Glucose addition and oven-heating of pork stimulate glycoxidation and protein carbonylation, while reducing lipid oxidation during simulated gastrointestinal digestion.

In the present study, it was investigated if glucose addition (3 or 5%) to pork stimulates glycoxidation (pentosidine, PEN), glycation (Maillard reaction products, MRP), lipid oxidation (4-hydroxy-2-nonenal, 4-HNE; hexanal, HEX; thiobarbituric acid reactive substances, TBARS), and protein oxidation (protein carbonyl compounds, PCC) during various heating conditions and subsequent in vitro gastrointestinal digestion. An increase in protein-bound PEN level was observed during meat digestion, which was significantly stimulated by glucose addition (up to 3.3-fold) and longer oven-heating time (up to 2.5-fold) of the meat. These changes were accompanied by the distinct formation of MRP during heating and digestion of the meats. Remarkably, stimulated glyc(oxid)ation was accompanied by increased protein oxidation, whereas lipid oxidation was decreased, indicating these reactions are interrelated during gastrointestinal digestion of meat. Glucose addition generally didn't affect these oxidative reactions when pork was packed preventing air exposure and oven-heated until a core temperature of 75 °C was reached.

The kinetics of glutathione in the gastrointestinal tract of weaned piglets supplemented with different doses of dietary reduced glutathione.

This study aimed to investigate the kinetics of dietary GSH in the gastrointestinal tract and the effect of GSH on the intestinal redox status of weaned piglets. Forty-eight piglets with an average age of 26 days and an average body weight of 7.7 kg were used in this study. The piglets were divided into three treatment groups including the control group with a basal diet (CON) and two GSH groups with a basal diet supplemented with 0.1% GSH (LGSH) and 1.0% GSH (HGSH), respectively. The basal diet did not contain any GSH. The experiment lasted for 14 days, with eight animals sampled from each group on d5 and 14. The parts of 0-5%, 5-75%, and 75-100% of the length of the small intestine were assigned to SI1, SI2, and SI3. The results showed that GSH almost completely disappeared from the digesta at SI2. However, no difference in the GSH level in mucosa, liver, and blood erythrocytes was found. The level of cysteine (CYS) in SI1 digesta was significantly higher in HGSH than CON and LGSH on d14, and similar findings were observed for cystine (CYSS) in SI3 digesta on d5. The CYSS level in HGSH was also significantly higher than LGSH in the stomach on d14, while no CYS or CYSS was detected in the stomach for control animals, indicating the breakdown of GSH to CYS already occurred in the stomach. Irrespective of the dietary treatment, the CYS level on d14 and the CYSS level on d5 and 14 were increased when moving more distally into the gastrointestinal tract. Furthermore, the mucosal CYS level was significantly increased at SI1 in the LGSH and HGSH group compared with CON on d5. Glutathione disulfide (GSSG) was recovered in the diets and digesta from the LGSH and HGSH group, which could demonstrate the auto-oxidation of GSH. It is, therefore, concluded that GSH supplementation could not increase the small intestinal mucosal GSH level of weaned piglets, and this could potentially relate to the kinetics of GSH in the digestive tract, where GSH seemed to be prone to the breakdown to CYS and CYSS and the auto-oxidation to GSSG.

In vitro and in vivo digestion of red cured cooked meat: oxidation, intestinal microbiota and fecal metabolites.

