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.     

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。     

Nitric oxide inhibits the formation of zinc protoporphyrin IX and protoporphyrin IX

The aim of this study was to elucidate the mechanism by which curing agents, especially nitrite, inhibit the formation of zinc protoporphyrin IX (ZPP) in dry-cured hams such as Parma ham. The oxidation–reduction potential of model solutions was increased by the addition of nitrite, but it was not clear whether the formation of ZPP is inhibited by the oxidizing property of nitrite. The effect of nitric oxide (NO) produced from nitrite on the formation of ZPP was examined. The amount of ZPP formed was decreased by the addition of NO donors. The amount of protoporphyrin IX (PPIX), which is the precursor of ZPP, was also decreased by the addition of NO donors. It is concluded that NO produced from nitrite inhibited the formation of PPIX and ZPP was therefore not formed in cured meat products with the addition of nitrite or nitrate.     

THE STUDY OF INGREDIENTS AND PROCESSING TECHNIQUES OF XUANWEI STYLE HAM

Xuanwei style ham is one of the famous dry-cured hams in the world. The influences of different ingredients and processing techniques on sensory properties, brine diffusing rate, chemical composition and hygienic quality of Xuanwei style cured ham were studied. Modified cured ham processing techniques were employed in this study. Massaging of green ham and injection of curing solution improved brine diffusing rate ( P <  0.05) and reduced curing time from the traditional 28 days to 21 days (weight of green ham about 13 kg). Ninety mg/kg sodium nitrite was used for improving cured ham color ( P <  0.05), and its residue still met the Chinese cured ham standard of 20 mg/kg. The hygienic and sensory qualities were not significantly affected by the addition of sodium ascorbate and polyphosphate ( P >  0.05).

PRACTICAL APPLICATIONS

The paper studied the Chinese famous Xuanwei dry-cured ham, modified curing ingredient and processing techniques of Xuanwei ham are beneficial for improving the ham quality and important to shorten the traditional cured ham processing procedure such as longer processing time and seasonal processing method to adapt to the new industrial procedures of manufacturing and marketing of Xuanwei cured ham.     

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.     

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%).     

云南干腌火腿加工工艺及其品质影响因素研究进展

云南干腌火腿是一种重要的传统发酵肉制品,以猪后腿为原料,经腌制、风干、发酵等加工而成。云南干腌火腿主要有宣威火腿、撒坝火腿、三川火腿、老窝火腿、诺邓火腿及鹤庆火腿等,本文对云南干腌火腿加工工艺及干腌火腿品质影响因素进行综述,从原料肉、腌制条件、色泽及风味等几方面做了详细讨论,旨在为干腌火腿品质改进提供一定的理论参考。     

Mass spectrometric evidence for a zinc-porphyrin complex as the red pigment in dry-cured Iberian and Parma ham

Extracts containing red pigment complexes from the two types of dry-cured hams, Italian Parma and Spanish Iberian ham, were obtained using water and acetone as extraction solvents followed by a crude purification with C18 column filtration. The purified extracts were then analyzed spectroscopically by recording absorption and fluorescence spectra (λ(ex)=420nm), which both indicate the presence of chemically identical red chromophores with properties similar to a complex of transition metals and protoporphyrin IX. Electrospray ionization mass spectrometry (ESI-MS) in the positive ion mode confirms the presence of identical chemical compounds. ESI-MS in the negative ion mode detects a cluster of seven isotopologue ions (that of m/z 623.2 as the most intense) with a pattern matching that of a Zn protoporphyrin IX complex. Based on mass spectral data it is concluded that a Zn-porphyrin complex constitutes a major chromophore in dry-cured Iberian ham as well as in Parma ham.     

传统腌腊肉制品挥发性风味物质的研究进展

本文以干腌火腿、腊肉、风鸭、发酵香肠和风干鱼制品为代表,对中国传统腌腊肉制品的挥发性风味物质进行了探讨。从总体上阐述了我国传统腌腊肉制品挥发性风味的形成机理,详尽列表总结分析了不同制品的特征风味化合物,分析它们之间的共性与异性,并从原料特性及加工工艺两个主要方面分析了差异性形成的可能原因。一方面,丰富了传统腌腊肉制品的形成机制,另一方面,为生产调控肉制品的风味提供理论依据。     

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.     

