Resolution of racemic γ-lactone flavors on Chiralpak AD by packed column supercritical fluid chromatography

The chiral separation by packed column supercritical fluid chromatography (pSFC) has pronounced advantages due to its enhanced efficiency and reduced solvent consumption. Seven racemic γ-lactones (γ-C₆–γ-C₁₂) which are important flavor and fragrance substances were successfully resolved in less than 15 min each using an analytical scale pSFC. Baseline or near baseline separations were achieved for γ-C₈–γ-C₁₂. The polysaccharide chiral stationary phase (CSP) of Chiralpak AD and the organic modifier of isopropanol were used. The separation conditions were optimized by systematically investigating the effect of varying the percentage of isopropanol in the mobile phase, column temperature, column pressure, and flow rate of the mobile phase. The optimum separations of the seven γ-lactones were achieved mostly at sub-critical conditions. For these compounds the length of the 4-alkyl chain exhibited a determined influence on the enantioselectivity. Among the series of γ-lactones studied, the largest selectivity factor and the largest resolution factor were both observed with the 4-pentyl derivative (γ-C₉).     

Solid-phase microextraction for headspace analysis of key volatile compounds in orange beverage emulsion

Headspace solid-phase microextraction (HS-SPME) gas chromatography was used to analyze target flavor compounds in orange beverage emulsion. The effects of SPME fiber (PDMS 100 μm, CAR/PDMS 75 μm, PDMS/DVB 65 μm and DVB/CAR/PDMS 50/30 μm), adsorption temperature (25–45 °C), adsorption time (5–25 min), sample concentration (1–100%), sample amount (5–12.5 g), pH (2.5–9.5), salt type (K₂CO₃, Na₂CO₃, NaCl and Na₂SO₄), salt amounts (0–30%) and stirring mode were studied to develop HS-SPME condition for obtaining the highest extraction efficiency and aroma recovery. For the head space volatile extraction, the optimum conditions were: CAR/PDMS fiber, adsorption at 45 °C for 15 min, 5 g of diluted beverage emulsion (1:100), 15% (w/w) of NaCl with stirring and original pH 4. The main volatile flavor compounds were: limonene, 94.9%; myrcene, 1.2%; ethyl butyrate, 1.1%; γ-terpinene, 0.41%; linalool, 0.36%; 3-carene, 0.16%; decanal, 0.12%; ethyl acetate, 0.1%; 1-octanol, 0.06%; geranial, 0.05%; β-pinene, 0.04%; octanal, 0.03%; α-pinene, 0.03%; and neral, 0.03%. The linearity was very good in the considered concentration ranges (R² ? 0.97). Average recoveries ranged from 88.3% to 121.7% and showed good accuracy for the proposed analytical method. Average relative standard deviation (RSD) for five replicate analyses was found to be less than 14%. The limit of detection (LOD) ranged from 0.06 to 2.27 mg/l for all volatile flavor compounds and confirmed the feasibility of the HS-SPME technique for headspace analysis of orange beverage emulsion. The method was successfully applied for headspace analysis of five commercial orange beverage emulsions.     

HS-SPME/GC–MS and chemometrics for the classification of Balsamic Vinegars of Modena of different maturation and ageing

Head space solid-phase microextraction (HS-SPME) coupled with GC–MS analysis has been applied for the determination of the characteristic volatile profile of Balsamic Vinegar of Modena (BVM) with the aim to distinguish the less matured products (matured in wooden barrels for at least 60 days) from the aged ones (aged in wooden barrels for at least 3 years). Coupling the HS-SPME/GC–MS analysis data with multivariate statistical techniques, such as Principal Components Analysis (PCA) and Classification Trees (CT), it has been possible to classify BVMs on the basis of different maturation and ageing. A matured BVM presents an aromatic profile characterised by high contents of 3-methyl-1-butanol, 4-ethyl-phenol, and 3-methyl-1-butanol acetate, while an aged BVM is characterised by a prevalence of ethyl acetate, ethyl acetoacetate, furans, 2,3-butanediol and 2,3-butanediol acetate. This work represents a first attempt to classify Balsamic Vinegars of Modena on the basis of their maturation and ageing.     

