Detection of potentially allergenic hazelnut (Corylus avellana) residues in food: a comparative study with DNA PCR-ELISA and protein sandwich-ELISA

Allergen detection is of increasing interest for food labeling purposes. A comparative study with a commercial hazelnut-specific PCR-ELISA and a sandwich-type ELISA detecting hazelnut protein was performed to investigate to what extent immunochemical and DNA-based techniques would correlate in the detection of trace amounts of potentially allergenic hazelnut residues. Both methods were highly sensitive and allowed the detection of even <10 ppm of hazelnut in complex food matrixes. The protein-ELISA was highly specific for hazelnut. However, some foods could lead to false-positive results at the 10 ppm level. The PCR-ELISA did not show any cross-reactions with non-hazelnut foods, thus reducing the probability of having false positives at the trace level. Forty-one commercial food products with and without hazelnut components on their labels were analyzed for the presence of hazelnut. Of the 27 products in which hazelnut components were detected, two samples were not identified by the protein-ELISA, and only one sample, namely one white chocolate having <1 ppm of hazelnut protein, was not detected by PCR-ELISA. The good correlation of the results of PCR-ELISA and protein-ELISA suggested that both PCR-based and immunochemical techniques are suitable for reliable detection of potentially allergenic hazelnut residues in foods at the trace level.     

Volumetric method for determing traces of organic chlorine in lipids.

The official AOAC method for determining chlorine in cake flour lipids has been modified to determine chlorine in lipids of animal carcasses cooled with chlorinated water. The sensitivity of the method has been improved 1000-fold to attain a detection limit of 2 ppm Cl in lipids. Recovery of the fine silver chloride precipitate was improved through centrifugation rather than filtration. The official method also did not take into consideration that when the silver chloride precipitate is exposed to light, it is converted to free chlorine and silver. Recovery of organic chlorine ranged from 84.6% at 4.1 ppm to 100.1% at 100 ppm. A number of samples of commercially chlorinated beef, pork, and chicken and laboratory-chlorinated chicken were analyzed by this method. In all cases where the level of chlorination was sufficient to result in a chlorine level in lipids in excess of 2 ppm, the chlorine content of the carcass lipids was measurable.     

Development of a real-time PCR and a sandwich ELISA for detection of potentially allergenic trace amounts of peanut (Arachis hypogaea) in processed foods

Hidden allergens in food products are, especially for peanut-allergic consumers, a serious problem because even low amounts (approximately 200 microg) of peanut can elicit allergic reactions. Undeclared peanut traces can be found in processed food products, because contaminations with peanut during production processes are frequent. To minimize the risk of such cross-contaminations, it is necessary to develop sensitive analytical methods for the detection of hidden allergens in foods. For this approach we developed two peanut-specific assays based on the detection of peanut protein by specific antibodies (sandwich ELISA) and by the detection of peanut-specific DNA (part of the coding region of Ara h 2) by a real-time PCR. Both tests did not show any cross-reactivity with 22 common food ingredients (cereals, nuts, legumes), and the limit of detection is <10 ppm peanut in processed foods. Thirty-three random samples of food products were tested for the presence of peanut to compare both assay types with each other and to evaluate the percentage of foods on the German market that are contaminated with peanut traces. We found that four products (13.3%) without peanut in the list of ingredients contained peanut protein in a range from 1 to 74 ppm peanut protein and that the results of both tests correlated well. The real-time PCR was able to detect one more positive sample than the sandwich ELISA. In conclusion, both assays are sensitive and specific tools for the detection of hidden allergens in processed foods.     

Fluoride content of foods made with mechanically separated chicken

The goal of the present study was to determine the extent to which foods made with mechanically separated chicken can contribute to total fluoride intake. Fluoride content of each blended sample was determined with a fluoride combination electrode following perchloric-acid-facilitated diffusion of hydrogen fluoride. Infant foods had the highest fluoride content followed by chicken sticks, luncheon meats, and canned meats. A single serving of chicken sticks alone would provide about half of a child's upper limit of safety for fluoride. Fluoride content of foods made with mechanically separated chicken was significantly correlated with calcium content, which is consistent with the possibility that the mechanical separation process was the source of the extra fluoride. Foods made with mechanically separated turkey were not a major source of fluoride.     

