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1.
Legislation requires labeling of foods containing allergenic ingredients. Here, we present a robust 10-plex quantitative and sensitive ligation-dependent probe amplification method, the allergen-multiplex ligation-dependent probe amplification (MLPA) method, for specific detection of eight allergens: sesame, soy, hazelnut, peanut, lupine, gluten, mustard, and celery. Ligated probes were amplified by polymerase chain reaction (PCR), and amplicons were detected using capillary electrophoresis. Quantitative results were obtained by comparing signals with an internal positive control. The limit of detection varied from approximately 5 to 400 gene copies, depending on the allergen. The method was tested using different foods spiked with mustard, celery, soy, or lupine flour in the 1-0.001% range. Depending on the allergen, sensitivities were similar or better than those obtained with qPCR. The allergen-MLPA method is modular and can be adapted by adding probe pairs for other allergens. The DNA-based allergen-MLPA method will constitute a complementary method to the traditional protein-based methods.  相似文献   

2.
Hazelnut is one of the most commonly consumed tree nuts, being largely used by the food industry in a wide variety of processed foods. However, it is a source of allergens capable of inducing mild to severe allergic reactions in sensitized individuals. Hence, the development of highly sensitive methodologies for hazelnut traceability is essential. In this work, we developed a novel technique for hazelnut detection based on a single-tube nested real-time PCR system. The system presents high specificity and sensitivity, enabling a relative limit of detection of 50 mg/kg of hazelnut in wheat material and an absolute limit of detection of 0.5 pg of hazelnut DNA (1 DNA copy). Its application to processed food samples was successfully achieved, detecting trace amounts of hazelnut in chocolate down to 60 mg/kg. These results highlight the adequacy of the technique for the specific detection and semiquantitation of hazelnut as potential hidden allergens in foods.  相似文献   

3.
Studies were conducted on the flavonoids (myricetin, quercetin, kaempferol, luteolin, and apigenin) contents of 62 edible tropical plants. The highest total flavonoids content was in onion leaves (1497.5 mg/kg quercetin, 391.0 mg/kg luteolin, and 832.0 mg/kg kaempferol), followed by Semambu leaves (2041.0 mg/kg), bird chili (1663.0 mg/kg), black tea (1491.0 mg/kg), papaya shoots (1264.0 mg/kg), and guava (1128.5 mg/kg). The major flavonoid in these plant extracts is quercetin, followed by myricetin and kaempferol. Luteolin could be detected only in broccoli (74.5 mg/kg dry weight), green chili (33.0 mg/kg), bird chili (1035.0 mg/kg), onion leaves (391.0 mg/kg), belimbi fruit (202.0 mg/kg), belimbi leaves (464.5 mg/kg), French bean (11.0 mg/kg), carrot (37.5 mg/kg), white radish (9.0 mg/kg), local celery (80.5 mg/kg), limau purut leaves (30.5 mg/kg), and dried asam gelugur (107.5 mg/kg). Apigenin was found only in Chinese cabbage (187.0 mg/kg), bell pepper (272.0 mg/kg), garlic (217.0 mg/kg), belimbi fruit (458.0 mg/kg), French peas (176.0 mg/kg), snake gourd (42.4 mg/kg), guava (579.0 mg/kg), wolfberry leaves (547.0 mg/kg), local celery (338.5 mg/kg), daun turi (39.5 mg/kg), and kadok (34.5 mg/kg). In vegetables, quercetin glycosides predominate, but glycosides of kaempferol, luteolin, and apigenin are also present. Fruits contain almost exclusively quercetin glycosides, whereas kaempferol and myricetin glycosides are found only in trace quantities.  相似文献   

