Hanne Christine Bertram

Cooking loss and juiciness of pork in relation to raw meat quality and cooking procedure

The study comprised two experiments with the aim to investigate the influence of raw meat quality and cooking procedure on cooking loss and juiciness of pork. The first experiment determined the cooking loss at 60, 70 and 80 °C centre temperature of 10 raw meat qualities (defined according to ultimate pH, drip loss, breed and rearing conditions) when cooked as steaks on a pan or as a roast in oven at a oven temperature of 90 or 190 °C. The differences in cooking loss between the raw meat qualities and the cooking procedures did decrease as the centre temperature increased and were almost negligble at 80 °C. Low water holding capacity (WHC) and low pH resulted in high cooking loss while no difference in cooking loss was observed between meat having medium or high WHC and pH. In the second experiment four raw meat qualities (standard, Duroc, low pH and heavy carcass weight) chosen from the first experiment to ensure a wide variation in cooking loss, were cooked in oven at 90 or 190 °C oven temperature. Juiciness was assessed three times during the chewing process. The results suggested that juiciness experienced initially in the chewing process depended only on the water content of the meat, whereas juiciness experienced later in the chewing process was determined by a combination of the water and intramuscular fat contents and the saliva production during chewing.

Continuous distribution analysis of T2 relaxation in meat-an approach in the determination of water-holding capacity

Continuous distribution analysis as well as traditional bi-exponential fitting analysis were carried out on NMR T(2) relaxation data measured 24 h post-mortem in 74 pork meat samples from M. longissimus dorsi. Areas of relaxation populations found by continuous distribution analysis and corresponding time constants were determined. Comparison of correlations to water holding capacity (WHC), determined by Honikels bag method [Honikel, K. O. (1998). Reference methods for the assessment of physical characteristics of meat. Meat Science, 49, 447-457], revealed superior performance of continuous distribution analysis (r=-0.85) compared with traditional bi-exponential fitting analysis (r=-0.77). Furthermore, the changes in continuous relaxation time distribution spectra were followed during loss of water from meat samples by centrifugation. It was found that the area of the slowest relaxation population (T(22)) decreased proportionally with the amount of expelled water. A physical explanation for the obtained correlation between continuous relaxation time distribution data and WHC is given upon the results from these data.

Comparative study of low-field NMR relaxation measurements and two traditional methods in the determination of water holding capacity of pork.

The correlation between transverse relaxation, T(2,) and water holding capacity (WHC) determined either by Honikels bag method (Honikel, 1998) or centrifugation has been investigated in meat samples from m. longissimus dorsi (LD) 24 h post mortem from 74 pigs. Bi-exponential analysis of the transverse relaxation, T(2), showed highly significant correlations between both the two NMR time constants (T(21),T(22)) and water holding capacity determined by both Honikels bag method (r(T(21))=-0.72 and r(T(22))=0.77) and centrifugation (r(T(21))-0.50 and r(T(22))=0.75). This shows that transverse relaxation measurement is an efficient method for determination of water holding capacity in pork. Significant correlations were also found between T(21) and T(22) measured 24 h post mortem and pH measured at various time post mortem. This indicates that transverse relaxation, T(2), reflects pH-induced structural changes occurring in muscles post mortem.

Aging-induced changes in microstructure and water distribution in fresh and cooked pork in relation to water-holding capacity and cooking loss - A combined confocal laser scanning microscopy (CLSM) and low-field nuclear magnetic resonance relaxation study.

