Jens K. S. Møller

Dittany (Origanum dictamnus) as a source of water-extractable antioxidants

Extracts of dittany (Origanum dictamnus L.) made with solvents of varying polarity were evaluated (i) by electron spin resonance (ESR) spectrometry using the spin trapping technique for their efficiencies as scavengers of free radicals, and (ii) by measurement of oxygen depletion in a methyl linoleate emulsion for their efficiencies as chain-breaking antioxidants. Aqueous extracts of dittany most efficiently scavenged hydroxyl radicals as generated by the Fenton reaction and this extract also most efficiently reduced oxygen consumption when initiated by metmyoglobin. Ethanol and acetone extracts of dittany showed less activity in both assays with methanol extract being intermediate. The high efficiencies found in the aqueous (and methanol) extracts of dittany were closely related to a high content of phenolic compounds in these extracts, while the amounts of phenolics in the ethanol and acetone extract were lower. In aqueous extracts, the antioxidative activity was confirmed in a turkey thigh meat homogenate, where development of thiobarbituric reactive substances was increasingly inhibited by increasing additions from 0.0018 mg dittany/g meat to efficient inhibition for 0.011 mg dittany/g meat and higher additions. Dittany should be further explored as a source of water-extractable antioxidants.

Colour formation in fermented sausages by meat-associated staphylococci with different nitrite- and nitrate-reductase activities.

Three Staphylococcus strains, S. carnosus, S. simulans and S. saprophyticus, selected due to their varying nitrite and/or nitrate-reductase activities, were used to initiate colour formation during sausage fermentation. During fermentation of sausages with either nitrite or nitrate added, colour was followed by L(∗)a(∗)b measurements and the content of nitrosylmyoglobin (MbFe(II)NO) quantified by electron spin resonance (ESR). MbFe(II)NO was rapidly formed in sausages with added nitrite independent of the presence of nitrite reducing bacteria, whereas the rate of MbFe(II)NO formation in sausages with added nitrate depended on the specific Staphylococcus strain. Strains with high nitrate-reductase activity showed a significantly faster rate of pigment formation, but other factors were of influence as well. Product stability for the sliced, packaged sausage was evaluated as surface colour and oxidation by autofluorescence and hexanal content, respectively. No significant direct effect of the Staphylococcus addition was observed, however, there was a clear correspondence between high initial amount of MbFe(II)NO in the different sausages and the colour stability during storage. Autofluorescence data correlated well with hexanal content, and may be used as predictive tools. Overall, nitrite- and nitrate-reductase activities of Staphylococcus strains in nitrite-cured sausages were of limited importance regarding colour development, while in nitrate-cured sausages strains with higher nitrate reductase activity were crucial for ensuring optimal colour formation during initial fermentation stages.

Effect of residual oxygen on colour stability during chill storage of sliced, pasteurised ham packaged in modified atmosphere

The critical level of residual oxygen to avoid light induced oxidative discoloration during chill storage of sliced, pasteurised ham packaged in modified atmosphere (20% carbon dioxide balanced with nitrogen in a 1:3 product to headspace volume ratio) was found to lie between 0.1 and 0.5% oxygen. In 0.5% oxygen light induced discoloration was significant, as detected by the tristimulus colorimetry redness parameter, when compared to the same product stored in the dark, while at 0.1 and 0.02% oxygen the colour was stable both in the dark and when exposed to light for up to 27 days in chill storage. Lipid oxidation, determined as 2-thiobarbituric acid-reactive substances, and total plate counts showed no difference between discoloured and colour stable products, although a trained panel in a triangle test could differentiate between the taste of ham from packages with 0.02 and 0.5% oxygen after 27 days of chill storage.

Electrospray ionization mass spectrometry fingerprinting of whisky: immediate proof of origin and authenticity

Authentic samples of whisky produced in Scotland and USA and counterfeit whisky samples commercialized in Brazil have been directly submitted to electrospray ionization mass spectrometry (ESI-MS) analysis in both the negative and positive ion modes to assess the potential of this technique for simple and rapid quality control and proof of authenticity of whisky samples. ESI in the negative ion mode yields the most characteristic whisky fingerprinting mass spectra in just a few seconds by direct infusion of the samples, detecting the most polar or acidic components of each sample in their deprotonated anionic forms. No pre-treatment of the sample, such as extraction or derivatization or even dilution, is required. The analysis of the ESI(-)-MS data both by simple visual inspection but more particularly by chemometric data treatment enables separation of the whisky samples into three unequivocally distinct groups: Scotch, American and counterfeit whisky, whereas single malt and blended Scotch whiskies are also distinguished to some extent. As indicated by ESI-MS/MS analysis, the diagnostic anions are simple sugars, disaccharides and phenolic compounds. Direct infusion ESI-MS therefore provides immediate chemical fingerprinting of whisky samples for type, origin and quality control, as demonstrated herein for American, Scottish and counterfeit samples, whereas ESI-MS/MS analysis of diagnostic ions adds a second dimension of fingerprinting characterization when improved selectivity is desired.

