Rabbit Monoclonal Antibody-Based Lateral Flow Immunoassay Platform for Sensitive Quantitation of Four Sulfonamide Residues in Milk and Swine Urine

Based on the available rabbit monoclonal antibody (RabMAb), a rapid and sensitive lateral flow immunoassay (LFA) platform has been developed for quantitative detection of four sulfonamide residues(SRs) of sulfadiazine (SD), sulfathiazole (STZ), sulfapyridine (SP), and sulfamethoxazole (SMX).Within the designed LFA competitive format assay, which was based on antigen-antibody properties, the hapten conjugate N¹-[4-(carboxymethyl)-2-thiazolyl] sulfanilamide linked to protein ovalbumin (TS-OVA) and goat anti-rabbit antibody were sprayed as capture and control reagents, respectively, and then the antibody was conjugated to colloidal gold particles as the detection reagent. With quantitative assessment aided by a colorimetric strip reader, the sensitivities of the established LFA method for SD, STZ, SP, and SMX were 0.91 ng mL^?1, 0.10ng mL^?1,0.12ng mL^?1, and 2.13ng mL^?1, and the half-maximum inhibition concentrations (IC₅₀) were 5.19 ng mL^?1, 1.25 ng mL^?1, 0.66 ng mL^?1, and 24.14 ng mL^?1, respectively. The recoveries at three spiked levels (5, 20, 50 ng mL^?1for SD, STZ, and SP; 20, 50, 100 ng mL^?1 for SMX) were in the range of 78.02–135.10% and 76.40–137.16% for milk and swine urine, respectively. More importantly, the detection performance of the established platform was consistent with that of in-parallel LC-MS/MS analysis. In conclusion, the proposed LFA platform has showed the potential for fast, sensitive and relatively accurate quantification of four sulfonamide residues in practical uses.     

对土壤及玉米植株中均三氮苯类除草剂的残留分析

土壤或玉米植株样品用Ｖ（甲醇）∶Ｖ（乙腈）＝１∶１提取,提取液用石油醚净化后,浓缩液过Ｃ１８小柱净化,最后用Ｎｏｖａ－ＰａｋＣ１８柱进行ＨＰＬＣ分析。回收率：氰草津为８２．４％～９９．８％,莠去津为８５．６％～１０２．３％,西草净为８９．１％～１０８．４％。     

Characterization of a New Monoclonal Antibody Against Mercury(II)

Monoclonal antibodies (mabs) were produced against mercury (II) and an enzyme immunoassay was developed for the detection of mercury (II) in water. Since mercury (II) ions are too small to elicit an immune response, they were coupled via glutathione (GSH) to the immunogenic carrier protein keyhole limpet hemocyanin (KLH). Several mice were immunised with this KLH-GSH-Hg immunoconjugate. Spleen cells of immunised mice were fused with myeloma cells. The resulting hybridoma cells were screened for the production of specific anti-Hg mabs. Five positive cells were detected. The hybridoma cell line K3C6 was adjusted to protein free medium; it produced mabs with high selectivity and sensitivity. A detection limit of 2.8 μg/L HgCl2 (= 2.1 μg/L Hg²⁺) was achieved with a non-competitive enzyme immunoassay (EIA). Cross-reactivities with other metals were below 1%. Measurement of spiked water samples with this EIA showed good correlation with results obtained by mass spectrometry with inductive coupled plasma (ICP-MS).     

Development of a Monoclonal Antibody-Based ELISA for the Detection of Sulfaquinoxaline in Chicken Tissues and Serum

Abstract

A monoclonal antibody (Mab) was produced by using sulfaquinoxaline-human serum albumin (SQX-HSA) conjugate as immunogen. The anti-SQX Mab exhibited negligible cross reactivity with other commonly used sulfonamides. Using this Mab, a competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed to detect SQX in chicken tissues and serum. The ciELISA showed a 50% inhibition (IC₅₀) value of 2.60 ng/mL. The recoveries of SQX from spiked chicken muscle, liver, and serum at levels of 5–50 µg/kg were 82.6–96.5%, 75.3–94.5%, and 69.7–89.3%, respectively. The coefficient variations (CVs) were 6.22–7.17%, 4.9–8.9%, and 1.20–10.15%, respectively. Detection limits were 1.29 µg/kg in muscle, 1.32 µg/kg in liver, and 2.44 µg/kg in serum.     

