Possible biological markers of the time of processing of dry-cured ham

Several muscle compounds (creatine, creatinine, hypoxanthine, inosine, inosine 5' monophosphate, xanthine, adenosine monophosphate, guanosine, and uridine) were studied as possible biological markers of a minimum dry-cured ham processing time. A correlation between the concentration of the compounds and the time of processing was found. The ratios for some of them were calculated to study their behaviour during processing. The Hx/Ino ratio substantially increased up to 5 months of curing and then remained constant (p<0.05). The Hx/Ino ratio might be considered as a potential biomarker of the minimum time of dry-cured processing (5 months). The Cn/Cr ratio increased during drying up to 9 months of ripening (p<0.05). However, although Cn/Cr ratios remained constant after 9 months of processing, variations between hams were observed due to the differences existing in the raw meats and small differences in processing conditions, making it difficult to consider Cn/Cr ratios as biomarkers of ripening time.     

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.     

Lipolysis in dry-cured ham: Influence of salt content and processing conditions

Fatty acid composition of neutral lipids (NLs), polar lipids (PLs) and free fatty acids (FFA) from the intramuscular fat of Semimembranosus and Biceps femoris muscles was analysed in 46 Iberian dry-cured hams processed with different amounts of salt (6% high salt batch – HS vs. 3% low salt batch – LS w/w) and different processing systems (traditional – T vs. modified – M).

Total amounts of saturated (SFA), monounsaturated (MUFA) and polyunsaturated (PUFA) fatty acids in NLs decreased in similar proportions during processing of the hams as well as SFA, MUFA and PUFA in the PL fraction, whereas the amounts of SFA, MUFA and PUFA of FFAs significantly increased in Semimembranosus and Biceps femoris muscles. The amount of total fatty acids (TFA), from NLs and PLs, decreased in both muscles throughout the processing. Such a decline was more intense in HS hams than in LS ones, which could be a sign of a promoting effect of sodium chloride on lipolysis. However, the increase in FFA content throughout processing was not more intense in HS hams. Processing conditions studied in this work did not affect the changes in the fatty acid content of each fraction.     

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.     

Changes of free amino acids and biogenic amines during extended ageing of Italian dry-cured ham

Sixty-two fresh hams were sub-grouped to undergo different processing times (15, 19 and 23 months), and corresponding dry-cured hams were analysed for changes in moisture, protein, NaCl, pH, proteolysis, free amino acids (FAAs) and biogenic amines (BAs) as related to the extended ageing. Dry-cured hams were influenced by ageing time, showing a decrease in moisture and water activity and an increase in pH, nonprotein nitrogen (NPN) and FAAs in more aged samples. The increase in FAAs and BAs progressively observed until the last sampling time might be enhanced by the moderate salt content (≈5 g NaCl/100 g muscle) and relatively high a_w (>0.90) of dried hams even at 23 months of processing. Among FAAs, arginine did not increase with ageing, which might be due to arginine hydrolysis to ammonia and ornithine, followed by decarboxylation to putrescine, i.e. molecules largely present in the more aged hams. Tyramine, the most abundant among BAs, putrescine and cadaverine showed a dependence on time and proteolysis indices (NPN and FAAs). In this respect, the practice of extending the standard ageing time of typical italian dry-cured ham (13-15 months), regarded as a tool for improving sensory property of this product, should be supported by further studies, mainly at the manufacturing level, to minimize FAA and BA generation.     

