Zinc Protoporphyrin-Rich Pork Liver Homogenates as Coloring Ingredients in Nitrite-Free Liver Pâtés.

This study aimed to investigate the coloring ingredient potential of liver homogenates that form Zn protoporphyrin (ZnPP), a natural red pigment, after anaerobic incubation. Liver homogenates were used to develop nitrite-free sterile pork liver pâtés. These homogenates were applied in the formulation of pâtés directly or after centrifugation to obtain a pellet that was highly concentrated in ZnPP. Both the whole homogenate and its insoluble fraction were adjusted to pH 7.5 before their use in the formulation of pâtés with and without antioxidant (0.5% ascorbate plus 0.1% tocopherol) addition. Pâtés formulated with the whole homogenate showed color and texture characteristics that were similar to those of the positive control with nitrite. However, high levels of the insoluble fraction also led to pâtés with improved color characteristics but with a two-fold softened texture. Therefore, the form and amount of ZnPP added played roles in the final appearance of the product. The ZnPP pigment was more stable than heme in the sterilization treatment, and antioxidant addition proved to be unnecessary. The ZnPP-rich ingredients allowed for the preparation of nitrite-free cooked liver pâtés with a stable red color and could thus be potentially applied in other uncured cooked meat products.

Utilization of Porcine Livers through the Formation of Zn-Protoporphyrin Pigment Optimized by a Response Surface Methodology.

There is a growing demand for clean-label products. This study aimed to obtain a food-grade coloring ingredient for meat products based on the formation of Zn-protoporphyrin from porcine livers, thus contributing to the development of nitrite-free products. First, the effects of sodium disulfite and acetic, ascorbic, and lactic acids on the formation of Zn-protoporphyrin and the total microbial count were studied. The combination of ascorbic and acetic acids resulted in a higher Zn-protoporphyrin content than acetic acid alone, and microbial levels were maintained (ca. 3 log CFU/mL). Second, a response surface methodology was used to maximize Zn-protoporphyrin while maintaining microbiological food standards. To that end, the effects of pH (4.2-5.4), incubation time (3-30 h), and temperature (25-50 °C) were studied. The selected conditions for Zn-protoporphyrin formation involved anaerobic incubation at pH 4.8 and 45 °C for 24 h. The safety was validated through challenge testing for relevant pathogens (Listeria monocytogenes, Salmonella spp., and Clostridium perfringens). A significant reduction (>6 log units) was observed in the selected conditions for L. monocytogenes and Salmonella, whereas C. perfringens spores remained at the inoculated levels. The optimized procedure is proven to be microbiologically safe, and may improve the color of nitrite-free meat products.

Innovative strategies to enhance the sensory quality of dry fermented sausages containing lactic ingredients by the addition of exogenous enzymes.

This study investigated the impact of the addition of exogenous enzymes (Accelerzyme CPG, Debitrase DBP20) or cellular preparations (FlavoGard), traditionally used in the cheese industry, to accelerate flavour development of dry fermented sausages with 6% of lactic derivatives content. Sausages were fermented to pH 5.0, dried for 32 days and vacuum packed stored under refrigeration for 60 days. Sausages were analysed for physicochemical parameters, technological microbiota and proteolysis after fermentation, drying/ripening and storage. Similar compositional results were obtained in all products (38-39% humidity in the final product; 38.2% fat and 40.7% protein as dry matter throughout the study). Debitrase application positively affected proteolysis by changing the free amino acid profile and increasing non-protein nitrogen and total free amino acids by 2.2 and 11.8-fold, respectively. Accelerzyme increased ripened cheese flavour and overall sensory quality from 5.1 to 5.8; Debitrase increased ripened cheese odour and flavour, bitterness, umami, adhesiveness, pastiness, and overall sensory quality from 5.0 to 5.9, and decreased acid and hardness. This study highlights the effects of adding some exogenous enzyme/bacterial preparations traditionally used in the cheese industry to enhance the flavour of dry fermented sausages with high content of lactic ingredients and increase its sensory quality.

Formation of Zn-protoporphyrin during the elaboration process of non-nitrified serrano dry-cured hams and its relationship with lipolysis.

