2-alkylcyclobutanones as irradiation dose indicators in irradiated ground beef patties

Alkylcyclobutanones have been recognized as chemical markers of irradiated lipid-containing foods since 1970. They are important because they are produced solely as a result of irradiation and not any other processing method. This study investigated the formation of 2-dodecylcyclobutanone (2-DCB) and 2-tetradec-5'-enylcyclobutanone (2-TDCB) in irradiated ground beef patties from commercial and noncommercial sources. Patties were irradiated using a (60)C source (gamma-irradiation) and electron beam irradiation, at five targeted absorbed doses of 0.5, 1.0, 2.5, 5.0, and 7.0 kGy. Commercially available irradiated patties were also studied. A supercritical fluid extraction (SFE) procedure was optimized and used for the extraction and isolation of the alkylcyclobutanones. Samples can be used for extraction without a prior cleanup step, which makes this procedure rapid and convenient to use. Identification and quantitation of the cyclobutanones were done by gas chromatography-mass spectroscopy. 2-DCB was detected in all of the irradiated samples (including commercial patties), and its concentration increased linearly with the irradiation dose. Electron beam irradiation produced a greater amount of 2-DCB compared to gamma-irradiation at dose levels >2.5 kGy. 2-TDCB was detected only at the two higher irradiation doses, whereas both marker compounds were not detected in the non-irradiated samples.     

Evaluation of 2-dodecylcyclobutanone as an irradiation dose indicator in fresh irradiated ground beef

Alkylcyclobutanones (2-ACBs) are radiolytic products formed when fatty acids are irradiated. These cyclobutanones are unique irradiation byproducts and therefore may serve as indicators of irradiation exposure. As only limited information exists about 2-ACB formation in retail meat products, reliable methods that can quantify 2-ACBs and thus estimate irradiation dose in commercial meat products are desired. The cyclobutanone studied in this experiment was 2-dodecylcyclobutanone (2-DCB), which is formed from palmitic acid. The formation of 2-DCB was evaluated in fresh irradiated ground beef patties at two fat levels. Patties containing 15% and 25% fat were irradiated by electron beam at 1.0, 2.0, 3.0, and 4.5 kGy. Commercially available 1-lb irradiated ground beef chubs with different fat levels were analyzed in order to estimate dose absorbed by these samples. The 2-DCB was extracted using supercritical fluid extraction (SFE) and analyzed by gas chromatography-mass spectroscopy (GC-MS) and was detected in all the irradiated samples. The concentration of 2-DCB increased linearly with dose with R2 = 0.9646 for 25% fat samples and R2 = 0.9444 for 15% fat samples. Further, there was no significant difference in 2-DCB concentrations between the two fat levels. The estimated doses applied to the commercial samples ranged between 1.38 and 1.55 kGy, values consistent with doses normally used in the industry (1.0-2.0 kGy). Our results show that 2-DCB can be used to monitor fresh irradiated beef and approximate the absorbed dose.     

Analysis of 2-alkylcyclobutanones with accelerated solvent extraction to detect irradiated meat and fish

A new analytical procedure has been developed to analyze 2-alkylcyclobutanones to detect gamma-ray-irradiated fat-containing foodstuffs. Samples were extracted with an accelerated solvent extraction system via hot and pressurized ethyl acetate in cells. A large amount of fat in the extract was precipitated and removed with filtration by standing at -20 degrees C after the addition of acetonitrile. The extract was further cleaned with a 1 g silica gel mini column, and the radiolytic compounds of 2-docecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutanone (2-TCB) were determined with gas chromatography with mass spectrometry (GC/MS). Sample preparation time before GC/MS was 7-8 h. At first, the procedure was evaluated with a recovery test in eight samples spiked with 2-DCB and 2-TCB at 20 ng/g, resulting in 70-105% recoveries with mostly less than 10% relative standard deviations. The procedure was further evaluated with beef, pork, chicken, and salmon samples irradiated with gamma-rays from 0.7 to 7.0 kGy at -19 degrees C. Both 2-DCB and 2-TCB in most samples were detected with good dose-response relations at all doses, while salmon was detected more than 2 kGy irradiation. The amounts of 2-alkylcyclobutanones produced reflected precursor fatty acids levels in samples, especially for the combination of 2-TCB and stearic acid. The results indicated that the production rate of 2-TCB to stearic acid was more obvious than that of 2-DCB to palmitic acid in frozen samples with gamma-ray irradiation.     