Thermal and photochemical degradation of myoglobin pigments in relation to colour stability of sliced dry-cured Parma ham and sliced dry-cured ham produced with nitrite salt

Lipophilic and hydrophilic extracts of the red pigments from Parma ham and nitrosylated pigment of dry-cured ham produced with nitrite salt were prepared with acetone/water (75/25 v/v %) solution and aqueous phosphate buffer, respectively. The spectral characteristics differed for both the lipophilic and the hydrophilic Parma ham pigment compared with the dry-cured ham produced with nitrite salt. The red lipophilic pigment(s) extractable from Parma ham was(were) found to be very stable towards thermal degradation in acetone/water (75/25 v/v %) solution for temperatures up to 70 °C in contrast to the lipophilic pigment(s) extractable from dry-cured ham produced with nitrite salt, which was(were) found to have an energy of activation of 99 kJ/mol for thermal degradation. In contrast, quantum yields for photodegradation of the lipophilic ham pigments exposed to 366 nm (420 nm) monochromatic light were larger for Parma ham than for nitrite-cured ham [1.6×10^–5 (6.9×10^–6) versus 1.6×10^–6 (2×10^–6) mol einstein^–1] as determined for acetone/water (75/25 v/v %) solution. In agreement with these findings for the extracted lipophilic pigments, sliced Parma ham showed better colour stability than sliced dry-cured ham produced with nitrite salt, when stored in the dark at low oxygen concentration, in contrast to a faster initial discolouration for Parma ham when exposed to light, as shown for chilled storage for 35 days under retail conditions for the two products each packed at two oxygen levels (0.4 and 21%).     

Hypoxanthine-based enzymatic sensor for determination of pork meat freshness

An enzyme sensor employing xanthine oxidase (XO), soluble or immobilised, in combination of an oxygen electrode has been developed and optimised to determinate the hypoxanthine (Hx) content in pork meat at different post-mortem times as measure of meat freshness. The amperometric signal obtained due to the oxygen depletion during the Hx oxidation was related with the consumed oxygen at 190 s in the soluble enzyme sensor or enzymatic rate at 10 s in the immobilised enzyme sensor. In both cases a linear relationship between the signal and the Hx concentration in the range 8.68–26.05 μM (R² = 0.999) and 15.63–127 μM. (R² = 0.995), respectively, was found. Both enzyme sensors exhibited very good working conditions and storage stability. A study of Hx oxidation was carried out in order to compare the Hx content measured by both sensors and those measured by high performance liquid chromatography (HPLC) obtaining a good agreement between both techniques. Therefore, the easy preparation and operation of both enzyme sensors suggests a reliable, rapid and an economical alternative for simple or multiple Hx measurements constituting a useful tool as quality control of meat freshness.     

Evaluation and selection of yeasts isolated from dry-cured Iberian ham by their volatile compound production

One hundred and seventeen yeast strains isolated from dry-cured Iberian ham from the four different protected designations of origin of Spain were investigated for their volatile compound production. The yeast strains were grouped into the two main yeast species usually found in this product (Debaryomyces hansenii and Candida zeylanoides) and 10 different biotypes by restriction mitochondrial DNA analysis. Yeast strains were grown in a designed model culture medium under conditions representative of dry-cured ham processing. Volatile compounds were extracted from this medium using solid-phase micro-extraction and were analysed by gas chromatography/mass spectrometry. Marked differences in volatile compound production were found between D. hansenii and C. zeylanoides and between the mitochondrial DNA patterns of these species. Two of the mitochondrial DNA patterns of D. hansenii exhibited the highest production of the volatile compounds involved in the dry-cured flavour. Consequently, these patterns of D. hansenii should be proposed as starter cultures for dry-cured ham.     

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.     

Chemical and structural changes in lipids during the ripening of Teruel dry-cured ham

The aim of this paper is to investigate the chemical and microstructural changes in intramuscular and subcutaneous fat during the processing of Teruel dry-cured ham by gas chromatography and electron microscopy techniques. This paper will contribute to the specific characterisation of a product included in the European Union list of special quality products, and provides a new perspective for the identification of changes related to flavour development. There seems to be a relationship between the degradation of phospholipids and the increase in the free fatty acid content, especially polyunsaturated fatty acids. The microstructural changes of the adipose tissue during the process would explain the availability of the fat to the lipolitic enzymes and the contribution to the typical flavour and taste of cured ham.     

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.     

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.     

