Biochemical changes during processing of traditional Jinhua ham

Jinhua ham is the most famous traditional meat product of China and one of the most famed dry-cured hams in the world. Its processing consists of six stages: green ham preparation, salting, washing and sun-drying and shaping, ripening, and post-ripening. Intense proteolysis and lipolysis occur during processing period. As a result, the content of free amino acids in final ham products is 14-16 times that of green ham, and 191 volatile compounds have been identified during processing, which make a major contribution to the flavor of Jinhua ham.     

基于智能感官与人工感官评价的中国三大干腌火腿风味特性分析

以我国三大干腌火腿金华火腿、宣威火腿和如皋火腿三个不同年份或等级的火腿为研究对象,分别采用人工感官评价和电子鼻、电子舌智能感官技术分别对香气与滋味进行研究。人工感官评价结果显示,肉香味、腌制味和油脂香是三大火腿的特征香气,如皋和金华火腿中2年/优级和3年陈/特级的干腌火腿较于1级/1年陈具有更加浓郁的肉香,而1年陈/1级火腿的酸味更为明显。电子鼻和电子舌数据的主成分分析(Principal component analysis,PCA)结果表明,不同干腌火腿样品均能实现良好区分。1年陈/1级与2年/优级、3年陈/特级干腌火腿气味轮廓相差较远,金华火腿与宣威2、3年陈的火腿香气和滋味轮廓均比较接近。采用软独立建模分类法(Soft independent modeling class analogy,SIMCA)构建了干腌火腿的判别模型,以三个产地最优干腌火腿为标准样品的等级鉴别模型能够实现对其他年份干腌火腿的有效判别。说明不同年份的火腿风味变化呈现一定规律,可利用智能感官技术对火腿进行快速有效的等级鉴别。     

Reduction of ochratoxin A in dry-cured meat products using gamma-irradiation

This study investigated the efficiency of gamma (γ)-irradiation in the reduction of ochratoxin A (OTA) present in dry-cured meat products prepared from intentionally contaminated raw materials from OTA-treated pigs. OTA concentrations determined in the samples (n = 24) ranged from 25.8 μg kg^–1 in bacon to 17.8 μg kg^–1 in smoked ham. After γ-irradiation at doses of 3, 7 and 10 kGy (i.e. the doses used in the food industry), a dose-depended OTA reduction was observed; however, it was not statistically significant. The mean OTA reduction achieved with 3-, 7- and 10-kGy γ-doses was approximated to 8.5%, 13.9% and 22.5%, respectively. The storage of irradiated samples (1 month, 4°C) did not significantly affect OTA levels. Based on the correlation between the OTA reduction level and basic chemical composition of dry-cured meat samples, OTA reduction may be linked to the samples’ fat content. The results indicate that γ-irradiation can reduce OTA levels in dry-cured meat products, but only to a limited extent due to the complexity of the matrix.     

Proteomic analysis of water soluble and myofibrillar protein changes occurring in dry-cured hams

The myofibrillar fraction of raw ham muscles and dry-cured hams with different ripening times was extracted in denaturing and reducing conditions and subjected to two-dimensional gel electrophoresis. The two-dimensional maps gave overall pictures of the already noted progressive disappearance of actin, tropomyosin and myosin light chains during ripening. In addition, two fragments from Myosin Heavy Chain proteolysis, marked as myosin chain fragments MCF1 and MCF2, were identified by immunodetection and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Furthermore, a new form of actin on two-dimensional gel was identified by MALDI-TOF peptide mapping. In 12-month-old dry-cured ham, most myofibrillar proteins were completely hydrolyzed. At this stage of ripening, in fact, in some Parma and S. Daniele dry-cured ham samples, myosin heavy chain fragments and other unidentified neo-formed spots were found. Some of the sarcoplasmic proteins in water extracts from pork meat markedly decreased in amount or disappeared totally, during ripening. Surprisingly, two-dimensional gel electrophoresis maps of the water soluble protein fraction from dry-cured ham showed the presence of two spots identified as tropomyosin α- and β-chain. This result suggests that some of the saline soluble myofibrillar proteins can disappear from this fraction because of salt solubilization and not due to complete enzyme action. Two-dimensional gel electrophoresis (2-DGE) has proved a powerful tool to evaluate the enzymatic susceptibility of meat proteins and the evolution of protein map fragmentation throughout ripening process as well as a means of obtaining a standard fingerprinting map characterizing the final product.     