4.
Toward the development of reliable qualitative and quantitative Polymerase Chain Reaction (PCR) detection methods of transgenic tomatoes, one tomato (Lycopersicon esculentum) species specific gene, LAT52, was selected and validated as suitable for using as an endogenous reference gene in transgenic tomato PCR detection. Both qualitative and quantitative PCR methods were assayed with 16 different tomato varieties, and identical amplified products or fluorescent signals were obtained with all of them. No amplified products and fluorescent signals were observed when DNA samples from 20 different plants such as soybean, maize, rapeseed, rice, and Arabidopsis thaliana were used as templates. These results demonstrated that the amplified LAT52 DNA sequence was specific for tomato. Furthermore, results of Southern blot showed that the LAT52 gene was a single-copy gene in the different tested tomato cultivars. In qualitative and quantitative PCR analysis, the detection sensitivities were 0.05 and 0.005 ng of tomato genomic DNA, respectively. In addition, two real-time assays employing this gene as an endogenous reference gene were established, one for the quantification of processed food samples derived from nontransgenic tomatoes that contained degraded target DNA and the other for the quantification of the junction region of CaMV35s promoter and the anti-sense ethylene-forming enzyme (EFE) gene in transgenic tomato Huafan No. 1 samples. All of these results indicated that the LAT52 gene could be successfully used as a tomato endogenous reference gene in practical qualitative and quantitative detection of transgenic tomatoes, even for some processed foods derived from transgenic and nontransgenic tomatoes.  相似文献   

5.
Lupine flour, protein, and fiber have become common ingredients in food products. The association of lupine-related allergic incidents with peanut allergy is a cause for concern as the latter may bring about severe reactions. In this study, a hybridization probe-based real-time PCR assay for the detection of lupine DNA in foods was developed. Particular attention was paid to the specificity of the method, which was verified by analysis of DNA extracts from more than 50 potential food ingredients such as legumes, cereals, seeds, nuts, spices, fruits, and meat. The limit of detection of the method was determined as 0.1 mg/kg. The successful detection of the presence/absence of lupine DNA in 20 samples proved the suitability of the assay for the analysis of frequently encountered food matrices.  相似文献   

6.
Antioxidant and antiproliferative activities of common vegetables   总被引：12，自引：0，他引：12  
Epidemiological studies have shown that consumption of fruits and vegetables is associated with reduced risk of chronic diseases. Increased consumption of fruits and vegetables containing high levels of phytochemicals has been recommended to prevent chronic diseases related to oxidative stress in the human body. In this study, 10 common vegetables were selected on the basis of consumption per capita data in the United States. A more complete profile of phenolic distributions, including both free and bound phenolics in these vegetables, is reported here using new and modified methods. Broccoli possessed the highest total phenolic content, followed by spinach, yellow onion, red pepper, carrot, cabbage, potato, lettuce, celery, and cucumber. Red pepper had the highest total antioxidant activity, followed by broccoli, carrot, spinach, cabbage, yellow onion, celery, potato, lettuce, and cucumber. The phenolics antioxidant index (PAI) was proposed to evaluate the quality/quantity of phenolic contents in these vegetables and was calculated from the corrected total antioxidant activities by eliminating vitamin C contributions. Antiproliferative activities were also studied in vitro using HepG(2) human liver cancer cells. Spinach showed the highest inhibitory effect, followed by cabbage, red pepper, onion, and broccoli. On the basis of these results, the bioactivity index (BI) for dietary cancer prevention is proposed to provide a simple reference for consumers to choose vegetables in accordance with their beneficial activities. The BI could be a new alternative biomarker for future epidemiological studies in dietary cancer prevention and health promotion.  相似文献   

7.
Myosmine has been regarded as a specific tobacco alkaloid until investigations pointed out that nuts and nut products constitute a significant source of myosmine. In the present study it is shown that the occurrence of myosmine is widespread throughout a large number of plant families. Using a method for extraction practicable for all examined foods, quantitative analysis through internal standard addition showed nanograms per gram amounts. Positively tested edibles were staple foods such as maize, rice, wheat flour, millet, potato, and milk and also cocoa, popcorn, tomato, carrot, pineapple, kiwi, and apples. No myosmine was detectable in other vegetables and fruits such as lettuce, spinach, cucumber, onion, banana, tangerines, and grapes. Myosmine is easily nitrosated giving rise to a DNA adduct identical to the esophageal tobacco carcinogen N-nitrosonornicotine. Therefore, the role of dietary myosmine in esophageal adenocarcinoma should be further investigated.  相似文献   