Occurrence of ochratoxin A in raw ham muscle,salami and dry-cured ham from pigs fed with contaminated diet

Pork meat-derived products can contribute to the overall ochratoxin A intake, either by carry-over effect, or by environmental mould population cross-contamination. In order to assess the role of these different contamination routes, a study was carried out with pigs challenged orally with OTA contaminated feed at subchronical level. After slaughtering, thighs and minced meat from control and treated groups were transformed into dry-cured hams and salami, respectively, which were analysed for OTA determination after ripening. From collected data, the carry-over in muscle was generally low, whereas a significant contribution to the OTA contamination in dry-cured hams was due to toxinogenic mould population growing on their surface during ripening. Finally, a survey of different types of dry-cured ham (n = 110), from the Italian market, was performed, showing the occurrence of OTA on the surface portion in 84 out of 110 samples with a median value of 0.53 μg/kg and in the inner core in 32 out of 110 samples with a median value lower than 0.1 μg/kg.     

Differentiation of yeasts growing on dry-cured Iberian ham by mitochondrial DNA restriction analysis, RAPD-PCR and their volatile compounds production

The efficiency of mitochondrial DNA (mtDNA) restriction analysis, RAPD-PCR and volatile compounds analysis to differentiate yeast biotypes involved in flavour development of dry-cured Iberian ham throughout the ripening process is evaluated. For this purpose, 86 yeasts isolated from Iberian hams in the main ripening stages at different industries of the four Protected Designations of Origin of this product, were used. The combination of mtDNA restriction analysis and RAPD-PCR using the primer (GACA)₄ showed a higher variability in the yeast species detected than obtained using only mtDNA restriction analysis. Only two species, Debaryomyces hansenii and Candida zeylanoides, were identified throughout the whole ripening process and a wide diversity of biotypes was found in these two species, with those of D. hansenii predominating. Clear differences between biotypes were detected in the generation of volatile compounds, with the biotype C2-2 of D. hansenii showing the highest concentrations of volatiles. The combined use of mtDNA restriction analysis and RAPD-PCR distinguishes yeast biotypes with different production of volatile compounds. In addition, analysis of the production profile of volatile compounds is needed to differentiate yeast strains of the same biotype recovered at different stages of ripening. Thus, the combination of these three methods could be very useful to select or monitor yeasts as starter cultures in dry-cured meat products.     

Accuracy of near infrared spectroscopy for prediction of chemical composition, salt content and free amino acids in dry-cured ham

The capability of near infrared (NIR) spectroscopy was examined for the purposes of quality control of the traditional Slovenian dry-cured ham “Kraški pršut.” Predictive models were developed for moisture, salt, protein, non-protein nitrogen, intramuscular fat and free amino acids in biceps femoris muscle (n = 135). The models' quality was assessed using statistical parameters: coefficient of determination (R²) and standard error (se) of cross-validation (CV) and external validation (EV). Residual predictive deviation (RPD) was also assessed. Best results were obtained for salt content and salt percentage in moisture/dry matter (R_CV² > 0.90, RPD > 3.0), it was satisfactory for moisture, non-protein nitrogen, intramuscular fat and total free amino acids (R_CV² = 0.75–0.90, RPD = 2.0–3.0), while not so for protein content and proteolysis index (R_CV² = 0.65–0.75, RPD < 2.0). Calibrations for individual free amino acids yielded R_CV² from 0.40 to 0.90 and RPD from 1.3 to 2.9. Additional external validation of models on independent samples yielded comparable results. Based on the results, NIR spectroscopy can replace chemical methods in quality control of dry-cured ham.     