Confocal laser scanning microscopy (CLSM) and low-field nuclear magnetic resonance (LF-NMR) relaxometry were combined to characterize microstructural changes and water distribution in fresh and cooked pork during an aging period of 14 days. At day 1 (24h postmortem) a few muscle fibres, which appear swollen, were observed in both fresh and cooked meat. An identical microstructure was still apparent after 14 days, however, the number of muscle fibres showing distinguished characteristics was found to increase throughout the aging period. Hence, it was apparent that during aging the individual fibres swell and disintegrate at different rates. Development in water-holding capacity (WHC) was followed during the aging period using gravimetric methods, and an increase in the WHC in the fresh meat was observed, which resembled the amount of extramyofibrillar water measured by NMR relaxometry (T(22) population). This was consistent with the CLSM images, as a substantial increase in the number of myofibrils that appeared swollen, capable of holding more water, was observed during aging. In the cooked meat the width of the T(21c) population, reflecting the myofibrillar water in the cooked meat, was seen to decrease during the entire storage period, which corresponds to the development of a more homogeneous structure. In the CLSM data a continuous degradation during the storage period was observed, which could resemble a shift to a more homogeneous structure. Comparison of CLSM of transverse sections of fresh and cooked pork revealed a pronounced shrinkage of muscle fibres upon cooking. This resulted in large gaps between the cooked muscle fibres, which also was visible as shrinkage at the level of the individual myofibrils. This pattern was also reflected in the NMR relaxation data. The cooking-induced shrinkage of the myofibrils occurred concomitantly with a decrease in the amount of intermyofibrillar water within the individual fibre and an increase in the larger extramyofibrillar spaces between fibres, i.e. water is expelled from the myofibrillar matrix upon cooking. Accordingly, the present study demonstrated that the use of CLSM together with NMR relaxometry can provide further information on the relationship between structural characteristics of meat and resultant water distribution.

An NMR-based metabonomic investigation on effects of milk and meat protein diets given to 8-year-old boys

The objective of the study was to investigate the ability of an NMR-based metabonomic approach, applied to biofluids, to explore and identify overall exogenous and endogenous biochemical effects of a short-time high intake of milk protein or meat protein given to prepubertal children, the aim being to compare relative differences and not an absolute quantification. A total of twenty-four 8-year-old boys were asked to take 53 g protein as milk (n 12) or meat daily (n 12). At baseline and after 7 d, urine and serum samples were collected and high-resolution 1H NMR spectra were acquired on these using a 800 MHz spectrometer. The milk diet reduced the urinary excretion of hippurate, while the meat diet increased the urinary excretion of creatine, histidine and urea. The NMR measurements on serum revealed minor changes in the lipid profile, which most probably should be ascribed to an increase in the content of SCFA in the blood after consumption of the milk diet. The meat diet had no effect on the metabolic profile of serum. The study for the first time demonstrates the capability of proton NMR-based metabonomics to identify the overall biochemical effects of consumption of different animal proteins. The urine metabolite profile is more susceptible to perturbations as a result of short diet interventions than the serum metabolite profile. The milk diet-induced reduction in urinary excretion of hippurate suggests alterations in gut microflora, which may be useful information for further studies elucidating the effects of bioactive components in milk.

NMR relaxometry and differential scanning calorimetry during meat cooking

By combining simultaneous nuclear magnetic resonance (NMR) T(2) relaxometry and differential scanning calorimetry (DSC) on pork samples heated to nine temperature levels between 25 and 75°C, the present study investigates the relationship between thermal denaturation of meat proteins and heat-induced changes in water characteristics. Principal component analysis (PCA) on the distributed (1)H NMR T(2) relaxation data revealed that the major changes in water characteristics during heating occur between 40 and 50°C. This is probably initiated by denaturation of myosin heads, which however, could not be detected in the DSC thermograms obtained directly on the meat. In contrast, the DSC thermograms revealed endothermic transitions at 54, 65 and 77°C, probably reflecting the denaturation of myosin (rods and light chain), sarcoplasmic proteins together with collagen and actin, respectively. Simultaneous modelling of DSC and NMR data by partial least squares regression (PLSR) revealed a correlation between denaturation of myosin rods and light chains at ∼53-58°C and heat-induced changes in myofibrillar water (T(2) relaxation time ∼10-60ms) as well as between actin denaturation at ∼80-82°C and expulsion of water from the meat. Accordingly, the present study demonstrates a direct relationship between thermal denaturation of specific proteins/protein structures and heat-induced changes in water mobility during heating of pork.

Functionality of myofibrillar proteins as affected by pH, ionic strength and heat treatment - a low-field NMR study.

Myofibrills were extracted from porcine muscle, and their water properties were characterized using low-field nuclear magnetic resonance (NMR) T(2) relaxometry. A T(2) relaxation pattern very similar to the pattern observed in intact meat and water contents comparable to the water content in meat were observed, implying that the myofibrillar structures are responsible for retaining the majority of water in meat. The effect of pH and ionic strength in the samples was investigated as pH was adjusted to 5.4, 6.2, and 7.0 and ionic strength to 0.29, 0.46 and 0.71 M, respectively. Even though there were interactions between pH and ionic strength, the water content in the samples increased significantly with increasing pH and ionic strength. Moreover, mean T(2) relaxation times likewise increased with increasing pH and ionic strength, which reveals that the increased water retention could be ascribed to a swelling of the myofibrils and thereby increased spacing between filaments. The present study demonstrates that NMR T(2) relaxometry is a potential tool to explore how processing factors such as pH and ionic strength affect the microstructure of meat.

