Detection of Pork in Processed Meat by an Enzyme-linked Immunosorbent Assay Using Antiswine Antisera

An antiserum to autoclaved porcine muscle extract was raised in sheep and tested for the detection of pork in mixtures of pork in mutton (P/ S) or beef (P/B) heated at 70, 100, and 120°C for 30 min using a competitive enzyme-linked immunosorbent assay (ELISA) method. The results indicated that the antiserum detected low percentages of pork (1%) mixed in beef and in mutton (0.5%) even when the meat mixtures were heated at 70, 100 or 120°C for 30 min, which correspond to heat treatments of commercially processed meat products. Regression analysis showed a high positive correlation (r > 0.99) between absorbance (OD) values and different percentages of experimental or commercially processed P/B meat mixtures. It was concluded that the combination of anti-porcine sera raised in sheep and the ELISA method can be used for the detection of low percentages of pork in processed meats.     

Inhibition of Heterocyclic Amine Formation in Beef Patties by Ethanolic Extracts of Rosemary

ABSTRACT: Heterocyclic amines (HCAs) are mutagenic compounds formed during cooking muscle foods at high temperature. Inhibition of HCAs by rosemary extracts were evaluated with beef patties cooked at 191 °C (375 °F) for 6 min each side and 204 °C (400 °F) for 5 min each side. Five rosemary extracts extracted with different solvents were used in this study: extract 100W (100% water), 10E (10% ethanol), 20E (20% ethanol), 30E (30% ethanol), and 40E (40% ethanol). The 5 extracts were directly added to beef patties at 3 levels (0.05%, 0.2%, and 0.5%) before cooking and HCA contents were extracted and quantified. All of the patties contained 2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline (MeIQx), and 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP). There was no statistical difference in the inhibition of HCAs in the 0.05%, 0.2%, and 0.5% rosemary extracts. All rosemary extracts significantly decreased the levels of MeIQx and PhIP at both cooking conditions. When cooking at 204 °C (400 °F) for 5 min each side, rosemary extracts 10E and 20E were superior to rosemary extracts 100W, 30E, and 40E in inhibiting HCA formation. Rosemary extract 20E showed the greatest inhibition of MeIQx (up to 91.7%) and PhIP (up to 85.3%). The inhibiting effect of rosemary extracts on HCA formation corresponded to their antioxidant activity based on a DPPH scavenging assay. Rosemary extract 10E and 20E contain a mixture of rosmarinic acid, carnosol, and carnosic acid. It is possible that these compounds might act synergistically in inhibiting the formation of HCAs.     

End-point Temperature (EPT) Affects N-Acetyl-β-D-Glucosaminidase Activity in Beef, Pork and Turkey

End-point tempcraturc (EPT) affected N-acetyl-β-D-glucosaminidase (NAGasc) activity in nonfrozcn and previously frozen cores and ground sampics of beef, pork, turkey breast and ground turkey leg muscle tissue. There was little loss of NAGase activity when heating the products to 40°C; however, as EPT increased from 40–70°C, them was (Pc0.05) loss of activity. At 70°C 90% of activity was lost in beef, 98% in pork, and 93 to 98% in turkey. Inactivation temperature values, IT50, (50% inactivation) were: beef, 59.8°C; pork, 53.4°C; turkey breast, 55.6°C; and turkey leg, 56.6°C.     

Heat and ultrafiltration extraction of broiler meat carnosine and its antioxidant activity

This study examined the effects of extraction and further ultrafiltration on the carnosine content, antioxidant activity and total iron content of chicken muscle extracts. Fresh breast meat had 7-fold higher carnosine than fresh thigh meat (2900 versus 419μg/g meat, respectively). Carnosine extracts of breast and thigh were prepared by heating at 60, 80 and 100°C, and ultrafiltration (UF) using a 5000MW cut-off. At increasing temperatures, protein concentrations decreased while carnosine, total iron and antioxidant activity increased. Antioxidant abilities of the 80 and 100°C-heated extracts were greater than that of the 60°C extract (p<0.05). The ultrafiltrate from the 80°C-heated extract had approximately 20% higher carnosine, but 40% lower protein and 10-30% lower iron than the 80°C-heated ultrafiltrate. However, compared in terms of carnosine concentration, the meat extracts had greater antioxidant activity than pure carnosine (p<0.05).     