Chromatographic methods for determination of macrolide antibiotic residues in tissues and milk of food-producing animals

Tylosin, an antibiotic developed specifically for agricultural use, and erythromycin are the main macrolide antibiotics used in animal production. Two-dimensional thin layer chromatography has been used for detection of tylosin in poultry meat, eggs, and milk and for erythromycin in poultry meat. Detection limits reported are, for tylosin, 0.1 ppm in poultry meat, 0.05 ppm in egg, and 0.01 ppm in milk, and for erythromycin, 0.25 ppm in poultry meat. Liquid chromatography (LC) has also been used for determination of tylosin in milk, blood, and tissues of animals. Samples (milk, blood serum, or tissue homogenates in water or pH 2.2 buffer) were deproteinized with acetonitrile, tylosin was partitioned into methylene chloride, and the extracts were concentrated and dissolved in acetonitrile. Chromatography was done on a reverse phase end-capped C18 column using 0.002-0.005 M ammonium dihydrogen phosphate-acetonitrile-methanol (10 + 60 + 30-5 + 80 + 15). Solvent composition was varied with the type of sample analyzed. The method will detect 0.1 ppm tylosin in tissues and less in milk and blood serum. The LC method was more sensitive than microbiological assays for detection of tylosin in tissues of treated swine; recoveries of tylosin by the LC method were frequently several-fold higher.     

Antigenic stability of pecan [Carya illinoinensis (Wangenh.) K. Koch] proteins: effects of thermal treatments and in vitro digestion

Rabbit polyclonal antibody-based inhibition ELISA as well as immunoblotting analyses of proteins extracted from variously processed pecans (cv. Desirable) indicate that pecan proteins are antigenically stable. Pecan antigens were more sensitive to moist heat than dry heat processing treatments. SDS-PAGE and immunoblotting analysis of the native and heat-denatured proteins that were previously subjected to in vitro simulated gastric fluid digestions indicate that stable antigenic peptides were produced. Both enzyme-to-substrate ratio and digestion time were influential in determining the stability of pecan polypeptides. The stable antigenic polypeptides may serve as useful markers in developing assays suitable for the detection of trace amounts of pecans in foods.     

规模化蛋鸡场现代化生产管理系统的建立与应用

为建立一个我国规模化蛋鸡场适用的生产管理系统，综合运用了多媒体技术、网络化管理、人工智能和自动化监控技术方法。通过人工神经网络与专家系统有机结合，改进鸡疾病专家系统；创建免疫接种备忘系统。由生产统计、生产计划、饲料配方、饲料库存、免疫接种和鸡病诊断专家系统构建成网络化软件系统。各子系统在数据库级别上达到企业数据资源的共享和利用，也可根据用户应用需求单独使用其中的子系统。同时，利用传感器、工控机和摄像机对鸡舍环境实行全方位的温湿度监测、移动摄像和鸡舍设备自动控制，实现鸡场远程管理。结果表明：系统高度集成，运     

Leak in evaporated milk by anodic stripping voltammetry and atomic absorption spectrophotometry: cooperative interlaboratory study

The results of a cooperative study on the determination of lead in evaporated milk, using a double blind referee technique, are reported. This study was designed to determine the normal variability of methods currently used for lead analysis by canned food industry laboratories. Twenty-three laboratories participated in this study. Each laboratory was instructed to use atomic absorption spectrophotometry (AOAC 25.065), anodic stripping voltammetry, or carbon rod atomic absorption spectrophotometry. Overall, the results appear to be in close agreement with the spiking levels. The coefficient of variation for all laboratories was 36.0% at the 0.15 ppm lead level and 16.8% at the 0.40 ppm lead level.     

