Solid phase microextraction-gas chromatography for quantifying headspace hexanal above freeze-dried chicken myofibrils

A method using solid phase microextraction (SPME) combined with gas chromatography/mass spectrometry (GC/MS) was developed and used to determine the oxidation of freeze-dried chicken myofibrils spiked with methyl linoleate. Freeze-dried chicken myofibrils were found to act as a significant reservoir for hexanal. Recovery of hexanal emissions from the headspace above spiked myofibrils was 95% using a 5 min sampling time, with a total analysis time of approximately 12 min/sample. The SPME-GC/MS working linear response was from 0.01 to 10 mg hexanal/L (r( 2) = 0.995). Freeze-dried chicken myofibrils with added methyl linoleate (0.6 mmol/g of protein) were stored at 50 degrees C at water activities of 0.30 and 0.75 for 0, 12, 27, and 50 h. Lipid oxidation was determined using SPME-GC/MS to measure headspace hexanal concentration, the thiobarbituric acid reactive substances assay (TBARS) to quantify malonaldehyde, and a conjugated diene assay. Lipid oxidation was influenced by storage time and water activity. A strong correlation (r = 0.938) existed between SPME-GC/MS and TBARS. The use of SPME-GC/MS was a sensitive and rapid method for detecting hexanal as an indicator of lipid oxidation in chicken myofibrils.     

Elisa to quantify hexanal-protein adducts in a meat model system.

Monoclonal antibodies (MAb) were produced to hexanal-bovine serum albumin conjugates. An indirect competitive ELISA was developed with a detection range of 1-50 ng of hexanal/mL. Hexanal conjugated to three different proteins was recognized, whereas free hexanal and the native proteins were not detected. The antibody cross-reacted with pentanal, heptanal, and 2-trans-hexenal conjugated to chicken serum albumin (CSA) with cross-reactivities of 37.9, 76.6, and 45.0%, respectively. There was no cross-reactivity with propanal, butanal, octanal, and nonanal conjugated to CSA. The hexanal content of a meat model system was determined using MAb and polyclonal antibody-based ELISAs and compared with analysis by a dynamic headspace gas chromatographic (HS-GC) method and a thiobarbituric acid reactive substances (TBARS) assay. Both ELISAs showed strong correlations with the HS-GC and TBARS methods. ELISAs may be a fast and simple alternative to GC for monitoring lipid oxidation in meat.     

Antioxidative activities of fractions obtained from brewed coffee

The antioxidative activity of column chromatographic fractions obtained from brewed coffee was investigated to find antioxidants and to assess the benefit of coffee drinking. The dichloromethane extract inhibited hexanal oxidation by 100 and 50% for 15 days and 30 days, respectively, at the level of 5 microg/mL. A GC/MS analysis of fractions, which exhibited oxidative activity, revealed the presence of antioxidative heterocyclic compounds including furans, pyrroles, and maltol. The residual aqueous solution exhibited slight antioxidative activity. The inhibitory activity (%) of the seven fractions from an aqueous solution toward malonaldehde formation from lipid oxidation ranged from 10 to 90 at a level of 300 microg/mL. The results indicate that brewed coffee contains many antioxidants and consumption of antioxidant-rich brewed coffee may inhibit diseases caused by oxidative damages.     

Far-infrared radiation increases the antioxidant properties of rice hull extract in cooked turkey meat

To determine the antioxidant effects of rice hull extract exposed to far-infrared radiation, the added extracts were compared with sesamol in cooked turkey breast. Rice hull extract showed antioxidant properties in cooked turkey breast by reducing lipid oxidation and volatile aldehydes. Far-infrared radiation increased significantly the antioxidant activities of rice hull extracts. Rice hull extract irradiated by far-infrared (FRH) had lower TBARS values and fewer volatile aldehydes (hexanal, pentanal, and propanal) than a non-irradiated extract (IRH) during the 3 days of aerobic storage. Addition of FRH at 0.2% (w/w) in turkey meat could reduce the amounts of volatile hexanal to 18-47% of the control during the storage. However, the antioxidant activities of rice hull extracts did not last as long as those of pure sesamol due to the relatively low concentration of phenolics, and the extracts had some peculiar odor. Addition of rice hull extracts also increased both a and b values of the samples due to its brown intensity.     

