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31.
酶联免疫吸附分析法测定水样中的阿特拉津 总被引:18,自引:0,他引:18
应用阿特拉津半抗原衍生物共价交联于载体蛋白分子上,制成免疫抗原,经免疫得到高质量的兔抗阿特拉津抗血清。经条件优选,建立了测定阿特拉津的酶联吸附分析竞争法,测定性范围为0.05-5.00μg/L,最低检出限仅为0.018-0.022μg/L,且精密度高,特异性强。 相似文献
32.
A fluorimetric assay for cortisol 总被引:2,自引:0,他引:2
Appel D Schmid RD Dragan CA Bureik M Urlacher VB 《Analytical and bioanalytical chemistry》2005,383(2):182-186
A simple, rapid and sensitive fluorimetric assay for the quantitative determination of cortisol is reported. The assay is
based on the formation of a fluorescent dye when cortisol is incubated with a mixture of sulfuric acid and acetic acid. The
fluorescence spectrum recorded for the resulting dye shows a maximum extinction at 475 nm and a maximum emission at 525 nm.
The solvent 2-methyl-4-pentanone was used for extraction and was found to act as a fluorescence amplifier. A limit of detection
of 2.7 μM was achieved, making it possible to forego solvent evaporation. The assay suffers minor interference from 11-deoxycortisol
which exhibits low fluorescence at λ
ex: 460 nm; λ
em: 505 nm. Typical standard deviations were below 4%. We validated the assay using a biotransformation with recombinant Schizosaccharomyces pombe which regioselectively hydroxylates 11-deoxycortisol to cortisol. The method described herein is suitable for preliminary
screening of microorganisms capable of steroid hydroxylation. 相似文献
33.
This paper presents a new simple and sensitive method for the micro-determination of protein containing SH– and –S–S– groups based on the single sweep polarographic wave of an electroactive derivative. In 0.04molL–1 Na3PO4 and 0.2% ascorbic acid solution, protein is heated in a boiling water bath for 15min, the reaction product giving a sensitive reduction wave at –0.70V (vs. SCE). The wave height is linearly proportional to the concentration of protein. The calibration curves of bovine serum albumin (BSA), human serum albumin (HSA), ovalbumin (OVA) and lysozyme (Lyso) are constructed under the optimal conditions. For BSA and HSA, the linear ranges and detection limits are 0.05–24mgL–1 and 0.02mgL–1, respectively. The method has been applied to the determination of protein in human serum samples with satisfactory results. The mechanism of the polarographic wave was also studied, and the results show that S2– ion is released from the protein molecule during the derivatization reaction, the wave being attributed to the reduction of HgS. 相似文献
34.
Joon Myong Song 《Analytica chimica acta》2004,507(1):115-121
In this work, we report Escherichia coli O157:H7 detection using antibody-immobilized capillary reactors, enzyme-linked immunosorbent assay (ELISA), and a biochip system. ELISA selective immunological method to detect pathogenic bacteria. ELISA is also directly adaptable to a miniature biochip system that utilizes conventional sample platforms such as polymer membranes and glass. The antibody-immobilized capillary reactor is a very attractive sample platform for ELISA because of its low cost, compactness, reuse, and ease of regeneration. Moreover, an array of capillary reactors can provide high-throughput ELISA. In this report, we describe the use of an array of antibody-immobilized capillary reactors for multiplex detection of E. coli O157:H7 in our miniature biochip system. Side-entry laser beam irradiation to an array of capillary reactors contributes significantly to miniaturized optical configuration for this biochip system. The detection limits of E. coli O157:H7 using the ELISA and Cy5 label-based immunoassays were determined to be 3 and 230 cells, respectively. This system shows capability to simultaneously monitor multifunctional immunoassay and high sensitive detection of E. coli O157:H7. 相似文献
35.
Surface plasmon resonance (SPR) has been successfully applied for the simple, rapid, and label-free assay of various biomolecules. This assay evaluates a novel wavelength modulation SPR biosensor for the detection of tetanus toxin. The wavelength modulation SPR biosensor is designed based on fixing the incident angle of light and measuring the reflected intensities in the resonance wavelength range spanning 400-800 nm simultaneously. Tetanus toxin (TeNT), one of the most potent toxins known, is synthesized as a 150 kDa single polypeptide chain. The SPR biosensor has been shown to be capable of directly detecting concentration of tetanus toxin as low as 0.028 Lf ml−1. Under selected experimental conditions, the SPR biosensor has a good reproducibility, sensitivity and reversibility. The results illustrate how wavelength modulation SPR biosensor can be used to detect biomolecular interactions. 相似文献
36.
Clinical applications of chemiluminescence 总被引:8,自引:0,他引:8
This article reviews the clinical applications of chemiluminescence in routine testing and surveys the diverse applications of chemiluminescence in clinical research. In routine clinical testing, chemiluminescent labels (acridinium ester, acridinium sulfonamide) and detection reactions for peroxidase and alkaline phosphatase enzyme labels (luminol and adamantyl 1,2-dioxetane-based reactions, respectively) are widely used in immunoassay and nucleic acid probe assays (e.g. hybridization protection assay, Hybrid Capture® assay). In clinical research the sensitivity, dynamic range and diversity of chemiluminescent assays has led to a vast range of applications, notably in protein and nucleic acid blotting, microarray-based assays, monitoring reactive oxygen species, and as detection reactions for substances separated by HPLC, capillary electrophoresis (CE), and flow-injection analysis. 相似文献
37.