Gas chromatographic determination of fenvalerate insecticide residues in processed tomato products and by-products

The results of a 5-laboratory collaborative determination of residues of the synthetic pyrethroid insecticide fenvalerate in tomato products are presented. Tomatoes from plants treated in the field at 2-4 day intervals (13 foliar applications) were processed into chopped fresh tomatoes, canned quarters, juice, paste, and the by-product skins plus seeds. Gas chromatographic analysis of the commodities for fenvalerate showed the fresh produce to contain 0.26 ppm, and the skins plus seeds contained 1.9 ppm. Residues were barely detectable in canned peeled quarters and juice, but averaged 0.12 ppm for paste, the concentration product of juice. High residues were associated with the skin content of the product. Five laboratories using modifications of the same analytical technique obtained good collaborative agreement.     

Enzyme-linked immunosorbent assay of benomyl and thiabendazole in some foods

Enzyme-linked immunosorbent assays were developed for benomyl as its decomposition product, methyl 2-benzimidazole carbamate, and thiabendazole in foods. Immunogens consisting of human serum albumin coupled to 2-succinamidobenzimidazole or 2-(2'-succinamido-4'-thiazolyl)benzimidazole were used in rabbits to raise antisera that were specific for the respective fungicides. Lower limits of quantitation of 0.35 ppm for benomyl and 0.03 ppm for thiabendazole were established without cleanup of the ethyl acetate extract. Recoveries of benomyl from 3 crops spiked at 0.5 to 10 ppm averaged 89% (range 73-109%) and of thiabendazole from 5 crops spiked at 0.1 to 2.0 ppm were 93% (range 81-105%).     

Survey of infant foods for Clostridium botulinum spores

A total of 236 samples of infant foods, including honey, dry cereal, nonfat dry milk, evaporated milk, canned formula, and canned baby food, were collected in the New York City area and tested for the presence of Clostridium botulinum spores. Methods for recovery of spores were validated using foods spiked with 4 spores/mL or g. None of the products contained C. botulinum spores, indicating that their incidence in these commercial foods is not widespread. This limited study did not identify any food types that could be suspected of being involved in the transmission of infant botulism.     

Differential detection of shrimp and crab for food labeling using polymerase chain reaction

Shrimp and crab are well-known as allergenic ingredients. According to Japanese food allergy labeling regulations, shrimp species (including prawns, crayfishes, and lobsters) and crab species must be differentially declared when ≥10 ppm (total protein) of an allergenic ingredient is present. However, the commercial ELISA tests for the detection of crustacean proteins cannot differentiate between shrimp and crab. Therefore, two methods were developed to discriminate shrimp and crab: a shrimp-PCR method with postamplification digestion and a crab-PCR method that specifically amplifies a fragment of the 16S rRNA gene. The sensitivity and specificity of both PCR methods were verified by experiments using DNA extracted from 15 shrimp species, 13 crab species, krill, mysid, mantis shrimp, other food samples (cephalopod, shellfish, and fish), incurred foods, and commercial food products. Both PCR methods could detect 5 pg of DNA extracted from target species and 50 ng of genomic DNA extracted from incurred foods containing 10 ppm (μg/g) total protein of shrimp or crab. The two PCR methods were considered to be specific enough to separately detect species belonging to shrimp and crab. Although false-positive and false-negative results were obtained from some nontarget crustacean species, the proposed PCR methods, when used in conjunction with ELISA tests, would be a useful tool for confirmation of the validity of food allergy labeling and management of processed food safety for allergic patients.     