Development of a Monoclonal Antibody-Based ELISA for the Detection of Oxytetracycline and 4-Epi-Oxytetracycline Residues in Chicken Tissues

In this study, a specific monoclonal antibody (Mab) against oxytetracycline (OTC) and its metabolite 4-epi-Oxytetracycline (4-epi-OTC) was produced. Based on this Mab, a sensitive and reliable method indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the detection of OTC and 4-epi-OTC from chicken meats. The ic-ELISA showed a 50% inhibition (IC₅₀) value of 2.01 ± 0.16 ng/ml and a detection limit of 0.13 ± 0.03 ng/ml. The recoveries from chicken muscles and livers spiked with OTC of 50–600 ng/g were 83.33–88.25% and 84.62–86.12%, respectively. The intra-assay coefficients of variation (CVs) were 4.73–9.31%, and the inter-assay CVs were 6.44–11.01%. The method showed a positive correlation with the traditional method HPLC (R² = 0.997) within a certain concentration of OTC used in this assay. The method developed in this study was simple and independent of specific expensive equipment. Thus, it could be useful as a convenient method to detect OTC residues.     

气相色谱-质谱法检测沉积物中自由态与束缚态多溴联苯醚和四溴双酚-A

建立了一种同时测定沉积物中不同赋存形态的多溴联苯醚(PBDEs)和四溴双酚A(TBBPA)的分析方法.样品由等体积的丙酮和正己烷混合溶剂抽提得到自由态目标物,再通过碱性水解反应释放束缚态目标化合物.通过调节酸度(pH值)实现PBDEs和TBBPA的分离和提取.PBDEs由复合硅胶柱净化,运用气相色谱-质谱(负离子化学源)-分时段选择离子监测技术测定;TBBPA经重氮甲烷衍生化反应后由酸性硅胶柱预纯化,运用气相色谱-质谱(电子轰击源)-分时段选择离子监测技术测定.8种低溴联苯醚(BDE28,-47,-66,-100,-99,-154,-153,-183),十溴联苯醚(BDE209)和TBBPA的检出限分别为0.6～12.5 pg/g,172 pg/g,4 2 pg/g.方法具有良好的准确度和精确度,回收率均在74%～106%之间,RSD≤10%.对东江沉积物样品的分析表明,本方法能够实现不同形态的PBDEs 和TBBPA的有效检测.     

Thermal Decomposition of Triazine Herbicides II. 6-Chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine (atrazine) and its metabolites

Thermal degradation of atrazine and its metabolites has been investigated using a thermogravimetric technique(TG) with the application of three types of crucibles: opened, Knudsen type and labyrinthtype, and non-isothermal DSC method, using hermetically closed and opened alumina sample pans. The great influence of decomposition conditions (the crucible type) on thermal degradation was observed. TG analysis showed that the degradation process of atrazine took place in three stages. The increase of amino groups in triazine ring increases the amount of non-volatile thermal degradation products by association. The presence of chlorine substituent facilitates the forming of products with low volatility. Hydroxyatrazine decomposes only in one stage process. The dealkylation process observed in hermetical sample pans (DSC) was two-stage and in open sample pans one-stage process. This revised version was published online in July 2006 with corrections to the Cover Date.     