Antioxidative activity of volatile extracts isolated from Angelica tenuissimae roots, peppermint leaves, pine needles, and sweet flag leaves

Volatile extracts were isolated from dried medicinal plants [Angelica tenuissimae roots (AT, Angelica tenuissima Nakai), peppermint leaves (PL, Mentha arvensis L.), pine needles (PN, Pinus sylvestris L.), and sweet flag leaves (SF, Acorus gramineus Rhizoma)] using steam distillation under reduced pressure, followed by continuous liquid-liquid extraction (DRP-LLE). The extracts were then analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The major volatile constituents of AT, PL, PN, and SF were 3-butylidene-4,5-dihydrophthalide (32 mg/g), menthol (18 mg/g), thunbergol (2.1 mg/g), and cis-asarone (37 mg/g), respectively. The inhibitory activity (%) of the extracts against hexanal oxidation ranged from 33 to 98% at a level of 50 microg/mL. Among the volatile extracts, PL and PN increased cell viabilities by 10 and 24%, respectively, at a dose of 1 microg/mL, compared to that of H2O2-treated brain neuroblastoma cells, SK-N-SH. However, a 20% reduction in the malonaldehyde level, an index for lipid peroxidation, was observed at only 1 microg/mL concentration of PN.     

Honey as a protective agent against lipid oxidation in ground turkey.

Lipid oxidation is a major deteriorative factor in meats. Sources of natural antioxidants that are as effective as commercially available antioxidants are desired. The objective of this research was to investigate honey as an inhibitor of lipid oxidation in ground poultry. The antioxidant content of different varieties of honey was investigated spectrophotometrically and honey's effectiveness in reducing oxidation of ground poultry determined by monitoring thiobarbituric acid reactive substances (TBARS). Buckwheat honey had the highest antioxidant content and acacia honey the lowest. Honeys of different floral sources differed in their protection against lipid oxidation. Buckwheat honey (5%, w/w) reduced TBARS approximately 70%, whereas acacia honey reduced TBARS approximately 34% at 3 days of storage at 4 degrees C. In comparison to butylated hydroxytoluene and tocopherol (0.02% of total fat), honey (at 5% of the weight of the meat) was much more effective at preventing oxidation. Honey has great potential as an antioxidant source and may result in greater acceptability of meat products and prevent negative health implications of oxidized meats.     

Antioxidant activity and characterization of volatile constituents of Taheebo (Tabebuia impetiginosa Martius ex DC)

Volatiles were isolated from the dried inner bark of Tabebuia impetiginosa using steam distillation under reduced pressure followed by continuous liquid-liquid extraction. The extract was analyzed by gas chromatography and gas chromatography-mass spectrometry. The major volatile constituents of T. impetiginosa were 4-methoxybenzaldehyde (52.84 microg/g), 4-methoxyphenol (38.91 microg/g), 5-allyl-1,2,3-trimethoxybenzene (elemicin; 34.15 microg/g), 1-methoxy-4-(1E)-1-propenylbenzene (trans-anethole; 33.75 microg/g), and 4-methoxybenzyl alcohol (30.29 microg/g). The antioxidant activity of the volatiles was evaluated using two different assays. The extract exhibited a potent inhibitory effect on the formation of conjugated diene hydroperoxides (from methyl linoleate) at a concentration of 1000 microg/mL. The extract also inhibited the oxidation of hexanal for 40 days at a level of 5 microg/mL. The antioxidative activity of T. impetiginosa volatiles was comparable with that of the well-known antioxidants, alpha-tocopherol, and butylated hydroxytoluene.     