Effect of gamma-irradiation on phenylalanine ammonia-lyase activity, total phenolic content, and respiration of mushrooms (Agaricus bisporus)

Ionizing treatments were applied at 0.5 kGy, 1.5 kGy, and 2.5 kGy to edible mature mushrooms (Agaricus bisporus, albidus) in order to assess the effect of the gamma-irradiation on some biochemical parameters. Irradiation at doses of 1.5 kGy and 2.5 kGy reduced significantly (p < or = 0.05) the rate of respiration of the mushrooms, compared to that of samples irradiated at 0.5 kGy and nonirradiated control samples (C). Ionizing treatments increased significantly (p < or = 0.05) the phenylalanine ammonia-lyase (PAL) activity and total phenols concentration between days 1 and 4. From days 3-4, to the end of the storage period (day 12), both PAL activity and total phenols in the irradiated samples (I) collapsed to lower values. In contrast, the activity of polyphenol oxidase (PPO) increased until days 7, 9, and 12 for samples treated at 0.5, 1, and 2 kGy, respectively. Color measurements showed a loss of whiteness (L value) during storage. After day 4, however, the effectiveness of gamma-irradiation became apparent, and highest L values were obtained for I only.     

Identification of irradiated prawn (Penaeus monodon ) using thermoluminescence and 2-alkylcyclobutanone analyses

Thermoluminescence (TL) and 2-alkylcyclobutanone (2-ACB) analyses were performed to identify irradiated prawns ( Penaeus monodon ). With the TL method, minerals were extracted from prawns using acid hydrolysis. The experimental results satisfied the evaluation criteria of European Norm (EN) 1788, even after low-dose irradiation (0.5 kGy) and a 60 day storage at -20 °C. With the 2-ACB method, 2-dodecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutanone (2-TCB) were successfully extracted from prawns by direct solvent extraction with purification using a conventional silica column and a sulfoxide column, which was used for 2-ACB for the first time. Both 2-ACB derivatives were absent from the non-irradiated samples but were identified in all irradiated samples by gas chromatography-mass spectrometry. Moreover, 2-DCB and 2-TCB production correlated with the applied dose (2.5-10 kGy), and the correlation did not diminish after 60 days of storage at -20 °C for any dose. Therefore, these two techniques provide rapid, simple, and promising methods for routine investigation of frozen prawns.     

Detection of gamma-irradiated sesame seeds before and after roasting by analyzing photostimulated luminescence, thermoluminescence, and electron spin resonance

Sesame seeds were irradiated using a (60)Co irradiator (0-4 kGy) and then roasted (220 degrees C for 10 min). To identify the irradiation treatment, physical detection methods like photostimulated luminescence (PSL), thermoluminescence (TL), and electron spin resonance (ESR) have been investigated before and after roasting. The photon counts of the irradiated samples (nonroasted and roasted) were higher than those of nonirradiated ones, making it possible to distinguish the two samples. The threshold values of nonroasted and roasted samples increased linearly with the irradiation dose, respectively. The TL for the nonirradiated nonroasted and roasted samples presented a lower peak at about 300 degrees C, but irradiated samples showed a higher peak at around 150 degrees C. The areas of TL glow curves were 15 times higher in nonroasted as compared with roasted samples. TL ratio [integrated area of TL 1 (the first glow)/TL 2 (the second glow)] obtained by the reirradiation step was 0 in nonirradiated samples and more than 0.15 in irradiated samples. The radiation-induced ESR signals originating from cellulose were determined in irradiated samples before and after roasting.     