无硝干腌火腿中红色素形成机制研究进展

干腌火腿经过长时间腌制发酵成熟,富含生物活性物质,具有良好的感官特性。在传统干腌火腿加工过程中常添加硝盐（硝酸盐或亚硝酸盐）以获得稳定的色泽,然而,由亚硝酸盐与仲胺反应生成的亚硝胺具有致癌风险,已引起人们的广泛关注。研究发现,国外的著名干腌火腿如帕尔玛火腿,在腌制过程中不添加硝盐,其红色的主要贡献者被证实为Zn-原卟啉IX。本文综述了添加硝盐与未添加硝盐干腌火腿中红色素的研究进展,重点探讨Zn-原卟啉IX在无硝干腌火腿加工过程中的形成机制,以期为干腌火腿加工过程的色泽调控提供科学理论依据。     

Zn-porphyrin formation in cured meat products: Effect of added salt and nitrite

Zn-porphyrin (Zn-pp) was quantified by fluorescence spectroscopy in the cured and dry cured meat products: Parma ham, Iberian ham, dry-cured ham with added nitrite, cooked ham with added nitrite, raw ham meat, raw bacon and Karree-Speck. The highest amount of Zn-pp was found in dry-cured Parma ham and Iberian ham, while the use of nitrite as curing agent was found to inhibit completely the formation of Zn-pp in meat products. A positive correlation between both Zn content and Fe content and the logarithmic transformed Zn-pp content (measured as fluorescence intensity I_fl) was found for the different cured and dry cured meat products, with correlation coefficients of 0.79 (p < 0.001) and 0.71 (p < 0.01), respectively. Log I_fl correlates best with the Zn content, indicating that the formation of Zn-pp is proportional to the Zn content. A model system with vacuum packed pork in brine with different added levels of sodium chloride with or without nitrite and Zn acetate was investigated in order to further elucidate the mechanism of Zn-pp formation. Zn-pp increased with time (up to 42 days investigated) in non-cured meat and for meat cured solely with NaCl lower than 9%. Addition of nitrite or Zn(II) in the curing brine was found to inhibit formation of Zn-pp confirming the observations from the various cured meat products. It is suggested that a chloride anion assisted dissociation of iron from myoglobin could be rate-determining for Zn-pp formation in meat products.     

Effect of Penicillium chrysogenum and Debaryomyces hansenii on the volatile compounds during controlled ripening of pork loins

During ripening of meat products such as dry-cured ham, the moulds and yeasts that proliferate on the surface may contribute to flavour development. However, their contribution to volatile components of dry-cured meat products is not known. One strain each of Penicillium chrysogenum and Debaryomyces hansenii, selected from dry-cured ham by their proteolytic activity, were tested to determine their effect on the volatile compounds during ripening. Sterile pork loins were inoculated and ripened for 106 days. Volatile compounds collected with a Solid Phase Micro-Extraction (SPME) fibre were analysed by GC/MS. Inoculation of pork loins with P. chrysogenum lead to a decrease in compounds attributed to lipid oxidation and to an increase of compounds derived from free amino acids. Inoculation with D. hansenii seemed to favour the formation of complex alcohols.     

