Mass spectrometry analysis of volatile compounds in raw meat for the authentication of the feeding background of farm animals

The authentication of the conditions of animal production, based on the analysis of meat commercial cuts, is a major challenge on both societal and analytical grounds. The aim of the present work was to propose a method for the extraction of the volatile compounds from ruminant raw muscles trimmed of fat and to assess by mass spectrometry-based techniques the relevance of these compounds for the authentication of the type of feeding offered to the animals. The first step of the study consisted of validating conditions of dynamic headspace (DH) extraction of volatile compounds that enabled us to minimize the appearance of heat-induced artifacts and to maximize the richness of the DH-gas chromatography-mass spectrometry profile (DH-GC-MS) of raw lamb muscle. An extraction temperature of 35 degrees C (vs 60 and 90 degrees C) and a sample mass of 6.25 g (vs 12.5, 25, and 50 g) were shown to be suitable. The second step aimed at identifying volatile compounds enabling us to discriminate muscle samples from 16 experimental lambs fed either concentrate (n = 8) or pasture (n = 8). Before, to carefully explore the information given by the DH-GC-MS signal, the MS spectra acquired along the chromatogram were summed and then converted in a virtual-DH-MS spectral fingerprint to have a quick overview of the discriminative potential of the volatile fraction. According to univariate (analysis of variance) and to multivariate (principal component analysis) data treatments performed on virtual-DH-MS fingerprints, the meat volatile fraction was relevant to reveal the type of feeding of the living animal. The detailed examination of the information given by the GC dimension showed that 33 volatile compounds among the 204 detected in the muscle by DH-GC-MS enabled us to discriminate the type of feeding of the lambs. The relevance of these results is discussed in light of previous studies performed on adipose tissues.     

Whole berries versus berry anthocyanins: interactions with dietary fat levels in the C57BL/6J mouse model of obesity

Male C57BL/6J mice received diets with either 10% of calories from fat (LF) or a high-fat diet [45% (HF45) or 60% (HF60) calories from fat] for 92 days (expt 1) or 70 days (expt 2). These were given with or without freeze-dried powders from whole blueberries (BB) or strawberries (SB) (expt 1) or purified anthocyanin extracts from BB or SB (expt 2). Body composition was determined utilizing Echo MRI. Berries added to the LF diet did not alter weight gain, final body weights, body fat, or protein (percent body weight) or diet (grams) or energy (kilocalories) intake. However, in both HF45- and HF60-fed mice, weight gain, final weights, body fat (percent), and epididymal fat weights increased and body protein decreased ( p < 0.01) compared to LF mice. In mice fed HF45 diet plus BB, body weight gains, body fat (percent of BW), and epididymal fat weights were significantly greater than those in the HF45-fed controls, whereas weights of mice fed SB HF were similar to those of HF controls. SB or BB feeding did not alter glucose tolerance, although glucose tolerance decreased with age and in HF45 versus LF mice. Baseline plasma glucose was lower in SB- versus HF45-fed mice. After 8 weeks, mice fed the HF60 diet plus purified anthocyanins from BB in the drinking water had lower body weight gains and body fat than the HF60-fed controls. Anthocyanins fed as the whole blueberry did not prevent and may have actually increased obesity. However, feeding purified anthocyanins from blueberries or strawberries reduced obesity.     

Volatile components in crabmeats of Charybdis feriatus

The volatile components of different meats (legs with claws, body, and carapace) of a popularly consumed edible crab in Asia, Charybdis feriatus, were investigated. Samples were extracted by simultaneous steam distillation-solvent extraction and analyzed by gas chromatography/mass spectrometry. Among 177 compounds detected, 130 were positively identified. Seventy-six compounds were previously reported in other crab species. A greater number of naphthalenes were detected in this crab compared with other crabs in the literature. Aromatic compounds, alcohols, and sulfur-containing compounds were the three predominant groups with >15 components. Carapace tissue contained a greater number of volatile components in each group, except for sulfur-containing compounds. Most of the common components in the leg meat and the body meat were found at similar levels (p > 0.05). Carapace tissue generally had the highest quantity of common components among the meats. The higher levels of volatile components present in the carapace tissue might account for its stronger flavor compared with the other meats. Furthermore, the higher number of aldehydes and lower number of sulfur-containing compounds detected in the carapace meat might contribute to its unique flavor.     

