Quantification of zinc-porphyrin in dry-cured ham products by spectroscopic methods Comparison of absorption, fluorescence and X-ray fluorescence spectroscopy

Zinc–protoporphyrin (Zn–pp), which has been identified as the major pigment in certain dry-cured meat products, was extracted with acetone/water (75%) and isolated from the following meat products: Parma ham, Iberian ham and dry-cured ham with added nitrite. The quantification of Zn–pp by electron absorption, fluorescence and X-ray fluorescence (XRF) spectroscopy was compared (concentration range used [Zn–pp] = 0.8–9.7 μM). All three hams were found to contain Zn–pp, and the results show no significant difference among the content of Zn–pp quantified by fluorescence, absorbance and X-ray fluorescence spectroscopy for Parma ham and Iberian ham. All three methods can be used for quantification of Zn–pp in acetone/water extracts of different ham types if the content is higher than 1.0 ppm. For dry-cured ham with added nitrite, XRF was not applicable due to the low content of Zn–pp (<0.1 ppm). In addition, XRF spectroscopy provides further information regarding other trace elements and can therefore be advantageous in this aspect. This study also focused on XRF determination of Fe in the extracts and as no detectable Fe was found in the three types of ham extracts investigated (limit of detection; Fe ? 1.8 ppm), it allows the conclusion that iron containing pigments, e.g., heme, do not contribute to the noticeable red colour observed in some of the extracts.     

Studies on the antioxidative activity of red pigments in Italian-type dry-cured ham

Aqueous phosphate buffer extracts and acetone/water extracts of pigments from Parma ham were assessed as antioxidants by (1) electron spin resonance spectroscopy using a spin probing technique to evaluate their efficiencies as scavengers of free radicals, and (2) by electrochemical measurement of oxygen depletion rate in an aqueous methyl linoleate emulsion to evaluate their efficiencies as chain-breaking antioxidant, and using both methods, compared with the effect of apomyoglobin and nitrosylmyoglobin. Aqueous phosphate extracts and acetone/water extracts of Parma ham pigment both scavenged a semi-stable nitroxide radical (Fremy's salt), and both extracts reduced the rate of oxygen consumption for lipid peroxidation (initiated by metmyoglobin) very efficiently. For apomyoglobin no antioxidative capacity was observed, and the heme moiety of the pigment(s) of Parma ham were concluded to have antioxidative properties. The more lipophilic pigment, as extracted by acetone/water, had the most significant effect, and its ability to inhibit lipid oxidation was further tested in a model food system based on cooked pork. The lipid oxidation was increasingly inhibited by increasing additions from 0.12 ppm to 0.24 ppm Parma ham pigment, and the pigment protected -tocopherol against degradation in a concentration dependent manner.     

Spectral characterisation of red pigment in Italian-type dry-cured ham. Increasing lipophilicity during processing and maturation

Spectroscopic studies of Parma ham during processing revealed a gradual transformation of muscle myoglobin, initiated by salting and continuing during ageing. Electron spin resonance spectra did, however, conclusively show that the pigment in dry-cured Parma ham at no stage is a nitrosyl complex of ferrous myoglobin as found in brine-cured ham and Spanish Serrano hams. Both near-infra red reflectance spectra of sliced ham and UV/visible absorption spectra of extract of hams, obtained with aqueous buffer or acetone, showed the presence of different red pigments at varying processing stages for both solvents. Especially, the pigment extracted with aqueous buffer exhibited unique spectral features different from those of well-known myoglobin derivatives. At the end of processing, the pigment(s) becomes less water extractable, while the fraction of red pigment(s) extractable with acetone/water (75%/25%) increases throughout the processing time up to full maturation at 18 months. The chemical identity of the 6th ligand of myoglobin could not be conclusively established, but possible candidates are discussed. The partition of the pigment(s) between pentane and acetone/water showed a strong preference for pentane, suggesting that only the heme moiety is present in the acetone/water extract, and that Parma ham pigment is gradually transformed from a myoglobin derivative into a non-protein heme complex, which was found to be thermally stable in acetone/water solution     

Identification of 2-dodecylcyclobutanone and linear-alkanes as irradiation markers in sliced dry-cured ham