Mechanisms explaining epidemiological associations between red (processed) meat consumption and chronic disease risk are not yet elucidated, but may involve oxidative reactions, microbial composition alterations, inflammation and/or the formation of toxic bacterial metabolites. First, in vitro gastrointestinal digestion of 23 cooked beef-lard minces, to which varying doses of nitrite salt (range 0-40 g/kg) and sodium ascorbate (range 0-2 g/kg) were added, showed that nitrite salt decreased protein carbonylation up to 3-fold, and inhibited lipid oxidation, demonstrated by up to 4-fold lower levels of 'thiobarbituric acid reactive substances', 32-fold lower 4-hydroxynonenal, and 21-fold lower hexanal values. The use of ascorbate increased the antioxidant effect of low nitrite salt levels, whereas it slightly increased protein carbonylation at higher doses of nitrite salt. The addition of a low dose of ascorbate without nitrite salt slightly promoted oxidation during digestion, whereas higher doses had varying antioxidant effects. Second, 40 rats were fed a diet of cooked chicken- or beef-lard minces, either or not cured, for three weeks. Beef, compared to chicken, consumption increased lipid oxidation (2- to 4-fold) during digestion, and gut protein fermentation (cecal iso-butyrate, (iso-)valerate, and fecal indole, cresol), but oxidative stress and inflammation were generally not affected. Cured, compared to fresh, meat consumption significantly increased stomach protein carbonylation (+16%), colonic Ruminococcaceae (2.1-fold) and cecal propionate (+18%), whereas it decreased cecal butyrate (-25%), fecal phenol (-69%) and dimethyl disulfide (-61%) levels. Fecal acetaldehyde and diacetyl levels were increased in beef-fed rats by 2.8-fold and 5.9-fold respectively, and fecal carbon disulfide was 4-fold higher in rats consuming cured beef vs. fresh chicken. Given their known toxicity, the role of acetaldehyde and carbon disulfide in the relation between meat consumption and health should be investigated in future studies.

Corrigendum: Phloroglucinol-Mediated Hsp70 Production in Crustaceans: Protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii.

[This corrects the article DOI: 10.3389/fimmu.2018.01091.].

Commercial luncheon meat products and their in vitro gastrointestinal digests contain more protein carbonyl compounds but less lipid oxidation products compared to fresh pork.

Because of the large diversity in processed meat products and the potential involvement of oxidation processes in the association between red and processed meat consumption and chronic diseases, the concentration of oxidation products after gastrointestinal digestion of commercial luncheon meat products was investigated. A broad spectrum of meat products (n = 24), displaying large variation in macro- and micronutrient composition and processing procedures, was digested in vitro by simulating digestion fluids of the human gastrointestinal tract. Lipid and protein oxidation was assessed in the meat products before digestion and in the corresponding digests by measurement of free malondialdehyde, 4-hydroxy-2-nonenal, hexanal and protein carbonyl compounds. Compared to an unprocessed cooked pork mince, that was included as a reference in the digestion experiment, levels of lipid oxidation products were low in the digests of most meat products. Only the digests of Parma ham had slightly higher or comparable levels as the reference pork. In contrast, protein carbonyl compounds were comparable or up to 6 times higher in the processed meat products compared to the reference pork. Particularly raw-cooked and precooked-cooked meat products and corresponding digests had higher protein carbonyl levels, but also lower protein contents and higher fat to protein ratios. In conclusion, most luncheon meat products and corresponding digests contained lower amounts of free lipid oxidation products, but more protein carbonyl compounds compared to the reference pork.

Untargeted Metabolomics to Reveal Red versus White Meat-Associated Gut Metabolites in a Prudent and Western Dietary Context.

SCOPE: To improve understanding of the epidemiological link between red and processed meat consumption and chronic diseases, more insight into the formation of metabolites during meat digestion is warranted. METHODS AND RESULTS: Untargeted mass-spectrometry-based metabolomics is applied to explore the impact of red and processed meat consumption (compared to chicken), combined with a prudent or Western dietary pattern. A pig feeding study (n = 32), as a sentinel for humans, is conducted in a 2 × 2 factorial design for 4 weeks. The luminal content of the small intestine and colon are collected to determine their metabolic fingerprints. Seventy-six metabolites (38 in the small intestine, 32 in the colon, and 6 in both intestinal compartments) contributing to the distinct gut metabolic profiles of pigs fed either chicken or red and processed meat are (tentatively) identified. Consumption of red and processed meat results in higher levels of short- and medium-chain acylcarnitines and 3-dehydroxycarnitine, irrespective of dietary context, whereas long-chain acylcarnitines and monoacylglycerols are associated with the red and processed Western diet. CONCLUSION: The identification of red and processed meat-associated gut metabolites in this study contributes to the understanding of meat digestion in a complex but controlled dietary context and its potential health effects.

