Heat-processed Foods Research Articles

Assessment of hydroxymethylfurfural intake in the Spanish diet

Estimation of hydroxymethylfurfural (HMF) exposure of the Spanish population from heat-processed food was performed. HMF levels in Spanish foods were obtained from the data previously published in peer-reviewed papers; in addition, it was identified which food categories contributed significantly to HMF exposure. The potential HMF exposure was calculated for three different scenarios by using individual food intakes and the minimum (scenario 1), median (scenario 2) and maximum (scenario 3) values of analytical data on the HMF content in foods. A mean HMF intake of 10 mg day−1 (corresponding to scenario 2) was obtained, which is only ten-fold lower than the tolerable daily intake. Coffee and bread are the most important food items that contribute nearly 85% to the total HMF daily exposure, although biscuits, breakfast cereals, beer, UHT milk and tomato products are also important for HMF exposure.

Application of the standard addition method for the determination of acrylamide in heat-processed starchy foods by gas chromatography with electron capture detector

A gas chromatography electron capture detector (GC-ECD) using the standard addition method was developed for the determination of acrylamide in heat-processed foods. The method entails extraction of acrylamide with water, filtration, defatting with n-hexane, derivatization with hydrobromic acid and saturated bromine-water, and liquid–liquid extraction with ethyl acetate. The sample pretreatment required no SPE clean-up and concentration steps prior to injection. The final extract was analyzed by GC-ECD. The chromatographic analysis was performed on polar columns, e.g. Supelcowax-10 capillary column, and good retention and peak response of the analyte were achieved under the optimal conditions. The qualification of the analyte was by identifying the peak with same retention time as standard compound 2,3-DBPA and confirmed by GC–MS. GC–MS analysis confirmed that 2,3-DBPA was converted to 2-BPA nearly completely on the polar capillary column, whether or not treated with triethylamine. A four-point standard addition protocol was used to quantify acrylamide in food samples. The limit of detection (LOD) was estimated to be 0.6μg/kg on the basis of ECD technique. Validation and quantification results demonstrated that the method should be regarded as a low-cost, convenient, and reliable alternative for conventional investigation of acrylamide.

Letter to the editor

Letter to the editor

Automated fluorimetry in quality control of pasteurized and ultra-high temperature-treated starch soup

Summary Automated monitoring of the microbiological quality of heat-processed foods by the resazurin reduction test was applied to microtitration plate incubator-fluorimeter technology. The appearance and disappearance of the fluorescing peak of resorufin was monitored on microtitration trays. Pasteurized or ultra-high temperature-treated starch-based soup was used as the model food system. Bacillus subtilis spores (ultra-high temperature treatment) and vegetative cells of Enterococcus faecalis (pasteurization) were inoculated into the soup before the heat treatment at levels which resulted in some survival. The timing of appearance of maximum fluorescence correlated with the number of bacteria in pre-incubated samples. Automated resazurin-reduction fluorimetry was compared with conventional plating, turbidometry and microcolony count by the direct epifluorescent filter technique. The results of the resazurin test correlated well with those of all the other methods tested. Fluorimetry had the advantage that the results could be read within 1–5h and the reproducibility was superior to the other methods.

Development of a novel bioassay for determining the available lysine contents of foods and feedstuffs

Lysine is an important indispensable amino acid, and describing the lysine content of a food or feedstuff provides useful information about nutritional value. However, when a food or feedstuff is subjected to heating the lysine present can be altered to nutritionally unavailable derivatives. These derivatives can revert back to lysine during the acid hydrolysis step used in amino acid analysis causing an overestimate of the lysine content. There have been many chemical methods developed to determine the reactive (unmodified) lysine content of foods and feedstuffs, but these do not take into account the incomplete absorption of lysine from the small intestine. There are also a number of animal-based assays for determining available lysine (the lysine that can be absorbed in a form that can be used for protein synthesis). The true ileal amino acid digestibility assay is commonly used to determine amino acid availability and is accurate for application to unprocessed foods and feedstuffs but is not accurate for lysine and possibly other amino acids when applied to heat-processed foods or feedstuffs. For such protein sources, assays such as the slope-ratio assay, indicator amino acid oxidation assay and the BIOLYSINEtrade mark assay (true ileal digestible reactive lysine assay) have been developed to determine available lysine. The present paper discusses the efficacy of the BIOLYSINEtrade mark assay as well as other assays for determining available lysine in processed foods and feedstuffs.