The usefulness of 2-dodecylcyclobutanone (2-DCB) and alkane hydrocarbons as irradiation markers in sliced dry-cured ham was evaluated by solid phase microextraction (SPME)-gas chromatography-mass spectrometry (GC–MS). To that aim, sliced dry-cured ham was irradiated with different irradiation doses and subsequently examined together with non-irradiated dry-cured ham used as a control sample. The SPME conditions utilized were selected as a consequence of performing an optimization process. The results found in this work reflected that the presence of 2-DCB may be used as an irradiation indicator in sliced dry-cured ham since it does not occur in non-irradiated samples. This confirms data reported in the literature on irradiated foods, other than ham. Likewise, the concentrations of alkanes C₁₄–C₁₇ increased considerably in samples irradiated with low doses, although a decrease was observed when high irradiation doses were used. As a consequence, the concentration of these compounds can also be used as an irradiation indicator when doses lower than 1.0 kGy are applied. These results are discussed.     

Changes in Zn-porphyrin and proteinous pigments in italian dry-cured ham during processing and maturation

Substitution of iron with zinc in myoglobin during maturation of Parma ham to yield zinc porphyrin extractable by 75% vol/vol acetone/water solution and detectable by fluorescence spectroscopy, was found to occur concomitant with protein modification in myoglobin. The content of zinc porphyrin increases throughout the whole processing and maturation of Parma ham, from I(fl) 0.1±0.06 for green ham to I(fl) 84.4±48.8 for fully matured Parma ham. In an aqueous extract of Parma ham with pH 6.0 protein alteration in myoglobin, as detected by size-exclusion chromatography, is initiated during the resting period following salting and seems to precede formation of zinc porphyrin. During maturation the results indicate that the modified myoglobin could undergo polymerization, and it is suggested that initial protein denaturation or degradation facilitates substitution of iron with zinc. The pigment polymerization may be a result of non-covalent protein association to zinc porphyrin in denatured or partly degraded myoglobin.     

Texture changes in dry-cured ham pieces by mild thermal treatments at the end of the drying process

The present study evaluated the effects of mild thermal treatments at the end of the drying process on physicochemical characteristics and instrumental and sensory texture in dry-cured ham. Experiment 1: effect of thermal treatments (4–46 °C) for 4 h and 24 h. Experiment 2: time effect (4–168 h) of thermal treatments at 30 °C and 36 °C. Both experiments were done on small dry-cured ham dices. Experiment 3: time effect (4–168 h) of thermal treatment at 30 °C on both instrumental and sensory texture of 4-cm-thick sections of dry-cured ham. The thermal treatment at 30 °C for 168 h on both dry-cured ham muscle dices (20 mm × 20 mm × 15 mm) and dry-cured ham sections (4 cm thick) decreased softness, adhesiveness and pastiness in BF muscle, without increasing hardness in SM muscle or affecting moisture, a_w and proteolysis index.     

Use of Magnetic Resonance Imaging for monitoring Parma dry-cured ham processing

Protocols were developed to apply Magnetic Resonance Imaging (MRI) to the dry-curing of Italian Parma ham. NMR relaxation analyses were performed on dry-cured hams at different processing stages to evaluate the ranges of variation of ¹H relaxation times T₁ and T₂ in representative ham muscle tissues, due to dehydration and salt uptake. MRI maps of the same ham sections were acquired, allowing T₁ and T₂ average values to be computed in selected Regions of Interest (ROI) inside muscle Semimembranosus, Semitendinosus and Biceps femoris. Chloride and moisture were determined by conventional chemical methods on the same ROIs, and MRI T₁ and T₁/T₂ ratio were selected in a model (R² = 0.90, P < 0.05) fitting the salt content of the analysed muscle cores. Short Time Inversion Recovery (STIR) sequences were also applied to green and cured hams, but on fresh samples only, a bright image, displaying a clear separation between lean and fat tissue, was obtained.     