Red and processed meat consumption within two different dietary patterns: Effect on the colon microbial community and volatile metabolites in pigs.

Pigs were fed either red and processed meat or chicken meat within either a prudent or a Western dietary pattern for four weeks (2 × 2 full factorial design). The colon microbial community and volatile organic compounds were assessed (either quantified or based on their presence). Results show that Lactobacilli were characteristic for the chicken × prudent dietary pattern treatment and Paraprevotella for the red and processed meat × prudent dietary pattern treatment. Enterobacteriaceae and Desulfovibrio were characteristic for the chicken × Western dietary pattern treatment and Butyrivibrio for the red and processed meat × Western dietary pattern treatment. Campylobacter was characteristic for chicken consumption and Clostridium XIVa for red and processed meat, irrespective of the dietary pattern. Ethyl valerate and 1-methylthio-propane were observed more frequently in pigs fed red and processed meat compared to chicken meat. The prevalence of 3-methylbutanal was >80% for pigs receiving a Western dietary pattern, whereas for pigs fed a prudent dietary pattern the prevalence was <35%. The concentration of butanoic acid was significantly higher when the prudent dietary pattern was given, compared to the Western dietary pattern, but no differences for other short chain fatty acids or protein fermentation products were observed.

Background Diet Influences TMAO Concentrations Associated with Red Meat Intake without Influencing Apparent Hepatic TMAO-Related Activity in a Porcine Model.

Red meat has been associated with an increased cardiovascular disease (CVD) risk, possibly through gut microbial-derived trimethylamine-N-oxide (TMAO). However, previous reports are conflicting, and influences from the background diet may modulate the impact of meat consumption. This study investigated the effect of red and white meat intake combined with two different background diets on urinary TMAO concentration and its association with the colon microbiome in addition to apparent hepatic TMAO-related activity. For 4 weeks, 32 pigs were fed chicken or red and processed meat combined with a prudent or western background diet. 1H NMR-based metabolomics analysis was conducted on urine samples and hepatic mRNA expression of TMAO-related genes determined. Lower urinary TMAO concentrations were observed after intake of red and processed meat when consumed with a prudent compared to a western background diet. In addition, correlation analyses between urinary TMAO concentrations and relative abundance of colon bacterial groups suggested an association between TMAO and specific bacterial taxa. Diet did not affect the hepatic mRNA expression of genes related to TMAO formation. The results suggest that meat-induced TMAO formation is regulated by mechanisms other than alterations at the hepatic gene expression level, possibly involving modulations of the gut microbiota.

Long-Chain n-3 PUFA Content and n-6/n-3 PUFA Ratio in Mammal, Poultry, and Fish Muscles Largely Explain Differential Protein and Lipid Oxidation Profiles Following In Vitro Gastrointestinal Digestion.

SCOPE: Muscle food characteristics (fatty acid profile, heme-Fe, intrinsic antioxidants) that relate to the formation of (patho)physiological oxidation products during gastrointestinal digestion are investigated. METHODS AND RESULTS: Muscles (n = 33) from 18 mammal, poultry, and fish species, of which some are mixed with lard to standardize their fatty acid profile, are digested in vitro. Lipid oxidation is assessed by thiobarbituric reactive substances (TBARS), n-3 PUFA derivative 4-hydroxy-2-hexenal and propanal, n-6 PUFA derivative 4-hydroxy-2-nonenal and hexanal, and protein oxidation by carbonylation. Digests of n-3 PUFA-rich fish demonstrated the highest n-3 PUFA oxidation, whereas digests of various poultry and rabbit muscles showed highest n-6 PUFA oxidation, which correlated significantly with the n-6/n-3 PUFA ratio. Without lard addition, lipid oxidation is significantly higher in chicken and pork loin digests versus beef and deer digests, whereas the opposite occurred when these muscles are mixed with lard. Protein carbonylation correlates significantly with levels of TBARS and the sum of hydroxy-alkenals in digests. The n-6/n-3 PUFA ratio correlates well with the 4-hydroxy-2-nonenal/4-hydroxy-2-hexenal ratio in digests. CONCLUSIONS: Muscular fatty acid profiles largely explain type and extent of lipid and protein oxidation during gastrointestinal digestion. Red meat only stimulates oxidation when digested with specific fat sources.