A review of the occurrence, formation and analysis of furan in heat-processed foods

In 2004, the US-FDA announced the findings of its exploratory surveys of furan in selected foods. In the 3 years since, there have been a number of publications on furan in foods. This review looks at furan in foods, its metabolism and toxicity. The important factor of heating is explored along with the precursors and chemical mechanisms that may be involved. Finally, prospects for limiting the occurrence, formation and retention of furan in foods are discussed.

Extractability, Stability, and Allergenicity of Egg White Proteins in Differently Heat-Processed Foods

Hen's egg white protein is a major cause of food allergy, and a considerable number of countries have introduced labeling directions for processed food products. To control compliance with these regulations, analytical assays for the detection of egg in manufactured foods have been developed. In this study, we have tested the performance of 3 commercially available kits for quantitative egg analysis using 6 model heat-processed foods. The 3 assays worked well under standard conditions with soluble egg white proteins, but only the kit using a denaturing-reducing extraction buffer detected egg in complex heat-treated food matrixes. The differently extracted food samples were further used to evaluate the stability and allergenicity of the egg white allergens ovalbumin, ovomucoid, ovotransferrin, and lysozyme with polyclonal anti-egg antibodies and sera of 6 patients with egg allergy. It could be shown that differences in egg protein extractability have a significant impact on the interpretation of study results.

Cyclodextrins Found in Enzyme- and Heat-Processed Starch-Containing Foods

Native and branched-type (glucosylated and maltosylated) cyclodextrins have been isolated and identified in different enzyme- and heat-processed starch-containing food products. Amylolytic enzyme-processed foods such as different beer samples, corn syrup of different dextrose equivalents, and thermally-processed food such as bread, contained minute amounts of different types of cyclodextrins. HPLC/MS Analyses of appropriately preconcentrated and purified food samples indicated the presence of parent beta- and gamma-cyclodextrins and all the three, alpha-, beta-, and gamma-branched cyclodextrins with different degrees of glycosylation. The data presented in this account are thought to be of practical importance in terms of both the analysis methods used for the cyclodextrins and the approval status of different cyclodextrin-containing food, cosmetic, and pharmaceutical products.

Some factors affecting the formation of furan in heated foods

Levels of furan in various foods were measured before and after heating under heating and laboratory conditions. The effect of contact with can coatings, sealing gaskets and the epoxidized oils used in gasket manufacture on furan formation was studied. The objective was to identify factors affecting furan formation. Furan present in heat-processed food samples persisted during cooking. Furan was shown to form in foods on heating, although it did not accumulate to a significant degree on heating in an open vessel. There were no interactions between foods and cans, can coatings or gaskets that had a significant influence on furan formation. Furan accumulated particularly in heat-processed canned and jarred foods because they are sealed containers that receive a considerable thermal load. Heating epoxidized oils used in sealing gaskets formed furan. At the levels used in gaskets, however, epoxidized oils should not affect the formation of furan in foods.

Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within the Bacillus cereus group

One hundred representative strains of Bacillus cereus were selected from a total collection of 372 B. cereus strains using two typing methods (RAPD and FT-IR) to investigate if emetic toxin-producing hazardous B. cereus strains possess characteristic growth and heat resistance profiles. The strains were classified into three groups: emetic toxin (cereulide)-producing strains ( n = 17), strains connected to diarrheal foodborne outbreaks ( n = 40) and food–environment strains ( n = 43), these latter not producing the emetic toxin. Our study revealed a shift in growth limits towards higher temperatures for the emetic strains, regardless of their origin. None of the emetic toxin-producing strains were able to grow below 10 °C. In contrast, 11% (9 food–environment strains) out of the 83 non-emetic toxin-producing strains were able to grow at 4 °C and 49% at 7 °C (28 diarrheal and 13 food–environment strains). non-emetic toxin-producing strains. All emetic toxin-producing strains were able to grow at 48 °C, but only 39% (16 diarrheal and 16 food–environment strains) of the non-emetic toxin-producing strains grew at this temperature. Spores from the emetic toxin-producing strains showed, on average, a higher heat resistance at 90 °C and a lower germination, particularly at 7 °C, than spores from the other strains. No difference between the three groups in their growth kinetics at 24 °C, 37 °C, and pH 5.0, 7.0, and 8.0 was observed. Our survey shows that emetic toxin-producing strains of B. cereus have distinct characteristics, which could have important implication for the risk assessment of the emetic type of B. cereus caused food poisoning. For instance, emetic strains still represent a special risk in heat-processed foods or preheated foods that are kept warm (in restaurants and cafeterias), but should not pose a risk in refrigerated foods.

The effect of thermal processing on the IgE reactivity of the non‐specific lipid transfer protein from apple, Mal d 3

Non-specific lipid transfer proteins (LTPs) are involved in allergy to fresh and processed fruits. We have investigated the effect of thermal treatment and glycation on the physico-chemical and IgE-binding properties of the LTP from apple (Mal d 3). Mal d 3 was purified from apple peel and the effect of heating in the absence and presence of glucose investigated by CD spectroscopy, electrospray and MALDI-TOF mass spectrometry. IgE reactivity was determined by RAST and immunoblot inhibition, SPT and basophil histamine release test. The identity and IgE reactivity of purified Mal d 3 was confirmed. Mild heat treatment (90 degrees C, 20 min) in the absence or presence of glucose did not alter its IgE reactivity. More severe heat treatment (100 degrees C, 2 h) induced minor changes in protein structure, but a significant decrease in IgE-binding (30-fold) and biological activity (100- to 1000-fold). Addition of glucose resulted in up to four glucose residues attached to Mal d 3 and only a 2- and 10-fold decrease of IgE-binding and biological activity, respectively. Only severe heat treatment caused a significant decrease in the allergenicity of Mal d 3 but glycation had a protective effect. The presence of sugars in fruits may contribute to the thermostability of the allergenic activity of LTP in heat-processed foods.

Innovative approach to low-level gluten determination in foods using a novel sandwich enzyme-linked immunosorbent assay protocol.

There is currently much call for a reliable enzyme-linked immunosorbent assay (ELISA) protocol for determining gluten in foods to serve as a basis for further Codex Alimentarius regulations. Given its ability to recognize the potential coeliac-toxic epitope QQPFP, which occurs repeatedly in alpha-, gamma- and omega-gliadins, hordeins and secalins, the monoclonal antibody R5 raised against a secalin extract may prove to be an essential tool for gluten analysis. This study was designed to develop a highly sensitive and specific sandwich ELISA to quantify low levels of wheat, barley and rye prolamins in foods for coeliacs. Simple sandwich ELISA based on the use of a single monoclonal antibody (R5) as both the coating and detection was developed. A quantitative cocktail gluten-extraction procedure for heat-processed foods was also tested. R5-ELISA was able to identify gliadins, hordeins and secalins with assay sensitivities of 0.78, 0.39 and 0.39 ng/ml, respectively. The assay's detection limit was 1.5 ng gliadins/ml (1.56 ppm gliadins, 3.2 ppm gluten). The system proved insensitive to the non-coeliac-toxic cereals maize, rice and oats, and was non-cultivar-dependent. It was also able to detect gliadins and hordeins in unprocessed and heat-processed wheat- and barley-based products, and to estimate the gluten content of hydrolysed foods. We present a new generation of a robust sandwich R5-ELISA with good reproducibility (8.7%) and repeatability (7.7%). Its gluten-detection limit of 3.2 ppm is lower than the existing threshold of 20-200 ppm. The ELISA, which is equally sensitive to barley, wheat and rye prolamins, is compatible with the quantitative cocktail extraction procedure for heat-processed foods. Along with the cocktail procedure, the Working Group on Prolamin Analysis and Toxicity is currently evaluating an R5-ELISA system as proposed by the Codex Alimentarius Commission.