This study assessed the ZnPP content, heme content, salt content, and instrumental color in the biceps femoris and semimembranosus muscles during the elaboration of Serrano dry-cured ham manufactured without the addition of nitrate and nitrite for 15 months. The effects of lipolysis and lipid oxidation on the content of Zn-protoporphyrin were also investigated in the biceps femoris. We found that the maximum formation of Zn-protoporphyrin occurred between end of resting and 6 months of processing, which coincides with temperature increase during processing and the end of salt equalization. Zn-protoporphyrin further increased in the biceps femoris until 9 months of processing but remained unchanged in the semimembranosus. Free fatty acid content increased till 6 months and then remained unchanged until the end of the process. These findings and those from an in vitro study reinforced the idea that the release of free fatty acids can promote the activity of the endogenous enzyme ferrochelatase and contribute to the formation of Zn-protoporphyrin from heme. However, the content of Zn-protoporphyrin decreased at the end of the processing, which may be due to the progression of lipid oxidation.

Effect of the sodium reduction and smoking system on quality and safety of smoked salmon (Salmo salar).

Excessive sodium (Na) intake has been associated with high blood pressure and cardiovascular diseases. Therefore, sodium reduction is a public health challenge worldwide. The aim of this study was to develop smoked salmon with a reduced Na content. Sodium chloride (NaCl) was replaced by potassium chloride (KCl) at 25% and 50% (molar replacement) and studied in combination with two smoking procedures (natural wood and liquid smoke) as well as two smoking temperatures (18-19 °C or 56 °C). Smoked salmon samples were characterized by physicochemical, sensory and microbiological analyses. No major differences were observed regarding physicochemical properties in the studied treatments. Smoked samples with 50% of NaCl replaced by KCl were slightly more bitter than those with 25% whereas samples with 25% of replacement did not show differences to those with non-reduced Na content (5 g of added NaCl per 100 g of salmon). Molar Na:K ratio decreased from 4,3 in controls to 1,4 and 0,6 in samples with a NaCl reduction level of 25% and 50% respectively. Microbiological assessment indicates that 2-week shelf-life would be appropriate and safe in terms of accomplishment of the EU regulation, taking into account foreseeable storage temperatures (up to 8 °C). Thus, it is possible to achieve a reduction of 25-50% of NaCl in smoked salmon by replacing NaCl by KCl and considerer this product as a "source" of K.

Co-extruded alginate as an alternative to collagen casings in the production of dry-fermented sausages: Impact of coating composition.

The performance of co-extruded alginate coatings containing no extra additives (A), polyglycerol esters of fatty acids (EA), or pea protein (PA) was assessed as an alternative to collagen casings (C) for the manufacturing of dry-fermented sausages (fuet) with no inoculation of moulds and without a fermentation step (NMNF) and fuet inoculated with Penicillium candidum and fermented (MF). Stuffing into collagen casings resulted in slower sausage drying kinetics compared with alginate coating. No significant differences in aw were observed among the studied casing types for NMNF and MF fuets and for the evolution of the technological and spoilage microorganisms. Fuets coated with A, EA, and PA showed lower pH values than fuets stuffed in collagen casings. No significant differences on sensory properties between casing types were observed. Therefore, alginate coatings would be a feasible alternative to collagen casing from a technological and safety point of view.

Effects of post mortem pH and salting time on Zinc-protoporphyrin content in nitrite-free Serrano dry-cured hams.

There is a growing demand for clean label products and thus the elimination of curing additives in various dry-cured meats is of interest while maintaining colour characteristics. This study was aimed to examine the effect of pH at 24 h post mortem (pHSM24h ≤ 5.4; 5.4 > pHSM24h < 5.9; pHSM24h ≥ 5.9) and salting time (standard vs reduced) on zinc-protoporphyrin content, heme content and other physicochemical parameters of Serrano dry-cured hams manufactured without the addition of curing agents. Results showed that in those hams with higher post mortem pH heme content was increased whereas ZnPP content and proteolysis index were decreased. Reduced salting time decreased salt content whereas ZnPP and heme contents remained unaffected. Lower post mortem pH and reduced salting time led to a higher content in various free fatty acids which, in turn, were found to correlate positively with ZnPP formation. However, the observed changes in heme and ZnPP contents had no effect on the instrumental color of the final product.

