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1.
A fluorescence polarization immunoassay (FPIA) based on a monoclonal antibody for the detection of parathion-methyl (PM) was developed and optimized. Fluorescein-labeled PM derivatives (tracers) with different structures were synthesized and purified by thin-layer chromatography. The influence of immunogen and tracer structures on the assay characteristics was investigated. PM concentration determinable by the FPIA ranged from 25 to 10000 ppb. The detection limit was 15 ppb. Methanol extracts of vegetable, fruit, and soil samples were diluted 1/10 for the analysis. Recovery in spiked samples averaged between 85 and 110%. The method developed is characterized by high specificity and reproducibility (CV ranged from 1.5 to 9.1% for interassay and from 1.8 to 14.1% for intra-assay). The FPIA method can be applied to the screening of food and environmental samples for PM residues without complicated cleanup.  相似文献   

2.
Development of a multianalyte enzyme-linked immunosorbent assay (ELISA) for detection of permethrin and aroclors 1248 or 1254 and implementation of the assay for analysis of soil/sediment samples are described. The feasibility of using the multianalyte ELISA to monitor aroclors 1254 and permethrin simultaneously was tested with permethrin and aroclor standards and with aroclor- and permethrin-containing soil/sediment and house dust samples. Comparison of the I?? and I?? values of the multianalyte with those of a single-analyte assay revealed similar results, and multianalyte ELISA determination of analyte amounts in soil/sediment dust samples yielded similar results to those of a single-analyte assay. A single-analyte assay of permethrin content in permethrin-containing dust samples showed that the ELISA can determine the analyte accurately in samples with dust matrix contents ranging from 6.25 to 100 mg as indicated by the good correlation between the results of the immunoassay and those of the gas chromatography analysis.  相似文献   

3.
A competitive-indirect enzyme-linked immunosorbent assay (CI-ELISA) was developed to quantify chlorimuron-ethyl in soil. The linear working range of the assay was from 1 to 1000 ng mL(-)(1). The assay had an I(50) value of 54 ng mL(-)(1), with a limit of detection of 2 ng mL(-)(1) and a limit of quantification of 27 ng mL(-)(1). Three soils were extracted using a carbonate buffer (pH 9.0) and the extracts spiked with chlorimuron-ethyl. Because of the effects of coextractants (matrix effects) from soil on the accuracy and precision of the ELISA, immunoaffinity chromatography (IAC) was used to purify chlorimuron-ethyl from soil extracts prior to analysis. The immunoaffinity columns, which had a total binding capacity of 1350 ng of chlorimuron-ethyl mL(-)(1) of immunosorbent, were prepared by binding anti-chlorimuron-ethyl antibodies to protein G Sepharose 4B. Although the matrix effects were largely removed using the affinity column, they could be completely removed by first passing the extract through a column containing epoxy-coupled 1,6-diaminohexane (EAH) Sepharose 4B to remove organic acids prior to IAC. Assay sensitivity was increased 100-fold using IAC to purify and simultaneously concentrate chlorimuron-ethyl from soil extracts. The purification strategy (EAH followed by IAC chromatography) removed matrix effects from all three soils and allowed for the accurate quantitation of chlorimuron-ethyl in soil extracts.  相似文献   

4.
Aqueous extracts from sixty almond samples representing various genotypes and interspecies hybrids of almond, including almond-peach, were analyzed for protein and peptide content using electrophoresis, Western immunoblotting, and enzyme-linked immunosorbent assay (ELISA). Nondenaturing nondissociating polyacrylamide gel electrophoresis (NDND-PAGE) of the aqueous extracts indicated that a single major storage protein (almond major protein -- AMP or amandin) dominated the total soluble protein composition. Denaturing SDS--PAGE analyses of the aqueous extracts revealed that the AMP was mainly composed of two sets of polypeptides with estimated molecular masses in the ranges of 38--41 kDa and 20--22 kDa, regardless of the source; however, distinct variations in the intensity and electrophoretic mobility of some bands were noted between samples. In addition to AMP, several minor polypeptides were also present in all the genotypes, and variations were seen in these as well. Regardless of the genotype, AMP was recognized in Western blots by rabbit polyclonal anti-AMP antibodies, mouse monoclonal anti-AMP antibodies (mAbs), and serum IgE from patients displaying strong serum anti-almond IgE reactivity. As with protein staining results, antibody reactivity also revealed common patterns but displayed some variation between samples. An anti-AMP inhibition ELISA was used to quantify and compare aqueous extracts for various samples. All samples (n = 60) reacted in this assay with a mean +/- standard deviation (sigma n) = 0.82 +/- 0.18 when compared to reference aqueous extract from Nonpareil designated as 1.0.  相似文献   