Analysis of volatile compounds of pineapple wine using solid-phase microextraction techniques

An analytical procedure based on headspace solid-phase microextraction (HS-SPME) method combined with GC–MS was developed for the extraction and quantification of volatile compounds from pineapple wine. Different sample preparation (SPME fibre type, addition of sodium chloride, extraction time and temperature) were evaluated to optimise the method. For the final method, 8 ml of pineapple wine were placed in a 15 ml headspace vial with addition of 1 g of NaCl; a polydimethylsiloxane SPME fibre was used for extraction at 30 °C for 30 min with continuous stirring. The volatile compounds have shown a good linearity in the range of concentrations studied with regression coefficients higher than 0.98, and the reproducibility expressed as relative standard deviation ranged from 4.2% (2-phenylethyl acetate) to 7.1% (ethyl benzoate). The values obtained for detection and quantification limits were low enough to permit the determination of volatiles in pineapple wine. Using this method, 18 volatiles were identified, including 13 esters, 4 alcohols and one acid. Ethyl octanoate, ethyl acetate, 3-methyl-1-butanol and ethyl decanoate were the major constituents. A tentative study to estimate the contribution of the identified compounds to the aroma of the wine, on the basis of their odour activity values (OAV), indicated that the compounds potentially most important to pineapple wine included ethyl octanoate, ethyl acetate and ethyl 2-methylpropanoate.     

Headspace-solid phase microextraction (HS-SPME) analysis of oxidized volatiles from free fatty acids (FFA) and application for measuring hydrogen donating antioxidant activity

Volatile compounds from thermally oxidized free fatty acids (FFA) at 93 °C for 200 min were analyzed by headspace-solid phase microextraction (HS-SPME)-gas chromatography (GC). Commercially available mixtures of FFA were used instead of selecting specific vegetable oils with various fatty acid compositions. As oxidation time increased, total volatiles and some individual volatiles including hexanal, 2-hexenal, 2-heptenal, 2,4-heptadienal, 2-octenal, and 2,4-decadienal increased linearly with 0.99 coefficient of determination (R²) at specific oxidation time against conjugated dienoic acid (CDA) or p-anisidine value (p-AV). Total volatiles showed the highest linearity (R²) of 0.99 and 2-heptenal showed the highest increasing slope in peak areas for 200 min oxidation. Not all oxidized volatiles increased linearly during oxidation. Availability of HS-SPME method for determining hydrogen donating antioxidant activity was tested using FFA containing serially diluted butylated hydroxytoluene (BHT) at 93 °C for 60 min. 2-Heptenal and total volatiles showed higher linearity against BHT concentration than hexanal. HS-SPME could be a useful method to determine the hydrogen donating antioxidant activity from FFA using total volatiles or 2-heptenal as oxidation markers.     

Prebiotic effects of yacon (Smallanthus sonchifolius Poepp. & Endl), a source of fructooligosaccharides and phenolic compounds with antioxidant activity

Montepulciano is one of the most famous and important red-berried grapes of Italy. This article presents and discusses a comparative study of aroma profile and phenolic content of the Montepulciano wine from the Marches and the Abruzzo regions. The volatile composition of wines was determined by using headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). The PDMS fibre was chosen. The dominating esters in Montepulciano wines were ethyl hexanoate, ethyl decanoate, and ethyl octanoate, whereas phenyl ethanol and 3-methyl-1-butanol were dominating alcohols.Phenolic compounds, namely gallic acid, p-coumaric acid, trans-ferulic acid, caffeic acid, trans-resveratrol, (+)-catechin and (−)-epicatechin, were examined using HPLC-MS with direct injection of wine samples. The total phenolic content of the analysed wines was in the range of 30.4-61.9 mg l⁻¹. The presence of high amounts of esters seems to characterise the volatiles of Montepulciano wines from the Marches, whereas a high level of alcohols was found in Montepulciano wines from Abruzzo. Moreover, multivariate chemometric techniques, such as cluster analysis and principal component analysis, supported this thesis. Headspace solid phase microextraction and gas chromatography-mass spectrometry were used to analyse 20 commercial wine samples (Montepulciano monovarietal red wines) from the Marches (10 samples) and Abruzzo (10 samples).     

The aroma of blueberries

The volatile components of bilberry, bog blueberry and cultivated high-bush blueberry (cv. Rancocas) were analysed by gas chromatography and mass spectrometry. Several new compounds not reported previously as blueberry volatiles were detected. These included methyl and ethyl 2-hydroxy-3-methylbutanoate, methyl and ethyl 3-hydroxy-3-methylbutanoate, 2-phenylethyl formate, methyl salicylate, farnesol, farnesyl acetate, vanillin, myristicine, 4-vinylphenol, 2-methoxy-5-vinylphenol, citronellol, hydroxycitronellol and some γ- and δ-lactones. The character impact compounds of bilberry were found to be the above-mentioned hydroxy esters together with 2-phenylethanol and its esters and the γ- and δ-lactones, whereas myristicine, citronellol, hydroxycitronellol, farnesol and farnesyl acetate were typical of the aroma of high-bush blueberry.     