Effects of Heat Treatment on Total Phenolic Contents,Antioxidant and Anti-Inflammatory Activities of Pleurotus Sajor-Caju Extract

Untreated and heat treated Pleurotus sajor-caju methanolic extracts were examined for total phenolic content, antioxidant and anti-inflammatory activities. Total phenolic content was determined by using the Folin-Ciocalteu method, and results were expressed as mg gallic acid equivalent per g of sample extract. Antioxidant activity was measured by using the DPPH free radical scavenging method while anti-inflammatory activity was measured by using the Griess assay. Methanolic extracts of fresh fruiting bodies were heated at 100 and 121°C for 15 and 30 min. The results showed that total phenolic contents of heat-treated samples did not increase significantly when compared to the control sample. Total phenolic contents were within the range of 0.06 ± 0.05 to 0.08 ± 0.05 mg gallic acid/equivalents g of dry weight. The samples heated at 100°C for 15 and 30 min showed the highest radical scavenging activity at 500 μg/ml with the lowest IC₅₀ values of 10.4 ± 3.11 and 11.67 ± 2.75 μg/ml, respectively. The results suggested that the increased antioxidant activity of P. sajor-caju after the heat treatment might have increased the health beneficial effects to humans.     

Effect of Temperatures on Fish Alkaline Protease, Protein Interaction and Texture Quality

The effect of the activity of fish alkaline protease and protein denaturation on texture changes in mullet muscle during heating were studied. The proteolytic enzyme and the urea denatured hemoglobin substrate were incubated at different pH values and various temperatures. The enzyme activity was optimal at pH 8 and at 65°C. The extent of protein-protein interactions in mullet actomyosin solutions increased with increasing temperature. The rate of protein-protein interactions increased sharply as temperature increased from 35°C to 45°C at pH 6. When mullet muscle was heated at various temperatures ranging from 55°-85°C, the shear force value increased (toughening) during the initial stage of heating and then decreased (softening). The toughening of fish flesh is probably due to muscle protein interactions, and the later tenderizing of fish flesh may be explained by alkaline protease activity (hydrolysis) masking the protein denaturation effect. When fish flesh was cooked at 100°C, no tenderizing effect was observed. This was probably due to loss of enzyme activity and only protein denaturation had significant effect on texture quality.     

Antioxidant activity of mechanically separated pork extracts

Utilization of synthetic carnosine as a food additive is limited by both regulatory and economic hurdles. Therefore, the potential of producing carnosine-containing antioxidant extracts from an underutilized skeletal muscle source, mechanically separated pork (MSP), was investigated. Carnosine-containing MSP extracts were capable of inhibiting lipid oxidation both in vitro and in salted ground pork. Heating (60-80°C) the MSP extract removed iron and increased in vitro antioxidant activity. Isolation of a low molecular weight fraction of the MSP extract by ultrafiltration was effective at decreasing iron but did not substantially increase in vitro antioxidant activity. Freeze dried extracts (untreated, 80°C, ultrafiltration permeate) were capable of inhibiting both TBARS and lipid peroxide formation in ground, salted pork stored at -15°C. While MSP extracts were capable of inhibiting lipid oxidation both in vitro and in salted, ground pork, their antioxidant activity was low suggesting that their use as a food additive would be impractical.     

Assessment of Previous Heat Treatment of Laboratory Heat-Processed Meat and Poultry using a Commercial Enzyme System

The APIZYM enzyme system was assessed with respect to: enzyme activity in aqueous and saline extract filtrates from raw and heat processed (60° and 71.1°C) beef; effect of rate of heating (0.2–0.3°C, 0.4–0.5°C, and 0.9–1.0°C/min) to internal temperatures of 66.7°, 67.8°, 68.9°, 70.0°, and 71.1°C and holding times of 0, 15, 30, 45, and 60 min at these temperatures on residual enzyme activity; reduction of enzyme assay incubation time; and reproducibility of the system. Results showed that slightly higher enzyme activity in heated samples was obtained with 0.9% saline extraction compared to water. Greater enzyme activity was observed in filtrates from pork samples heat-processed at a fast rate than at a slow rate. Enzyme activity could be detected after 2 hr incubation, but 4 hr were needed for the enzymes to react with their respective substrates. The system was found to be reproducibile. The results of this study also indicated the potential of using leucine aminopeptidase as an indicator enzyme for determining the adequacy of heat treatment of meat and poultry products.     