Impact of Red versus White Meat Consumption in a Prudent or Western Dietary Pattern on the Oxidative Status in a Pig Model.

Human diets contain a complex mixture of antioxidants and pro-oxidants that contribute to the body's oxidative status. In this study, 32 pigs were fed chicken versus red and processed meat in the context of a prudent or Western dietary pattern for 4 weeks, to investigate their oxidative status. Lipid oxidation products (malondialdehyde, 4-hydroxy-2-nonenal, and hexanal) were higher in the chicken versus red and processed meat diets (1.7- to 8.3-fold) and subsequent in vitro (1.3- to 1.9-fold) and in vivo (1.4 to 3-fold) digests ( P < 0.001), which was presumably related to the higher polyunsaturated fatty acid content in chicken meat and/or the added antioxidants in processed meat. However, diet had only a marginal or no effect on the systemic oxidative status, as determined by plasma oxygen radical absorbance capacity, malondialdehyde, glutathione, and glutathione peroxidase activity in blood and organs, except for α-tocopherol, which was higher after the consumption of the chicken-Western diet. In conclusion, in contrast to the hypothesis, the consumption of chicken in comparison to that of the red and processed meat resulted in higher concentrations of lipid oxidation products in the pig intestinal contents; however, this was not reflected in the body's oxidative status.

Combined Consumption of Beef-Based Cooked Mince and Sucrose Stimulates Oxidative Stress, Cardiac Hypertrophy, and Colonic Outgrowth of Desulfovibrionaceae in Rats.

SCOPE: High red meat and sucrose consumption increases the epidemiological risk for chronic diseases. Mechanistic hypotheses include alterations in oxidative status, gut microbiome, fat deposition, and low-grade inflammation. METHODS AND RESULTS: For 2 weeks, 40 rats consumed a diet high in white or red meat (chicken-based or beef-based cooked mince, respectively), and containing corn starch or sucrose in a 2 × 2 factorial design. Lard was mixed with lean chicken or beef to obtain comparable dietary fatty acid profiles. Beef (vs chicken)-fed rats had higher lipid oxidation products (malondialdehyde, 4-hydroxy-2-nonenal, and hexanal) in stomach content and blood, and lower blood glutathione. Sucrose (vs corn starch)-fed rats showed increased blood lipid oxidation products and glutathione peroxidase activity, higher liver weight and malondialdehyde concentrations, and mesenterial and retroperitoneal fat accumulation. Beef-sucrose-fed rats had increased cardiac weight, suggesting pathophysiological effects on the cardiovascular system. The colonic microbiome of beef-sucrose-fed rats showed an outgrowth of the sulfate-reducing family of the Desulfovibrionaceae, and lower abundance of the Lactobacillus genus, indicating intestinal dysbiosis. Blood C-reactive protein, a marker for inflammation, was not different among groups. CONCLUSIONS: Consumption of a cooked beef-based meat product with sucrose increased oxidative stress parameters and promoted cardiac hypertrophy and intestinal dysbiosis.

Enhancing the health potential of processed meat: the effect of chitosan or carboxymethyl cellulose enrichment on inherent microstructure, water mobility and oxidation in a meat-based food matrix.