5.
Aflatoxins spiked at three different levels (6.5, 13.0, and 19.5 microg/kg) in tahini, a sesame butter, were analyzed by using three different methods: high-performance liquid chromatography (HPLC), fluorometry, and enzyme-linked immunosorbent assay (ELISA). An immunoaffinity column was used for cleanup and purification of extracts prior to detection by HPLC and fluorometry. All methods were statistically evaluated for accuracy, precision, and simple correlations. Additionally, 14 tahini samples randomly obtained from Turkish retail markets were analyzed using an immunoaffinity column cleanup procedure coupled with the HPLC detection method. The fluorometric determination method involving an immunoaffinity column cleanup step was found to be highly correlated with the HPLC method (r = 0.978). Both methods were found to be effective due to their high recoveries and low variance for the prediction of total aflatoxin contamination in tahini samples. The ELISA method, due to its high variation in replicates, was found to be applicable only as a screening method. The survey study demonstrated the need for control of aflatoxin contamination of foodstuffs involving sesame seeds as an ingredient.  相似文献   

6.
The ferrous oxidation-xylenol orange (FOX) assay was adapted for quantifying lipid hydroperoxides (LOOHs) in plant extracts. Excised pieces of several fruit and vegetable species were exposed to 83 kJ m(-2) day(-1) of biologically effective ultraviolet B irradiance (UV-B(BE)) for 10-12 days to induce cellular oxidation. The LOOH and thiobarbituric acid reactive substance (TBARS) concentrations of these plant tissues were assessed with the FOX and iodometric assays for the former and a modified TBARS assay for the latter. There was generally good agreement between the FOX and iodometric methods both prior to and following the UV exposure. However, the iodometric assay appeared to have some difficulty in consistently quantifying lower LOOH levels (<11 microM), whereas the FOX assay measured LOOH concentrations as low as 5 microM. All tissues exhibited UV-induced increases in TBARS, indicating a marked degree of cellular oxidation in the exposed tissue segments. Compared with the iodometric assay, the FOX method consistently generated less variable LOOH values. The presence of authentic linoleic acid-OOHs in spiked avocado and spinach samples (11 microM) was identified with liquid chromatography-mass spectrometry techniques, which validated corresponding FOX assay results. The FOX method is inexpensive, is not sensitive to ambient O2 or light levels, and can rapidly generate LOOH measurements. The physiological value of the FOX assay resides in its ability to measure initial rather than more advanced fatty acid oxidation; hence, early membrane-associated stress events in plant tissue can be detected.  相似文献   

7.
A pre-lysis buffer washing procedure was introduced to DNA extraction from a forest soil with high organic matter and iron oxide contents. Sodium phosphate of 0.1 M (pH 7.5) was used as a buffer to wash soil samples when subsequent lysis buffer was phosphate, and 20 mM EDTA (pH 7.5) was used when subsequent lysis buffer included EDTA. Initial experiments were not successful because the DNA extracts could not be amplified by polymerase chain reaction (PCR). The consideration of introducing a pre-lysis washing procedure was based on the idea that the washing should promote soil dispersion and homogeneity, decrease DNA adsorption by soil components (e.g. iron oxides), and remove covalent cations and those easily-dissolving organic compounds from the soil samples. Results revealed that humic substance content decreased by 31%, but DNA yield increased by 24% in the DNA extracts of the pre-lysis washing procedures, compared to the non-washing procedures. DNA extracted by the pre-washing procedure needed less purification for subsequent 18S and 16S rDNA PCR amplifications. It was recommended that the pre-lysis buffer washing should be used for DNA extraction from those difficult environmental samples, such as the forest soil with high contents of organic matter and iron oxides.  相似文献   