Innovative approach to low-level gluten determination in foods using a novel sandwich enzyme-linked immunosorbent assay protocol

Objectives There is currently much call for a reliable enzyme-linked immunosorbent assay (ELISA) protocol for determining gluten in foods to serve as a basis for further Codex Alimentarius regulations. Given its ability to recognize the potential coeliac-toxic epitope QQPFP, which occurs repeatedly in α-, γ- and ω-gliadins, hordeins and secalins, the monoclonal antibody R5 raised against a secalin extract may prove to be an essential tool for gluten analysis. This study was designed to develop a highly sensitive and specific sandwich ELISA to quantify low levels of wheat, barley and rye prolamins in foods for coeliacs. Methods Simple sandwich ELISA based on the use of a single monoclonal antibody (R5) as both the coating and detection was developed. A quantitative cocktail glutenextraction procedure for heat-processed foods was also tested. Results R5-ELISA was able to identify gliadins, hordeins and secalins with assay sensitivities of 0.78, 0.39 and 0.39 ng/ml, respectively. The assay's detection limit was 1.5 ng gliadins/ml (1.56 ppm gliadins, 3.2 ppm gluten). The system proved insensitive to the non-coeliac-toxic cereals maize, rice and oats, and was non-cultivar-dependent. It was also able to detect gliadins and hordeins in unprocessed and heat-processed wheat- and barleybased products, and to estimate the gluten content of hydrolysed foods. Conclusion We present a new generation of a robust sandwich R5-ELISA with good reproducibility (8.7%) and repeatability (7.7%). Its gluten-detection limit of 3.2 ppm is lower than the existing threshold of 20-200 ppm. The ELISA, which is equally sensitive to barley, wheat and rye prolamins, is compatible with the quantitative cocktail extraction procedure for heat-processed foods. Along with the cocktail procedure, the Working Group on Prolamin Analysis and Toxicity is currently evaluating an R5-ELISA system as proposed by the Codex Alimentarius Commission.

Formation of the comutagenic beta-carboline norharman in a simple tryptophan-containing model system at low temperature (40 degrees C-80 degrees C).

The formation of the comutagenic tryptophan derivate norharman in heat-processed food is a well known phenomenon. As a first step to investigate the possible formation of norharman during food storage particularly in regard to fortified food we developed a model system and studied the non-enzymatic formation of the comutagen at temperatures below 100 degrees C. With standard conditions (4 h, 80 degrees C) iron and copper (Fe2+/Cu2+) was required for the formation of norharman. Addition of glucose to the reaction mixture increased the norharman formation and a plateau was reached with 400 microM tryptophan. Decreasing the reaction temperature to 40 degrees C led to a significant formation of norharman after 72 hours of incubation indicating that the formation of norharman can take place at temperatures that are close to ranges occurring during food storage.

Effect of urea treatment on recovery of staphylococcal enterotoxin A from heat-processed foods.

The effects of urea treatment on the potential reactivation of heat-damaged antigenic components of staphylococcal enterotoxin A (SEA) were examined with cooked foods, including mushrooms, ham, bologna, salami, and turkey. The thermal stability of purified SEA spiked into foods and native SEA produced by Staphylococcus aureus in foods was also examined. Food samples containing either spiked or native SEA were thermally processed by autoclaving or retorting. This was followed sequentially by toxin extraction, urea treatment, dialysis, reconstitution, and SE assays with the reversed passive latex agglutination and/or enzyme immunoassay kit. The results indicate that (i) urea treatment did not result in any reactivation of heat-inactivated antigenic components of SEA in any of the foods tested, (ii) the serological components of purified SEA were destroyed (> or = 96%) by autoclaving at 121.1 degrees C for 5 to 15 min or by retorting at an F0 of 4 to 18, and (iii) the immunological property of the native SEA was approximately threefold-more heat resistant than that of the purified SEA. The study suggested that the current urea method is not suitable for the detection of heat-denatured SEA in the thermally processed foods.