The addition of dietary fibers can alleviate the deteriorated textural properties and water binding capacity (WBC) that may occur when the fat content is lowered directly in the formulas of comminuted meat products. This study investigated the effects of the addition of chitosan or carboxymethyl cellulose (CMC) (2% w/w) to a model meat product. Both dietary fibers improved the water-binding capacity (WBC), while chitosan addition resulted in a firmer texture, CMC lowered the hardness. Chitosan addition resulted in a 2-fold reduction of lipid oxidation products, whereas CMC had no significant effect on oxidation. The effect of chitosan addition on lipid oxidation was evident both in the meat system and after simulated in vitro gastrointestinal digestion. Low-field nuclear magnetic resonance (NMR) relaxometry revealed that the fibers impacted the intrinsic water differently; the addition of chitosan resulted in a faster T2 relaxation time corresponding to water entrapped in a more dense pore network. Coherent anti-Stokes Raman scattering (CARS) microscopy was for the first time applied in a meat product to study the microstructure, which revealed that the two fibers exerted different effects on the size and entrapment of fat droplets in the protein network, which probably explain the mechanisms by which chitosan reduced lipid oxidation in the system.

Phloroglucinol-Mediated Hsp70 Production in Crustaceans: Protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii.

The halophilic aquatic bacterium, Vibrio parahaemolyticus, is an important aquatic pathogen, also capable of causing acute hepatopancreatic necrosis disease (AHPND) in shrimp resulting in significant economic losses. Therefore, there is an urgent need to develop anti-infective strategies to control AHPND. The gnotobiotic Artemia model is used to establish whether a phenolic compound phloroglucinol is effective against the AHPND strain V. parahaemolyticus MO904. We found that pretreatment with phloroglucinol, at an optimum concentration (30 µM), protects axenic brine shrimp larvae against V. parahaemolyticus infection and induced heat shock protein 70 (Hsp70) production (twofolds or more) as compared with the control. We further demonstrated that the Vibrio-protective effect of phloroglucinol was caused by its prooxidant effect and is linked to the induction of Hsp70. In addition, RNA interference confirms that phloroglucinol-induced Hsp70 mediates the survival of brine shrimp larvae against V. parahaemolyticus infection. The study was validated in xenic Artemia model and in a Macrobrachium rosenbergii system. Pretreatment of xenic brine shrimp larvae (30 µM) and Macrobrachium larvae (5 µM) with phloroglucinol increases the survival of xenic brine shrimp and Macrobrachium larvae against subsequent V. parahaemolyticus challenge. Taken together, our study provides substantial evidence that the prooxidant activity of phloroglucinol induces Hsp70 production protecting brine shrimp, A. franciscana, and freshwater shrimp, M. rosenbergii, against the AHPND V. parahaemolyticus strain MO904. Probably, phloroglucinol treatment might become part of a holistic strategy to control AHPND in shrimp.

Validity and Reproducibility of a Food Frequency Questionnaire for Dietary Factors Related to Colorectal Cancer.

Dietary factors play a major role in the development of colorectal cancer. This study evaluated the reproducibility and validity of a 109-food item Food Frequency Questionnaire (FFQ) to measure the consumption of foods and nutrients related to the development of colorectal cancer in a population aged ≥50 years in Flanders, Belgium. A semi-quantitative FFQ was administered two times in a period of two weeks to evaluate reproducibility (FFQ1 and FFQ2). The validity of the FFQ was assessed by comparing FFQ1 against the 3-day diary method (3 DD). A total of 162 respondents (mean age 57.5 years) provided data for the FFQ, of whom 156 also participated in the validity assessment. Mean differences in the intake of foods and nutrients between FFQ1 and FFQ2 were, overall, small and statistically insignificant. However, a higher estimation was observed by FFQ1 as compared to the 3-DD method for the majority of food groups and nutrient intake in the validity assessment. A systematic mean difference (g/day) was observed for eight food groups in the Bland-Altman agreement test; the largest was for fruit intake. Regarding the nutrients, a systematic mean difference was observed in calcium, fat, and vitamin D intake. Overall, the reproducibility of the FFQ was good, and its validity could be satisfactory for estimating absolute food and nutrient intakes and ranking individuals according to high and low intake categories.