金华火腿质量与原料选择的关系研究

中式火腿如金华火腿与西方地中海地区的干腋火腿都属于高档的肉类制品。金华火腿质量取决于两个主要方面：一是独特的加工方法；二是严格的原料选择。本文从选料着手，分析在金华火腿加工过程中原料的质量对火腿成熟的作用，以及最终对产品质量的影响。     

Proteolytic activity of Penicillium chrysogenum and Debaryomyces hansenii during controlled ripening of pork loins

The role of micro-organisms on the ripening process of dry-cured ham, particularly with respect to proteolysis, is not clear. This is partially due to the lack of an adequate system to study changes on a sterile control meat product for long ripening times. Using a meat system based on sterile pork loins ripened under aseptic conditions for 106 days, the contribution to the proteolysis of two micro-organisms isolated from dry-cured ham has been established. Changes were studied by SDS-PAGE of sarcoplasmic and myofibrillar proteins, capillary zone electrophoresis (CZE) of low ionic strength-soluble nitrogen compounds, and HPLC of free amino acids. Debaryomyces hansenii Dh345 did not show any significant proteolytic activity. However, Penicillium chrysogenum Pg222 showed high proteolytic activity on myofibrillar proteins resulting in an increase in soluble nitrogen compounds. For this, P. chrysogenum Pg222 should be considered to be used as starter culture in meat products made using long ripening times.     

云南干腌火腿加工过程中肌肉组织理化特性和氧化稳定性研究

云腿是我国一类重要的传统肉制品。研究通过对加工过程中的云腿肌肉组织进行理化指标测定,初步确定了云腿的理化特性和氧化稳定性。结果显示随着加工工艺的进行,云腿肌肉组织中pH缓慢上升,aw值逐渐下降。股二头肌和半膜肌中的食盐含量分别增加到7.94%和6.75%。水分含量到了平衡期之后快速下降,蛋白质和脂肪含量相对增加。肌内脂肪和蛋白质氧化持续进行,但是总体上处于较低的氧化水平。TBARS值先增加后降低,半膜肌中的蛋白质氧化程度强于股二头肌。     

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.     

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.     

Biogenic amine content during the manufacture of dry-cured lacón, a Spanish traditional meat product: Effect of some additives

The content of nine biogenic amines (agmatine, tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine) was determined throughout the manufacture of dry-cured lacón, a traditional dry-salted and ripened meat product made in the north-west of Spain from the fore leg of the pig following a similar process to that of dry-cured ham. The effect of the use of additives (glucose, sodium nitrite, sodium nitrate, sodium ascorbate and sodium citrate) on the biogenic amine content during manufacture was also studied. Tryptamine and spermine were the main biogenic amines in fresh meat, while tryptamine and cadaverine were the most abundant at the end of the manufacturing process. During ripening the total amine content increased significantly (P<0.05) in the batches made both without and with additives. The use of additives significantly (P<0.05) increased the total amine content and the content of tryptamine, tyramine and histamine. The total biogenic amine content at the end of the manufacturing process was low as expected for a product in which there is little active microbial metabolism during manufacture.     

Lipolytic and proteolytic properties of dry-cured boneless hams ripened in modified atmospheres

We studied proteolytic and lipolytic properties of dry-cured boneless ham (porcine quadriceps femoris) made with chilled (10°C, 48 h) or frozen/thawed meat (frozen at -20°C frozen for 90 days and followed by thawing at 10°C for 48 h) were determined. Dry-cured meats were stored in modified atmosphere packages (100% N(2) and a mixture of 75% N(2)+25% CO(2)) at 15°C with the intention of reducing ripening space. Results showed that dry-cured hams made with frozen/thawed raw meat had more salt, volatile fatty acids and free fatty acid content after salting and smoking. Whereas, samples prepared with chilled meats contained more nitrogenous compounds (water-soluble nitrogen, non-protein nitrogen, and free amino acids). Volatile and free fatty acid contents in all samples significantly increased with storage. Acetic acid was the predominant volatile fatty acid. To confirm lipolytic activity in dry-cured ham stored in modified atmospheres, we calculated the lipolytic coefficient. The lipolytic coefficients of all samples were positive values and significantly (P<0.05) increased with storage indicating lipolysis in samples were still active. Furthermore, nitrogenous compounds in dry-cured ham significantly (P<0.05) increased with storage indicating proteolysis in samples were not affected by modified atmosphere storage. Aerobic, anaerobic and lactic acid bacteria counts in dry-cured meats were stable to modified atmospheres storage for 20 weeks at 15°C. Flavor, texture and color score in sensory evaluation for dry-cured ham made with chilled meat were significantly higher than that made with frozen/thawed meat. All samples had high overall acceptance scores in sensory evaluation. Results in this study suggested that dry-cured boneless ham stored in modified atmospheres for 20 weeks at 15°C was another feasibility to ripen the meat without affecting lipolysis, proteolysis, microbiology and sensory quality.     