High performance liquid chromatography method for the simultaneous determination of α-, γ- and δ-tocopherol in vegetable oils in presence of hexadecyltrimethylammonium bromide/n-propanol in mobile phase

A rapid methodology by RP-HPLC, 10 min of total analysis time, for the analysis of α-, γ- and δ-tocopherol in vegetable oils, in presence of retinol, retinyl acetate and α-tocopherol acetate was developed. Hexadecyltrimethylammonium bromide 0.1 M/n-propanol 65% (v/v) are used as mobile phase. Tocopherols were detected using UV and fluorescence detection. The quantification limits for α-, γ- and δ-tocopherols were 0.28, 0.12 and 0.23 mg L⁻¹, respectively. The method yielded satisfactory results when it was applied to vegetable oils and thus, it is suitable for their quality control.     

Development of a solid phase microextraction protocol for the GC–MS determination of volatile off-flavour compounds from citral degradation in oil-in-water emulsions

The conditions for headspace solid phase microextraction (HS-SPME) analysis of volatile off-flavour compounds in citral emulsion were determined. Type of SPME phase (65 μm PDMS/DVB, 100 μm PDMS and 75 μm CAR/PDMS), adsorption temperature and salt concentration were significant factors affecting total peak area in the gas chromatogram and optimised in one factor experiments. Then, adsorption temperature (30–50 °C), adsorption time (20–40 min), and salt concentration (0–6 M) were studied to develop HS-SPME condition for obtaining the highest extraction efficiency. PDMS/DVB in 65 μm was the optimum fiber because of high adsorption efficiency and good reproducibility. The optimal condition was adsorption at 50 °C for 40 min and 6 M salt added to sample. Good Linearity, high recovery, good reproducibility and low limit of detection (LOD) for all off-odour compounds according to the optimised SPME conditions indicated that the SPME procedure was applicable for the analysis of the degraded citral products in headspace volatile of emulsion.     

Stereoisomeric flavour compounds LXIX: stereodifferentiation of δ(γ)-lactones C8–C18 in dairy products,margarine and coconut

The enantiomeric distributions of δ- and γ-lactones from dairy products are determined by enantioselective multidimensional gas chromatography. By using heptakis(2,3-di-O-acetyl-6-O-TBDMS)-β-cyclodextrin as the chiral main column, the simultaneous stereodifferentiation of the δ- and γ-lactones C₁₃–C₁₈ is reported for the first time. Genuine lactones of dairy products preferably occur as (R)-enantiomers. Flavoured margarine and coconut are analysed and discussed. Reference compounds are synthesized as racemates and separated by enantioselective HPLC into the enantiomers. Chiral compounds of characteristic enantiomeric ratios are detected irrespective of the manufacturing processes used.     

Effect of pasteurisation on ascorbic acid,dehydroascorbic acid,tocopherols and fatty acids in pooled mature human milk

Ascorbic and dehydroascorbic acids (vitamin C), tocopherols (vitamin E) and unsaturated fatty acids are heat-sensitive and therefore, their concentrations in human milk could be affected by pasteurisation. Here we determined the concentrations of ascorbic acid plus dehydroascorbic acid, ascorbic acid alone, and α- and γ-tocopherols, and the percentages of fatty acids in samples of human milk after pasteurisation by a slow (62.5 °C, 30 min) or fast heating (100 °C, 5 min) procedure. Both methods led to a significant decrease in the concentrations of ascorbic acid plus dehydroascorbic acid (12% and 29%), ascorbic acid (26% and 41%), α-tocopherol (17% and 34%) and γ-tocopherol (13% and 32%), respectively. However, milk fatty acids, including the polyunsaturated long-chain fatty acids, were unaffected by the two methods. On the basis of these observations, we recommend that human milk be treated using a slow pasteurisation. In addition, we propose ascorbic acid as a marker of the degree of heat treatment.     