8.
Fruit pulp and seeds from the jujube plant possess nutritional and medicinal properties. The bioactive components have been shown to vary both with cultivar and with growing conditions. Most studies report the components of varieties from China. We measured free amino acid, individual phenolic, and total phenolic content, and antioxidative activities in three jujube fruit pulp extracts from Boeun-deachu, Mechu, and Sanzoin cultivars and two seed extracts (Mechu and Sanzoin) from plants grown in Korea. In g/100 g dry weight, total free amino acid content measured by ion-exchange chromatography ranged from 5.2 to 9.8 in the pulp and from 4.0 to 5.3 in the seed. Total phenolic content measured by Folin-Ciocalteu ranged from 1.1 to 2.4 in the pulp and from 3.6 to 4.6 in the seed. Flavonoids were measured by HPLC and ranged from 0.7 to 1.8 in the pulp and from 3.2 to 4.0 in the seed. Flavonoids were identified by HPLC elution position and UV/vis and mass spectra. Fruits contained the following flavonoids: procyanidin B2, epicatechin, quercetin-3-O-rutinoside (Q-3-R), quercetin-3-O-galactoside (Q-3-G), kaempferol-glucosyl-rhamnoside (K-G-R), and two unidentified compounds. Seeds contained the following flavonoids: saponarin, spinosin, vitexin, swertish, 6'-hydroxybenzoylspinosin (6'-HBS), 6'-feruloylspinosin (6'-FS), and one unidentified substance. Dimensions and weights of the fresh fruit samples affected phenolic content. The distribution of the individual flavonoids among the different samples varied widely. Data determined by the FRAP antioxidative assay were well correlated with total phenolic content. In a departure from other studies, data from the DPPH free radical assay were not correlated with FRAP or with any of the measured compositional parameters. Because individual jujube flavonoids are reported to exhibit different health-promoting effects, knowledge of the composition and concentration of bioactive compounds of jujube products can benefit consumers.  相似文献   

9.
松华坝库区土壤氮磷累积特征与流失风险研究   总被引:1,自引:0,他引:1  
分析了松华坝水库库区土壤的N,P累积特征。结果表明,(1)整个库区土壤(0—20 cm)全氮含量平均值为1.79 g/kg;NH4—N含量平均值为13.55 mg/kg;NO3—N含量平均值为7.29 mg/kg。全氮含量依次为:粮田>果园>菜地;NH4—N含量依次为:果园>粮田>菜地;NO3—N含量依次为:果园>菜地>粮田。(2)整个库区土壤(0—20 cm)全磷平均值为1.60 g/kg;速效磷含量平均值为10.36 mg/kg。全磷含量依次为:粮田>果园>菜地;速效磷含量依次为:菜地>粮田>果园。(3)入库河流的全N含量最初为Ⅱ类水,但是在入库口全N含量为Ⅴ类水。入库河流的全P含量最初为Ⅱ类水,全P含量最高为Ⅲ类水,入库口水质全P下降为Ⅲ类水。  相似文献   

10.
The N-methylcarbamate pesticide carbofuran is a very important insecticide used worldwide. In the present work, the validation of a monoclonal antibody-based enzyme immunoassay (ELISA) to determine this compound in fruits and vegetables is described. The immunoassay is a competitive heterologous ELISA in the antibody-coated format, with an I(50) value for standards in buffer of 740 ng/L and with a dynamic range between 200 and 3100 ng/L. For recovery studies, peppers, cucumbers, strawberries, tomatoes, potatoes, oranges, and apples were spiked with carbofuran at 10, 50, and 200 ppb. After liquid extraction, analyses were performed by ELISA on extracts purified on solid-phase extraction (SPE) columns and crude, nonpurified extracts. Depending on the crop, mean recoveries in the 43.9--90.7% range were obtained for purified samples and in the 90.1--121.6% range for crude extracts. The carbofuran immunoassay performance was further validated with respect to high-performance liquid chromatography (HPLC) with postcolumn derivatization and fluorescence detection (EPA Method 531.1). Samples were spiked with carbofuran at several concentrations and analyzed as blind samples by ELISA and HPLC after SPE cleanup. The correlation between methods was very good (y = 0.90x + 2.66, r(2)() = 0.958, n = 25), with HPLC being more precise than ELISA (mean coefficients of variation of 4.1 and 11.5%, respectively). The immunoassay was then applied to the analysis of nonpurified extracts of the same samples. Results also compared very well with those obtained by HPLC on purified samples (y = 1.02x + 10.44, r(2)() = 0.933, n = 29). Therefore, the developed immunoassay is a suitable method for the quantitative and reliable determination of carbofuran in fruits and vegetables even without sample cleanup, which saves time and money and considerably increases the sample throughput.  相似文献   