Zinc-protoporphyrin content in commercial Parma hams is affected by proteolysis index and marbling.

The contents of zinc-protoporphyrin (ZnPP) and heme in twenty-four sliced Parma hams made without the addition of curing agents were determined. Expressed on a dry weight basis, ZnPP averaged 45 mg/kg and ranged from 23 to 85 mg/kg. The heme content averaged 37 mg/kg on a dry matter basis and ranged from 17 to 73 mg/kg. A Principal Component Analysis (PCA) and Partial Least Squares (PLS) regression analyses were carried out to examine the existing correlations between these pigments and various physicochemical parameters in the final product. PCA showed the existence of associations between ZnPP, sensory redness and salt content. PLS suggests that the conversion of ZnPP from heme is facilitated in those hams with a higher proteolysis index and higher marbling.

Effect of nitrate and nitrite on Listeria and selected spoilage bacteria inoculated in dry-cured ham.

The effect of nitrate and the combination of nitrate/nitrite on Listeria innocua (as surrogate of Listeria monocytogenes). And two selected spoilage microorganisms (Proteus vulgaris and Serratia liquefaciens) was studied in dry-cured ham. Hams were manufactured with different concentrations of curing agents: KNO3 (600 and 150mg/kg) alone or in combination with NaNO2 (600 and 150mg/kg). The addition of 500mg/kg of sodium ascorbate was also evaluated in a batch with 600mg/kg of nitrate and nitrite. The target microorganisms were inoculated by injection in semimembranosus, biceps femoris and in the shank, prior to curing. P. vulgaris and S. liquefaciens were controlled by temperature and aw, respectively, and no effect of nitrate/nitrite was observed. The presence of nitrite in the curing mix reduced L. innocua in semimembranosus, which population was 1.5logcfu/g lower at the end of resting (p<0.05), while at the end of the process it was more frequently detected in the no- and low-nitrite added hams. None of the treatments was able to control Listeria in deeper areas of ham. The addition of sodium ascorbate to the curing mix containing the highest amount of nitrate and nitrite did not show any effect on the microorganisms studied.

Relevance of nanocomposite packaging on the stability of vacuum-packed dry cured ham.

In this study effects of a novel high barrier multilayer polyamide film containing dispersed nanoclays (PAN) on the stability of vacuum packed dry-cured ham were investigated during 90days refrigerated storage in comparison with non-modified multilayer polyamide (PA) and a commercial high barrier film. Characteristic bands of the mineral in FT-IR spectra confirmed the presence of nanoclays in PAN, enhancing oxygen transmission barrier properties and UV protection. Packaging in PAN films did not originate significant changes on colour or lipid oxidation during prolonged storage of vacuum-packed dry-cured ham. Larger oxygen transmission rates in PA films caused changes in CIE b* during refrigerated storage. Ham quality was not affected by light exposition during 90days and only curing had a significant benefit on colour and TBARS, being cured samples more stable during storage in all the packages used. Packaging of dry-cured ham in PAN was equivalent to commercial high barrier films.

Sensory characterisation and consumer acceptability of potassium chloride and sunflower oil addition in small-caliber non-acid fermented sausages with a reduced content of sodium chloride and fat.

The effect of the simultaneous reduction of fat proportion (from 20% to 10% and 7%) and added salt (from 2.5% to 1.5%) and the subsequent addition of 0.64% KCl and sunflower oil (1.5% and 3.0%) on the physicochemical, instrumental colour and texture, sensory properties and consumer acceptability of small caliber non-acid fermented sausages (fuet type) was studied. This simultaneous reduction of fat and salt increased weight loss, moisture, water activity (aw), redness, instrumental texture parameters (hardness, chewiness and cohesiveness), sensory attributes (darkness, hardness, elasticity) and the consumer acceptability. The subsequent addition of 0.64% KCl to the leanest batch decreased the aw and barely affected instrumental texture parameters and consumer acceptability. Subsequent sunflower oil addition decreased hardness, chewiness and cohesiveness and increased crumbliness and oil flavour which may decrease the consumer acceptability. The simultaneous reduction of fat and NaCl with the addition of 0.64% KCl was the preferred option by the consumers.

