酸改性纳米TiO_2降解模拟污染物的研究

用溶胶-凝胶法制备了酸改性纳米二氧化钛,并用XRD、UV-vis等测定技术对所制得的粉体试样进行了表征,同时以甲基橙为模拟污染物,评价了改性后纳米二氧化钛的光催化性能,发现其光催化活性大大高于未改性二氧化钛粉体。并从酸改性的样品中筛选出了光催化活性最好的粉体,以光催化降解苯酚为模型反应,确立了降解的最优条件。     

鲫鱼(Carassius auratus)卵黄蛋白原的ELISA检测

采用Sephadex G-200柱层析从鲫鱼(Carassius auratus)成熟鱼卵中提取卵黄脂磷蛋白并制备抗卵黄脂磷蛋白抗血清.用DEAE-cellulose阴离子交换层析和Sephacryl S-300分子筛凝胶层析相结合的两步层析从经17b-雌二醇诱导的雌鱼血浆中提纯卵黄蛋白原.免疫印迹反应显示卵黄脂磷蛋白和卵黄蛋白原都与抗卵黄脂磷蛋白抗血清有特异的反应.以抗卵黄脂磷蛋白抗血清作为抗体,卵黄蛋白原作为竞争抗原建立了间接竞争法酶联免疫吸附反应(ELISA)方法来检测雄鱼血液中卵黄蛋白原含量.该方法可测的最低浓度为7.8ng/mL,工作范围为0.39~25mg/mL,批内和批间误差分别为6.5%和7.5%.     

Specific antibody for pesticide residue determination produced by antibody-pesticide complex

A new method for specific antibody production was developed using antibody (Ab)-pesticide complex as a unique immunogen. Parathion (PA) was the targeted pesticide, and rabbit polyclonal antibody (Pab) and mouse monoclonal antibody (Mab) were used as carrier proteins. The Ab-PA complexes were generated by conjugating the two antibodies with an excessive dosage of PA. It was shown that the sensitivity of homologous enzyme-linked immunosorbent assay (ELISA) using the new antibodies was similar to that using original antibodies. However, the new mouse Pab had not only similar positive recognizing spectrum as the original Mab, but also a significantly improved sensitivity in heterologous ELISA when some recognizable competitors were applied. IC₅₀ value of ELISA based on a combination of the new mouse Pab and hapten 9 was 0.24 ng/mL, which was 445.54 times as that of the homologous ELISA. An Ab-pesticide complex may be a suitable alternative immunogen for producing highly specific antibody to improve the immunoassay sensitivity.     

ELISA AND GC/MS ANALYSIS OF PESTICIDE RESIDUES IN NORTH CAROLINA1

ABSTRACT: Studies were conducted to analyze the presence of 11 pesticide residues in 12 surface waters in the Piedmont and coastal plain regions of North Carolina. Samples were assayed using enzyme‐linked immunosorbent assays (ELISAs). All ELISA results of one part per billion (ppb, μg/L) or greater were confirmed using gas chromatography/mass spectrometry (GCIMS). ELISA detection limits were approximately an order of magnitude higher than GCJMS methods. Of the 5,035 analytical results from 742 surface water samples, atrazine was detected in approximately 45 percent of the samples, five of which were at or above the Maximum Contaminant Level (MCL) of 3‐ppb. Metolachlor was detected in 64 percent of the samples. Aldicarb, 2,4‐D, chlorpyrifos, and chlorothalonil were also detected, and each was found in less than 12 percent of the samples. The remaining pesticides, including carbaryl, acetochior, methomyl, carbofuran, and alachlor, were not detected during the study period.     

Rapid assays for environmental and biological monitoring

Abstract

Rapid, inexpensive, sensitive, and selective enzyme‐linked immunosorbent assays (ELISAs) now are utilized in environmental science. In this laboratory, many ELISAs have been developed for pesticides and other toxic substances and also for their metabolites. Compounds for which ELISAs have recently been devised include insecticides (organophosphates, carbaryl, pyrethroids, and fenoxycarb), herbicides (s‐triazines, arylureas, triclopyr, and bromacil), fungicides (myclobutanil), TCDD, and metabolites of naphthalene and toluene. New rapid assays have been developed for mercury.     

水体中微囊藻毒素-LR的间接竞争ELISA检测

在自制包被完全抗原浓度为5μg/mL、单克隆抗体工作稀释度为1∶3 000、酶标二抗鼠工作稀释度为1∶3 000、微囊藻毒素-LR浓度在0.001～30μg/L、显色底物为邻苯二胺,采用间接竞争酶联免疫吸附试验对水体中的微囊藻毒素-LR进行检测,结果表明,该方法与高效液相色谱的检测结果相关系数大于0.99,多次重复实验相对标准偏差小于10%,最低检测限能达到0.01μg/L,定量检测区间为0.01～3μg/L,该方法对[4-精氨酸]微囊藻毒素能特异性识别,对来自实际水样中的干扰有相当的耐受力.     