Effect of 60Co-irradiation on postharvest quality and selected enzyme activities of Hypsizygus marmoreus fruit bodies

Hypsizygus marmoreus fruit bodies were exposed to different doses of (60)Co gamma-irradiation, stored at 4 degrees C and 65-70% relative humidity, and various physiological changes associated with postharvest deterioration, as well as the activities of selected enzymes widely considered to play a role in the process of senescence, were monitored over a subsequent storage period of 25 days. Exposure to 0.8 kGy irradiation was clearly beneficial in maintaining the postharvest appearance of the mushroom sporophores compared to non-irradiated samples and fruit bodies exposed to higher doses (1.2-2.0 kGy) of irradiation. Samples treated with 0.8 kGy also exhibited smaller initial declines in soluble protein, smaller increases in reducing sugar content, and lower levels of malondialdehyde accumulation during the early storage period. Smallest increases in proteinase activity were recorded in samples dosed with 0.8 and 2.0 kGy, and levels of superoxide dismutase were significantly higher in samples exposed to 0.8 kGy compared with non-irradiated controls. Large initial increases in catalase activity were detected in samples irradiated with 0.8, 1.2, and 1.6 kGy and, although enzyme levels gradually decreased in all samples during further storage, residual levels after 25 days were still severalfold higher in irradiated samples compared with controls. The data increase the current understanding of the effects of gamma-irradiation on the biochemical changes associated with postharvest senescence and should lead to more targeted strategies for reducing postharvest quality loss in H. marmoreus and other mushrooms.     

Effects of Electron-Beam Radiation on Nutritional Parameters of Portuguese Chestnuts ( Castanea sativa Mill.)

Chestnuts are a widely consumed fruit around the world, with Portugal being the fourth biggest producer in Europe. Storage of these nuts is an important step during processing, and the most widely used fumigant was banned in the European Union under the Montreal Protocol because of its toxicity. Recently, radiation has been introduced as a cheap and clean conservation method. Previous studies of our research group proved that γ radiation had no negative effect on the nutritional value of chestnuts; in fact, storage time had a much bigger influence on the chestnut quality. In the present study, we report the effect of a less ionizing radiation, electron beam, with doses of 0, 0.5, 1, 3, and 6 kGy in the nutritional value of chestnuts (ash, energy, fatty acids, sugars, and tocopherols), previously stored at 4 °C for 0, 30, and 60 days. The storage time seemed to reduce fat and energetic values but reported a tendency for higher values of dry matter. With regard to fatty acids, there was a higher detected quantity of C20:2 in non-irradiated samples and four fatty acids were only detected in trace quantities (C6:0, C8:0, C10:0, and C12:0). γ-Tocopherol decreased during storage time but did not alter its quantity for all of the radiation doses (as like α-, β-, and δ-tocopherol); in fact, these compounds were present in higher concentrations in the irradiated samples. Sucrose and total sugars were lower in non-irradiated samples, and raffinose was only detected in irradiated samples. Electron-beam irradiation seems to be a suitable methodology, because the effects on chemical and nutritional composition are very low, while storage time seems to be quite important in chestnut deterioration.     

Evaluation of 2-alkylcyclobutanones in irradiated cured pork products during vacuum-packed storage

The 2-Alkylcyclobutanones (2-ACBs) content was determined in three Italian cured pork products (salame Milano, coppa, and pancetta) irradiated at different targeted irradiation doses (2, 5, and 8 kGy) during vacuum-packed storage. Among 2-ACBs, three different compounds were investigated, namely, 2-dodecylcyclobutanone, 2-tetradecylcyclobutanone, and 2-(tetradec-5'-enyl)cyclobutanone. 2-ACBs were absent from the nonirradiated samples, whereas their content increased with irradiation dose. Their presence was recorded occasionally at 2 kGy and constantly at higher irradiation doses (5 and 8 kGy). The plot of 2-ACBs content against targeted irradiation doses showed an exponential relationship. The effect of vacuum-packed storage time on the 2-ACBs content was dependent on the irradiation dose. During vacuum-packed storage for up to 60 days, the 2-ACBs content remained unchanged in the cured pork products irradiated at 2 and 5 kGy, whereas a significant increase was observed in the pork products irradiated at 8 kGy.     

Oil quality and sensory evaluation of almond (Prunus amygdalus) stored after electron beam processing

The changes in the lipid fraction and the deterioration of its quality were studied in almonds (Prunus amygdalus) of the variety Guara after treatment with accelerated electrons at doses of 3, 7, and 10 kGy, during a storage period of 5 months. In almond oil, the most significant difference from the nutritional point of view was seen in the fatty acid linolenic (18:3), which shows at 3 kGy a maintenance of the initial content during the whole storage period, whereas, at 7 and 10 kGy, the content in 18:3 disappears from the first moment. The quality indices of the oil (K(232), K(270)) decreased at all doses and remained stable during the time of storage. The peroxide value did not show changes at the doses of 3 and 7 kGy, in non-irradiated samples, but significantly increased when the maximum dose of 10 kGy was applied. These changes were reflected in the sensory analysis, in which the tasters did not find sensory differences between the controls and those irradiated at doses of 3 or 7 kGy, whereas almonds irradiated at 10 kGy exhibited a rancid flavor and a significant decrease in general quality.     