Antioxidant activity of ginger extract in sunflower oil

The antioxidant activity of dichloromethane extract from ginger was evaluated during 6 months of storage of refined sunflower oil at 25 and 45 °C. Free fatty acid (FFA) content, peroxide value (POV) and iodine value (IV) were used as criteria to assess ginger extract as an antioxidant. After 6 months of storage at 45 °C, sunflower oil containing 1600 and 2400 ppm ginger extract showed lower FFA contents (0.083 and 0.080%) and POVs (24.5 and 24.0 meq kg^?1) than the control sample (FFA contents 0.380%, POV 198.0 meq kg^?1). Sunflower oil containing 200 ppm butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) showed FFA contents of 0.089 and 0.072% and POVs of 26.5 and 24.7 meq kg^?1 respectively after 6 months of storage at 45 °C. Similarly, after 6 months of storage at 45 °C, IVs of sunflower oil containing 1600 and 2400 ppm ginger extract were 80 and 92 respectively, higher than that of the control sample (53). However, IVs of sunflower oil treated with 200 ppm BHA and BHT were 94 and 96 respectively after 6 months of storage at 45 °C. These results illustrate that ginger extract at various concentrations exhibited very strong antioxidant activity, almost equal to that of synthetic antioxidants (BHA and BHT). Ginger extract also showed good thermal stability and exhibited 85.2% inhibition of peroxidation of linoleic acid when heated at 185 °C for 120 min. Therefore the use of ginger extract in foods is recommended as a natural antioxidant to suppress lipid oxidation. © 2003 Society of Chemical Industry     

Lactate Dehydrogenase Activity as an Endpoint Heating Indicator in Cooked Beef

Lactate dehydrogenase (LDH, E.C. 1.1.1.27) activity, specifically LDH5 isoenzyme, declined significantly when bovine Semimembranosus (SM) slurries were heated from 57° to 66°C. When SM whole muscle roasts were brine pumped and oven-cooked, total LDH and specific activities declined proportionately from 57° to 63°C resulting in almost no activity at 63°± 0.5°C. LDH activities were consistently higher in younger cattle and lower in older cattle for SM, Infraspinatus (IN) and Longissimus dorsi (LD) muscles and were lowest overall in the IN. LDH colorimetric and fluorescent assays were more accurate indicators for predicting beef roast heating temperature endpoints between 60° and 63°C than the USDA-FSIS method.     

Thermal Properties of Beef Loaf Produced in Foodservice Systems

There is a lack of information on thermal and other properties of foods, especially food mixtures. Physical properties of beef loaf prepared following formula and procedures used in alternate foodservice systems were determined: heat capacity (0.88 cal/g°C uncooked, 0.91 heated to 60°C) moisture content (72.1% uncooked, 66.2% heated), thermal conductivity (0.40 w/cm°C uncooked, 0.47 heated), fat (17.6% uncooked, 13.0% heated) and density (1.00 g/cm³ uncooked and 0.70 heated). The surface heat transfer coefficient for a forced convection oven was also determined (62 w/ m²°C.) Times to heat beef loaves to specified ending temperatures in a forced convection oven at 163°C and 176°C were calculated. There was less than 1 min difference in calculated heating time required to reach the desired end temperature. The actual times to reach ending temperature show very good agreement with the calculated times.     

Stability of hydroxycinnamic acids and caffeine from green coffee extracts after heating in food model systems

This study was designed to characterize the stability of hydroxycinnamic acids and caffeine obtained from green coffee beans in the form of lyophilized extract (GCE) during heating after supplementation to model systems with saccharose, potato starch, egg white protein, and sunflower oil. Also the addition of iron ions was used. Systems were prepared as a mixture of GCE with a single substance or in a more complex matrix. Heating was carried out at 180 °C for 0.5 and 1 h. Because of the saccharose content, some systems were heated only at 110 °C. The losses of hydroxycinnamic acids in heated systems ranged from 18 to 84 %, and the caffeine from 1.5 to 10 %. The presence of sunflower oil in the systems had the greatest influence on hydroxycinnamic acids degradation. However, in case of the system of each of the examined substances with the coffee preparation, an increased degradation of hydroxycinnamic acids resulting from the introduction of ferrous ions into the systems was observed. Earlier results concerning antioxidant activity of systems containing GCE allow to conclude that the degradation of hydroxycinnamic acids was weakly related to the decrease in antioxidant activity.     