DNA adductomics to study the genotoxic effects of red meat consumption with and without added animal fat in rats.

Digestion of red and processed meat has been linked to the formation of genotoxic N-nitroso compounds (NOCs) and lipid peroxidation products (LPOs) in the gut. In this study, rats were fed a meat based diet to compare the possible genotoxic effects of red vs. white meat, and the interfering role of dietary fat. To this purpose, liver, duodenum and colon DNA adductomes were analyzed with UHPLC-HRMS. The results demonstrate that the consumed meat type alters the DNA adductome; the levels of 22 different DNA adduct types significantly increased upon the consumption of beef (compared to chicken) and/or lard supplemented beef or chicken. Furthermore, the chemical constitution of the retrieved DNA adducts hint at a direct link with an increase in NOCs and LPOs upon red (and processed) meat digestion, supporting the current hypotheses on the causal link between red and processed meat consumption and the development of colorectal cancer.

Short-term beef consumption promotes systemic oxidative stress, TMAO formation and inflammation in rats, and dietary fat content modulates these effects.

A high consumption of red and/or processed meat is associated with a higher risk to develop several chronic diseases in which oxidative stress, trimethylamine-N-oxide (TMAO) and/or inflammation are involved. We aimed to elucidate the effect of white (chicken) vs. red (beef) meat consumption in a low vs. high dietary fat context (2 × 2 factorial design) on oxidative stress, TMAO and inflammation in Sprague-Dawley rats. Higher malondialdehyde (MDA) concentrations were found in gastrointestinal contents (up to 96% higher) and colonic tissues (+8.8%) of rats fed the beef diets (all P < 0.05). The lean beef diet resulted in lower blood glutathione, higher urinary excretion of the major 4-hydroxy-nonenal metabolite, and higher plasma C-reactive protein, compared to the other dietary treatments (all P < 0.05). Rats on the fat beef diet had higher renal MDA (+24.4% compared to all other diets) and heart MDA (+12.9% compared to lean chicken) and lower liver vitamin E (-26.2% compared to lean chicken) (all P < 0.05). Rats on the fat diets had lower plasma vitamin E (-23.8%), lower brain MDA (-6.8%) and higher plasma superoxide dismutase activity (+38.6%), higher blood glutathione (+16.9%) (all P < 0.05) and tendency to higher ventral prostate MDA (+14.5%, P = 0.078) and prostate weight (+18.9%, P = 0.073), compared to rats on the lean diets. Consumption of the beef diets resulted in higher urinary trimethylamine (4.5-fold) and TMAO (3.7-fold) concentrations (P < 0.001), compared to the chicken diets. In conclusion, consumption of a high beef diet may stimulate gastrointestinal and/or systemic oxidative stress, TMAO formation and inflammation, depending on the dietary fat content and composition.

Meat: The balance between nutrition and health. A review.

Fresh and processed meats provide high biological value proteins and important micronutrients. On the other hand, a working group of IARC recently classified processed meat as 'carcinogenic to humans' and red meat as 'probably carcinogenic to humans' for colorectal cancer, appealing to critically consider the future role of meat in a healthy diet. This manuscript first evaluates the contribution of meat consumption to the supply of important micronutrients in the human food chain, and the extent to which this can be improved by primary production strategies, and impacts on human health. Secondly, the IARC hazard analysis of the carcinogenicity of red and processed meat consumption is discussed, arguing that having more insight in the mechanisms of the association offers opportunities for mitigation. It is advocated that the benefits and risks associated with red and processed meat consumption should not necessarily cause dilemmas, if these meats are consumed in moderate amounts as part of balanced diets.

Reducing Compounds Equivocally Influence Oxidation during Digestion of a High-Fat Beef Product, which Promotes Cytotoxicity in Colorectal Carcinoma Cell Lines.