Glutathione peroxidase activity, TBARS, and alpha-tocopherol in meat from chickens fed different diets.

This study investigated the effect of feeding broilers with diets differing in dietary fat source (lard, sunflower oil, olive oil) and vitamin E (basal vs supplemented with 200 mg of alpha-tocopheryl acetate/kg) on meat lipid oxidative stability. The diets differed by their degree of unsaturation and included the natural antioxidant alpha-tocopherol (vitamin E). Glutathione peroxidase (GSHPx) activity was measured in raw meat and ranged from 3.62 to 8.06 nmol NADPH/min/mg protein. The enzyme activity was influenced by the degree of unsaturation of the diet. Capillary gas chromatography analyses showed that dietary alpha-tocopherol accumulated in the muscle tissue and contributed to a better oxidative stability of the raw and cooked meat. Thigh meat alpha-tocopherol levels ranged from 2.73 to 3.62 microg/g in unsupplemented chickens whereas levels from 8.69 to 13.37 microg/g were observed in the thigh meat from alpha-tocopherol supplemented animals. The inclusion of olive oil and alpha-tocopherol in the animal diet gave lower thiobarbituric acid reactive substance (TBARS) values and lower GSHPx activity. High correlations were found between the parameters studied. The results suggest that the glutathione peroxidase activity could be used as an indicator of the meat oxidative stability. A negative relationship was observed between GSHPx activity and tissue alpha-tocopherol levels, and a positive relationship was evidenced between TBARS and antioxidant enzyme activity.     

Effect of a hay-based diet or different upland grazing systems on milk volatile compounds

The effect of animal feeding on milk volatile organic compounds (VOCs) of metabolic origin was tested on a hay-based diet (H), a highly diversified pasture under continuous grazing (CG), or a less diversified pasture under rotational grazing (RG). Individual milk of 24 Montbe?liarde cows (8 per treatment) were sampled after 2 weeks. Pasture-derived milk was richer (p < 0.05) in camphene, sabinene, β-caryophyllene, and skatole than H milk. Neither milk yield nor fat content affected the majority of VOCs measured. Skatole increased slightly with milk yield, while indole and cineole decreased slightly with milk fat content but with poor regression (R(2) < 0.54). Multivariate analysis showed that, on the basis of those VOCs of metabolic origin whose concentration differed between treatment (dimethyl-sulfone, skatole, toluene, undecanoic acid, 1-octadecene, benzeneacetaldehyde, octanoic acid, and 2-pentanone-4-hydroxy-4-methyl), it was possible to obtain good discriminations among feeding systems. This study is promising for a future use of VOCs of metabolic origin to trace animal feeding systems.     

Hypolipidemic and Body Fat–Lowering Effects of Giant Embryo Brown Rice (Seonong 17 and Keunnunjami) in High‐Fat‐Fed Mice

The effects of giant embryo brown rice Seonong 17 and Keunnunjami, in comparison with that of ordinary normal brown rice, on the body weight and lipid profile in mice under a high‐fat diet condition were investigated. The animals were fed with experimental diets for eight weeks: normal control, high fat (HF), and high fat supplemented with normal brown rice, Seonong 17 rice, or Keunnunjami rice powder. Their body and fat weights, lipid profiles, lipogenic enzyme activities, and adipokine concentrations were determined. Significant differences between the animal groups were analyzed with Tukey's test (P < 0.05). A marked increase in the body weight and fat, plasma triglyceride and total cholesterol concentrations, and free fatty acid level were found in HF mice relative to the control group. On the other hand, diet supplementation with rice, particularly the giant embryo rice samples, significantly suppressed this high‐fat‐induced body weight gain and hyperlipidemia via regulation of lipogenesis and adipokine production. Both the Seonong 17 and Keunnunjami exhibited greater hypolipidemic and body fat–lowering effects than the normal brown rice. The results demonstrate that these giant embryo mutants may be beneficial as biomaterials for functional foods with antiobesity and hypolipidemic effects.     