11.
符明明  贾萌萌  胡文友  黄标 《土壤》2016,48(5):887-892
设施栽培条件下土壤性质变化剧烈,其对作物吸收硒(Se)的影响值得研究。本文通过对南京市典型设施蔬菜种植基地土壤及对应植物样品进行取样分析,探讨高强度利用模式下土壤Se的存在形态以及不同蔬菜类型对Se吸收的差异,分析蔬菜Se含量与土壤Se存在形态及土壤性质之间的关系,并对长期设施栽培条件下蔬菜Se的摄入健康风险进行评价。结果表明,不同蔬菜可食部分对Se的富集能力表现为叶菜类(平均含Se量为60μg/kg,干重)根茎类(30μg/kg)茄果类(26μg/kg)。蔬菜可食部分Se含量随土壤有机质及有机结合态Se含量的增加而降低。研究区域有机肥的大量施入引起的有机结合态Se含量的增加可能是降低Se有效性的最重要原因。  相似文献   

12.
The N-methylcarbamate pesticide carbaryl is one of the most important insecticides used worldwide. In the present work, the validation of a monoclonal antibody-based enzyme immunoassay (ELISA) for the determination of this compound in fruits and vegetables is described. The immunoassay is a competitive heterologous ELISA in the antibody-coated format, with an I(50) value for standards in buffer of 101.0 +/- 26.9 ng/L and with a dynamic range between 31.6 and 364.0 ng/L. For recovery studies, peppers, cucumbers, strawberries, tomatoes, potatoes, oranges, and apples were spiked with carbaryl at 10, 50, and 200 ppb. After liquid extraction, analyses were performed by ELISA on both extracts purified on solid-phase extraction (SPE) columns and crude, nonpurified extracts. Depending on the crop and the fortification level, recoveries in the 59.0--120.0% range were obtained for purified samples and in the 70.0--137.7% range for crude extracts. The carbaryl immunoassay performance was further validated with respect to high-performance liquid chromatography (HPLC) with postcolumn derivatization and fluorescence detection (EPA Method 531.1). Samples were spiked with carbaryl at several concentrations and analyzed as blind samples by ELISA and HPLC after SPE cleanup. The correlation between methods was excellent (y = 1.04x + 0.71, r(2) = 0.992, n = 33), with HPLC being more precise than ELISA (mean coefficients of variation of 5.2 and 12.0%, respectively). The immunoassay was then applied to the analysis of nonpurified extracts of the same samples. Results also compared very well with those obtained by HPLC on purified samples (y = 1.28x - 0.59, r(2) = 0.987, n = 33) while maintaining similar precision. Therefore, the developed immunoassay is a suitable method for the quantitative and reliable determination of carbaryl in fruits and vegetables even without sample cleanup, which saves time and money and considerably increases sample throughput.  相似文献   

13.
A method for the direct determination of lead in evaporated milk and in fruit juice with no prior sample digestion was successfully collaborated by 13 laboratories. The anodic stripping voltammetric (ASV) method studied consisted of adding 0.2 mL aliquots of evaporated milk or 0.3 mL aliquots of fruit juice to 2.9 mL of a dechelating reagent, Metexchange. The reagent-sample mixture is then analyzed for lead by ASV with no further sample preparation. Each collaborator received 24 samples, 2 each at 5 different levels (0.07-0.70 ppm for spiked evaporated milk and 0.09-0.87 ppm for spiked apple juice) along with duplicate practice samples of labeled lead content at each of 2 levels for each sample type. All unknowns were coded with random numbers. Approximately 69% of the reporting laboratories had never analyzed either evaporated milk or fruit juice for lead. Average time between receipt of samples and reporting of results was 1.6 days for all laboratories. The pooled variations between duplicate determinations for apple juice and evaporated milk were 0.00059 and 0.00043, respectively. The method was adopted official first action for both fruit juice and evaporated milk.  相似文献   