干腌肉块复合腌制新技术研究

采用低温、复合腌制剂、干燥、真空包装、发酵成熟等工艺过程,进行低咸度腌肉制品安全性研究。通过正交试验筛选出的最佳复合腌制剂配方(NaCl3%、Nisin600mg/kg、VC2mg/kg、亚硝酸钠40mg/kg),能够避免腌制失败的风险。加工过程中,肉块的pH(5.75-6.25)变化较小,抑菌效果不显著;而水分活度(0.960-0.809)明显下降,是腌肉安全贮藏的重要栅栏因子。发酵成熟的干腌肉块含水分53.93%,食盐含量降至6.0%,表现出干腌肉制品的特征风味。在发酵成熟阶段,蛋白质水解和轻度的脂肪氧化,对干腌肉块风味品质的形成有重要作用。     

Red Pigment of Parma Ham and Bacterial Influence on its Formation

The red pigment of Parma ham was compared with the myoglobin derivatives present in meat and meat products. Spectral patterns of 75% acetone extracts and electron spin resonance spectra from Parma ham differed from those of the myoglobin derivatives. Staphylococci isolated from Parma ham generated red myoglobin derivative from metmyoglobin. Model fermented sausage prepared by inoculation with the isolates developed a more desirable red color without nitrite or nitrate treatment. The red pigment of Parma ham and the model sausage appeared to be the same myoglobin derivative. The reddening of Parma ham is probably caused by the action of bacteria.     

Inhibition of Listeria monocytogenes using natural antimicrobials in no-nitrate-or-nitrite-added ham

Consumer demand for foods manufactured without the direct addition of chemical preservatives, such as sodium nitrite and organic acid salts, has resulted in a unique class of "naturally" cured meat products. Formulation with a natural nitrate source and nitrate-reducing bacteria results in naturally cured processed meats that possess traits similar to conventionally cured meats. However, previous research has shown that the naturally cured products are more susceptible to pathogen growth. This study evaluated Listeria monocytogenes growth on ham manufactured with natural curing methods and with commercially available clean-label antimicrobials (cultured sugar and vinegar blend; lemon, cherry, and vinegar powder blend) and assessed impacts on physicochemical characteristics of the product. Hams made with either of the antimicrobials supported L. monocytogenes growth similar to that in the traditionally cured control (P > 0.05). Hams made with prefermented celery juice powder had the lowest residual nitrite concentrations (P < 0.05), and when no antimicrobial was added, L. monocytogenes growth was similar to that of the uncured control (P > 0.05). Aside from residual nitrite and nitrate concentrations, few physicochemical differences were identified. These findings show that ham can be produced with natural curing methods and antimicrobials to provide similar L. monocytogenes inhibition and physicochemical traits as in traditionally cured ham.     

A Zn-porphyrin complex contributes to bright red color in Parma ham

The Italian traditional dry-cured ham (Parma ham) shows a stable bright red color that is achieved without the use of nitrite and/or nitrate. In this study we examined the pigment spectroscopically, fluoroscopically and by using HPLC and ESI-HR-MASS analysis. Porphyrin derivative other than acid hematin were contained in the HCl-containing acetone extract from Parma ham. A strong fluorescence peak at 588 nm and a weak fluorescence peak at 641 nm were observed. By HPLC analysis the acetone extract of Parma ham was observed at the single peak, which eluted at the same time as Zn-protoporphyrin IX and emitted fluorescence. The results of ESI-HR-MS analysis showed both agreement with the molecular weight of Zn-protoporphyrin IX and the characteristic isotope pattern caused by Zn isotopes. These results suggest that the bright red color in Parma ham is caused by Zn-protoporphyrin IX.     

Incubation of curing brines for the production of ready-to-eat, uncured, no-nitrite-or-nitrate-added, ground, cooked and sliced ham

Salt concentration, vegetable juice powder (VJP) concentration and temperature were investigated to determine necessary conditions for incubation of curing brines including VJP and a starter culture containing Staphylococcus carnosus prior to production of naturally cured, no-nitrate/nitrite-added meat products. Subsequently, incubated brines were utilized to produce no-nitrate/nitrite-added sliced ham in which quality characteristics and residual nitrite concentrations were measured to determine feasibility of brine incubation for nitrate conversion prior to injection. Two ham treatments (one with VJP and starter culture; one with pre-converted VJP) and a nitrite-added control were used. No differences (P>0.05) were found for color in the VJP treatments. Control sliced ham was redder after 42 days of storage, retaining significantly (P<0.05) greater a* (redness) than either of the VJP treatments. Residual nitrite concentration was greater (P<0.05) in the control hams during the first week of storage. While the nitrite-added control retained greater red color and initially had more residual nitrite than the VJP treatments, the two VJP treatments did not differ from each other.     