Effect of long-term dietary supplementation of high-gamma-linolenic canola oil versus borage oil on growth, hematology, serum biochemistry, and N-6 fatty acid metabolism in rats

Dietary supplementation of a high-gamma-linolenic acid canola oil (HGCO) containing approximately 36% (w/w) of gamma-linolenic acid (GLA, 18:3n-6) from the seeds of a genetically transformed canola strain, was assessed for its long-term biological effects. Growing Sprague-Dawley rats (n = 30) were fed a purified AIN93G diet containing 5, 10, or 15% (w/w) of HGCO as the fat source. For comparison, a separate group of rats (n = 10) was given the diet containing 15% (w/w) of borage oil (BO), which contained 22% (w/w) of GLA. After 12 weeks of feeding, the growth, relative organ weights, hematology, and serum biochemistry were found to be similar among rats fed the 5, 10, and 15% HGCO diets. The GLA levels in plasma and liver phospholipids (PL) were also similar. However, the levels of GLA in peripheral tissues (muscle PL and adipose triacylglycerols) were significantly higher in rats fed the 10 and 15% HGCO diets than those fed the 5% HGCO diet. When the above biologic parameters were compared between the 15% HGCO and 15% BO dietary groups, there were no significant differences except for lower final body weights and higher tissue levels of GLA, dihomo-gamma-linolenic acid (20:3n-6) and arachidonic acid (20:4n-6) in the 15% HGCO dietary group as compared with the 15% BO dietary group. This is due to a higher GLA content and possibly a more favorable stereospecific distribution of GLA in HGCO. Overall, long-term (12-week) feeding with diets containing up to 15% HGCO resulted in no adverse effects on growth, organ weight, hematology and serum biochemistry as compared to the diet containing 15% BO, suggesting that HGCO may be a safe alternative source of GLA.     

Hypoglycemic and Antioxidative Effects of Instant Cooked Giant Embryonic Rice in High‐Fat‐Fed Mice

The effect of instant cooked rice made from giant embryo mutant or ordinary normal rice on the glucose metabolism and antioxidative defense system in mice under a high‐fat (HF) diet condition was investigated. The animals were randomly divided and given experimental diets for seven weeks: normal control diet, HF diet, and HF diet supplemented with instant normal white, normal brown, giant embryonic white, or giant embryonic brown rice. At the end of the experimental period, the HF mice showed a marked increase in the blood glucose level, insulin concentration, and plasma and erythrocyte lipid peroxidation and a significant decrease in the glycogen level relative to the control group. However, diet supplementation with instant cooked giant embryonic and normal brown rice counteracted this high‐fat‐induced hyperglycemia and oxidative stress through regulation of the glucose‐regulating and antioxidant enzyme activities. The giant embryonic brown rice was the most effective in improving the glucose metabolism and antioxidant defense status in mice under the HF diet condition. The results demonstrate that the giant embryo rice mutant may be useful in the development of instant cooked rice with strong hypoglycemic and antioxidative properties.     

Effect of dietary conjugated linoleic acid and monounsaturated fatty acid content on pig muscle and adipose tissue lipase and esterase activity

Three levels (0%, 1%, and 2%) of conjugated linoleic acid (CLA) were combined with two levels (low and high) of monounsaturated fatty acids (MUFA) for pig feeding. The activity of neutral lipase (NL), acid lipase (AL), phospholipase (PL), neutral esterase (NE), and acid esterase (AE) was measured in extracts from muscle and subcutaneous adipose tissues. The addition of CLA in the diet only affected the lipolytic activity in muscle, whereas differences in MUFA content of pig diets were mainly responsible for the lipolytic enzyme modifications observed in adipose tissue. Nevertheless, a significant effect of the interaction CLA x MUFA on the activity of several lipolytic enzymes was observed in both tissues. The effect of either linoleic acid (LA) or CLA on the activity of muscle and adipose lipolytic enzymes was determined by in vitro assays. Remarkable inhibitory or activation effects were detected depending on the enzyme and kind of tissue.     

