, and 22 aroma active volatiles were observed in Jasmine, Basmati and Jasmati cooked rice samples. 2-Acetyl-1-pyrroline was aroma active in all three rice types, but the sulphur-based, cooked rice character impact volatile, 2-acetyl-2-thiazoline was aroma active only in Jasmine rice. Five additional sulphur volatiles were found to have aroma activity: dimethyl sulphide, 3-methyl-2-butene-1-thiol, 2-methyl-3-furanthiol, dimethyl trisulphide, and methional. Other newly-reported aroma active rice volatiles were geranyl acetate, ß-damascone, ß-damascenone, and ɑ-ionone, contributing nutty, sweet floral attributes to the aroma of cooked aromatic rice. The first two principal components from the principal component analysis of sulphur volatiles explained 60% of the variance. PC1 separated Basmati from the other two cultivars and PC2 completely separated Jasmine from Jasmati cultivars.

Chemical and behavioral analysis of the cuticular hydrocarbons from Asian citrus psyllid, Diaphorina citri.

Huanglongbing (HLB) is the most destructive disease of citrus worldwide. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the vector of the phloem-inhabiting bacterium, Candidatus Liberibacter asiaticus, which is presumed to cause HLB in Florida citrus. Laboratory and field studies were conducted to examine the behavioral responses of male and female D. citri to their cuticular extracts. In olfactometer assays, more male D. citri were attracted to one, five, or 10 female cuticular extract equivalent units than blank controls. The results were confirmed in field studies in which clear or yellow traps baited with 10 female cuticular extract equivalent units attracted proportionately more males than clear traps baited with male cuticular extract or unbaited traps. Analyses of cuticular constituents of male and female D. citri revealed differences between the sexes in chemical composition of their cuticular extracts. Laboratory bioassays with synthetic chemicals identified from cuticular extracts indicated that dodecanoic acid attracted more males than clean air. Traps baited with dodecanoic acid did not increase total catch of D. citri as compared with blank traps at the dosages tested; however, the sex ratio of psyllid catch was male biased on traps baited with the highest lure loading dosage tested (10.0 mg).

Historical review of citrus flavor research during the past 100 years.

Citrus juices are a complex mixture of flavor and taste components. Historically, the contributions of taste components such as sugar (sweet) and acid (sour) components were understood before impactful aroma volatiles because they existed at higher concentrations and could be measured with the technologies of the 1920s and 1930s. The advent of gas chromatography in the 1950s allowed citrus researchers to separate and tentatively identify the major citrus volatiles. Additional volatiles were identified when mass spectrometry was coupled to capillary GC. Unfortunately, the major citrus volatiles were not major influences of citrus flavor. The major aroma impact compounds were found at trace concentrations. With the advent of increasingly more sensitive instrumental techniques, juice sample size shrank from 2025 L in the 1920s to 10 mL today and detection limits fell from percent to micrograms per liter. Currently gas chromatography-olfactometry is the technique of choice to identify which volatiles in citrus juices possess aroma activity, determine their relative aroma strength, and characterize their aroma quality but does not indicate how they interact together or with the juice matrix. Flavor equations based primarily on nonvolatiles and other physical measurements have been largely unsuccessful. The most successful flavor prediction equations that employ instrumental concentration values are based on a combination of aroma active volatiles and degrees Brix (sugar) values.

A comparison of citrus blossom volatiles.

The objective of this study was to identify the major volatiles and their relative concentrations in intact grapefruit, sweet orange, sour orange, mandarin, lemon, lime and pummelo blossoms. Volatiles from freshly picked blossoms were collected and concentrated using static headspace solid-phase microextraction and then separated and identified using GC-MS. Seventy volatiles were detected, 66 identified, of which 29 were identified for the first time in citrus blossoms. Major volatiles consisted of linalool, beta-myrcene, alpha-myrcene, limonene, (E)-ocimene, methyl anthranilate and indole. In terms of total volatiles: pummelo >> grapefruit approximately = sweet orange > sour orange approximately = mandarin approximately = lemon-lime > Volkamer lemon > Kaffir lime. Principal component analysis of blossom volatiles demonstrated that there were three widely separated, tightly clustered groups which consisted of mandarin, lemon-lime and pummelo. Other cultivars of possible mixed parentage produced non-overlapping values within the boundaries of these three clustered groups. The first two Eigenvectors explained 83% of the total variance. Linalool, limonene and myrcene had the highest loading values. Those cultivars requiring insect pollination such as pummelo produced highest levels of total volatiles as well as highest levels of known honeybee stimulants such as 1-hexanol and linalool.

Impact of thermal and nonthermal processing technologies on unfermented apple cider aroma volatiles.