14.
The measurement of antioxidant capacity in fruits differs from that of other biological samples due to their low pH and very low lipophilic antioxidant capacity. In this report, we present a modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method for fruits and compare its performance with the other commonly used antioxidant methods of 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The antioxidant capacity and reaction kinetics of four phenolic compounds, two antioxidant standards, and five fruits were also investigated. The modified ABTS method prepared at a pH of 4.5 with sodium acetate buffer is highly stable and easily applied to fruit samples as compared to the standard (pH 7.4) version. The measured antioxidant capacity of samples varied with the assay method used, pH, and time of reaction. Traditional antioxidant standards (trolox, ascorbic acid) displayed stable, simple reaction kinetics, which allowed end point analysis with all of assays. Of the phenolic compounds examined, chlorogenic and caffeic acids exhibited the most complex reaction kinetics and reaction rates that precluded end point analysis while gallic acid and quercetin reached stable end points. All fruit extracts exhibited complex and varied kinetics and required long reaction times to approach an end point. Because the antioxidant capacity of fruit extracts is a function of the array of individual antioxidants present, accurate comparisons among fruit samples require that reaction times be standardized and of sufficient length to reach steady state conditions and that more than one assay be used to describe the total antioxidant activity of fruit samples.  相似文献   

15.
Aflatoxin B1 (B1), T-2 toxin (T2), and ochratoxin A (OA) were assayed in a single extract from barley grain by using competitive enzyme linked immunosorbent assays (ELISAs) with monoclonal antibodies. B1 and T2 monoclonal antibodies were conjugated to horseradish peroxidase for direct competitive ELISA while an indirect competitive ELISA was used for OA determination. The competitive ELISA detected 0.1 ng/mL of B1, 10 ng/mL of T2, or 1 ng/mL of OA. Acetonitrile-0.5% KCl-6% H2SO4 (89 + 10 + 1) extracts of barley grain either were diluted 1:10 for direct assay or were subjected to a simple liquid-liquid cleanup procedure to concentrate the extract 10:1 before assay. For cleanup, water was added to the acetonitrile extract to partition water-soluble interfering substances, and then the mycotoxins were re-extracted with chloroform. The chloroform extract was evaporated to dryness and redissolved in Tris HCl buffer for ELISA. The mean recoveries from barley spiked with 4-60 ng/g of B1, 50-5000 ng/g of T2, and 5-500 ng/g of OA were, respectively, 93.8, 80.6, and 95.8%. The mean within-assay, inter-assay, and subsample coefficients of variation by ELISA of barley grain colonized with toxigenic fungi were less than 12% for B1 and OA but as high as 17% for T2.  相似文献   

16.
3,5,6-三氯-2-吡啶醇(TCP)的化学结构稳定、水溶性高、运移能力强,对水体具有潜在的污染风险。在有机质含量低、大孔隙度高、导水性好的紫色土地区,风险更加显著。为研究TCP在紫色中的迁移规律,该研究依据紫色土的典型耕作类型,在中国科学院盐亭国家紫色土农业生态试验站采集3组土样(小麦-玉米轮作的坡地、水稻-油菜轮作的水旱农田和萝卜-白菜套种的菜地),通过批量平衡法研究紫色土对TCP的吸附特征,并采用稳定流场饱和均质土柱的易混合置换试验研究TCP的动态迁移过程,最后对其迁移动态进行模拟。结果表明:紫色土对TCP的吸附特征呈线性,在坡地、水旱农田和菜地中的吸附系数分别为1.94、1.22和1.02 L/kg,且黏土含量和矿物组成是主要影响因子;TCP的出流平衡浓度分别为初始浓度的77%(坡地)、84%(水旱农田)和92%(菜地),相应的平衡时刻分别为2.88PV、4PV和6.5PV,表明TCP对环境的污染风险较高;用非平衡两点对流弥散模型模拟TCP在3种耕作条件下迁移,表明TCP以瞬时吸附为主,其水动力弥散系数和分形系数在坡地、水旱农田和菜地中依次减小,但一阶动力学常数依次增大。研究结果为探索TCP在紫色土壤中的迁移机制和预测防止TCP对环境的污染提供了参考依据。  相似文献   