Low intramuscular fat (but high in PUFA) content in cooked cured pork ham decreased Maillard reaction volatiles and pleasing aroma attributes.

The influence of intramuscular fat content (high - HI versus low - LI) and fatty acid composition on pork cooked cured ham flavour was analysed by gas chromatography-olfactometry using nasal impact frequency (GC-O/NIF) and quantitative descriptive analysis (QDA). Potential relationships were studied by principal component analysis (PCA). Sixteen and fourteen odourants were identified by GC-O/NIF in LI and HI cooked hams, respectively. The two ham types differed in lipid oxidation odourants: polyunsaturated fatty acid (PUFA) derivatives hexanal, 1-octen-3-one and (E,E)-2,4-decadienal were higher in LI ham; while monounsaturated fatty acid (MUFA) derivative decanal was higher in HI. HI samples resulted in higher values for odour-active aroma compounds from Maillard reaction, which are related to roast flavour and a higher overall flavour liking. In summary, our results suggest that Maillard derived odour-active aroma compounds were partially inhibited in LI samples (high in PUFA), resulting in lower positive sensory ratings.

Optimisation of stir-bar sorptive extraction (SBSE), targeting medium and long-chain free fatty acids in cooked ham exudates.

The purpose of our research was to optimise the extraction conditions of the stir-bar sorptive extraction (SBSE) targeting the identification of lipid compounds particularly medium and long-chain free fatty acids in cooked cured pork ham exudates. The analytical conditions of extraction (including sample volume, extraction time, stirring speed, pH and dilution of the sample) were checked using the Simplex method approach. As a result of the SBSE optimisation, improved detection limits and linear ranges for hexanoic, heptanoic, octanoic, nonanoic, decanoic, dodecanoic and tetradecanoic fatty acids were obtained. When comparing results with those obtained by the commonly used SPME methodology, optimisation of SBSE achieved better results for volatile compounds of low volatility, such as medium and long-chain free fatty acids, whereas compounds with high volatility and polarity were only detected by SPME. SBSE also confirmed its potential as a tool to help identify undesirable contaminants/residues in meat products.

Effect of temperature, high pressure and freezing/thawing of dry-cured ham slices on dielectric time domain reflectometry response.

Dielectric Time Domain Reflectometry (TDR) is a useful technique for the characterization and classification of dry-cured ham according to its composition. However, changes in the behavior of dielectric properties may occur depending on environmental factors and processing. The effect of temperature, high pressure (HP) and freezing/thawing of dry-cured ham slices on the obtained TDR curves and on the predictions of salt and water contents when using previously developed predictive models, was evaluated in three independent experiments. The results showed that at temperatures below 20 °C there is an increase of the predicted salt content error, being more important in samples with higher water content. HP treatment caused a decrease of the reflected signal intensity due to the major mobility of available ions promoting an increase of the predicted salt content. Freezing/thawing treatment caused an increase of the reflected signal intensity due to the microstructural damages and the loss of water and ions, promoting a decrease of the predicted salt content.

Effect of reducing and replacing pork fat on the physicochemical, instrumental and sensory characteristics throughout storage time of small caliber non-acid fermented sausages with reduced sodium content.

The effect of pork fat reduction (from 44% to 20% final fat content) and its partial substitution by sunflower oil (3% addition) on the physicochemical, instrumental and sensory properties throughout storage time of small caliber non-acid fermented sausages (fuet type) with reduced sodium content (with partial substitution of NaCl by KCl and K-lactate) and without direct addition of nitrate and nitrite (natural nitrate source used instead) was studied. Results showed that sausages with reduced fat (10% initial fat content) and with acceptable sensory characteristics can be obtained by adding to the shoulder lean (8% fat content) during the grinding, either 3.3% backfat (3% fat content) or 3% sunflower oil, both previously finely comminuted with lean. Furthermore, sunflower oil showed to be suitable for partial pork backfat substitution in very lean fermented sausages, conferring desirable sensory properties similar to those of sausages with standard fat content. The sensory quality of the sausages was maintained after three-month cold storage in modified atmosphere.

Salt uptake and water loss in hams with different water contents at the lean surface and at different salting temperatures.