Antioxidative activity of volatile chemicals extracted from beer

Volatile chemicals obtained from a commercial beer by liquid-liquid continuous extraction were evaluated for antioxidant activity. The inhibitory ability of this extract toward the conversion of hexanal to hexanoic acid was monitored over a 35-day period. The volatile extract demonstrated >99% effectiveness at inhibiting hexanal oxidation at 50 microg/mL, comparable to that of the natural antioxidant alpha-tocopherol (vitamin E). Volatile compounds contained in the extract were isolated and identified by gas chromatography-mass spectrometry (GC-MS). From the volatile constituents identified in beer extract, phenylethyl alcohol, maltol, and 2-furanmethanol were examined for antioxidative activities. At a concentration of 500 microg/mL, maltol and 2-furanmethanol demonstrated approximately 95 and 100% inhibition of hexanal oxidation over 35 days, respectively. Phenylethyl alcohol did not show any appreciable level of inhibition of hexanal oxidation. Heterocyclic compounds, some of which are known to possess antioxidative activities, were also identified in the volatile extract.     

Effect of electron beam irradiation and storage at 5 degrees C on thiobarbituric acid reactive substances and carbonyl contents in chicken breast meat infused with antioxidants and selected plant extracts

This study evaluated the effectiveness of synthetic and natural antioxidants, green tea, commercial grape seed extracts/combinations, and TBHQ, with varying concentrations of lipid oxidation of nonirradiated and irradiated chicken breast meats stored at 5 degrees C for 12 days. Fresh boneless and skinless chicken breast meats were vacuum-infused with varying concentrations of antioxidants: green tea, grape seed extracts alone/in combination, and TBHQ. The irradiation dosage was 3.0 kGy. Carbonyl values of raw chicken meat and thiobarbituric acid reactive substances (TBARS) values of raw and cooked chicken meat were determined for 0-12 days at 5 degrees C storage. TBARS values for 0-12 days of storage at 5 degrees C ranged from 1.21 to 7.3 and 1.22 to 8.51 mg malondialdehyde/100 g chicken for nonirradiated and irradiated raw chicken, respectively. TBARS values of cooked chicken ranged from 2.19 to 35.83 and 2.45 to 45.72 mg malondialdehyde/100 g chicken for nonirradiated and irradiated chicken, respectively. Irradiation increased TBARS values of both controls and plant extracts. The carbonyl content in meat lipid ranged from 1.7 to 2.9 and 1.7 to 4.41 micromol acetophenone/10 g of nonirradiated and irradiated chicken meat, respectively, and meat protein ranged from 1.4 to 2.07 and 1.41 to 2.72 micromol/10 g meat. Infusion of chicken meat with selected plant extracts is an effective method to minimize lipid oxidation and volatiles developments caused by irradiation.     

Inhibition of lipid oxidation in pork bundles processing by superheated steam frying

The effect of superheated steam treatment on the oxidative stability of lipids in packaged Zousoon (pork bundles) was investigated. The aroma quality of Zousoon samples was evaluated by sensory analysis and chromatographic analysis of volatiles. Results of this study indicated that oxidation of lipids occurred in pan-fried Zousoon after prolonged storage. Significant amounts of highly volatile compounds such as formaldehyde, acetaldehyde, acetone, and hexanal in Zousoon were identified by a modified method of cysteamine derivatization followed by gas chromatography-mass spectrometry (GC-MS) analysis. Superheated steam was found to be effective in suppressing lipid oxidation in canned Zousoon as compared with Zousoon fried by the conventional method in a frying pan. The superheated steam-fried samples had relatively low thiobarbituric acid reactive substance (TBARS) and peroxide (POV) values before and after storage, whereas samples prepared by pan frying had relatively high TBARS and POV values before and after storage. Superheated steam-fried Zousoon had superior lipid stability to that prepared by the conventional pan-frying method.     

Temperature effect on lactose crystallization, maillard reactions, and lipid oxidation in whole milk powder

Whole milk powder with an initial water content of 4.4% (w/w) and a water activity of 0.23 stored in hermetically sealed vials for up to 147 days below (37 and 45 degrees C) and above (55 degrees C) the glass transition temperature (T(g) determined to have the value 48 degrees C) showed a strong temperature dependence for quality deterioration corresponding to energies of activation close to 200 kJ/mol for most deteriorative processes. The glass transition was found not to cause any deviation from Arrhenius temperature dependence. Lactose crystallization, which occurred as a gradual process as monitored by isothermal calorimetry, is concluded to liberate bound water (a(w) increase to 0.46) with a modest time delay (approximately 2 days at 55 degrees C) and with concomitant surface browning as evidenced by an increasing Hunter b-value. Browning and formation of bound hydroxymethyl-furfural determined by HPLC seem to be coupled, while formation of another Maillard reaction product, furosine, occurred gradually and was initiated prior to crystallization. Initiation of lipid oxidation, as detected by lipid-derived radicals (high g-value ESR spectra), and progression of lipid oxidation, as detected by headspace GC, seem not to be affected by lactose crystallization and browning, and no indication of browning products acting as antioxidants could be determined.     