应用HS-SPME和GC/MS技术检测舍饲合作猪肌肉中的风味物质

用顶空固相微萃取和气-质联用技术分析检测了舍饲合作猪背最长肌的挥发性风味物质。结果表明:在60℃样品前处理条件下,共鉴定出50种风味化合物,其中有效成分43种;80℃时共鉴定出168种风味化合物,其中有效成分98种。检测结果还表明,醛类物质、含硫含氮化合物以及呋喃类物质可能是影响合作猪肉风味的重要挥发性物质。     

Incorporation of alpha-tocopherol and linoleic acid in fresh lambs by feeding chemically treated dietary supplements containing dl-alpha-tocopheryl acetate and sunflower oil

The effects of feeding chemically treated dietary supplements (CTDS) containing sunflower oil and dl-alpha-tocopheryl acetate (TA) on alpha-tocopherol content and fatty acid profile in edible tissues of lambs were estimated. Compared with lambs fed control diet (CD), lambs fed CD plus 250 IU of either TA or CTDS increased serum alpha-tocopherol. The CTDS-fed lambs further increased serum alpha-tocopherol by 29% over those fed CD plus 250 IU of TA. Lambs supplemented with TA or CTDS increased alpha-tocopherol in muscle and adipose tissues as compared with lambs fed CD. The CTDS-fed lambs had higher levels of alpha-tocopherol in gluteus medius (7.55 vs 6.05 mug/g), psoas major (7.43 vs 6.02 mug/g), and subcutaneous fat (12.6 vs 9.98 mug/g) compared with the TA-fed lambs. Feeding lambs CTDS also substantially increased levels of linoleic acid in the adipose tissues while decreasing the content of palmitic and oleic acids.     

Effect of some fat replacers on the release of volatile aroma compounds from low-fat meat products

The effect of fat content and carbohydrate fat-replacers on the release of volatile odor compounds from beefburger, salami, and frankfurter has been investigated. The reduction in fat content in any of the three meat products studied resulted in a tendency toward an increase in the quantities of volatiles released in the headspace. Tapioca starch and maltodextrin appear to delay the release of certain classes of compounds selectively; for instance, tapioca starch appears to slow the release of some Maillard products while maltodextrin has a similar effect on terpenes. In contrast, oat fiber decreases the release of most of the compounds analyzed. Thus, the addition of carbohydrate fat-replacers to low-fat meat products could assist the flavor qualities of low-fat meat products by slowing down the release of odor compounds.     

Polyphenolic transmission to Segureno lamb meat from ewes' diet supplemented with the distillate from rosemary (Rosmarinus officinalis) leaves

The aim of the present work is to study whether the introduction of rosemary plant byproduct, from plant steam distillation, in daily Segurena sheep feeding allows the transfer of active antioxidant components to lamb meat, without detriment to the animal productivity. For this, 36 Segurena ewes were assigned randomly to three homogeneous groups. One group was fed a basal diet as a control and the diet of the other two groups was modified by substituting 10 or 20% of the control diet (respectively) with distilled rosemary leaves. Chromatographic analysis allowed the identification of 11 polyphenolic components previously identified in the rosemary and basal diet pellets, respectively. Among them, rosmarinic acid, carnosol, and carnosic acid were the phenolic components that had a significantly increased presence ( P < 0.05) in the lamb meat from sheep mothers fed this aromatic herb, when compared to the control group. The incorporation of this byproduct into the animal diet favored the antioxidant capacity of these lamb meat samples. Fresh meat produced on rosemary had higher total ferric reducing antioxidant power (FRAP) ( P < 0.05), greater ability to reduce ABTS*+, and lower IC50 (DPPH*) ( P < 0.05) values when compared to the control group. Because no statistically significant differences were detected among the results obtained from the lamb meat belonging to the different ewe groups fed rosemary leaf extract (10 or 20%), it can be concluded that the incorporation of distilled rosemary leaves at a rate of 10% of the ewes' diet should be enough to improve the lamb meat antioxidant status.     

Influence of feeding soybean oil on conjugated linoleic acid content in beef

Forty-eight steers were used to study the influence of feeding soybean oil (SO) on the conjugated linoleic acid (CLA) content of beef. Steers were fed either a control diet containing 954 g/kg of dry matter (DM) corn-based concentrate (CTL) or a control diet supplemented with SO at 20 (SO2) or 40 g/kg (SO4) of diet DM for 105 days. Adipose tissue samples were collected from the M. longissimus dorsi (LD) and from the M. semitendinosus (ST) on days 0 and 63 of the experiment. Adipose and muscle tissue samples were collected from the LD and ST immediately after slaughter. Feeding 40 g/kg of DM as SO increased the proportions of trans-C(18:1) in beef lipid as compared to CTL and SO2 treatments. The C(18:2) cis-9, trans-11 isomer of CLA as a proportion of total fat was not different in adipose and muscle across treatments. Supplementing SO increased C(18:2) trans-10, cis-12 CLA in adipose tissue of the LD. Supplementing high-grain finishing diets with SO is not an effective strategy to enhance the C(18:2) cis-9, trans-11 isomer of CLA in beef.     