Concentration of bisphenol A in highly consumed canned foods on the U.S. market

Metal food and drink cans are commonly coated with epoxy films made from phenolic polymers produced from bisphenol A (BPA). It is well established that residual BPA monomer migrates into can contents during processing and storage. While a number of studies have reported BPA concentrations in foods from foreign markets and specialty foods on the U.S. market, very few peer-reviewed data for the BPA concentrations in canned food from the U.S. market were available. This study quantified BPA concentrations in 78 canned and two frozen food products from the U.S. market using an adaptation of a previously reported liquid chromatography-tandem mass spectrometry method. The tested products represented 16 different food types that are from the can food classifications that constitute approximately 65% of U.S. canned food sales and canned food consumption. BPA was detected in 71 of the 78 canned food samples but was not detected in either of the two frozen food samples. Detectable BPA concentrations across all foods ranged from 2.6 to 730 ng/g. Large variations in BPA concentrations were found between different products of the same food type and between different lots of the same product. Given the large concentration ranges, the only distinguishable trend was that fruits and tuna showed the lowest BPA concentrations. Experiments with fortified frozen vegetables and brine solutions, as well as higher BPA concentrations in canned food solids over liquid portions, clearly indicated that BPA partitions into the solid portion of foods.     

Volatile organic compounds in foods: a five year study

A purge and trap procedure was used with gas chromatography-mass spectrometry determination to analyze 70 foods for volatile organic compounds (VOCs). The results from analyses over a 5 year period (1996-2000) are reported. VOCs were found in at least one sample of all foods tested, although no single compound was found in each of the foods. The total amount of VOCs found in a single food item over the 5 year period ranged from 24 to 5328 ppb, with creamed corn (canned) the lowest and cheddar cheese the highest. Benzene was found in all foods except American cheese and vanilla ice cream. Benzene levels ranged from 1 to 190 ppb, with the highest level found in fully cooked ground beef. Benzene was found in 12 samples of cooked ground beef, with an average of 40 ppb. Benzene levels above 100 ppb were also seen in at least one sample each of a cola (138 ppb), raw bananas (132 ppb), and cole slaw (102 ppb). This compares to a maximum contaminant level of 5 ppb set by the U.S. EPA for drinking water.     

Determination of fluoride in deboned meat

A rapid analytical procedure was investigated for measuring the fluoride content of deboned meat, using a specific ion electrode. The method involves a defatting procedure followed by chelation with EDTA before final solubilization and measurement. Fluoride content is determined by comparison to a standard curve on 2-cycle semi-logarithmic graph paper of fluoride ion vs. millivolt readings. The method is applicable to hand and mechanically deboned pork, beef, and poultry within a range of from 0.7 to 25.0 ppm fluoride. Higher concentrations may be measured by analyzing additional standards and extending the curve into a third cycle. The curve is not linear below 0.7 ppm. Samples less than 0.7 ppm may be estimated by extrapolation. Studies with fortified samples indicate average recoveries of 92 +/- 4.2%. The standard deviation of duplicates within the laboratory is 0.5 ppm in the range stated above.     

Detection and stability of the major almond allergen in foods

Almond major protein (AMP or amandin), the primary storage protein in almonds, is the major allergen recognized by almond-allergic patients. A rabbit antibody-based inhibition ELISA assay for detecting and quantifying AMP in commercial foods has been developed, and this assay, in conjunction with Western blotting analyses, has been applied to the investigation of the antigenic stability of AMP to harsh food-processing conditions. The ELISA assay detects purified AMP at levels as low as 87 +/-16 ng/mL and can detect almond at between 5 and 37 ppm in the tested foods. The assay was used to quantify AMP in aqueous extracts of various foods that were defatted and spiked with known amounts of purified AMP or almond flour. In addition, AMP was quantified in commercially prepared and processed almond-containing foods. Neither blanching, roasting, nor autoclaving of almonds markedly decreased the detectability of AMP in subsequent aqueous extracts of almonds. Western blots using both rabbit antisera and sera from human almond-allergic patients confirm a general stability of the various peptides that comprise this complex molecule and show that the rabbit antibody-based assay recognizes substantially the same set of peptides as does the IgE in sera from almond-allergic patients.     