8.
Because of the individual biological effects and the uncertain or missing information on levels of tocopherols (T) and tocotrienols (T3) in foods frequently consumed in Hawaii, 79 food items (50 in duplicate) were analyzed for alpha-, beta-, gamma-, and delta-tocopherol (alphaT, betaT, gammaT, and deltaT) and alpha-, beta-, gamma-, and delta-tocotrienol (alphaT3, betaT3, gammaT3, and deltaT3) in addition to alpha-tocopheryl acetate (alphaTac). Foods from local markets were stored according to usual household habits, freeze-dried, homogenized, and extracted three times with hexane containing butylated hydroxytoluene as a preservative and tocol as an internal standard. A normal-phase high-pressure liquid chromatography system was applied with fluorescence and photodiode array detection that resulted in baseline separation of all eight analytes and the internal standard tocol (To). The sum of all E vitamer concentrations, or total E vitamers (TEV), in all foods analyzed ranged an average from 0.6 to 828 mg/kg (T < or = 542 mg/kg and T3 < or = 432 mg/kg) and showed the following ranges: oils, 497-828 mg/kg (mainly alphaT and gammaT); margarines, 359-457 mg/kg (mainly gammaT); salad dressings, 20-291 mg/kg (mainly gammaT, except alphaT when soy oil was the main ingredient); cookies, 54-138 mg/kg (mainly gammaT); snacks, 101-220 mg/kg (mainly gammaT); nuts, 22-201 mg/kg (mainly alphaT); vegetables, 2-152 mg/kg (mainly alphaT); pasta, 24-90 mg/kg; cereals, 4-56 mg/kg (mainly betaT3 followed by alphaT); fish, 2-39 mg/kg (mainly alphaT); fried tofu, 64 mg/kg (mainly gammaT); breads, 20-22 mg/kg (mainly betaT3); fat-free mayonnaise, 5 mg/kg (mainly alphaT); poi (fermented taro root), 2 mg/kg (mostly alphaT); and fruits, 2 (papaya) to 13 mg/kg (canned pumpkin) with alphaT predominating. Cereals fortified with alphaTac ranked third and eighth among all foods assayed regarding alphaT and TEV levels, respectively. As compared to the few data available in the literature, our values agreed with some (corn flakes, mango fruit, fat-free mayonnaise, dry-roasted macadamia nuts, dry-roasted peanuts, mixed nuts, spaghetti/marinara pasta sauce, oils, and red bell pepper) but differed for many other items. Our results provide new information on the E vitamer content in foods, emphasize the vast differences of bioactivities of individual E vitamers, and confirm the need for analyses of foods consumed in specific study populations.  相似文献   

9.
This study investigated the production of polyclonal (pAB) antibodies and the first time production of monoclonal (mAB) antibodies against the mycotoxin alternariol, and their implementation in enzyme immunoassay (EIA) for the rapid determination of alternariol in foods. Both EIAs were highly sensitive, with detection limits (IC??) of 35 ± 6.9 pg/mL (mAb EIA) and 59 ± 16 pg/mL (pAb EIA). Food products (n = 109; apple and tomato products, white wine) from German retail shops were analyzed. At a detection limit of 1-2 μg/kg, alternariol at 1-13 μg/kg was found with high frequency in apple (67%) and tomato (93%) products. Tomatoes with visible signs of Alternaria infection, stored at room temperature for up to 4 weeks, contained alternariol at levels up to 50 mg/kg, as determined by EIA and HPLC-FLD. It is concluded that the alternariol immunoassays present a versatile screening tool which could facilitate food control for Alternaria toxins.  相似文献   