Postmortem meat quality and sex affect textural properties and protein breakdown of dry-cured ham

Texture measurements by instrumental texture profile analysis (TPA) and protein degradation analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) were performed on 30 dry-cured hams resulting from four different post-mortem meat qualities categories (PSE, RSE, RFN and DFD). The main differences were observed in dry-cured hams from PSE and RFN meat qualities. Penetration force (80%), hardness, springiness, cohesiveness and chewiness, were significantly lower (P<0.05) in PSE than in RFN quality classes. The rate of the ripening process was affected as a higher proteolysis and absence of fragments at 150 and 85 KDa in PSE in relation to RFN classes, and with an intermediate proteolysis of RSE and DFD classes. The effect of sex was observed as a significant (P<0.05) low hardness in the hams obtained from female pigs. The duration of the ripening, for a better uniformity in dry-cured ham production, should be adapted to the initial pH and to drip loss parameters of the raw material.     

Amino acid and nucleotide contents and sensory traits of dry-cured products from pigs with different genotypes

The free amino acid and nucleotide contents of dry-cured ham, shoulder and loin from two genetic lines selected from pigs according to the paternal allele (homozygous AA and heterozygous AG) of the insulin-like growth factor-II gene were studied by HPLC. Their influence on the flavor and taste characteristics was also studied. The increase of lean content caused by the IGF-II mutation could affect proteolysis during the ripening process and therefore the sensory characteristics. The lower intramuscular fat content in the AA ham batch had a positive effect on the free amino acid content. However, similar flavor traits between ham batches were found, but the AG loin batch showed greater value. The enhancing effect of the IMP on the overall flavor intensity was limited by the amino acid and the IMF contents in dry-cured ham and loin, while in dry-cured shoulder, the IMP could be the reason for the significant differences in after taste and cured flavor scores.     

Effect of pressure and holding time on colour,protein and lipid oxidation of sliced dry-cured Iberian ham and loin during refrigerated storage

The effect of high pressure (HP) treatments (200 MPa 15 min, 200 MPa 30 min, 300 MPa 15 min, 300 MPa 30 min) on colour, lipid and protein oxidation in sliced vacuum-packed dry-cured Iberian ham and loin during refrigerated storage (90 days, + 4 °C) was evaluated. Pressure level and holding time increased the extent of lipid oxidation in both products. Dry-cured ham showed a higher susceptibility to lipid oxidation than dry-cured loin since HP treatment increased TBA-RS values in dry-cured ham samples while HP treatment decreased TBA-RS values in dry-cured loin samples. However, HP treatment did not affect protein oxidation in both meat products. On the other hand, HP treatment affected instrumental colour since non-pressurized dry-cured meat products showed higher redness than pressurized ones. Regarding changes under storage, after 90 days of refrigerated storage lipid and protein oxidation increased while redness decreased in both HP treated and non-treated dry-cured meat products. Changes induced by HP were only noticeable after HP treatment, as storage reduced the initial differences between HP treated and non-treated samples. Therefore, the lack of differences in long stored dry-cured ham and loin HP treated and non-treated indicates that the application of HP (200–300 MPa/15–30 min) could not affect the quality of dry-cured meat products.Industrial relevanceDry-cured meat products are the meat-based products with the highest sensory quality in Spain and have a high projection in exterior markets. High pressure processing is effective in controlling pathogen and spoilage microorganisms in meat and meat products although it can promote color and oxidation changes that modify sensory characteristics. The study aimed the evaluation of pressure and holding time on color changes and protein and lipid oxidation at vacuum packed slices of Iberian dry-cured ham and loin during subsequent extended chilled storage. High pressure treatment of dry-cured Iberian ham and loin induce changes after treatment although initial differences are not maintained along refrigerated storage.     