Use of computed tomography to study raw ham properties and predict salt content and distribution during dry-cured ham production

Varying salt content in hams of equal brand is a major challenge for Norwegian dry-cured ham producers. This study was thus undertaken to test existing computed tomography (CT) calibration models for salt on entire hams, regarding predictability of salt content at different processing times including final ham and to study salt distribution during processing of dry-cured ham. Twenty-six hams were scanned by computed tomography (CT) 11 times during dry-curing for this purpose. However, previously established calibration models had to be adjusted as they overestimated salt in dry samples. Prediction of ultimate salt content was more accurate approaching the end of the dry-curing process (RMSEP = 0.351-0.595% salt). Inclusion of remaining weight loss improved the prediction accuracy in un-dried samples by approximately 0.1% NaCl. The prediction errors were sufficiently low to be of practical interest.     

Effects of high pressure application (400 and 900 MPa) and refrigerated storage time on the oxidative stability of sliced skin vacuum packed dry-cured ham

The effect of high pressure processing at 400 MPa and 900 MPa on the oxidative stability of sliced and vacuum packaged commercial dry-cured ham was determined by analyzing the antioxidant enzyme activities, TBARS levels (thiobarbituric acid reactive substances), vitamin E content and physicochemical characteristics during refrigerated storage for 50 days in different light conditions. In dry-cured ham pressurized at 400 MPa color changes and sensory analyses were also assessed. The high pressure process at 900 MPa produced a decrease in superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) activities and increased vitamin E content. In contrast, pressurization at 400 MPa, increased SOD activity, and showed no effect on vitamin E content and GSHPx activity. In general the physicochemical parameters determined (fat, moisture and collagen) were unaffected by pressurization. Treatment at 400 MPa increased the instrumental color measurement of lightness (L* values, CIELAB). This level of pressure also modified the hardness, chewiness, saltiness and color intensity. These changes of the sensory attributes in dry-cured ham were significant, but small.     

Beliefs and attitudes of butchers and consumers towards dry-cured ham

The objectives were to evaluate the attitude of butcher and consumer towards soft and pasty texture problems in dry-cured ham and to assess the butcher and consumer behaviour when selecting and purchasing this product. Ninety-nine butchers and 200 consumers were interviewed with 17-question and 20-question surveys, respectively. The texture problems (softness and pastiness) were highly important for both butchers and consumers. The characteristics most frequently considered “important” or “very important” by butchers were processing time, smell/aroma and texture as selection criteria and salty taste and aged/matured flavour as sensory indicators of dry-cured ham quality. According to butchers’ opinion, salty taste and their own advice are the factors most frequently considered “important” or “very important” as affecting the consumer’s decision to purchase. For the consumer, the sensory attributes were considered more important than extrinsic characteristics when purchasing dry-cured ham.     

Influence of high pressure application on the nutritional, sensory and microbiological characteristics of sliced skin vacuum packed dry-cured ham. Effects along the storage period

The effect of high pressure processing at 600 MPa on sliced and vacuum packaged commercial dry-cured ham was determined by analyzing the induced color changes, the physicochemical characteristics, the microbiological count and the changes in sensory attributes and also its effect on lipid oxidation measured through TBARS (thiobarbituric reactive substances), antioxidant enzyme activities and the content of fatty acid from total, free and phospholipid fractions. This effect was also studied during 50 days refrigerated storage with different light conditions. The high pressure processing at 600 MPa modified the color of dry-cured ham producing an increase in lightness L*-parameter. Many sensory attributes were also modified resulting in an increase in hardness, chewiness, brightness, odor intensity and saltiness, while reducing color intensity. High pressure produced a reduction in the aerobic count. The oxidative stability of the pressurized dry-cured ham was not altered as observed from the absence of differences in fatty acid contents and antioxidant enzyme activities. Initially pressurization produced a decrease in TBARS levels, however this effect was not found after 50 days of refrigerated storage.

Industrial relevance

The application of high hydrostatic pressure (HHP) in food technology is a useful tool to combine a minimal processing that increases the shelf-life of the food products, maintaining their safety and nutritional properties and with minimal changes in flavor or taste. This processing technology is very important to increase the safety of the sliced foods which are prone to the microbiological deterioration, as cured meat products and cooked ham. Pressurization of sliced dry-cured ham at 600 MPa has been shown effective reducing spoilage associated microorganisms. The present study was carried out to evaluate the oxidative stability, nutritional quality, safety and sensory attributes of dry-cured ham pressurized at 600 MPa. This is very important because the hypothesis that HHP does not affect to sensorial characteristics needs to be confirmed in different kind of foods. Also, the possible causes of the observed effects should be explained by the study of physic–chemical parameters. In this way the oxidative stability is an important issue because some authors claimed that HHP can increase oxidation in food containing lipids.     