Preventing lipid oxidation during recovery of functional proteins from herring (Clupea harengus) fillets by an acid solubilization process

It has previously been found that a process based on solubilization at pH 2.7 gives high yields of herring muscle proteins with good functionality. In this study, the development of lipid oxidation during acid processing of herring mince was studied. It was tested how modifications of the process conditions and/or additions of antioxidants could prevent lipid oxidation during the actual process and then during ice storage of the protein isolates. Processing parameters evaluated were prewash of the mince, exposure time to pH 2.7, inclusion or exclusion of a high-speed centrifugation, and addition of antioxidants. Antioxidants tested were erythorbate (0.2%, 9.3 mM), sodium tripolyphosphate (STPP; 0.2%, 5.4 mM), ethylenediaminetetraacetic acid (EDTA; 0.044%, 1.5 mM), and milk proteins (4%). The first three antioxidants were added in the prewash or during the homogenization step, whereas milk proteins were added to the final precipitate. At time 0, all isolates were analyzed for pH, moisture content, and thiobarbituric reactive substances (TBARS). Selected isolates were also analyzed for lipid and protein content. Stability during ice storage was followed in terms of odor, TBARS, and color (a/b values). Extensive lipid oxidation took place using the "control" process without high-speed centrifugation. This was not significantly (p < or = 0.05) affected by a prewash or varied exposure time to pH 2.7. Including high-speed centrifugation (20 min, 10,000g) significantly (p < or = 0.05) reduced TBARS values, total lipids, a values and b values. Erythorbate alone, or in combination with STPP/EDTA, significantly (p < or = 0.05) reduced lipid oxidation during processing if added in the prewash or homogenization step. During ice storage, better stability was gained when antioxidants were added in both of these steps and when EDTA was used instead of STPP.     

Antioxidant properties of aroma compounds isolated from soybeans and mung beans

Aroma compounds contained in the extracts of soybean and mung bean that possess antioxidant activity were identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major aroma constituents of soybeans were 1-octen-3-ol (13.699 ppm), maltol (1.662 ppm), phenylethyl alcohol (1.474 ppm), hexanol (1.430 ppm), and gamma-butyrolactone (1.370 ppm). The major aroma constituents of mung beans were hexanol (3.234 ppm), benzyl alcohol (2.060 ppm), gamma-butyrolactone (1.857 ppm), 2-methyl-2-propenal (1. 633 ppm), and pentanol (1.363 ppm). The major aroma chemicals of soybeans and mung beans were examined for antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol showed potent antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited the oxidation of hexanal by 100%, 93%, 84%, and 32%, respectively, for a period of 40 days at the 500 microg/mL level. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited malonaldehyde (MA) formation from cod liver oil by 91%, 78%, 78%, and 78%, respectively, at the 160 microg/mL level. The antioxidative activity of eugenol was comparable to that of the natural antioxidant alpha-tocopherol (vitamin E).     

Loss of redness (a*) as a tool to follow hemoglobin-mediated lipid oxidation in washed cod mince