新型三嗪桥连的双1,8-萘酰亚胺衍生物的合成及其光物理行为

以1,8-萘酰亚胺和三聚氯氰为主要原料, 合成了两种由三嗪环桥连的双1,8-萘酰亚胺化合物3 和5. 采用紫外-可见光谱和荧光光谱等手段考察了两种化合物在不同溶剂中的光物理行为. 与参比化合物N-丁基-1,8-萘酰亚胺相比, 在二氯甲烷、三氯甲烷和甲醇等极性溶剂中, 化合物3和5除了在短波区(λ<400 nm)存在1,8-萘酰亚胺的特征荧光发射峰外, 在长波区(>450 nm)均产生一个较强的新荧光发射峰, 表现出分子内激基缔合物的光物理行为. 与化合物5相比, 由于化合物3特殊的构象异构, 其荧光强度发生严重的猝灭. 在非极性溶剂甲基环己烷中, 化合物5 由于存在较强的分子间氢键作用而聚集, 受激后形成了较稳定的分子间激基缔合物, 但未观察到明显的分子内激基缔合物的形成. 在甲苯溶剂中, 化合物3和5与甲苯分子形成了激基复合物, 并未形成分子内激基缔合物. 进一步研究3和5的固态激发态性质, 发现化合物3和5的固体薄膜受激后分别在465和469 nm处出现激基缔合物的特征荧光发射峰.     

Analytical Separation of Copper(II), Cobalt(II) and Nickel(II) Using Polymer Bound Anthrantic Acid

Abstract

The preferential complexing tendency of different nietal ions towards chelating agents anchored on a polymer has been used for separation of transition metals. the anthranilic acid group was anchored on the polymeric cellulose back-bone by successive coupling with trifunctional reagent cyanuric chloride, in diozane medium, at pH 7 and 9–10, respectively. This polymer bound chelating agent was used to separate copper(II), nickel(II) and cobalt(II) in the concentration range 1.0–0.1 mmol/L. the separation of a mixture of two components was quantitative using column chromatography.     

高分子希夫碱金属配合物的研究进展 (Ⅰ)高分子担载希夫碱金属配合物的制备与性能

综述了近十几年来有关高分子担载希夫碱金属配合物的研究进展。此类配合物所采用的合成方法主要有：（ａ）含乙烯侧基的配体或金属配合物共聚；（ｂ）小分子配体或配合物锚连于高分子载体。其主要性能为结合分子氧和催化活性。     

Disappearance of Dislodgable Residues of Methomyl from Leaves and Vegetables in Tamilnadu (India

Abstract

The dislodgable residues of methomyl from leaves and vegtables were determined by a single-step methanol extraction followed by reversed-phase HPLC. The residues on the bhendi vegetables and leaves were several times higher than on the brinjal vegetables and leaves. The pesticide, however, dissipates rapidly under the South Indian climatic conditions resulting in undetectable residues within three days of spraying.     

大口径毛细管气相色谱法测定果蔬中15种有机氯农药残留量

果蔬样品中１５种有机氯农残用丙酮－石油醚提取,经含５％Ｈ２Ｏ的浓硫酸磺化离心净化,ＧＣ－ＥＣＤ测定。色谱柱为ＨＰ－５０＋大口径毛细管柱,方法的回收率为８５．７～１０８．５％,变异系数４．８～１２．６％。     

Amperometric Immunosensor for the Detection of 2,4-Dichlorophenoxyacetic Acid (2,4-D) in Water

ABSTRACT

An amperometric immunosensor for the determination of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) in water has been developed using sequential injection analysis techniques. The system is based on a rapid competitive enzyme immunoassay employing an alkaline phosphatase-labeled monoclonal antibody directed against the herbicide and an immunoreactor with 2,4-D immobilized via bovine serum albumin either to Eupergit in a column or directly to the surface of a glass capillary. The detection limit of the immunosensor at 0.1 μg 2,4-D/l without enrichment of the analyte makes automatic measurements of 2,4-D in drinking and ground water feasible.     

Corn Silage Supplemented with Pomegranate (Punica granatum) and Avocado (Persea americana) Pulp and Seed Wastes for Improvement of Meat Characteristics in Poultry Production

In the present study, pomegranate peels, avocado peels, and seed vacuum microwave extraction solid by-products were supplemented in corn silage in order to investigate the effects on meat quality and growth rate in broiler chicken. There were 50 broilers, divided in two groups, treated with experimental or usual feed for 43 days (group A: 25 broilers fed with avocado and pomegranate by-products and group B: 25 broilers fed with corn-silage used as control). The results showed that broiler chickens fed with a diet supplemented with a mixture of pomegranate avocado by-products (group A) showed significant differences in chicken leg meat quality, significantly improving the level of proteins and fatty acids content in breast and leg meat, respectively. More specific ω3 and ω6 fatty acids content were three times higher than in group B. Moreover, a protective effect on the decomposition of polyunsaturated fatty acids, induced by free radicals and presented in chicken meat, is based on the evaluation of lipid peroxidation by measuring thiobarbituric acid reactive substances. Pomegranate peels, avocado peels, and seed by-products appeared to have a slight reduction on meat production, while it was found to improve the qualitative chicken meat characteristics. Regarding the production costs, it was calculated that the corn-silage supplementation, used in this study, lead to a 50% lower cost than the commercial corn-silage used for the breeding of broilers.     