Arachidonic acid and long-chain n-3 polyunsaturated fatty acid contents in meat of selected poultry and fish species in relation to dietary fat sources

Arachidonic acid (AA) content, long-chain n-3 polyunsaturated fatty acid (PUFA) equivalent [LCE; calculated as 0.15 x linolenic acid (LA) + eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA)], and PUFA n-6/PUFA n-3 ratio were determined in meat [breast meat (BM), thigh meat (TM), and fillets (F), respectively] within four sets of chickens, five sets of turkeys, one set of common carp, and four sets of rainbow trout, fed either commercial diet or diets with manipulated PUFA n-3 and PUFA n-6 contents. AA content was within the range of 20 mg/100 g (F of rainbow trout fed the diet with linseed oil, LO) to 138 mg/100 g (TM of chickens fed restrictively the diet based on maize to the age of 90 days). AA content in BM of turkeys fed the diet with LO or fish oil (FO) did not differ (P > 0.05) from that of rainbow trout F. LCE was in the range of 16 mg/100 g (BM of turkeys fed a commercial feed mixture) to 681 mg/100 g (F of rainbow trout fed a commercial feed mixture). With regard to BM, only turkeys fed the diet with LO deposited more (P < 0.01) LCE (71 mg/100 g) as compared to all other poultry sets except turkeys fed the diet with FO (123 mg/100 g). Apart from all fish samples, also both BM and TM of turkeys fed the diet with either LO or FO met the recommended value of the PUFA n-6/PUFA n-3 ratio (<4). AA content in the tissue increased significantly (P < 0.001) with increasing dietary LA in both all chicken tissues and all turkey tissues, which is contrary to the suggested strong metabolic regulation of the AA formation. When all tissues within all animal species were taken as a one set, both AA percentage and EPA + DHA percentage in the tissue (Y, %) decreased (P < 0.001) with increasing fat content in the tissue (X, %), according to the equation Y = 4.7 - 0.54X (R (2) = 0.41) and Y = 6.0 - 0.33X (R (2) = 0.35), respectively. AA content in chicken BM, chicken TM, and turkey BM, respectively, decreased linearly (P < 0.01) with increasing live weight reached at the slaughter age.     

Consumption of oxidized oil increases oxidative stress in broilers and affects the quality of breast meat

A total of 120 4-week-old broiler chickens were allotted to 12 pens and fed one of three diets including control, oxidized diet (5% oxidized oil), or antioxidant-added diet (500 IU vitamin E) for 2 weeks. Blood samples were collected at the end of feeding trial, and breast muscles were sampled immediately after slaughter. Breast meats were also collected 24 h after slaughter and used for meat quality measurements. Oxidative stress in blood, lipid and protein oxidation, and sarcoplasmic reticulum Ca2(+)-ATPase (SERCA) activity of breast muscle were determined. The oxidized diet increased oxidative stress in blood and increased carbonyl content in breast meat compared with the other two dietary treatments (P < 0.05). Lipid oxidation of breast muscles with the antioxidant-supplemented diet was lower than that with the oxidized and control diet groups (P < 0.05). Meat from birds fed the oxidized diet showed higher drip loss after 1 and 3 days of storage and greater 0-1 h post-mortem pH decline (P < 0.05). Significant differences in specific SERCA activity in breast muscles from birds fed control and oxidized diets (P < 0.05) were detected. This suggested that dietary oxidized oil induced oxidative stress in live birds and increased lipid and protein oxidation in breast muscle. Decrease in SERCA activity in breast muscles due to oxidative stress in live animals accelerated post-mortem glycolysis, which sped the pH drop after slaughter and increased drip loss, indicating that oxidation of diet can cause PSE-like (pale, soft, and exudative) conditions in broiler breast muscles.     