Evaluation of hamburgers and hot dogs for the presence of mutagens

Food products derived from heat-treated (fried, broiled, baked) meat may contain traces of mutagenic heterocyclic amine contaminants, some of which are proven carcinogens in rodents. To confirm their presence and range in Canadian foods, and estimate the average human intake of these types of mutagens from frequently consumed heat-processed foods, several commercially prepared fried-beef patties (hamburgers) and hot dogs (weiners) were analysed for their mutagenic capacity. The mutagenicity of the extracts was tested in the Salmonella/microsome assay using strain TA98 with metabolic activation. 16 samples of hamburgers and 14 samples of hot dogs, randomly obtained from ‘fast food’ commercial establishments; or street vendors, were used in this survey. The mutagenic activity of these samples ranged from very low to 1042 revertants/g equivalent for the hamburgers and from non-detectable to 4875 revertants/g equivalent for the hot dogs. The average values were 199 and 424 revertants/g for the hamburgers and hot dogs, respectively. The wide range in mutagenicity was found even for the same type of product obtained from the same outlet at different times. This indicates possible inconsistency in cooking procedures during the preparation of these products. It also shows the difficulty in accurately assessing the intake of mutagenic heterocyclic amines from hamburgers and hot dogs prepared in ‘fast food’ outlets.

Stability of Beefy Meaty Peptide to Pasteurization and SterilizationTemperatures

Beefy meaty peptide (BMP) is an octapeptide found in meat with desirable sensory properties. The purpose of this study was to examine the stability of BMP after heating under pasteurization and sterilization conditions. BMP was heated to 71°C for 15 s and 121°C for 20 min; and then evaluated with high performance liquid chromatography and mass-spectrometry. Results indicated that BMP was relatively stable to these heat processes. Less than 10% of BMP was denatured under the sterilization condition. The results indicate BMP is a heat-resistant peptide and it may be used in heat-processed food.

A centrifugation/quadrant plate technique for the simplified differential-bacteriological examination of adequately heat-processed foods

Foods manufactured or prepared, stored and distributed with due care for microbiological safety are likely to contain very low numbers of potentially hazardous bacteria. A centrifugation/quadrant plate method is described that allows the estimation of levels of contamination between 0·4 and 100 cfu/g of Enterobacteriaceae, Staphylococcus aureus, Bacillus cereus and other toxinogenic Bacillus spp., Streptococcus pyogenes and Listeria monocytogenes. When preceded by an adequate repair procedure, the method is reliable, convenient, rapid and inexpensive.

Formation of methoxypyrazines in reactions of 2(1H)-pyrazinones with naturally occurring methylating agents

Methoxypyrazines are formed in low yield when 2(1H)-pyrazinones are heated with fruit pectin, betaine, trigonellin, S-methylmethionine, choline, and lecithin. The methylation reactions provide a chemical link to nonenzymic methoxypyrazine formation in heat-processed foods

Methylreductic acid and hydroxymethylreductic acid: oxygen radical-forming agents in heated starch

Two compounds forming oxygen radicals were isolated from heated starch by monitoring the potency to form 8-hydroxydeoxyguanosine from deoxyguanosine during purification procedures. These compounds were identified as hydroxymethylreductic acid and methylreductic acid. The former compound was mutagenic to Salmonella strains TA100, TA102 and TA104 and the latter compound induced sister-chromatid exchanges in human NL3 cells. Hydroxymethylreductic acid was found to be a novel compound. Considerable amounts of these compounds were detected in various heat-processed foods.