17.
Determination of cutin-bound residues of chlorothalonil by immunoassay   总被引:6,自引:0,他引:6  
An indirect competitive enzyme-linked immunosorbent assay (ELISA) was used to determine photochemically cutin-bound residues of chlorothalonil in enzymatically isolated tomato and apple fruit cuticles. The samples were spiked, irradiated, exhaustively extracted, and depolymerized with boron trifluoride complex resulting in a soluble depolymerisate. With this procedure, the ELISA could be calibrated with free target molecules for the quantification of originally bound chlorothalonil residues. In fruit cuticles that were irradiated for 8 h by simulated sunlight, 0.030 and 0.068 mg/g photoinduced cutin-bound residues of wax-free cuticles (calculated as chlorothalonil) were determined for tomatoes and apples, respectively. For the used antibody mAb chl. 4/11, cross-reactivities with derivatives of chlorothalonil simulating different types of cuticle-bound residues are given and discussed.  相似文献   

18.
上海市郊旱作农田土壤养分资源状况   总被引:8,自引:1,他引:8  
研究了上海市郊旱作农田土壤养分特征,结果表明:自80年代以来经过近20年的土壤耕作演替,上海市郊蔬菜、瓜果旱作土壤总体养分状况表现为有机质、氮素、磷素和钾素含量均很丰富。土壤氮、磷养分积累明显,较80年代第二次土壤普查有大幅度增加,一方面施用过多肥料造成浪费,另一方面带来潜在的非点源污染,对周围的环境造成威胁。各区域由于受土壤质地、土壤类型、气候条件、作物种类、耕作方式等不同土壤养分有所差异。不同耕作方式对农田土壤养分的含量影响较大,其水平高低依次为:大棚土壤、露天菜地土壤、传统自留地土壤。大棚瓜、菜地土壤有酸化趋势,菜园土壤有效磷含量远高于稻麦田土壤,这主要和施肥水平有关。  相似文献   

19.
An enzyme-linked immunosorbent assay (ELISA) for fipronil was developed by using polyclonal antibodies (pABs) or monoclonal antibodies (mABs), and its suitability of the determination of this analyte in spiked water samples was studied. The pABs-based assay showed I50 = 17.95 ppb, I90 = 203.40 ppb, and I10 = 0.066 ppb, whereas the mABs-based assay showed I50 = 5.99 ppb, I90 = 485.40 ppb, and I10 = 0.074 ppb. The recoveries of fipronil from tap water samples by pABs-based ELISA were 93.00-124.00% in the range of 0-500 ng/mL, and those obtained from the samples by mABs-based ELISA were 94.70-108.00%. Different types of water from pool, river, and sea were spiked at different levels (ranging form 0.1 to 10 microg/L) and were assayed by the indirect ELISA with mABs. The recoveries of fipronil by this ELISA were in the range of 80-120%. The results demonstrate that this assay is suitable for the quantitative detection of fipronil at trace levels in water samples.  相似文献   

20.
Liquid chromatographic determination of acifluorfen in soil and water   总被引:1,自引:0,他引:1  
An analytical method based on the use of a liquid chromatograph equipped with a UV detector was developed for the determination of acifluorfen in soil and water. Acifluorfen was extracted from soil in methanol-0.10N NaOH (80 + 20 v/v) and from water by partition with dichloromethane. Solvent partitioning and solid-phase extraction were used to separate acifluorfen from major interfering sample components. Average recoveries from soil at 1, 0.1, and 0.01 ppm fortification levels were 95.1 +/- 3.4, 92.6 +/- 2.9, and 73.9 +/- 3.0%, respectively. Recoveries from water spiked at levels from 0.01 to 1 ppm averaged 96.5 +/- 5.4%. Method limits of detection were 0.006 ppm in soil and 0.003 ppm in water.  相似文献   

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