Effect of gamma and e-beam radiation on the essential oils of Thymus vulgaris thymoliferum, Eucalyptus radiata, and Lavandula angustifolia

The microbiological contamination of raw plant materials is common and may be adequately reduced by radiation processing. This study evaluated the effects of gamma- and e-beam ionizing radiations (25 kGy) on three plants used as food or as medicinal products (Thymus vulgaris L., Eucalyptus radiata D.C., and Lavandula angustifolia Mill.) as well as their effects on extracted or commercial essential oils and pure standard samples. Comparison between irradiated and nonirradiated samples was performed by GC/FID and GC/MS. At the studied doses, gamma and e-beam ionizing radiation did not induce any detectable qualitative or quantitative significant changes in the contents and yields of essential oils immediately after ionizing radiation of plants or commercial essential oils and standards. As the maximum dose tested (25 kGy) is a sterilizing dose (much higher than doses used for decontamination of vegetable drugs), it is likely that even decontamination with lower doses will not modify yields or composition of essential oils of these three plants.     

Determination of 2-alkylcyclobutanones with electronic impact and chemical ionization gas chromatography/mass spectrometry (GC/MS) in irradiated foods

The use of a column containing 60 g of silica gel for cleanup and the use of isobutane as a reactant reagent for chemical ionization-mass spectrometric analysis of the saturated and monounsaturated alkyl side-chain 2-alkylcyclobutanones (2-ACBs; specifically induced by irradiation from fat in foods until the proof of contrary) has improved both the sensibility and the selectivity of the method when applied for the detection of irradiated foods. The quality of the chromatograms obtained was improved, allowing the detection of food samples (avocados) irradiated at low doses (0.1 kGy) or irradiated ingredients included in low proportions (less than 5%, wt/wt) in nonirradiated culinary foods. These analytical modifications for the detection of 2-ACBs on the official EN 1785 method enable an extension of its current field of application using common equipment of food quality control laboratories.     

Combined effect of ascorbic acid and gamma irradiation on microbial and sensorial characteristics of beef patties during refrigerated storage

The present study was undertaken to evaluate the effect of ascorbic acid concentrations (0.03 to 0.5%) and irradiation doses (0.5 to 4 kGy) on microbial growth, color coordinates (L, a, and b), and sensory characteristics (taste and odor) of beef patties during storage at 4 +/- 1 degrees C. Ascorbic acid was also compared to citric acid at a similar pH value in order to differentiate the effects of ascorbic acid from those of pH reduction. Results showed significant reduction (p< or = 0.05) of aerobic plate counts (APCs) and total coliforms, and a significant interaction (p< or = 0.05) between ascorbic acid and irradiation dose was observed. The irradiation treatment had detrimental effects on redness, yellowness, and hue angle values of meat. However, incorporation of ascorbic acid into the meat before irradiation resulted in significant (p< or = 0.05) stabilization of color parameters. The color improvement obtained with ascorbic acid was not related to the pH reduction. Also, no significant detrimental effect on taste or odor was found in irradiated samples containing ascorbic acid.     

ESR法定量检测干果类辐照食品

以葵花籽、核桃、开心果和榛子为试验材料,研究在0、1.0、3.0、5.0和10.0kGy剂量范围内4种样品辐照剂量与信号强度的关系以及相关性.结果表明:4种样品在辐照前后ESR波谱有明显区别,其信号强度与辐照剂量均呈正相关.辐照后样品的ESR强度和谱形都发生变化,应用ESR法能够鉴定4种食品是否经过辐照.其中核桃和开心...     