10.
通过野外调查取样与实验测定相结合的方法,对兰州市农业生态系统土壤-蔬菜体系49个样点进行研究,分析探讨了研究区蔬菜与土壤的铅污染水平以及蔬菜铅富集效应特性。结果表明：①蔬菜种植地土壤铅平均含量为26.30 mg.kg-1,合格率为93.62%,在不同县区存在显著差异,其大小顺序为红古区〉榆中县〉永登县;②蔬菜铅平均含量为0.59 mg.kg-1,合格率为60.78%,在不同县区存在显著差异,其大小顺序为永登县〉榆中县〉红古区;③蔬菜铅富集系数为（1.69±2.99）%,永登县蔬菜的铅富集能力显著高于榆中县和红古区,不同种类蔬菜铅富集能力在不同县区具有差异性;④土壤全钾除与白菜和甘蓝有正相关性以外,与其他蔬菜均为负相关性,其中与白菜、番茄、土豆显著相关,与胡萝卜为极显著相关;⑤土壤速效钾与各种蔬菜的铅富集能力均为正相关性,其中与胡萝卜、甜菜、甘蓝为显著正相关,与番茄、土豆、菜花为极显著正相关;⑥土壤pH值与胡萝卜、甜菜铅富集系数呈显著负相关,与菜花呈极显著负相关。研究表明,兰州市不同蔬菜产区的各类蔬菜对土壤铅的富集具有明显的差异性,同时不同形态的钾元素及土壤pH值对蔬菜铅富集能力具有显著的影响。  相似文献   

11.
Soils under intensive agricultural practices such as those for growing vegetables in plastic greenhouses are an important anthropogenic source of nitrous oxide (N2O). Nitrous oxide emissions and measures to mitigate them through fertilizer N management have been less frequently studied than open field systems. The objectives of this study were to measure N2O emissions from vegetables under greenhouse conditions in Southern China and to investigate the effect of reducing the amount of applied synthetic N fertilizer compared with local practice. Results indicate that the average N2O‐N flux during the growth of four vegetables (tomato, cucumber, celery and a second tomato crop) was 117.4 ± 9 μg N/m2/h, and the annual emission rate was 8.1 ± 0.6 kg/N/ha for local farms. Temperature was important with much lower emissions during the celery‐growing season when soil and air temperatures were frequently <10 °C. Nitrous oxide emissions from the greenhouse vegetables were seven times greater than from the rice–wheat system in the same area and soils. Reducing the amount of applied synthetic N fertilizer by 40% relative to local farmers’ normal usage could reduce annual cumulative N2O emissions by 33% without any impact on crop yields.  相似文献   

12.
The objective of this study was to establish analytical approaches to quantify 1,3-beta-glucan (1,3-beta-G) in foods. Six food categories including legumes, cereals, tubers, vegetables, fruits, and mushrooms and 17 total items were tested. An extraction procedure was designed to prepare food cold-water soluble, hot-water soluble, cold-alkaline soluble, and hot-alkaline soluble fractions. A fluorescence microassay based on aniline blue dye, which bound specifically to 1,3-beta-G, was developed to measure its content in the food fractions. Curdlan was used as standard to construct the 1,3-beta-G calibration curve, and a linear correlation within a 14 microg/mL concentration range was obtained. This microassay displayed selectivities among various 1,3-beta-G species. Biologically active ones such as pachyman and yeast glucan possessed much stronger fluorescent signals than others such as laminarin and barley glucan. Possible fluorescent interference from food proteins was estimated. This assay tolerated up to 50% of bovine serum albumin in 10 microg/mL curdlan. Analysis of the four food fractions showed that besides the well-known lentinan-containing shiitake, popular foods such as celery, chin-chian leaves, carrot, and radish contained nearly 20% 1,3-beta-G in their total sugar. Soybean dry weight contained 0.8% 1,3-beta-G, which was twice the amount compared to shiitake. Snow mushroom dry weight had the highest 1,3-beta-G content, at 2.5%, and was rich in both water (0.67%) and alkaline soluble (1.87%) forms. In conclusion, this dye-binding fluorescence microassay in conjunction with the extraction procedure can be applied in the prescreening of potential foods rich in functional 1,3-beta-G.  相似文献   