Quantitative analysis of volatiles in transesterified coconut oil by headspace-solid-phase microextraction-gas chromatography–mass spectrometry

A simple and fast headspace-solid-phase microextraction (HS-SPME) method coupled with gas chromatography–mass spectrometry (GC–MS) was developed for the analysis of volatile compounds in transesterified coconut oil by applying solvent dilution. Solvent dilution conditions (solvent type and solvent amount) and HS-SPME sampling parameters (adsorption temperature and time) were optimised through monitoring the adsorption result of the selected volatiles (octanoic acid esters) in transesterified coconut oil samples. The incubation of methanol (800 μl)-diluted oil (200 μl) sample at 60 °C for 30 min led to the best result. The method was further validated by determining the calibration linear range, correlation coefficient (R²), accuracy, precision, limit of detection and limit of quantification through spiking standards into the blank matrix consisting of coconut oil and methanol. This method may also be applicable for detection and determination of volatile compounds in other transesterified oil samples.     

Optimization of HS-SPME conditions for quantification of 2-acetyl-1-pyrroline and study of other volatiles in Pandanus amaryllifolius Roxb.

HS-SPME coupled with GC–FID was applied for the quantification of 2-acetyl-1-pyrroline (2AP) in Pandanus amaryllifolius, the best available natural source of (2AP). Conditions for qualitative and quantitative analysis of aroma volatiles with special reference to 2AP from P. amaryllifolius have been optimized using HS-SPME/GC–FID. About 10 mg powdered leaf material at 80 °C temperature for 35 min was found to be the most effective combination for maximum release of 2AP. Volatile analyses led to the identification of 31 compounds, of which 21 are reported for the first time in P. amaryllifolius. 2AP contents and other volatiles were found to vary in small and large growth forms.     

Influence of germination techniques on sprout yield,biosynthesis of ascorbic acid and cooking ability,in chickpea (Cicer arietinum L.)

Effects of germination time and illuminations on sprout yield, biosynthesis of ascorbic acid, cooking ability and moisture accumulation in chickpeas were significant (p ? 0.01). Green light had the highest promoting effect on the ascorbic acid level (40.59 mg/100 g) as compared to other illuminations but significantly reduced the sprout yield (188.6 g) as compared to dark, fluorescence and γ-rays illuminations with significantly high sprout yield (196 g) and imbibing moisture (51%). Cooking time was reduced by 43% due to γ-rays in un-soaked seed. Cooking time increased in all treated chickpea samples after 24 h germination and thereafter decreased significantly. Red light significantly increased the cooking time (68.44 min) followed by fluorescent (64.5 min), yellow (61.8 min) and green light (60.9 min). The results indicated that germination of chickpea under green light was an effective process in enhancing ascorbic acid content while dark, fluorescence and γ-rays were effective in promoting sprout growth and to some extent biosynthesis of ascorbic acid.     

Monitoring the oxidation of almond oils by HS-SPME–GC–MS and ATR-FTIR: Application of volatile compounds determination to cultivar authenticity

In this study, the oxidative process of Spanish and American almond oils was monitored by using headspace solid-phase microextraction/gas chromatography–mass spectrometry (HS-SPME/GC–MS) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). Oxidative stability of samples was evaluated after thermal treatment at 100 °C for 1, 3, 5, 7, 10, 15 and 20 days, in order to accelerate the lipid oxidation process. Variations observed in bands of infrared spectra were used to monitor the progress of the oxidation process of almond oils. HS-SPME–GC–MS was used for isolation and determination of volatile compounds in oxidised almond oils. The following aldehydes were identified in all samples: hexanal, (E)-2-heptenal, (E)-2-octenal, nonanal, (E)-2-nonenal, (E,E)-2,4-nonadienal and (E,E)-2,4-decadienal. Determination of volatile compounds resulting from lipid oxidation from three different almond cultivars (Spanish Guara and Marcona and Butte from USA) was performed to obtain a set of parameters for discrimination between Spanish and American cultivars. Successful discrimination was obtained in samples kept under thermal treatment for 7 days, by using HS-SPME/GC–MS combined with multivariate stepwise linear discriminant analysis (LDA).     