Aroma composition and microbial quality of identical lots of apple cider treated by pulsed electric field (PEF), ultraviolet irradiation (UV), or thermal pasteurization stored at 4 degrees C were compared at 0 and 4 weeks. Conditions were optimized to achieve identical 5 log reductions in Escherichia coli K12 for each treatment. PEF and thermal pasteurization maintained acceptable microbial quality for 4 weeks, but UV samples fermented after 2 weeks. Twenty-eight volatiles were quantified using gas chromatography-mass spectrometry (GC-MS) and odor activity values (OAV) determined. OAVs of 69:hexyl acetate, 41:hexanal, 25:2-methylbutyl acetate, 23:2-methyl ethyl butyrate, and 14:2-(E)-hexenal were observed for the control cider. Significant differences (p < 0.05) in the levels of these odorants were observed between treated apple ciders only after 4 weeks of storage. Thermal samples lost 30% of the major ester and aldehyde volatiles during storage with significant decreases (p < 0.05) in butyl acetate, 2-methylbutyl acetate, hexanal, and 2-(E)-hexenal. In UV cider, hexanal and 2-(E)-hexenal were completely lost after 4 weeks of storage. Microbial spoilage in UV cider after 4 weeks of storage was chemically confirmed by the detection of the microbial metabolite 1,3-pentadiene. PEF cider lost <2% of its total ester and aldehydes after 4 weeks of storage and was preferred by 91% of the sensory panel over thermally treated cider.

Sulfur volatiles in guava (Psidium guajava L.) leaves: possible defense mechanism.

Volatiles from crushed and intact guava leaves (Psidium guajava L.) were collected using static headspace SPME and determined using GC-PFPD, pulsed flame photometric detection, and GC-MS. Leaf volatiles from four common citrus culitvars were examined similarly to determine the potential component(s) responsible for guava's protective effect against the Asian citrus psyllid (Diaphorina citri Kuwayama), which is the insect vector of Huanglongbing (HLB) or citrus greening disease. Seven sulfur volatiles were detected: hydrogen sulfide, sulfur dioxide, methanethiol, dimethyl sulfide (DMS), dimethyl disulfide (DMDS), methional, and dimethyl trisulfide (DMTS). Identifications were based on matching linear retention index values on ZB-5, DB-Wax, and PLOT columns and MS spectra in the case of DMDS and DMS. DMDS is an insect toxic, defensive volatile produced only by wounded guava but not citrus leaves and, thus, may be the component responsible for the protective effect of guava against the HLB vector. DMDS is formed immediately after crushing, becoming the major headspace volatile within 10 min. Forty-seven additional leaf volatiles were identified from LRI and MS data in the crushed guava leaf headspace.

Fresh squeezed orange juice odor: a review.

Fresh orange juice is a highly desirable but unstable product. This review examines analytical findings, odor activity, and variations due to cultivar, sampling methods, manner of juicing, plus possible enzymatic and microbial artifacts. Initial attempts to characterize orange juice odor were based on volatile quantitation and overemphasized the importance of high concentration volatiles. Although over 300 volatiles have been reported from GC-MS analytical studies, this review presents 36 consensus aroma active components from GC-olfactometry studies consisting of 14 aldehydes, 7 esters, 5 terpenes, 6 alcohols, and 4 ketones. Most are trace (microg/L) components. (+)-Limonene is an essential component in orange juice odor although its exact function is still uncertain. Total amounts of volatiles in mechanically squeezed juices are three to 10 times greater than hand-squeezed juices because of elevated peel oil levels. Elevated peel oil changes the relative proportion of several key odorants. Odor active components from solvent extraction studies differ from those collected using headspace techniques as they include volatiles with low vapor pressure such as vanillin. Some reported odorants such as 2,3-butanedione are microbial contamination artifacts. Orange juice odor models confirm that fresh orange aroma is complex as the most successful models contain 23 odorants.

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.

Thermal oxidation of 9'-cis-neoxanthin in a model system containing peroxyacetic acid leads to the potent odorant beta-damascenone.

The potent odorant beta-damascenone was formed directly from 9'-cis-neoxanthin in a model system by peroxyacetic acid oxidation and two-phase thermal degradation without the involvement of enzymatic activity. Beta-damascenone formation was heavily dependent on pH (optimum at 5.0) and temperature, occurring over the two sequential phases. The first was incubation with peroxyacetic acid at 60 degrees C for 90 min, and the second was at above 90 degrees C for 20 min. Only traces of beta-damascenone were formed on application of only one of the two phases. Formate and citrate solutions produced a much better environment for beta-damascenone formation than acetate and phosphate. About 7 microg/L beta-damascenone was formed from 5.8 mg/L 9'-cis-neoxanthin under optimal experimental condition. The detailed pathway by which beta-damascenone is formed remains to be elucidated.

Identification of aroma active compounds in orange essence oil using gas chromatography-olfactometry and gas chromatography-mass spectrometry.