13.
A rotation experiment was conducted in a greenhouse with three vegetable crops on red yellowish soil (RYS) and silt loamy soil (SLS) to study Cd accumulation in pak choi ( Brassica chinensis L.), tomato ( Lycopersicon esculentum), and radish ( Raphanus sativus L.). Critical Cd concentrations in the two soils were evaluated for these vegetables based on human dietary toxicity. Cadmium was added as Cd(NO 3) 2 at a rate of 0-7.00 mg Cd kg (-1) soil. Shoot growth was not inhibited by Cd except for radish grown on RYS. A small amount of Cd stimulated growth of the vegetables. Cadmium concentration in edible parts of the vegetables generally increased with Cd concentration in soils but was higher in RYS than SLS. The distribution of Cd in pak choi and tomato decreased in the order root > shoot > fruit, but the order was shoot > root for radish. When Cd content in the edible parts reached maximum contaminant levels for safety food standards, the soil total Cd concentrations were 0.327 and 0.120 mg kg (-1) in RYS and 0.456 and 0.368 mg kg (-1) in SLS for pak choi stem and radish, respectively, whereas ammonium acetate-extractable Cd was 0.066 and 0.089 mg kg (-1) in RYS and 0.116 and 0.092 mg kg (-1) in SLS for pak choi leaf and tomato, respectively, based on food safety standards.  相似文献   

14.
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.  相似文献   

15.
Catechins, compounds that belong to the flavonoid class, are potentially beneficial to human health. To enable epidemiological evaluation of these compounds, data on their contents in foods are required. HPLC with UV and fluorescence detection was used to determine the levels of (+)-catechin, (-)-epicatechin, (+)-gallocatechin (GC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECg), and (-)-epigallocatechin gallate (EGCg) in 24 types of fruits, 27 types of vegetables and legumes, some staple foods, and processed foods commonly consumed in The Netherlands. Most fruits, chocolate, and some legumes contained catechins. Levels varied to a large extent: from 4.5 mg/kg in kiwi fruit to 610 mg/kg in black chocolate. (+)-Catechin and (-)-epicatechin were the predominant catechins; GC, EGC, and ECg were detected in some foods, but none of the foods contained EGCg. The data reported here provide a base for the epidemiological evaluation of the effect of catechins on the risk for chronic diseases.  相似文献   

16.
Almond ( Prunus dulcis ) has been widely used in all sorts of food products (bakery, pastry, snacks), mostly due to its pleasant flavor and health benefits. However, it is also classified as a potential allergenic seed known to be responsible for triggering several mild to life-threatening immune reactions in sensitized and allergic individuals. Presently, eight groups of allergenic proteins have been identified and characterized in almond, namely, PR-10 (Pru du 1), TLP (Pru du 2), prolamins (Pru du 2S albumin, Pru du 3), profilins (Pru du 4), 60sRP (Pru du 5), and cupin (Pru du 6, Pru du γ-conglutin), although only a few of them have been tested for reactivity with almond-allergic sera. To protect sensitized individuals, labeling regulations have been implemented for foods containing potential allergenic ingredients, impelling the development of adequate analytical methods. This work aims to present an updated and critical overview of the molecular characterization and clinical relevance of almond allergens, as well as review the main methodologies used to detect and quantitate food allergens with special emphasis on almond.  相似文献   

17.
Among allergenic foods, soybean is known as a food causing adverse reactions in allergenic patients. To clarify the validity of labeling, the specific and sensitive detection method for the analysis of the soybean protein would be necessary. The p34 protein, originally characterized to be p34 as an oil-body associated protein in soybean, has been identified as one of the major allergenic proteins and named Gly m Bd 30K. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of the soybean protein in processed foods was developed using polyclonal antibodies raised against p34 as a soybean marker protein and the specific extraction buffer for extract. The developed sandwich ELISA method was highly specific for the soybean protein. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed ELISA were 0.47 ng/mL (equivalent to 0.19 microg/g in foods) and 0.94 ng/mL (equivalent to 0.38 microg/g in foods), respectively. The recovery ranged from 87.7 to 98.7%, whereas the intra- and interassay coefficients of variation were less than 4.2 and 7.5%, respectively. This study showed that the developed ELISA method is a specific, precise, and reliable tool for the quantitative analysis of the soybean protein in processed foods.  相似文献   