Potential of human saliva for nuclear magnetic resonance-based metabolomics and for health-related biomarker identification.

In the present study, the ability of (1)H nuclear magnetic resonance (NMR) for metabolic profiling of human saliva samples was investigated. High-resolution (1)H NMR spectra were obtained, and signals were assigned to various metabolites mainly representing small organic acids and amino acids. In addition, the use of human saliva for metabolomic studies was evaluated, and multivariate data analysis revealed that the 92 morning and night samples from 46 subjects could be discriminated with a predictability of 85%. The diurnal effect on the salivary metabolite profile were ascribed to changes in intensities of several metabolites including trimethylamine oxide (TMAO), choline, propionate, alanine, methanol, and N-acetyl groups. No effects of gender and body mass index (BMI) on the salivary metabolite profile were detected. The relationships between the salivary metabolome and glycated hemoglobin, systolic and diastolic blood pressure were investigated; however, no significant correlations could be established.

Effect of freezing temperature, thawing and cooking rate on water distribution in two pork qualities.

The effects of freezing temperature (-20 versus -80°C) in combination with long-term freezer storage (-20°C) on water mobility and distribution in pork of two qualities (normal and high ultimate pH) were explored using low-field NMR T(2) relaxometry. A marked effect of freezing temperature on the characteristics of intra- and extramyofibrillar water (T(2) relaxation times) in the thawed pork was demonstrated, implying that the freezing temperature in combination with prolonged freezer storage affects the distribution and chemical-physical state of water in the thawed meat. Determination of technological properties (thawing and cooking loss) revealed that the observed T(2) variations related to water distribution and water properties, which were found to be consistent with the degree of structural damage assessed by light microscopy, also were reflected in the technological quality of the meat. Low freezing temperature in combination with prolonged freezer storage was associated with increased thawing and cooking loss. In addition, pH in the fresh meat had a pronounced effect on the distribution of myofibrillar water, as a more homogenous pore size distribution was evident in meat with high pH compared with normal pH. A clear effect of cooking rate on the T(2) relaxation characteristics in the cooked pork was also demonstrated, probably reflecting a cooking rate-induced effect on the myofibrillar structures. The effect of cooking rate on water distribution resulted in a significantly lower cooking loss upon a slow cooking rate (0.5°C/min from 25 to 65°C and 0.3°C/min from 65 to 80°C) compared with a fast cooking rate (1°C/min).

NMR-based metabonomic studies reveal changes in the biochemical profile of plasma and urine from pigs fed high-fibre rye bread.

This study presents an NMR-based metabonomic approach to elucidate the overall endogenous biochemical effects of a wholegrain diet. Two diets with similar levels of dietary fibre and macronutrients, but with contrasting levels of wholegrain ingredients, were prepared from wholegrain rye (wholegrain diet (WGD)) and non-wholegrain wheat (non-wholegrain diet (NWD)) and fed to four pigs in a crossover design. Plasma samples were collected after 7 d on each diet, and 1H NMR spectra were acquired on these. Partial least squares regression discriminant analysis (PLS-DA) on spectra obtained for plasma samples revealed that the spectral region at 3.25 parts per million dominates the differentiation between the two diets, as the WGD is associated with higher spectral intensity in this region. Spiking experiments and LC-MS analyses of the plasma verified that this spectral difference could be ascribed to a significantly higher content of betaine in WGD plasma samples compared with NWD samples. In an identical study with the same diets, urine samples were collected, and 1H NMR spectra were acquired on these. PLS-DA on spectra obtained for urine samples revealed changes in the intensities of spectral regions, which could be ascribed to differences in the content of betaine and creatine/creatinine between the two diets, and LC-MS analyses verified a significantly lower content of creatinine in WGD urine samples compared with NWD urine samples. In conclusion, using an explorative approach, the present studies disclosed biochemical effects of a wholegrain diet on plasma betaine content and excretion of betaine and creatinine.