Influence of partial replacement of NaCl with KCl, CaCl2 and MgCl2 on lipolysis and lipid oxidation in dry-cured ham

Sodium intake above nutritional recommendations may involve harmful consequences to health such as the increased risk of cardiovascular diseases. Dry-cured ham constitutes a product with a relatively large amount of sodium. Thus, to obtain a healthier product for consumers with reduced sodium content, two formulations containing KCl alone (formulation II) or mixed with CaCl2 and MgCl2 (formulation III) have been proposed to partially replace NaCl. Lipolysis and lipid oxidation occurring in hams processed with these formulations have been studied since they have direct influence on the final flavor. No significant differences in acid lipase activity or lipid oxidation were found at the end of the process between the alternative formulations and formulation I (control with 100% NaCl). Differences in some free fatty acids, generated along the processing, were detected among treatments and at the end of dry-curing. Data suggests a slight trend towards a major lipolysis during treatment III.     

Consumers’ acceptance of innovations in dry-cured ham: Impact of reduced salt content, prolonged aging time and new origin

The objective of this study was to evaluate effects of information about reduced salt content, prolonged aging time and new origin on the acceptance of dry-cured ham. The study was carried out in Norway and origins of tested hams were Norway and Spain. Consumers’ acceptance of hams was investigated in blind, expected and informed conditions. Results showed that ratings in the informed condition changed in the direction of the expectations and significant assimilation effects occurred for two products. Two consumer clusters were identified. Consumers in the first cluster were more open to trying new kinds of food and this attitude was exemplified by a relatively high expected score for dry-cured ham with reduced salt level, long aging and Spanish origin. Consumers in the second cluster were more sceptical to new food and new dishes. This was reflected in a relatively high expected score for the traditional Norwegian ham with high salt level and short aging time.     

Inactivation of Serratia liquefaciens on dry-cured ham by high pressure processing

To quantify the inactivation of Serratia liquefaciens exerted by high pressure processing (HPP), slices of dry-cured ham were inoculated and processed combining different levels of technological parameters: pressure (347–852 MPa), time (2.3–15.8 min) and temperature (7.6–24.4 °C) according to a central composite design. Bacterial inactivation, as logarithmic reduction, indicated that S. liquefaciens was relatively sensitive to HPP. Six log reductions were achieved in a total of 10 trials combining pressures of 600 MPa or above with different holding times and temperatures. The inactivation of S. liquefaciens was analysed through the multiple regression analysis to generate a second order polynomial equation. Pressure and time were the two factors which significantly determined the inactivation of S. liquefaciens on dry-cured ham. Temperature did not significantly affect the lethality of the process. The response surface methodology was used to determine optimum process conditions to maximize the inactivation of S. liquefaciens in the experimental range tested. The maximum inactivation of S. liquefaciens in dry-cured ham was achieved by combining a pressure of 650 MPa with a holding time of 8 min. Combinations above these values (i.e. 750 MPa for 13 min) would not significantly improve the lethality of the process.     

Dynamics and characterization of yeasts during ripening of typical Italian dry-cured ham

The evolution of the yeast population during manufacturing and ripening of dry-cured Parma ham was investigated. Contamination levels ranged from 10(5) to 10(7) cfu/g on muscle surface, 10(4) to 10(6) cfu/g on covering fat and exceeded 10(7) cfu/g on spreadable fat mince ("sugna"). Two hundred and sixty one yeast isolates underwent identification test, showing that the predominant species of yeast population during the whole maturing process were Debaryomyces hansenii, Candida zeylanoides, Debaryomyces maramus, and to a lesser extent, Candida famata and Hyphopichia burtonii. The species Candida catenulata, Candida guilliermondii, Candida edax and other genera like Cryptococcus and Wingea were occasionally found. The yeast counts and species distribution changed according to the stage of processing and to the ham sampling location. At the end of the cold phase, the washing procedure was effective in lowering the yeast count in muscle and fat surface layers, but during the next ageing stages, yeast colonization of unskinned ham muscle increased again, though species distribution changed if compared to previous manufacturing phases. The ripening steps taken into account from the end of the cold phase to the final outcome, were always characterized by more than one yeast species, suggesting that yeasts other than Debaryomyces spp. could play a remarkable role on the sensory and safety properties of typical Italian dry-cured ham.