Green Tea Prevents Obesity by Increasing Expression of Insulin-like Growth Factor Binding Protein-1 in Adipose Tissue of High-Fat Diet-Fed Mice

It is known that green tea has the ability to prevent obesity, but the underlying molecular mechanism is not fully understood to date. A preventive mechanism of green tea on obesity in C57BL/6 mice fed a high-fat (HF) diet was investigated by evaluating the expression levels of obesity-related proteins in mesenteric white adipose tissue by using protein array. An increase in the expression level of insulin-like growth factor binding protein (IGFBP)-1 by green tea was found in the white adipose tissues of both control and HF diet-fed mice by protein array and confirmed by Western blot. Moreover, the expression level was negatively correlated with adipose tissue weight. In 3T3-L1 adipocytes, treatment with green tea and its major polyphenol, (-)-epigallocatechin gallate, induced the expression of IGFBP-1 in a dose-dependent manner by Western blot. In conclusion, IGFBP-1 in adipose tissue is a novel molecule target for the prevention of obesity by green tea.     

Antihyperlipidemic and body fat-lowering effects of silk proteins with different fibroin/sericin compositions in mice fed with high fat diet

The effect of silk protein with different fibroin/sericin compositions on body weight and lipid metabolism in high fat-fed mice was investigated. The animals were given experimental diets for 6 weeks: normal control (NC), high fat (HF) and high fat diet supplemented with F100 (pure fibroin, HF-F100), F81 (81:19 fibroin/sericin, w/w, HF-F81) or F50 (50:50 fibroin/sericin, w/w, HF-F50). The silk protein-fed mice showed markedly reduced body weight and enhanced lipid profile relative to the HF group. In general, the amount of body fat, triglyceride and total plasma cholesterol levels, atherogenic index and free fatty acid level tended to decrease, while the HDL-cholesterol level increased, with increased amount of sericin in the diet. This hypolipidemic effect was partly due to increased fecal lipid excretion, inhibition of lipogenesis and regulation of adipokine production. These findings illustrate that silk protein, particularly sericin, may be beneficial in the prevention of high fat diet-induced hyperlipidemia and obesity.     

Aroma components of cooked tail meat of American lobster (Homarus americanus)

Key aroma components of cooked tail meat of American lobster (Homarus americanus) were studied by gas chromatography-olfactometry (GCO) techniques. Components of low and intermediate volatility were evaluated by aroma extract dilution analysis of solvent extracts prepared by direct solvent extraction-high vacuum distillation and vacuum steam distillation-solvent extraction, whereas headspace volatile components were assessed by GCO of decreasing headspace (static and dynamic modes) samples. Forty-seven odorants were detected by all techniques. 3-Methylbutanal (chocolate, malty), 2,3-butanedione (buttery), 3-(methylthio)propanal (cooked potato), 1-octen-3-one (mushroom), 2-acetyl-1-pyrroline (popcorn), and (E,Z)-2,6-nonadienal (cucumber), were identified as predominant odorants by all four isolation methods. The highly volatile compounds methanethiol (rotten, sulfurous) and dimethyl sulfide (canned corn) were detected by headspace methods only. These eight odorants along with three unknown compounds with crabby, amine, fishy odors were found to predominate in the overall aroma of cooked lobster tail meat.     

Gas chromatographic determination of electron capture sensitive volatile industrial chemical residues in foods, using AOAC pesticide multiresidue extraction and cleanup procedures

Electron capture (EC) gas chromatographic (GC) parameters have been developed for determining some of the more volatile industrial chemicals that can be determined by the AOAC multiresidue method for organochlorine and organophosphorus pesticides with modified GC operating conditions. Retention times relative to pentachlorobenzene are reported for 143 industrial chemicals, pesticides, and related compounds on OV-101 GC columns at 130 degrees C. Also reported for most of the compounds are recoveries from fortified samples carried through the AOAC extraction and cleanup procedures for fatty and/or nonfatty foods, Florisil elution characteristics, and GC relative retention times on mixed OV-101 + OV-210 columns at 130 degrees C. Our laboratory has used the modified EC/GC parameters with the AOAC multiresidue extraction/cleanup procedures to determine many volatile halogenated industrial chemical contaminants in foods, chiefly in samples of fresh-water fish. Other modifications of the AOAC method are described to improve the tentative identification and quantitative measurement of these volatile residues.