Instrumental measurement of redness loss (decrease in a* value) was evaluated as a tool to follow hemoglobin (Hb)-mediated lipid oxidation in fish muscle. Two washed cod mince model systems were used (prepared at pH 6.5 and 5.5), both fortified with 15 micromol/kg of trout Hb and adjusted to pH 6.5 and 81% moisture. The rate of oxidation was varied through pH alterations (pH 6.1 and 6.9) and addition of an antioxidative cod muscle press juice. During ice storage, TBARS, painty odor, and a* values were followed. In all "oxidizing" samples, a* values correlated well with TBARS and painty odor development; r = -0.95 and -0.77, respectively. In press juice containing samples, the correlation was lower (0.55 for a vs TBARS) because there was a slight a* value decrease even in the absence of measurable lipid oxidation. a* values distinguished between "oxidizing" and stable samples within 1 day, before any lipid oxidation products could be chemically detected. It was confirmed in an aqueous phosphate buffer model system that the redness loss corresponded to a buildup of brownish met-Hb at the expense of oxy- and deoxy-Hb. The a* value data were best used as a lipid oxidation index by calculating the rate of decrease (k value) in the "initial phase" of the redness loss (before accumulation of lipid oxidation products) or in the "differentiation phase" (during the exponential raise in TBARS/painty odor). Calibration to lipid oxidation products must, however, be made for each specific sample type. Washing method, pH, Hb-type, etc., all affected both k values and absolute a* readings. Small yellowness (b*) increases also occurred along with a value losses, possibly the result of polymerized Schiff bases.     

Chemical and functional characterization of Italian propolis obtained by different harvesting methods

The composition and antioxidant activity of Italian poplar propolis obtained using three harvesting methods and extracted with different solvents were evaluated. Waxes, balsams, and resins contents were determined. Flavones and flavonols, flavanones and dihydroflavonols, and total phenolics were also analyzed. To characterize the phenolic composition, the presence of 15 compounds was verified through HPLC-MS/MS. The antioxidant activity was evaluated through 1,1-diphenyl-2-picrylhydrazyl radical and reducing power assays. The ability of propolis to inhibit lipid oxidation was monitored by analyzing hydroperoxide and TBARS formation in lipids incorporated into an oil-in-water (O/W) emulsion. Acetone shows the highest extraction capacity. Wedge propolis has the highest concentration of active phenolic compounds (TP = 359.1 ± 16.3 GAEs/g; TFF = 5.83 ± 0.42%; TFD = 7.34 ± 1.8%) and seems to be the most promising for obtaining high-value propolis more suitable to prepare high-quality dietary supplements (TBARS = 0.012 ± 0.009 mmol std/g; RP = 0.77 ± 0.07 TEs/g).     

Volatile composition of sunflower oil-in-water emulsions during initial lipid oxidation: influence of pH.

The formation of odor active compounds resulting from initial lipid oxidation in sunflower oil-in-water emulsions was examined during storage at 60 degrees C. The emulsions differed in initial pH, that is, pH 3 and 6. The volatile compounds were isolated under mouth conditions and were analyzed by gas chromatography/sniffing port analysis. The lipid oxidation rate was followed by the formation of conjugated hydroperoxide dienes and headspace hexanal. The initial pH affected the lipid oxidation rate in the emulsions: the formation of conjugated diene hydroperoxides and the hexanal concentration in the static headspace were increased at pH 6. Pentanal, hexanal, 3-pentanol, and 1-octen-3-one showed odor activity in the emulsions after 6 days of storage, for both pH 3 and 6. Larger amounts of odor active compounds were released from the pH 6 emulsion with extended storage. It was shown that this increased release at pH 6 was not due to increased volatility because an increase in pH diminished the static headspace concentrations of added compounds in emulsions.     

Rapid Gas Chromatographic Technique for Quantifying 2‐Acetyl‐1‐Pyrroline and Hexanal in Rice (Oryza sativa,L.)

The aroma of rice plays a role in its consumer acceptability. The popcorn‐like smell of aromatic rice stemming primarily from its 2‐ acetyl‐1‐pyrroline (2‐AP) content is considered desirable by many consumers. Conversely, hexanal has been correlated with off odors in rice that develop from lipid oxidation. A rapid method for 2‐AP and hexanal quantification suitable for use in breeding programs, large‐scale research efforts, and quality assurance programs is needed. While developing such a method, sample preparation (degree of milling, particle size), solvent extraction time and temperature, and gas chromatographic parameters were studied. Particle size had no influence on 2‐AP or hexanal recovered. One extraction solubilized ≈80% of the 2‐AP and 56% of the hexanal present in milled rice. The optimum extraction method was assessed to require 0.3 g of ground brown or milled rice in methylene chloride held at 85°C for 2.5 hr. The complete gas chromatographic run requires ≈25 min, and 50 samples can be analyzed per day. The optimized method's linear response (R² = 0.99) and reproducibility was demonstrated. The stability of 2‐AP and hexanal in frozen milled rice and in refrigerated methylene chloride extracts was excellent for at least six months. Milled and unmilled commercial and breeders' aromatic rice samples contained 10–1,104 ng/g of 2‐AP and 148–2,541 ng/g of hexanal. Genotype had the greatest effect on the 2‐AP and hexanal content of two lines grown over four years and in four states.     