Optimisation of colour stability of cured ham during packaging and retail display by a multifactorial design

A multifactorial design, including (1) percent residual oxygen, (2) oxygen transmission rate of packaging film (OTR), (3) product to headspace volume ratio, (4) illuminance level and (5) nitrite level during curing, was established to investigate factors affecting light-induced oxidative discoloration of cured ham (packaged in modified atmosphere of 20% carbon dioxide and balanced with nitrogen) during 14 days of chill storage. Univariate statistical analysis found significant effects of all main factors on the redness (tristimulus a-value) of the ham. Subsequently, Response Surface Modelling of the data further proved that the interactions between packaging and storage conditions are important when optimising colour stability. The measured content of oxygen in the headspace was incorporated in the model and the interaction between measured oxygen content in the headspace and the product to headspace volume ratio was found to be crucial. Thus, it is not enough to keep the headspace oxygen level low, if the headspace volume at the same time is large, there will still be sufficient oxygen for colour deteriorating processes to take place.

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.

Optimizing colour quality of modified atmosphere packed sliced meat products by control of critical packaging parameters.

To study the influence of different packaging and storage parameters on the colour stability of modified atmosphere packed, cured, cooked ham, a multiplicative analysis of variance model (GEMANOVA) was developed. The critical parameters investigated were % residual-O(2), product to headspace volume ratio (P/H volume ratio), temperature, light intensity and oxygen transmission rate (OTR). The model illustrated that all the investigated parameters interacted, but especially % residual-O(2) and P/H volume ratio - i.e., the absolute O(2) content, influenced the degree of discoloration. The complex interactions of the parameters justified the selected model, as it emphasised the necessity of evaluating the parameters simultaneously instead of considering them individually. The importance of absolute O(2) content was further validated through an industrial experiment including three different kinds of sliced meat products.

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.

Formation of amino acid (L-leucine, L-phenylalanine) derived volatile flavour compounds by Moraxella phenylpyruvica and Staphylococcus xylosus in cured meat model systems

A bacterial strain isolated from Danish immersion curing brine, Moraxella phenylpyruvica 0100, and a commercial meat starter culture, Staphylococcus xylosus DD34, were tested for their ability to form characteristic volatile compounds in minimal medium with the added amino acid L-leucine or L-phenylalanine under different environmental conditions (pH 5.5 and 6.0; 0 and 210 ppm nitrate; pre-incubation with and without agitation) and compared with respect to their ability to form volatile compounds in cured meat extracts and vacuum-packed cured meat cuts. The characteristic cured meat aroma precursors/compounds 3-methylbutanal and 3-methylbutanol were found to be formed in cured meat extracts and vacuum-packed cured meat cuts inoculated with M. phenylpyruvica. These volatiles are most probably formed by metabolic conversion of the amino acid L-leucine by M. phenylpyruvica, as they were also produced in minimal media with added L-leucine inoculated with this organism. The characteristic L-phenylalanine derived compound, benzaldehyde, formed by M. phenylpyruvica in minimal medium in the presence of nitrate (210 ppm), was not produced in any noticeable amount in cured meat extracts or vacuum-packed cured meat inoculated with M. phenylpyruvica. In contrast, benzacetaldehyde, which has been described as a possible metabolic product of the microbial conversion of L-phenylalanine, was found to be a characteristic volatile compound formed in cured meat extracts and vacuum-packed cured meat inoculated with M. phenylpyruvica, indicating an alternative metabolic pathway for L-phenylalanine by this organism in a cured meat environment. Even though S. xylosus was able to form volatile compounds characteristic of cured meats (3-methylbutanal, 3-methylbutanol) in minimal media with added L-leucine, this bacterial strain seemed not to be able to produce these characteristic volatiles in the studied cured meat systems. The present data imply that M. phenylpyruvica, in particular, is a potential meat starter for ensuring superior flavour development in cured meat.

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.