Enzyme Profile of Raw and Heat-Processed Beef, Pork and Turkey Using the "Apizym" System

A semi-quantitative micromethod enzyme system (APIZYM) designed for the detection of 19 individual enzymes was used to determine the presence and level of activity of these enzymes in raw and heat processed (60 and 71.1°C) beef, pork and turkey muscle tissue. Filtrates were obtained from 0.9% saline extracts of the raw and heat processed samples. Heat processing the product to 60°C greatly decreased the number of enzymes that could be detected. Samples heated to 71.1°C exhibited little enzymatic action, except for minimal activity of leucine aminopeptidase. Thus, indicating the possibility of using the APIZYM system as a new and/or improved method for determining the end-point temperature to which meat and poultry products have been heat processed.     

Functional properties and antioxidative activity of protein hydrolysates from toothed ponyfish muscle treated with viscera extract from hybrid catfish

Functional properties and antioxidative activity of a protein hydrolysate prepared from toothed ponyfish (Gazza minuta) muscle, using viscera extract from hybrid catfish (Clarias macrocephalus × Clarias gariepinus), with a degree of hydrolysis (DH) of 70%, were investigated. The protein hydrolysate had a good solubility. It was soluble over a wide pH range (3–9), in which more than 77% solubility was obtained. The emulsifying activity index of the protein hydrolysate decreased with increasing concentration (P < 0.05). Conversely, the foaming abilities increased as the hydrolysate concentrations increased (P < 0.05). Protein hydrolysate exhibited the increases in 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH), 2,2‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulphonic acid) (ABTS) radical scavenging activities, ferric reducing power (FRAP) and metal chelating activity as hydrolysate concentration increased (P < 0.05). ABTS radical scavenging activity of protein hydrolysate was stable when heated at 100 °C for 180 min and subjected to a wide pH range (1–11). Therefore, protein hydrolysate from the muscle of toothed ponyfish produced by viscera extract from hybrid catfish can be used as a promising source of functional peptides with antioxidant properties.     

Effect of Temperature on Collagen Extractability and Kramer Shear Force of Restructured Beef

Collagen extractability and Kramer shear force were measured for raw and heated restructured beef products made with trimmed (epimysium removed) and untrimmed clods (triceps brachii, infraspinatus, and supraspinatus). Collagen extractability was significantly (p < 0.001) higher for untrimmed samples heated at 50°C than for trimmed samples. Kramer shear forces were significantly higher (p < 0.01) for untrimmed than trimmed when raw or heated at 35°C, 45°C, WC, and 55°C. Collagen extractabilities showed no distinct changes with temperature for trimmed samples. Collagen extractability of untrimmed samples increased then decreased as heating temperature increased. Kramer shear forces decreased between 55°C and 60°C for both trimmed and untrimmed samples.     

Antioxidant Properties of Aqueous Extract of Roasted Hulled Barley in Bulk Oil or Oil‐in‐Water Emulsion Matrix

Antioxidant properties of the aqueous extracts of hulled barley (Hordeum vulgare L.) that had been roasted at 210 °C for 20 min were determined in bulk oil and oil‐in‐water (O/W) emulsions. Bulk oils were heated at 60, 100, and 180 °C, and O/W emulsions were oxidized under riboflavin photosensitization. The content of phenolic compounds was analyzed by high‐performance liquid chromatography, and in vitro antioxidant assays were also conducted. The major phenolics contained in the aqueous extract of roasted hulled barley (AERB) were p‐coumaric, ferulic, protocatechuic, chlorogenic, 4‐hydroxybenzoic, and vanillic acids. Depending on the concentration and oxidation temperature, AERB had antioxidant or prooxidant properties in bulk oil. At 60 °C, AERB at a concentration of 0.5% acted as a prooxidant, whereas at 1.0% it acted as an antioxidant. At 100 °C, AERB acted as an antioxidant irrespective of concentration. In 180 °C conditions, 0.5% AERB acted as a prooxidant, whereas other concentrations of AERB acted as antioxidants. In the case of riboflavin photosensitized O/W emulsions, AERB showed antioxidant properties irrespective of concentration. Antioxidant abilities of AERB are affected by the food matrix, including bulk oil and O/W emulsions, and concentrations of AERB, even though diverse phenolic compounds may display high antioxidant properties in in vitro assays.     