Antioxidative activity of heterocyclic compounds found in coffee volatiles produced by Maillard reaction

Typical heterocyclic compounds substituted with various functional groups found in Maillard reaction products were examined for antioxidant activity. Pyrroles exhibited the greatest antioxidant activity among all heterocyclic compounds tested. All pyrroles inhibited hexanal oxidation by almost 100% at a concentration of 50 microg/mL over 40 days. Addition of formyl and acetyl groups to a pyrrole ring enhanced antioxidative activity remarkably. Pyrrole-2-carboxaldehyde, 2-acetylpyrrole, 1-methyl-2-pyrrolecarboxaldehyde, and 2-acetyl-1-methylpyrrole inhibited hexanal oxidation by >80% at 10 microg/mL. Unsubstituted furan exhibited the greatest antioxidant activity among furans tested. Addition of all functional groups used in this study to furan decreased antioxidative activity. The antioxidant activity of thiophene increased with the addition of methyl and ethyl groups, but the addition of formyl or acetyl groups to thiophene decreased antioxidant activity. Thiazoles and pyrazines were ineffective antioxidants at all concentrations tested. Reaction of all heterocyclic compounds with hydrogen peroxide resulted in the formation of various oxidized products.     

Impact of tween 20 hydroperoxides and iron on the oxidation of methyl linoleate and salmon oil dispersions

To determine the role of surfactant hydroperoxides on the oxidative stability of fatty acids, the oxidation of methyl linoleate micelles and salmon oil-in-water emulsions was measured as a function of varying Tween 20 hydroperoxide concentrations. Increasing Tween 20 hydroperoxide concentrations from 3.5 to 14.7 micromol hydroperoxide/g Tween 20 decreased the lag phase of headspace hexanal formation but did not increase the total amount of hexanal formed in methyl linoleate/Tween 20 micelles. In the micelle system, Fe(2+) decreased the lag phase of hexanal formation but increased total hexanal concentrations only in micelles with the highest Tween 20 hydroperoxide concentrations (14.7 micromol hydroperoxide/g surfactant). Increasing Tween 20 surfactant hydroperoxide concentrations also increased the oxidation of salmon oil-in-water emulsions as determined by lipid hydroperoxides and headspace propanal. In both the micelle and emulsion systems, the prooxidant effect of Fe(2+) decreased with increasing Tween 20 hydroperoxide concentrations. These data show that surfactant hydroperoxides such as those in Tween 20 could decrease the oxidative stability of lipids in food emulsions.     

Development of an enzyme-linked immunosorbent assay for the detection of dicamba

A competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) was developed to quantitate the herbicide dicamba (3,6-dichloro-2-methoxybenzoic acid) in water. The CI-ELISA has a detection limit of 2.3 microg L(-1) and a linear working range of 10--10000 microg L(-1) with an IC(50) value of 195 microg L(-1). The dicamba polyclonal antisera did not cross-react with a number of other herbicides tested but did cross-react with a dicamba metabolite, 5-hydroxydicamba, and structurally related chlorobenzoic acids. The assay was used to estimate quantitatively dicamba concentrations in water samples. Water samples were analyzed directly, and no sample preparation was required. To improve detection limits, a C(18) (reversed phase) column concentration step was devised prior to analysis, and the detection limits were increased by at least by 10-fold. After the sample preconcentration, the detection limit, IC(50), and linear working range were 0.23, 19.5, and 5-200 microg L(-1), respectively. The CI-ELISA estimations in water correlated well with those from gas chromatography-mass spectrometry (GC-MS) analysis (r(2) = 0.9991). This assay contributes to reducing laboratory costs associated with the conventional GC-MS residue analysis techniques for the quantitation of dicamba in water.