The salt uptake homogeneity is crucial in assuring quality in dry-cured hams. The aim of this study was to evaluate the effect of the water contents at the lean surface before salting and of the temperature during salting on the salt uptake. Pieces of loin stored at 3°C for 3 days before salting absorbed less salt through a surface that has been dried during storage. A group of raw hams were subjected to different pre-salting storage times (0, 3 and 6 days) and another group subjected to different set room temperatures during salting (-1.0, 0.5 and 4.0°C). The duration of storage before salting and the temperature during salting had a negative and a positive effect on the average salt absorption, respectively. The most important effects appeared after 6 days of storage and at 4°C. No significant differences in salt uptake homogeneity were found between storage times and between salting temperatures.

The effect of high pressure and residual oxygen on the color stability of minced cured restructured ham at different levels of drying, pH, and NaCl.

Color stability of minced cured restructured ham was studied by considering the effects of high pressure (HP) (600 MPa, 13°C, 5 min), raw meat pH24 (low, normal, high), salt content (15, 30 g/kg), drying (20%, 50% weight loss), and residual oxygen level (0.02%-0.30%). Raw hams were selected by pH24 in Semimembranosus, mixed with additives, frozen, sliced, and dried by the Quick-Dry-Slice® (QDS) process followed by HP treatment or not (control). Packaging and storage simulated industrial packaging: modified atmosphere containing 80% N2, 20% CO2, and residual O2 in one of three intervals: <0.1%, 0.1%-0.2%, or 0.2%-0.3%, and retail storage conditions: chill storage, 12 h light, 12 h darkness. HP improved the stability of the redness of 20% QDS hams, while the stabilizing effect on 50% QDS hams was smaller, concluding that water has the dominating role. Raw meat pH24, salt content, and residual oxygen level had varying effects on the stability of the red color.

Identification of proteomic biomarkers in M. Longissimus dorsi as potential predictors of pork quality.

Meat quality traits have low heritability and large environmental influences. To predict, improve and manage meat quality, proteomic biomarkers are superior to genetic markers. The objectives of this research were (1) to find associations between proteome profiles of longissimus muscle at slaughter and meat quality accuracies of prediction of traits ranged from 20 up to 80%. Differentially expressed proteins related to drip loss and ultimate pH were identified by NanoLC-FTMSMS. The proteins highlight biological mechanisms that may explain how these traits develop biologically and how they are related to each other.

Effect of the type of fat on the physicochemical, instrumental and sensory characteristics of reduced fat non-acid fermented sausages.

Four batches of reduced fat non-acid fermented sausages were manufactured with pork-ham lean, and the addition of no fat (Lean), 5% pork backfat (BF), 5% sunflower oil (SO) and 5% diacylglycerols (DAGs). The effect of the type of fat as pork-fat substitute on some physicochemical parameters, instrumental color and texture and sensory attributes of the sausages was studied. Results showed that reduced fat non-acid fermented sausages containing less than 12.5% of fat (BF, SO and DAGs) had a good overall sensory quality. This means a fat reduction of more than 70% compared with the average fat content of standard fermented sausages of similar characteristics. Sausages with SO showed higher sensory ratings in desirable ripened odor and flavor attributes and improved texture defined by lower hardness and chewiness (both sensory and instrumental) and higher crumbliness. Sausages with DAGs showed a similar behavior to that of BF, so they could be a good alternative to produce healthier reduced fat non-acid fermented sausages.

NIR technology for on-line determination of superficial a(w) and moisture content during the drying process of fermented sausages.

Three different NIR equipment were evaluated based on their ability to predict superficial water activity (a(w)) and moisture content in two types of fermented sausages (with and without moulds on surface), using partial least squares (PLS) regression models. The instruments differed mainly in wavelength range, resolution and measurement configuration. The most accurate equipment was used in a new experiment to achieve robust models in sausages with different salt contents and submitted to different drying conditions. The models developed showed determination coefficients (R(2)(P)) values of 0.990, 0.910 and 0.984, and RMSEP values of 1.560%, 0.220% and 0.007% for moisture, salt and a(w) respectively. It was demonstrated that NIR spectroscopy could be a suitable non-destructive method for on-line monitoring and control of the drying process in fermented sausages.