Evolution of nitrate and nitrite during the processing of dry-cured ham with partial replacement of NaCl by other chloride salts

Nitrate and nitrite are commonly added to dry-cured ham to provide protection against pathogen microorganisms, especially Clostridium botulinum. Both nitrate and nitrite were monitored with ion chromatography in dry-cured hams salted with different NaCl formulations (NaCl partially replaced by KCl and/or CaCl(2), and MgCl(2)). Nitrate, that is more stable than nitrite, diffuses into the ham and acts as a reservoir for nitrite generation. A correct nitrate and nitrite penetration was detected from the surface to the inner zones of the hams throughout its processing, independently of the salt formulation. Nitrate and nitrite achieved similar concentrations, around 37 and 2.2 ppm, respectively in the inner zones of the ham for the three assayed salt formulations at the end of the process, which are in compliance with European regulations.     

Creatine and creatinine evolution during the processing of dry-cured ham

Dry-curing of ham involves many biochemical reactions that depend on the processing conditions. The aim of this study was to evaluate the effect of the dry-cured processing on the concentration of creatine, creatinine and the creatinine/creatine ratio. Dry-cured hams under study were salted using three different salt mixtures (100% NaCl; NaCl and KCl at 50% each; and 55% NaCl, 25% KCl, 15% CaCl₂ and 5% MgCl₂) in order to observe its influence on creatinine formation but no significant differences were found between them at any time of processing. However, significant differences between different post-salting times (20, 50 and 80 days) and the ripened hams (7, 9 and 11 months of ripening) were observed. Results showed that creatine and creatinine remain stable once the ripening period is reached. These results were confirmed when analysing dry-cured ham samples submitted to extreme conditions of temperature and time (20, 30, 40 and 70 °C during 0, 20, 40 and 60 min) as well as commercial dry-cured hams with more than 12 months of processing.     

High pressure treatment of dry-cured Iberian ham. Effect on radical formation,lipid oxidation and colour

The effect of high-pressure treatment (200, 400, 600 and 800 MPa) on radical formation in solid samples of dry-cured Iberian ham and in ham slurries was evaluated using the spin-trapping technique and electron spin resonance (ESR) spectroscopy. In addition, hexanal was, as a secondary lipid oxidation product, assessed by gas chromatography. The effect of pressure treatments on ESR signals determined in the spin-trapping assay following pressure treatment in solid samples of dry-cured Iberian ham was not statistically significant. However, the lowest level of pressure applied initiated radical formation and promoted lipid peroxidation, whereas intermediate to high levels of pressure seemed to promote further reaction and disappearance of free radicals. Pressure significantly affected hexanal content in the same way as seen for formation of free radicals. Regarding the slurries prepared from dry-cured ham, pressure significantly increased the tendency of radical formation as seen from the ESR signals and significantly increased the hexanal content. The different pattern between solid ham and ham slurries points toward an initiation mechanism associated with the membrane phospholipids for oxidation. In addition, surface colour (L*, a* and b* values and percent reflectance values) of non-pressurized and pressurized (200–800 MPa) dry-cured Iberian ham samples was assessed. Non-pressurized samples showed a higher lightness than pressurized samples, and redness significantly decreased with pressure treatment.     

High pressure treatments on the inactivation of Salmonella Enteritidis and the physicochemical, rheological and color characteristics of sliced vacuum-packaged dry-cured ham

The effect of high pressure (HP) on Salmonella Enteritidis in sliced dry-cured ham stored under temperature abuse (8°C) during 60d was investigated. After treatment, reductions of S. Enteritidis were 1.06, 2.54 and 4.32 log units in ham treated at 400, 500 and 600MPa for 5min at 12°C, compared to non-pressurized samples. After 60d, counts of S. Enteritidis in ham treated at 400 and 500MPa were 2.56 and 2.66 log units lower than in non-treated ham, whereas the pathogen was only detected after enrichment in ham treated at 600MPa. Lipid oxidation increased with storage and pressurization, whereas total free amino acid contents were similar in HP and control samples after 60d. Dry-cured ham treated at the highest pressures exhibited lower shear resistance, whereas the maximum force to compress the sample was slightly changed. Color (L*, a* and b*) varied with pressurization and storage. Changes induced by HP in dry-cured ham were attenuated during storage.     