Heating and Texture Profiles of Packaged Pasteurized Beef Loin Steaks from Precooked Roasts

Precooked beef loin steaks obtained from waterbath cooked roasts, were oasteurized usinge three different temoeratures (65° 85° and 100°C) and rimoved after reaching internal temperatures of 60°, 70° and 80°C. Temperatures of the steaks were monitored at three locations during heating and cooling. Steaks heated to internal temperatures close to the cooking water temperatures had the least differences in temperature between layers. Total cooking losses increased (P<0.05) as internal endpoint temperatures and pasteurization water temperature increased. Texture profiles of steaks cut from precooked roasts were similar to pasteurized steaks.     

Antioxidant activity and β-glucan contents of hydrothermal extracts from maitake (Grifola frondosa)

Maitake (Grifola frondosa) was hydrothermally extracted at 4 different temperatures (121, 130, 140, and 150°C) for 30 and 60 min, and the total phenolic content (TPC), antioxidant activity, and β-glucan content of the extracts was evaluated. The highest TPC was detected in the extract at 150°C treated for 60 min with 13.61 mg gallic acid equivalents (GAE)/g. The antioxidant activity of the maitake extracts, evaluated by determining DPPH radical scavenging activity (RSA), ABTS RSA, reducing power, and tyrosinase inhibitory activity increased with increasing treatment temperature and time. The content of β-glucan was the highest (5.13%) in the extract prepared at 140°C for 30min. These results suggest that hydrothermal extraction could be used as a tool to increase the antioxidant activity of maitake extracts.     

Effect of some factors used to the chicken meat preservation and processing on the protease activity

The obtained results indicated that the cathepsin activity was higher by about 60% in the extract from thigh than from breast muscles. Freezing and defrosting (not stored) of chicken meat did not influence the breast muscle cathepsin activity while they caused a decrease of activity of about 20% in the case of thigh muscles. The increase in cathepsin activity was noticed in both kinds of muscles during storage at ?20 °C up to 4 months (45.6% and 19.4% for thigh and breast muscles respectively). The activity of cathepsin in extract from 5 months stored meat reached 80% in case of breast muscles and 83% in case of thigh muscles in relation to control sample respectively. The cathepsin activity significantly increased during heating of breast muscles up to 60 °C, but in case of thigh muscles it was slightly higher than at 50 °C. The heating of cured chicken breast muscles up to 60 °C caused a non significant growth in cathepsin activity opposite to raw muscles. The cathepsin activity from all cured samples heated up to 70 °C were several times lower in relation to control samples. The cathepsin activity of both thigh and breast muscles were resistant to gamma radiation. The investigated factors caused changes in the activity of cathepsin but none of them caused its total inactivation. The changes of cathepsin activity depended on the kind of muscles and the kind and the value of acting factors.     

TENSILE PROPERTIES OF BEEF SEMITENDINOSUS MUSCLE AS AFFECTED BY HEATING RATE AND END POINT TEMPERATURE

The effects of heating rate and end point temperature on tensile properties of beef semi-tendinosus muscle were studied. Samples of meat heated in glass tubes in a water bath to simulate oven roasting at 93 °C had lower (P < 0.01) shear values than those heated to simulate oven roasting at 149 °C. Samples heated to 60 and 70°C had lower (P < 0.001) shear values than samples heated to 50°C. Instron breaking strength values varied (P < 0.001) with end point temperature, but were not significantly affected by the rate of heating. Significant (P < 0.001) interactions between the end point temperature and heating rate, as well as between the end point temperature and fiber direction suggested that Instron breaking strength measurements may be more sensitive to changes in tenderness than Warner Bratzler shear measurements.