18.
Furocoumarins represent a family of natural food constituents with phototoxic and photomutagenic properties. They are found mainly in plants belonging to the Rutaceae and Umbilliferae such as celery, carrots, and parsnips. Parsnips (Pastinaca sativa L.) have become more and more popular as a vegetable, e.g., as a constituent of or ingredient in baby food. Previous work has shown that microbial infection of parsnip roots can result in a dramatic increase in furocoumarin levels. In this study, freshly harvested parsnips were stored as whole roots, pieces (cubes), or homogenate at +4 degrees C or -18 degrees C over various time periods under standard conditions. It was found that furocoumarin concentrations (sum of five furocoumarins: angelicin, isopimpinellin, 5-methoxypsoralen, 8-methoxypsoralen, and psoralen) in freshly harvested parsnips, analyzed by HPLC after extraction with diethyl ether and sequential solid phase (reversed-phase and silica) extraction, was generally lower than 2.5 mg/kg, and storage of parsnips in any form investigated at -18 degrees C over up to 50 days did not lead to a marked increase in furocoumarin levels. In contrast, storage of whole parsnips, but not of cubes or homogenate, at +4 degrees C resulted in a marked biphasic increase of furocoumarin concentrations after 7 and 38 days of storage up to levels of about 40 mg/kg. A dramatic increase in furocoumarin concentrations up to 566 mg/kg was observed when whole parsnips obtained from the market were kept at room temperature over 53 days, resulting in a visible microbial (mold) infection. Baby food products from the German market containing parsnips as an ingredient or constituent showed furocoumarin levels < or =0.41 mg/kg, suggesting that properly stored roots/preparations have been used. It is recommended that, after harvesting, parsnips be kept at -18 degrees C or under other conditions that prevent microbial infections.  相似文献   

19.
硒对胡萝卜含硒量、产量及品质的影响   总被引：9，自引：1，他引：9  
采用不同浓度Na2SeO3进行胡萝卜叶面喷施的田间试验,研究胡萝卜对硒的吸收、转化特点和硒对胡萝卜产量、品质等的影响。结果表明,叶面施硒可明显提高胡萝卜中各形态硒含量,随施硒量的增加,有机硒转化率上升;其总硒、有机硒含量在最大处理浓度(500.mg/kg)呈最高,分别是CK的15.9和26.6倍。施硒胡萝卜产量较对照降低,但未达显著水平。营养品质中除Vc下降,P变化不一外,总糖、胡萝卜素、粗纤维、Ca、Mg、Fe、K在一定浓度下有不同程度提高。综合分析硒对胡萝卜含硒量、产量、各品质因素以及对人、畜食用的安全性,施硒量以不超过1.0mg/kg为宜。  相似文献   

20.
Numerous in vitro and in vivo studies have suggested that dietary anthocyanins and ellagitannins or ellagic acid might have beneficial health effects. Epidemiological evidence on the disease-preventing potential of these polyphenols is lacking, due to the absence of reliable data on their contents in foods. In this study was analyzed the content of anthocyanins and ellagitannins (as ellagic acid equivalents after acid hydrolysis) in foods consumed in Finland, including berries, fruits, vegetables, and processed products, using high-performance liquid chromatographic (HPLC) methods. Anthocyanins were detected in 41 of 54 selected food items. The total anthocyanin content varied in berries from 1 to 611 mg/100 g, in fruits from 2 to 66 mg/100 g, and in vegetables from 3 to 75 mg/100 g of fresh weight as the weight of the aglycone. Ellagitannins were screened in 33 food items, but were detected only in 5 species of berries, that is, in cloudberry, raspberry, rose hip, strawberry, and sea buckthorn, the content ranging from 1 to 330 mg/100 g. The results underscore the superiority of berries, especially dark blue or red berries, as excellent sources of anthocyanins and certain berries of the Rosaceae family as the major source of ellagitannins in the Finnish diet.  相似文献   

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