Identification of irradiated cashew nut by electron paramagnetic resonance spectroscopy

Cashew nut samples were irradiated at gamma-radiation doses of 0.25, 0.5, 0.75, and 1 kGy, the permissible dose range for insect disinfestation of food commodities. A weak and short-lived triplet (g = 2.004 and hfcc = 30 G) along with an anisotropic signal (g perpendicular = 2.0069 and g parallel = 2.000) were produced immediately after irradiation. These signals were assigned to that of cellulose and CO 2 (-) radicals. However, the irradiated samples showed a dose-dependent increase of the central line (g = 2.0045 +/- 0.0002). The nature of the free radicals formed during conventional processing such as thermal treatment was investigated and showed an increase in intensity of the central line (g = 2.0045) similar to that of irradiation. Characteristics of the free radicals were studied by their relaxation and thermal behaviors. The present work explores the possibility to identify irradiated cashew nuts from nonirradiated ones by the thermal behaviors of the radicals beyond the period, when the characteristic electron paramagnetic resonance spectral lines of the cellulose free radicals have essentially disappeared. In addition, this study for the first time reports that relaxation behavior of the radicals could be a useful tool to distinguish between roasted and irradiated cashew nuts.     

Volatile sulfur compounds in irradiated precooked turkey breast analyzed with pulsed flame photometric detection

Ionizing radiation is an effective processing technology for pathogen inactivation on various foods. However, the generation of off-odor is a concern for some irradiated meats. This study was conducted to investigate volatile sulfur compounds of precooked ready-to-eat turkey breast as functions of radiation dose and subsequent storage. Precooked turkey breast was exposed to 0, 1, 2, 3, 4, and 5 kGy of gamma radiation and stored for 14 days at 5 degrees C. Volatile sulfur compounds were extracted using solid phase microextraction (SPME), followed by gas chromatographic separation and pulsed flame photometric detection. Irradiation dramatically increased concentrations of hydrogen sulfide, sulfur dioxide, methanethiol, and dimethyl disulfide. The rate of increase was higher at low doses (0-2 kGy) than at higher doses of 3-5 kGy. Carbon disulfide was the only volatile sulfur compound that was reduced by irradiation. Concentrations of all volatile sulfur compounds decreased in both irradiated and nonirradiated samples stored at 5 degrees C.     

电子鼻结合HS-SPME-GC-MS分析辐照对甲鱼预制菜挥发性风味成分的影响

为了探究辐照对甲鱼预制菜挥发性风味成分的影响,采用0、4.7、7.1、9.9 kGy剂量⁶⁰Co-γ射线辐照处理甲鱼预制菜,通过感官评定并利用电子鼻结合顶空固相微萃取-气相色谱-质谱联用技术(HS-SPME-GC-MS)分析辐照前后挥发性风味成分的变化。结果表明,5 kGy以内剂量辐照对甲鱼预制菜的感官品质无明显影响,高于5 kGy会影响其气味和滋味,进而可能产生异味。辐照对甲鱼预制菜特征气味引起的差异主要表现在传感器响应值较高的芳香成分和有机硫化物、氮氧化合物、甲烷、醇类和醛酮类物质,利用主成分分析(PCA)和线性判别分析(LDA)能够有效区分经不同剂量辐照处理的甲鱼预制菜。4.7 kGy组与对照组气味差异较小,7.1、9.9 kGy组与对照组差异相对较大。不同剂量辐照后甲鱼预制菜挥发性成分的种类增加,醛类、酸类相对含量降低,烃类、芳香族类、酮类、酯类相对含量升高,醇类、含氮含硫及杂环类相对含量先降低后升高。相对气味活度值(ROAV)分析结果表明,壬醛、癸醛、辛醛、己醛、庚醛、1-辛烯-3-醇和2-戊基呋喃是甲鱼预制菜的关键风味成分,苯甲醛、苯乙醛、(E,E)-2, 4-癸二烯醛对其风味具有重要修饰作用。辐照后辛醛、己醛、1-辛烯-3-醇对甲鱼预制菜总体风味的贡献率降低,庚醛、2-戊基呋喃的贡献率先降低后增加,苯乙醛的贡献率增加并成为关键风味成分。因此,建议采用不超过5 kGy剂量的⁶⁰Co-γ射线辐照处理甲鱼预制菜,在杀菌的同时最大限度保持其原有风味。本研究结果为辐照技术在甲鱼预制菜杀菌保鲜中的应用提供了科学依据。     