Characterization and differentiation of Italian Parma,San Daniele and Toscano dry-cured hams: A multi-disciplinary approach

This study aimed at characterizing the sensory quality of Italian PDO dry-cured Parma, San Daniele and Toscano hams, applying a multi-disciplinary approach. Ham sensory profile as well as physico-chemical, aromatic, morphological and textural characteristics was investigated. There was a great difference between Toscano ham and Parma and San Daniele hams, which were more similar even though differentiated. Toscano ham showed higher scores for pork-meat odor, saltiness, dryness, fibrousness and hardness; accordingly, this ham was described by a high NaCl content and by high values of instrumental hardness, cohesiveness, gumminess and chewiness. Parma ham was characterized by a cured flavor, whereas San Daniele ham showed a wider fatty area and higher pH values. Parma and San Daniele hams were also described by higher values of sweetness, RGB color values and water activity. Sensory characteristics evaluated by trained assessors were correlated to instrumental measures, indicating that instrumental devices can be effectively applied for dry-cured ham characterization.     

巴马火腿红色色素的研究进展

综述了巴马火腿红色色素研究的最新进展。巴马火腿在腌制时不添加硝酸盐或亚硝酸盐 ,经过长期的加工过程 ,巴马火腿肉具有稳定的红色。巴马火腿的红色色素与添加硝酸盐或亚硝酸盐腌制的普通火腿的红色色素具有不同的吸收光谱、颜色稳定性和抗氧化性。根据巴马火腿红色色素的吸收光谱、荧光光谱、高压液相色谱和高分辨率电喷雾电离质谱 ,证实了巴马火腿的红色色素是锌 原卟啉IX。     

Consumers’ preferences for Iberian dry-cured ham and the influence of mast feeding: An application of conjoint analysis in Spain

This paper analyzes consumers’ preferences for Iberian dry-cured ham, one of the most typical and highly prized meat products in Spain. The data were obtained by a survey carried out between April and May 2006 with a sample of 417 consumers in Extremadura (SW Spain). Conjoint Analysis was used to estimate the relative importance of the main attributes affecting preferences for Iberian ham and to create consumer segments with similar preference profiles. Results have shown that Price and Type of ham are the most important attributes for the choice of ham. Simulation analyses determined the surcharge that consumers are willing to pay for an Iberian mast-fed ham instead of an Iberian ham, thus identifying an ideal cluster for Iberian mast-fed ham.     

Irradiated Chinese Rugao ham: Changes in volatile N-nitrosamine, biogenic amine and residual nitrite during ripening and post-ripening

N-nitrosamines, biogenic amines and residual nitrite are harmful substances and often present in cured meat. The effects of gamma-irradiation (γ-irradiation) on these chemicals in dry-cured Chinese Rugao ham during ripening and post-ripening were investigated. Rugao hams were irradiated at a dose of 5kGy before ripening and were then ripened in an aging loft. Although γ-irradiation degraded tyramine, putrescine and spermine, on the other hand, it promoted the formation of spermidine, phenylethylamine, cadaverine and tryptamine. Residual nitrite was significantly reduced by γ-irradiation. N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine (NDEA) and N-nitrosopyrrolidine (NPYR) were found in Chinese Rugao ham during ripening and post-ripening but could be degraded with γ-irradiation. The results suggest that γ-irradiation may be a potential decontamination measure for certain chemical compounds found in dry-cured meat.     

Relations between compositional traits and sensory qualities of French dry-cured ham

The left hams of 20 pig carcasses were processed by dry-curing for either 179 (n = 10) or 273 days (n = 10). The Biceps femoris was taken at the end of the processing and submitted to chemical and sensory analyses. The following chemical traits were determined: pH, moisture, salt content, fatty acid content and composition of lipid classes (triglycerides, phospholipids, free fatty acids), nitrogen fractions (soluble and insoluble protein, non-protein nitrogen, free amino acids) and volatile compounds. Sensory analysis was performed by a panel of 12 trained members, who scored texture traits (firmness, dryness, fibrousness, mellowness), taste (salty, acid), intensity of aromas (typical of dry ham, fresh meat, fat, cured meat, rancid, metal), persistence of dry ham aroma, persistence of taste after deglutition, persistence of astringency, persistence of metallic taste. The sensory qualities of dry-cured ham showed noticeable changes between days 179 and 273. Firmness, dryness and intensities of aromas typical of dry-cured ham and cured meat were enhanced with time. Neither pH, moisture nor salt content played a noticeable role in determining the sensory quality of dry-cured ham. Among the volatile compounds, the levels of several ketones and 1-butanol were significantly correlated with the aromas of dry ham and cured meat, while rancid aroma was related to aldehydes, ethylacetate, 2,3-pentanedione and nonane. Therefore, the aromas of dry ham and cured meat appeared to be determined mainly by the products of lipid oxidation. However, they were related also to unidentified volatile compounds, whose identification would be of particular interest.