Comparison of paired-ion liquid chromatographic method with AOAC fluorometric and microbiological methods for riboflavin determination in selected foods

A paired-ion liquid chromatographic (LC) technique coupled with fluorometric detection to determine riboflavin in various food matrices is described. Chromatograms of many foods showed 2 peaks of interest due to presence of riboflavin and flavin mononucleotide (FMN). Relatively high levels of FMN were found in raw beef, corned beef, chicken liver, and canned mushrooms. When riboflavin and FMN contents were summed, LC values were comparable to those obtained by the AOAC standard procedures. The LC technique was sensitive, rapid, and simple, yielding a mean standard deviation of 3.1% which was comparable to the AOAC fluorometric method (3.0%) and better than the AOAC microbiological assay (9.6%). Mean spike recoveries were 91.8% for LC compared to 90.5% and 89.6% for the AOAC fluorometric and microbiological methods, respectively.     

病死猪酶解及超声波预处理工艺优化

病死畜禽资源化利用是解决病死畜禽污染的一条重要途径。为探索病死猪肉酶解工艺条件,该文以猪肉为原料,以胰蛋白酶为试验用酶,以水解度为指标,选取加酶量、底物浓度、pH值、温度作为试验因素,通过单因素试验初步确定了胰蛋白酶的水解条件,并分析了各因素对酶解反应的影响规律。应用Box-Behnken中心组合设计建立数学模型,以水解度为响应值,进行了四因素三水平的响应面优化试验,确定了最佳酶解工艺条件,并通过响应面模型的曲面图直观地分析了各影响因素之间的交互作用。在此基础上,探索频率为20 k Hz,功率为500 W的超声波预处理猪肉20 min对酶解效果的影响,并应用扫描电子显微镜在微观结构上对其原因进行了分析。结果表明,各因素对酶解反应的影响大小依次为:加酶量温度pH值底物浓度,在试验范围内得到的酶解最佳工艺条件为:加酶量为1.15%(质量分数)、底物质量浓度为80.5 g/L、pH值为7.96、温度为40.6℃,酶解1 h的预测水解度可达16.74%,验证试验水解度为16.77%,表明试验结果与软件分析结果相符,最佳水解时间为6 h,此时的水解度为28.91%。超声波预处理后,最佳水解时间为4 h,水解度达到32.86%,因此超声波预处理能缩短水解周期2 h,提高水解度4个百分点。由此可见,应用超声波预处理可以提高酶解效率,缩短工作时间。     

Analysis of the early stages of lipid oxidation in freeze-stored pork back fat and mechanically recovered poultry meat

An analytical method that can detect low levels of oxidation in food earlier than a sensory panel would be a valuable tool for food manufacturers as well as research institutes. Two model matrixes, pork back fat and mechanically recovered poultry meat (MRPM), were freeze-stored in air at -20 degrees C for 26 weeks. Peroxide value, thiobarbituric acid reactive substances, volatiles analyzed with dynamic headspace gas chromatography-mass spectrometry (GC-MS) and a gas-sensor array technique (electronic nose), chemiluminescence, and front-face fluorescence were evaluated against sensory analysis with regard to detection of early oxidation and correlation with sensory data. Fluorescence and GC-MS could detect oxidative changes in pork back fat earlier than the sensory panel and the electronic nose at the same time. The three methods were highly correlated with sensory attributes (r = 0.8-0.9). GC-MS gave the best results with regard to detection of small oxidative changes in MRPM.     

辐照对冷却猪、鸡肉中细菌ATP发光强度的影响

采用ATP发光法检测辐照对冷却猪、鸡肉ATP发光强度的影响,并对其他试验因素是否对样品ATP发光强度产生影响进行了分析。结果表明:ATP标准品的浓度与发光强度呈显著的正相关,ATP标准品发光强度与辐照剂量表现为显著的负相关;辐照冷却肉的细菌ATP发光强度随辐照剂量的变化趋势呈现为倒"S"形特征,其中6.0kGy辐照时发光强度最大,4.0、8.0kGy剂量辐照时强度最小;无菌水、灭菌样品未对ATP发光强度造成干扰,而未辐照大肠杆菌浓度与ATP发光强度之间呈显著线性正相关,相关系数达到0.9437。