Papaverine (1-(3,4-dimethoxybenzyl)-6,7-dimethoxyisoquinoline, PAP) is a member of the benzylisoquinoline sub-group of the opium alkaloids. It has been widely used for treating diseases like pulmonary arterial embolism and renal or biliary colic. In this paper, a specific conjugate of mono-demethylated papaverine-O-carboxylmethyl ether (MDMPAP-O-CME) and bovine serum albumin (BSA) was synthesized and used as the complete antigen (PAP-BSA), with which we successfully obtained a high-titer anti-PAP polyclonal antibody (pAb) by immunization of rabbits. The anti-PAP pAb showed high affinity to papaverine with an affinity constant (Kaff) of 7.3 × 107 L/mol. With this antibody, we established a sensitive immunochemical method for the determination of papaverine based on indirect competitive enzyme-linked immunosorbent assay (ELISA). The optimal concentrations of the coated antigen (PAP-OVA) and purified pAb used in the ELISA were 5 and 1.2 μg/mL, respectively. The cross reactivity of other benzylisoquinoline derived substances, including 1-(3,4-dihydroxybenzyl)-7-hydroxy-6-methoxy-isoquinoline (6-methoxy-papaveroline, MPAPO), morphine (MP) and codeine (CD) were all lower than 1%. The linear range of the calibration curve was 0.1-1000 ng/mL. Normal human serum samples were spiked with known amount of papaverine and measured by the ELISA. Recoveries were between 102% and 105%. Papaverine content in a commercial papaverine hydrochloride injection sample was also determined using the established ELISA. Compared with the results given by the control experiment of HPLC, the recoveries of ELISA to detect injection samples were 102-110%. The limits of detection for synthetic serum samples and injection samples of papaverine hydrochloride were 0.25 and 0.06 ng/mL, respectively. 相似文献
38.
R.-L. Yun Y.-M. Chang C.-H. Lin K.-H. Hu C.-M. Shu 《Journal of Thermal Analysis and Calorimetry》2006,85(1):107-113
In industrial processes, information
on the safety property of chemicals is essentially crucial for safe handling
during unit operations. Ensuring the safe use of combustible or flammable
substances in processes is unlikely without detailed investigations of their
flammability characteristics and related hazards. We studied 3-methyl pyridine
(3-picoline), e.g., flammability limits (LFL/UFL), maximum explosion pressure
(P
max), maximum
explosion pressure rise (dP/dt)max, minimum oxygen concentration
(MOC), vapor deflagration index (K
g),
and characterized the influence of inert steam (H2O)
on critical parameters for 3-picoline/water mixtures at 270°C, 1 atm,
various oxygen concentrations, and vapor mixing ratios (100/0, 30/70, 10/90
and 5/95 vol.%) with a 20-L-Apparatus in simulated conditions, respectively.
The results showed that the flammability characteristics
of 3-picoline(aq) all increased with the oxygen concentration.
However, as the composition of inert steam increased, the flammability parameters
and the degree of fire and explosion hazards were significantly reduced, instead.
This study elucidated the flammability properties of 3-picoline mixed with
inert steam. The conclusions could be applied to proactively prevent the relevant
processes from incurring fire and explosion accidents. 相似文献
39.
化学发光单孵多层免疫技术检测粪样中轮状病毒 总被引:3,自引:0,他引:3
以酶联免疫吸附分析(ELISA)夹心测定粪样中轮状病毒(RV)实验为模型,将传统又孵式免疫操作变成单孵式,即在包被RV抗体Ab的微孔中同时加入含RV的粪样上清液和根过氧化物酶(HRP)标记抗RV抗原共同孵育,其结果在载体表面形成多层酶免疫复合物,以HRP催化鲁米诺-对碘苯酚-过氧化氢化学体系作为最终信号检测系统,从而建立起化学发光单孵多层免疫技术(CL-SIMIT)。其灵敏度是ELISA的16倍, 相似文献
40.
Yunhui Zhai Xijun Chang Yuemei Cui Ning Lian Shoujun Lai Hong Zhen Qun He 《Mikrochimica acta》2006,154(3-4):253-259
A rapid, selective method that utilize 4-(2-Pyridylazo)-resorcinol (PAR)-modified nanometer SiO2 (nanometer SiO2–PAR) as a new solid-phase extractant for preconcentration of trace mercury (II) has been developed. The adsorption property
of nanometer SiO2–PAR for metal ions was studied by selectively extracting different metal ions from aqueous solutions. The results revealed
an excellent affinity of the nanometer SiO2–PAR for mercury (II) in presence of interfering metal ions at pH 4. The main parameters of solid-phase extraction such as
shaking time, elution and sample dilution effect were studied. The extractant shows rapid kinetic sorption, and the adsorption
equilibrium of mercury (II) on nanometer SiO2–PAR was achieved in less than 2 min. The adsorbed mercury (II) was easily eluted by 4 mL of 6 mol L−1 HCl. The maximum preconcentration factor was 50. The maximum static adsorption capacity was 276 μmol g−1 at pH 4. The detection limit (3σ) was 0.43 μg L−1 for cold vapor atomic absorption spectrometry (CVAAS), and the relative standard deviation of the eight replicate determinations
was 2.4% for the determination of 2.0 μg of Hg(II) in 100 mL water sample. The method was applied to the determination of
trace mercury (II) in sample solutions with satisfactory results. 相似文献