Microstructural changes in Teruel dry-cured ham during processing

Dry-cured ham, "Protected Designation of Origin (PDO)" Teruel, is a high quality meat product processed using a traditional dry-curing procedure. The objective of this paper was to study the microstructure of the Biceps femoris and Semimembranosus muscles during "PDO Teruel" dry-cured ham processing using electron microscopy techniques (Cryo-SEM, SEM and TEM). The Semimembranosus and Biceps femoris muscles from raw ham show the typical structure of muscle tissue in meat. The muscle fibres appear firmly attached to one another by the endomysial connective tissue; the myofibrils inside the cells are strongly attached to each other and to the sarcolemma. In raw ham, the typical structural elements that constitute the sarcomere can be observed by TEM. Important microstructural changes are observed following salting; reflected in the fact that the Z-disks inside the myofibrils are no longer in line and there is important degradation of the cell membranes in this stage. At the end of the process, an accumulation of solutes, which could be products from proteolysis occurring mainly in the last stage of ripening, could be observed in both muscles. Thus the proteolysis and dehydratation phenomena that take place during "PDO Teruel" dry-cured ham processing occur at an ultrastructural level, and this is the basis to produce a product with the sensory features appreciated by consumers.     

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%).     

Effect of irradiation on colour and lipid oxidation of dry-cured hams from free-range reared and intensively reared pigs

The effect of irradiation dose (0, 5 and 10 kGy) of vacuum-packaged Iberian dry-cured ham slices from pigs fed on concentrate (CON) or free-range reared (FRG) was studied in relation to TBA-RS, hexanal content and instrumental colour changes. TBA-RS values increased after irradiation in the two sets of hams and the increase was dose-dependent. FRG samples showed higher TBA-RS values than CON samples in all treatments, although differences in the rate of formation of thiobarbituric acid-reactive substances were different in the two types of hams, being higher in CON samples than in FRG ones. Irradiation of dry-cured ham slices significantly increased hexanal contents in both sets of hams. Irradiation produced statistically significant increases in vacuum-packed dry-cured ham slices lightness (L^⁎-value), yellowness (b^⁎-value) and chroma (C^⁎-value). Irradiation resulted in significantly lower hue angle (h°) values and higher a^⁎-values in both sets of hams indicating a redder colour of irradiated samples than non-irradiated, and these changes were greater in FRG samples than in CON samples. Differences in composition characteristics of raw material could play an important role in the irradiation-induced changes on colour and lipid oxidation of vacuum-packaged dry-cured ham slices.Industrial relevanceIberian ham is dry-cured meat product with a high sensory quality due to pig fattening feature and the characteristics of its processing. Both factors lead to a product with characteristic cured red colour of the lean, an intense, characteristic and pleasant flavour, unique in dry-cured hams, derived from intense lipid oxidative phenomena. New commercialization formats of dry-cured Iberian hams include vacuum-packed sliced ham. Although the low Aw, NaCl and nitrite/nitrate contents make dry-cured ham not adequate for the growth of pathogens and spoilage microorganisms (cocci gram+ and yeast), those may be present on the surface of whole hams and reach the sliced product during the final boning, slicing and packaging operations even with the observation of strict hygienic procedures despite the implementation of modern technologies and HACCP systems, constituting a permanent risk of contamination. Dry-cured Iberian ham, as a result of its chemical characteristics, is a product with a long shelf-life at refrigeration temperature, however the risk of spoilage or pathogen growth could increase in the case of temperature abuse for a long stored period. Finally, due to health concerns, salting and curing process of dry-cured Iberian hams tend to a reduction in levels of NaCl and nitrite used. That could increase the risk associated to microorganism presence in dry-cured sliced hams. The use of irradiation could minimizes this risk, however irradiation could have important effects on overall quality of dry-cured Iberian products in which colour and extent of lipid oxidation and lipid oxidation-derived volatile compounds play an important role in consumer acceptation of the product.