Development of a quantitative method for headspace analysis of methylsulfanyl-volatiles from kiwifruit tissue

Minor fruit volatiles are likely to be missed using sampling techniques optimized for the extraction of major compounds. This can be a disadvantage if these minor compounds contribute to characteristic fruit flavors. In this comparative study, headspace solid-phase microextraction (HS-SPME) and dynamic headspace sampling (DHS) parameters were systematically optimized to ensure highest extraction yields of methylsulfanyl-volatiles from kiwifruit tissue samples. A significant “salting-out” effect from the fruit matrix was observed using both sampling techniques after (NH₄)₂SO₄ saturation. HS-SPME at optimized conditions (polydimethylsiloxane-divinylbenzene-coated fiber, (NH₄)₂SO₄ saturation, 5 min equilibration and 20 min sampling at 40 °C) was faster and more convenient to use than DHS for qualitative purposes. Despite this, the qualitative and quantitative methylsulfanyl-volatile profile was improved using optimized DHS ((NH₄)₂SO₄ saturation; sampling time 20 h; flow rate 30 mL min^− 1) compared with HS-SPME, making this the more sensitive and preferred method for quantitative studies. The optimization strategies for increasing headspace extraction yields of trace compounds presented in this study can easily be applied to tissue samples from other fruit.     

Comparison of headspace-SPME-GC–MS and LC–MS for the detection and quantification of coumarin,vanillin, and ethyl vanillin in vanilla extract products

A headspace-solid phase micro-extraction (HS-SPME) GC–MS method has been developed for the determination of coumarin, vanillin and ethyl vanillin in vanilla products. Limits of detection ranged from 1.33 to 13.2 ng mL⁻¹. Accuracy and precision data for the method were measured and compared to those obtained using LC-ESI-MS. A survey of 24 commercially available vanilla products was completed using both techniques. No coumarin was detected in any of the samples. Examination of the GC–MS chromatograms revealed the presence of 18 other flavor related compounds in the samples. The method validation and sample analysis data using HS-SPME-GC–MS were comparable to those obtained using the LC–MS method. Because the two methods are conceptually different from one another, both methods would not be subject to the same interferences. This would allow them to be used as confirmatory methods for each other.     

Optimization of ethanol extraction and further purification of isoflavones from soybean sprout cotyledon

Isoflavones from cotyledons of soybean sprouts were extracted with aqueous ethanol and further concentrated to obtain a product with a high concentration of isoflavone. The ethanol concentration, extraction time and reaction temperature were optimized by using response surface methodology (RSM). Isoflavones in aqueous ethanol were concentrated by a three-step procedure comprised of solid phase extraction (SPE) with Diaion HP-20 and Amberlite-XAD-2 adsorption columns, acid hydrolysis, and liquid–liquid extraction. The maximum amount of isoflavone in aqueous ethanol extracts (11.6 mg/g solid) was obtained when isoflavones in cotyledons (2.18 mg/ g solid) were extracted with 80–90% (v/v) aqueous ethanol above 90 °C for more than 100 min. The isoflavone extracts, obtained by SPE with a Diaion HP-20 column contained 100 mg/g solid. The liquid–liquid extraction (LLE) with ethyl ether further concentrated the extracts up to 229 mg/g solid, retaining 63% of the initial isoflavones.     

Constituents of Sideritis syriaca. ssp. syriaca (Lamiaceae) and their antioxidant activity

The aerial parts of Sideritis syriaca ssp. syriaca (Lamiaceae) were extracted, after defatting, with diethyl ether, ethyl acetate and n-butanol. The antioxidant activities of the extracts were evaluated through in vitro model systems, such as 1,1-diphenyl-2-picryl hydrazyl (DPPH) and Co(II) EDTA-induced luminol chemiluminescence. In both model systems the ethyl acetate extract was the most effective. Phytochemical analysis of ethyl acetate extract showed the presence of two new isomeric compounds (1 and 1′), identified as 1-rhamnosyl, 1-coumaroyl, dihydrocaffeoyl, protocatechuic tetraester of quinic acid, as well as chlorogenic acid (2), apigenin 7-O-glucoside (3), apigenin (4), 4′-O-methylisoscutellarein 7-O-[6′′′-O-acetyl-β-D-allopyranosyl-(1 → 2)-β-d-glucopyranoside] (5), isoscutellarein 7-O-[6′′′-O-acetyl-β-D-allopyranosyl-(1 → 2)-β-d-glucopyranoside] (6), 4′-O-methylisoscutellarein 7-O[β-d-allopyranosyl-(1 → 2)-β-d-glucopyranoside] (7) and 4′-O-methylisoscutellarein 7-O-[β-d-allopyranosyl-(1 → 2)-6′′-O-acetyl-β-d-glucopyranoside] (8). The above compounds were identified by spectroscopic methods.