Preparation of Monoclonal Antibody and Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Fluorescence-Linked Immunosorbent Assay for Detecting 3-Amino-5-methylmorpholino-2-oxazolidinone (AMOZ) in Edible Animal Tissue

To monitor the illegal used of furaltadone, a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and fluorescence-linked immunosorbent assay (FLISA) based on a monoclonal antibody (mAb) were developed for the detection of 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), the major metabolite of furaltadone in animal tissues. The highly specific mAb, which was very sensitive to a nitrophenyl derivative of AMOZ (2-NP-AMOZ) with IC₅₀ values of 0.11 and 0.09 ng/mL for ic-ELISA and FLISA, respectively, was selected for the development of immunoassays. For both the ic-ELISA and FLISA for AMOZ-spiked experiments, acceptable recovery rates of 81.1–105.3% and coefficients of variation of 4.7–9.8% were obtained. In addition, results from both ic-ELISA and FLISA methods for spiked samples’ data showed excellent correlation coefficients ranging from 0.9652 to 0.9927. Meanwhile, the proposed ic-ELISA and FLISA for thirty spiked samples were confirmed by standard LC-MS/MS with high correlation coefficients of 0.9911 and 0.9921, respectively. These results suggest that the developed ic-ELISA and FLISA are valid and cost-effective tools for high-throughput monitoring methods for AMOZ residues in animal tissues.     

Solvent (ionic liquid) impregnated resin-based extraction coupled with dynamic ultrasonic desorption for separation and concentration of four herbicides in environmental water

A new method was developed for the determination of monolinuron, propazine, linuron, and prebane in environmental water samples. The solvent (ionic liquid) impregnated resin (IL-SIR)-based extraction coupled with dynamic ultrasonic desorption (DUSD) was applied to the separation and concentration of the analytes. The high performance liquid chromatography (HPLC) was applied to the determination of the analytes. The ionic liquid [C₆MIM][PF₆] was immobilized on Diaion HP20 resin by immersing the resin in ethanol solution containing [C₆MIM][PF₆]. The effect of extraction parameters, including pH value of sample solution, salt concentration in sample and extraction time, and elution conditions, including the concentration of ethanol in elution solvent, the flow rate of elution solvent and the ultrasonic power, were examined and optimized. The limits of detection and quantification for the analytes were in the range of 0.15-0.29 μg L⁻¹ and 0.51-0.98 μg L⁻¹, respectively. Some environmental water samples were analyzed and the analytical results were satisfactory.     

纳米零价铁在水相反应中的表面化学和晶相转化

纳米零价铁在水相中的表面化学特性和晶相等性质变化,将影响其反应活性及环境归趋等.总结近期课题组关于纳米零价铁在水相中表面化学和晶相转化的研究进展,为纳米零价铁污染控制化学提供基础理论.重点探讨水中有无溶解氧、不同水力学条件复氧（静态和扰动）、重金属共存、无机阴离子共存对纳米零价铁颗粒表面化学特性和晶相转变的影响.同时也研究高分子电解质表面修饰后,颗粒在水相中表面及晶相的演变及对重金属去除性能的影响.研究表明,纳米零价铁与水相中的水分子、溶解氧、重金属离子及无机阴离子反应,零价铁失去电子演变为氧化铁、羟基氧化铁等;环境条件对颗粒结构性能产生影响,从而影响污染物去除效率及其在环境中的归趋.未来研究将重点探讨结构性能动态变化与不同污染物之间反应性能的影响,建立纳米颗粒的结构与性能之间关系模型,为纳米零价铁材料的环境应用提供理论依据.