Analysis of gamma-irradiated melon, pumpkin, and sunflower seeds by electron paramagnetic resonance spectroscopy and gas chromatography-mass spectrometry

Seeds of melon (Citrullus lanatus var. sp.), pumpkin (Cucurbita moschata), and sunflower (Heliantus annus) were gamma-irradiated at 1, 3, 5, and 10 kGy and analyzed by electron paramagnetic resonance (EPR) and gas chromatography-mass spectrometry (GC-MS) according to EN1787:2000 and EN1785:2003, respectively. Distinguishable triplet signals due to the presence of induced cellulose radicals were found at 2.0010-2.0047 g in the EPR spectra. The gamma-irradiated radiolytic markers of 2-dodecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutanone (2-TCB) were identified in all irradiated seed samples. Both the free radicals and the alkylcyclobutanones were found to increase with irradiation dose. In general, linear relationships between the amount of radicals and irradiation dosage could be established. Studies at an ambient temperature (20-25 degrees C) in a humidity-controlled environment showed a complete disappearance of the cellulosic peaks for irradiated samples upon 60 days of storage. Such instability behavior was considered to render the usefulness of using EPR alone in the determination of irradiated seed samples. On the other hand, 2-DCB and 2-TCB were also found to decompose rapidly (>85% loss after 120 days of storage), but the radiolytic markers remained quantifiable after 120 days of postirradiation storage. These results suggest that GC-MS is a versatile and complimentary technique for the confirmation of irradiation treatment to seeds.     

Determination of monounsaturated alkyl side chain 2-alkylcyclobutanones in irradiated foods

The 2-alkylcyclobutanones (2-ACBs) are formed from triglycerides by irradiation treatment and may be used as markers for this type of food processing. This paper describes a detection method for the analysis of monounsaturated alkyl side chain 2-ACBs, which is formed upon irradiation from monounsaturated fatty acids which frequently are the most abundant fatty acids in foods. The estimated radioproduction yields of the cis-2-(dodec-5'-enyl)-cyclobutanones (cis-2-dDeCB) and the cis-2-(tetradec-5'-enyl)-cyclobutanones (cis-2-tDeCB) were 1.0 +/- 0.5 and 0.9 +/- 0.2 nmol.mmol(-1) precursor fatty acid.kGy(-1), respectively, being similar to that of saturated 2-ACBs. The stability study of the s- and mu-2-ACBs in poultry meat samples irradiated at 10 kGy and stored for 3-4 weeks at 4 degrees C and 25 degrees C showed that these compounds undergo some transformation, their amounts being reduced by about 50%. These storage losses did not depend on the saturation state of the alkyl side chain. The EI-MS detection limit of 2-tDeCB is 3 times higher (0.6 pmol) than that of 2-dodecylcyclobutanone (0.2 pmol). Consequently, when the oleic acid content of the analyzed food exceeds the content of palmitic acid by a factor of 3, it would be of an advantage to apply 2-tDeCB as a marker for detection of the irradiation treatment.     

Policosanol content and composition in perilla seeds

Policosanols, long-chain alcohols, have many beneficial physiological activities. Contents and compositions in perilla seeds (Perilla frutescens) produced in Korea and China were determined. Waxy materials were extracted from perilla seeds using hot hexane. Yield of the waxy materials from perilla seeds was 72.1 mg/100 g of dry weight. Contents and compositions of the waxy materials and policosanols were identified and quantified by TLC, HPLC, and GC. Major components of the waxy materials from Korean and Chinese perilla seeds were policosanols (25.5 and 34.8%, respectively), hydrocarbons (18.8 and 10.5%), wax esters, steryl esters and aldehydes (53.0 and 49.8%), acids (1.7 and 2.1%), and triacylglycerols (1.0 and 2.9%), determined by HPLC. For comparison, waxy materials of sesame seeds were also analyzed. Yield of the waxy materials from sesame seeds were 8.6 mg/100 g. Less than 5% policosanols were detected in the waxy materials extracted from sesame seeds produced in Korea and China. Wax esters or steryl esters accounted for 93-95% of the sesame waxy materials. Policosanols in the perilla seeds were composed of 67-68% octacosanol, 16-17% hexacosanol, 6-9% triacontanol, and others.