Using GC-MS and GC-flame ionization detection (FID)/olfactometry, 95 volatile components were detected in orange essence oil, of which 55 were aroma active. In terms of FID peak area the most abundant compounds were: limonene, 94.5%; myrcene, 1%; valencene, 0.8%; linalool, 0.7%, and octanal, decanal, and ethyl butyrate, 0.3% each. One hundred percent of the aroma activity was generated by slightly more than 4% of the total volatiles. The most intense aromas were produced by octanal, wine lactone, linalool, decanal, beta-ionone, citronellal, and beta-sinensal. Potent aroma components reported for the first time in orange essence oil include: E-2-octenal, 1-octen-3-ol, Z-4-decenal, E,E-2,4-nonadienal, guaiacol, gamma-octalactone, and m-cresol. Over 20 compounds were identified for the first time in orange essence oil using MS, however, most did not exhibit aroma activity.

GC-olfactometric characterization of aroma volatiles from the thermal degradation of thiamin in model orange juice.

Model orange juice solutions containing 0.024 mM thiamin hydrochloride were stored for up to 8 weeks at 35 degrees C in amber glass containers. Volatiles were evaluated, primarily, using gas chromatography (GC) with olfactometry but also with flame ionization detector, pulsed-flame photometer detector (PFPD) (sulfur specific), and MS detection. Both 2-methyl-3-furanthiol (MFT) and its dimer, bis(2-methyl-3-furyl) disulfide (MFT-MFT) were identified thus confirming that thiamin could serve as the precursor to these potent off-flavors in thermally degraded citrus juices. Thirteen aroma active components were observed. MFT and MFT-MFT were observed after only a few days storage, and produced 33% of the total aroma activity after 7 d storage. Both compounds were observed olfactometrically earlier than they could be detected using PFPD. Other aroma-active compounds included 4,5-dimethylthiazole (skunky, earthy), 3-thiophenethiol (meaty, cooked), 2-methyl-4,5-dihydro-3(2H)-thiophenone (sour-fruity, musty, green), 2-acethylthiophene (burnt), 2-formyl-5-methylthiophene (meaty), and 2-methyl-3-(methyldithio) furan (meaty).

Gas chromatographic-olfactometric characterization of aroma compounds in two types of cashew apple nectar.

Cashew apple nectar is a secondary product from the production of cashew nuts and possesses an exotic tropical aroma. Aroma volatiles in pasteurized and reconstituted (from concentrate) Brazilian cashew apple nectars were determined using GC-MS and split, time-intensity GC-olfactometry (GC-O)/GC-FID. Methional, (Z)-1,5-octadien-3-one, (Z)-2-nonenal, (E,Z)-2,4-decadienal, (E,E)-2,4-decadienal, beta-damascenone, and delta-decalactone were identified for the first time in cashew apple products. These compounds plus butyric acid, ethyl 3-methylbutyrate, 2-methylbutyric acid, acetic acid, benzaldehyde, homofuraneol, (E)-2-nonenal, gamma-dodecalactone, and an unknown were the most intense aroma volatiles. Thirty-six aroma volatiles were detected in the reconstituted sample and 41 in the pasteurized sample. Thirty-four aroma active components were common to both samples. Ethyl 3-methylbutyrate and 2-methylbutyric acid were character impact compounds of cashew apple (warm, fruity, tropical, sweaty). Using GC-pFPD, 2-methyl-3-furanthiol and bis(2-methyl-3-furyl) disulfide were identified for the first time in cashew apple. Both were aroma active (meaty).

Aroma composition changes in early season grapefruit juice produced from thermal concentration.

Differences in aroma components and total volatiles between a single unpasteurized Marsh grapefruit juice and its 65 Brix concentrate reconstituted to 10 Brix were examined using GC-olfactometry (GC-O) and GC-FID. Total volatiles (FID) in the reconstituted concentrate were reduced to less than 5% of initial values, but 57% of total aroma (GC-O) remained. Forty-one aroma-active compounds were observed in unpasteurized single strength juice, whereas 27 components were found in the unflavored reconstituted concentrate. Aroma-active compounds were classified into grapefruit/sulfury, sweet/fruity, fresh/citrusy, green/fatty/metallic, and cooked/meaty groups. Five of six components in the sweet/fruity and 14 of 18 green/fatty/metallic components survived thermal concentration. However, only 4-mercapto-4-methyl-2-pentanone in the grapefruit/sulfury group, and linalool and nootkatone from the fresh/citrusy group, were found in the reconstituted concentrate. Methional was the only aroma compound in the cooked/meaty category found in both juice types. beta-Damascenone and 1-p-menthen-8-thiol were found only in the reconstituted concentrate. 4-Mercapto-4-methyl-2-pentanol was found for the first time in grapefruit juice.