间接竞争ELISA方法测定水中2，4-D的研究

以2,4-D-BSA为包被抗原,采用自行制备的2,4-D单克隆特异性抗体6D11建立了水中2,4-D的间接竞争 ELISA检测方法．本研究比较了包被抗原2,4-D-BSA浓度分别为240ng·mL-1、120ng·mL-1和60ng·mL-1的反应体系和竞争反应时间为60min和15min的间接竞争ELISA剂量-反应曲线,确定了当包被抗原浓度为60 ng·mL-1、竞争反应时间为15min时,剂量-反应曲线的IC50值较低．采用上述实验条件分别测定了由PBS缓冲溶液、饮用水、清华大学地下水和圆明园福海地表水配制的2,4-D标准溶液的剂量．反应曲线,发现实际水样的基质效应对检测结果的影响较大;采用实际水样和PBS缓冲溶液配水在含有5％乙醇的PBS缓冲体系中反应的方法,基本上消除了基质效应对检测结果的干扰．采用上述优化试验条件,测定2,4-D浓度分别为0．5mg·L-1、0．125mg·L-1和0．03mg·L-1的加标样品,测定数据的准确度符合痕量有机污染物定量检测对准确度的要求,但是平行样品测定数据之间的变异系数较大,需要进一步改进检测方法,用于实际水样的检测．     

A sensitive monoclonal antibody-based enzyme-linked immunosorbent assay for chlorpyrifos residue determination in Chinese agricultural samples

A monoclonal antibody-based competitive antibody-coated enzyme-linked immunosorbent assay (ELISA) was developed and optimized for determining chlorpyrifos residue in agricultural products. The IC₅₀ and IC₁₀ of this ELISA were 3.3 ng/mL and 0.1 ng/mL respectively. The average recoveries in six agricultural products were between 79.5% and 118.0%, with the intra-assay coefficient of variation being less than 8 %. The limit of detection for all tested products was 30 ng/g. To the best of our knowledge, this assay has the best specificity among all the published research on ELISAs for chlorpyrifos.     

剑尾鱼卵黄蛋白原的ELISA检测

以卵黄脂磷蛋白(lipovitellin,Lv)抗血清为抗体,以纯化的卵黄蛋白原(vitellogenin,Vtg)为抗原,建立了间接酶联免疫吸附反应(ELISA)方法检测剑尾鱼(Xiphophorus helleri)雄鱼整体匀浆液中ρ(Vtg).结果表明,该方法的检测灵敏度为7.8 ng/mL,批内变异系数为5.094%,批间变异系数为5.540%,工作范围为32.5～2 000 ng/mL,在该范围内,标准曲线具有良好的线性和重复性.由于该方法可直接在1块或在不同的酶标板上准确地进行比较,因而可利用ELISA方法测定经诱导雄鱼整体匀浆液中ρ(Vtg)水平.结果显示,50 μg/L 17-β雌二醇(E₂)诱导21 d的雄性剑尾鱼有明显的Vtg产生;10 μg/L E₂诱导剑尾鱼亦有Vtg产生,但较50 μg/L E₂诱导组低;1 μg/L E₂诱导组及对照组剑尾鱼没有Vtg产生,检测孔OD450/对照OD₄₅₀(P/N)值小于2.1.      

The acute lethality and endocrine effect of 1,2,3,7,8-PeCDD in juvenile goldfish （ Carassius auratus ） in vivo

1,2,3,7,8-PeCDD was administrated to juvenile goldfish （Carassius auratus） by peritoneal injections to explore the acute lethality and endocrine effects of 1,2,3,7,8-PeCDD in vivo. The value of acute median lethal dosage （LD50） of 1,2,3,7,8-PeCDD was determined in acute lethality tests. The endocrine effect of 1,2,3,7,8-PeCDD, whose exposed concentrations were determined based on the LD50 （1.84 mg/kg）, was studied by measuring the plasma vitellogenin （Vtg） content in juvenile male goldfish with enzyme-linked immunosorbent assays （ELISA）. Due to its significant induction of the plasma Vtg after one week＇s exposure in vivo in the 1/2 LD50 and LD30 groups, 1,2,3,7,8-PeCDD might be one of the important contributors to the estrogenic effect of PCDDs in the environment. The values of 1/2 LD50 and LD30 were within the range of the effective dosages of 1,2,3,7,8-PeCDD, indicating that there was a certain relationship between the estrogenic effective dosages and the LD50.