We studied the formation of malondialdehyde, 4-hydroxy-nonenal, and hexanal (lipid oxidation products, LOP) during in vitro digestion of a cooked low-fat and high-fat beef product in response to the addition of reducing compounds. We also investigated whether higher LOP in the digests resulted in a higher cyto- and genotoxicity in Caco-2, HT-29 and HCT-116 cell lines. High-fat compared to low-fat beef digests contained approximately 10-fold higher LOP concentrations (all P < 0.001), and induced higher cytotoxicity (P < 0.001). During digestion of the high-fat product, phenolic acids (gallic, ferulic, chlorogenic, and caffeic acid) displayed either pro-oxidant or antioxidant behavior at lower and higher doses respectively, whereas ascorbic acid was pro-oxidant at all doses, and the lipophilic reducing compounds (α-tocopherol, quercetin, and silibinin) all exerted a clear antioxidant effect. During digestion of the low-fat product, the hydrophilic compounds and quercetin were antioxidant. Decreases or increases in LOP concentrations amounted to 100% change versus controls.

Supra-nutritional levels of α-tocopherol maintain the oxidative stability of n-3 long-chain fatty acid enriched subcutaneous fat and frozen loin, but not of dry fermented sausage.

BACKGROUND: Meat products enriched with n-3 fatty acids are more prone to oxidation. The aim was to investigate whether supra-nutritional levels of α-tocopherol can enhance the colour and lipid oxidative stability of n-3 fatty acids enriched dry fermented sausages, frozen loins and subcutaneous fat. Pigs were fed a diet supplemented with 18 g kg(-1) fish oil and 50, 150 or 300 mg kg(-1) α-tocopheryl acetate. The control group received 12 g kg(-1) soy oil and 150 mg kg(-1) α-tocopheryl acetate. RESULTS: α-Tocopherol levels of the frozen loin, dry fermented sausage and subcutaneous fat were elevated as a result of the dietary α-tocopherol supplementation. Lipid oxidation occurred to the same extend in the n-3 fatty acid enriched frozen loins when compared to the control group. In the subcutaneous fat enriched with n-3 fatty acids reduced lipid oxidation was found when comparing 50 mg kg(-1) versus 150 and 300 mg kg(-1) . However, in the dry fermented sausages no such effect was observed and higher TBARS values were found in the n-3 fatty acid enriched sausages compared to the control group. Colour parameters of the loin and subcutaneous fat were not affected, whereas some significant differences in the dry fermented sausages were found. The colour stability of the frozen loins was not affected by the dietary treatments. CONCLUSION: Supra-nutritional levels of α-tocopherol maintain the oxidative stability of n-3 fatty acid enriched frozen loins and subcutaneous fat, but not of dry fermented sausages. © 2016 Society of Chemical Industry.

Determination of α-tocopherol by reversed-phase HPLC in feed and animal-derived foods without saponification.

BACKGROUND: The analysis of α-tocopherol in feed and animal-derived foods usually involves a saponification step. However, since saponification often leads to losses of α-tocopherol, a method for the determination of α-tocopherol in feed and in animal-derived foods was developed without a saponification step. RESULTS: In this method, α-tocopherol is extracted with hot ethanol and the co-extracted fat is removed by centrifugation. Removal of the fat fraction is made possible by the addition of water, to achieve an ethanol:water ratio of 40:7, followed by cooling on ice before centrifugation. This procedure allows removal of the fat fraction, while α-tocopherol is retained. Matrices differing in gross composition and α-tocopherol content were analyzed: fresh pork, cooked ham, subcutaneous fat, liver, egg yolk, milk and a compound pig feed. Higher α-tocopherol concentrations were found for this novel method compared to a conventional method with saponification, particularly for subcutaneous fat (P < 0.05). Recoveries were higher (P < 0.05) for the novel method (82-103%), compared to the saponification method (66-90%; for subcutaneous fat < 25%). CONCLUSION: Determining α-tocopherol in feed and animal-derived foods using pure ethanol without saponification results in higher extraction yields and recoveries compared to the saponification method.