Micronutrients in Italian ham: A survey of traditional products

The study provides original analytical data on the micronutrient profile of some traditional Italian hams, representative of the major ham categories produced in Italy: 4 dry-cured hams (Modena, Nazionale, Parma, San Daniele), 3 cooked hams (Cotto, Scelto, Alta Qualità), 1 smoked ham (Speck). Data on macronutrients (protein, lipid, moisture), energy, trace elements (Fe, Zn, Cu, Mn, Se), B vitamins (B1, B2, PP, B6, B12) and vitamin E level in the 80 Italian hams sampled are reported. Smoked and dry-cured ham were the richest sources of Fe, Zn and Se and, among vitamins, dry-cured ham had the highest level of B2, PP, B6 and B12; cooked ham provided the lowest energy intake. The contribution of ham to micronutrients recommended dietary allowances was estimated: a ham portion (50 g) was a good source especially of Zn and Se providing over 12% of RDA of both; among B vitamins, dry-cured ham greatly contributed to B1 and B6 vitamins RDA (both over 30%).     

Factors affecting dry-cured ham consumer acceptability

The objectives of the present study were (1) to compare the relative importance of price, processing time, texture and intramuscular fat in purchase intention of dry-cured ham through conjoint analysis, (2) to evaluate the effect of dry-cured ham appearance on consumer expectations, and (3) to describe the consumer sensory preferences of dry-cured ham using external preference mapping. Texture and processing time influenced the consumer preferences in conjoint analysis. Red colour intensity, colour uniformity, external fat and white film presence/absence influenced consumer expectations. The consumer disliked hams with bitter and metallic flavour and with excessive saltiness and piquantness. Differences between expected and experienced acceptability were found, which indicates that the visual preference of consumers does not allow them to select a dry-cured ham that satisfies their sensory preferences of flavour and texture.     

A note on the relationships between cured-cooked and dry-cured ham processing yields

Right and left hams from 353 pigs slaughtered at around 100 kg body weight were processed into cured-cooked hams and dry-cured hams, respectively. Weights and yields at various stages of each process, carcass lean content and fresh meat quality traits were registered. Technological yield of cured-cooked processing (saleable cooked ham weight/defatted–deboned fresh ham weight) was more closely correlated to ultimate pH (r = 0.51, p < 0.001) than to carcass leanness (r = −0.13, p < 0.05) whereas the reverse situation – r = 0.15 (p < 0.01) and r = −0.62 (p < 0.001), respectively – was found for technological yield of dry-cured processing (saleable dry ham weight/trimmed fresh ham weight). The correlation between the two technological yields was significantly positive but of fairly moderate magnitude (r = 0.36). The correlation between the overall yields (saleable processed ham weight/entire fresh ham weight) of the two processes revealed to be very close to zero (r = −0.01).     

The effect of NaCl-free processing and high pressure on the fate of Listeria monocytogenes and Salmonella on sliced smoked dry-cured ham

NaCl is an important multifunctional ingredient applied in dry-cured ham elaboration. However, its excessive intake has been linked to serious cardiovascular diseases causing a recent increase in the development of reduced salt products. In the present study Listeria monocytogenes and Salmonella, food-borne pathogens which can cross-contaminate post processed products, were spiked with < 100 CFU/g on slices of both standard (S) and NaCl-free processed (F) (elaborated with KCl + potassium lactate instead of NaCl) smoked dry-cured ham. Although L. monocytogenes and Salmonella counts decreased faster in S ham, pathogens were present in both types of non-pressure treated ham during the entire refrigerated storage period (112 days). Pressurisation at 600 MPa for 5 min caused the elimination of both pathogens in S ham after 14 days. In contrast, Salmonella and L. monocytogenes were present in F ham until days 28 and 56, respectively, indicating that the NaCl-free processed dry-cured ham had lower stability than standard smoked dry-cured ham.