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设计了一种测定人血清中癌胚抗原的毛细管电泳-化学发光检测均相免疫分析新方法。采用四苯硼酸钠增强luminol-H2O2-HRP体系化学发光的原理,化学发光检测经毛细管电泳分离的,用辣根过氧化物酶(HRP)标记的癌胚抗体及免疫复合物。测定癌胚抗原的线性范围2.0~80.0μg/L(R=0.9921),检出限为0.1μg/L(绝对检出限为0.75 fg)。 相似文献
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根据某些氨基酸和蛋白质在碱性条件下具有能显著增强鲁米诺-铁氰化钾体系化学发光信号的特性,对毛细管电泳-化学发光法直接分离测定生物分子的可行性进行了研究.电泳所用缓冲溶液为2.5 mmol/L、含2.0 mmol/L鲁米诺的硼砂缓冲溶液;氧化试剂为2.0×10-4 mol/L铁氰化钾.在优化条件下,L-精氨酸和胃蛋白酶可直接实现分离,其检出限(S/N=3)分别为1.3×10-4 mol/L和7.1×10-7 mol/L.通过对市售的3种L-精氨酸标准样品进行分离分析研究,证明该方法可应用于L-精氨酸标准样品的纯度分析. 相似文献
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基于诺氟沙星对联吡啶钌在铂电极上的电致发光信号有增敏作用,与毛细管电泳结合,建立了一种测定诺氟沙星的电化学发光分析新方法。研究了工作电极电位、磷酸盐缓冲液浓度及其pH值、进样电压和进样时间等实验参数对诺氟沙星测定的影响。在优化的实验条件下,其浓度线性范围为0.02~10μmol/L;检出限(3σ)为0.0048μmol/L;峰面积的相对标准偏差为2.6%(1.0μmol/L,n=11)。本法可直接用于尿液中诺氟沙星(NFLX)含量的测定。回收率为92.7%~97.9%,结果满意。 相似文献
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毛细管电泳-化学发光法测定人血清中的异烟肼 总被引:2,自引:0,他引:2
基于碱性介质中异烟肼对laminol-K3Fe(CN)6化学发光体系的增敏作用,设计了一个经毛细管电泳(CE)分离,在线化学发光检测异烟肼的新方法.研究并优化了毛细管电泳分离及化学发光检测的条件.在优化的实验条件下,该方法测定异烟肼的线性范围为4.0×10-7~1.0×10-5g/mL(R2=0.9984),检出限(3σ)为1×10-8g/mL,对6.0×10-6g/mL的异烟肼进行6次平行测定,其相对标准偏差为4.0%.方法已用于血清中异烟肼的测定. 相似文献
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基于诺氟沙星对联吡啶钌在铂电极上的电致发光信号有增敏作用,与毛细管电泳结合,建立了一种测定诺氟沙星的电化学发光分析新方法.研究了工作电极电位、磷酸盐缓冲液浓度及其pH值、进样电压和进样时间等实验参数对诺氟沙星测定的影响.在优化的实验条件下,其浓度线性范围为0.02~10 μmol/L;检出限(3σ)为0.0048 μmol/L;峰面积的相对标准偏差为2.6%(1.0 μmol/L,n=11).本法可直接用于尿液中诺氟沙星(NFLX)含量的测定.回收率为92.7%~97.9%,结果满意. 相似文献
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一种毛细管电泳-化学发光联用系统的改进及应用 总被引:2,自引:1,他引:2
在毛细管电泳 化学发光联用研究工作的基础上 ,提出了一种简单可靠的联用检测接口 ,并对两端电极池作了改进。高电位池的改进 ,解决了支持缓冲液中鲁米诺电解造成的发光信号漂移问题 ;低电位池的改进 ,使发光反应中排出的废液不混入电极缓冲溶液。从而保持化学发光信号的基线稳定 ,避免发光试剂的损耗并延长缓冲溶液的使用时间。整个体系简单易行 ,采用此联用系统和鲁米诺 铁氰化钾间接化学发光法 ,分离了绿原酸和芸香苷。本法的检出限达到 1.4× 10 -5mol/L ,比毛细管电泳 紫外吸收法降低约 2个数量级 相似文献
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Dungchai W Siangproh W Lin JM Chailapakul O Lin S Ying X 《Analytical and bioanalytical chemistry》2007,387(6):1965-1971
A micro-magnetic chemiluminescence (CL) enzyme immunoassay with high sensitivity, selectivity, and reproducibility was developed
for the determination of the tumor marker, carcinoembryonic antigen (CEA) in human serum. A sandwich scheme assay has been
utilized with fluorescein isothiocyanate antibody (FITC)-labeled anti-CEA antibody and alkaline phosphate (ALP)-labeled anti-CEA
antibody being used in the CL detection. The CL signal produced by the emission of photons from 4-methoxy-4-(3-phosphate-phenyl)-spiro-(1,2-dioxetane-3,2′-adamantane)
(AMPPD) was directly proportional to the amount of analyte present in a sample solution. The influences of the reaction time
of antigen with antibody, the reaction time of substrate with label, the dilution ratio of ALP-labeled anti-CEA antibody,
the concentration of FITC-labeled anti-CEA antibody, and other relevant variables upon the CL signal were examined and optimized.
The CL responses depended linearly on the CEA concentration over the range from 2 to 162 ng mL−1 in a logarithmic plot. Assay sensitivity as low as 0.69 ng mL−1 was achieved. A coefficient of variance of less than 13% was obtained for intra- and inter-assay precision. This method has
been successfully applied to the analysis of CEA in human serum. According to the procedure based on spiked standards, the
recoveries obtained were 80–110%. Comparison experiments were carried out with the commercially available CEA chemiluminescence
immunoassay. Satisfactory results were obtained according to a paired t-test method (t value < t
critical at the 95% confidence level). 相似文献
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Determination of clenbuterol by capillary electrophoresis immunoassay with chemiluminescence detection 总被引:2,自引:0,他引:2
A competitive immunoassay for clenbuterol (CLB) based on capillary electrophoresis with chemiluminescence (CL) detection was established. The method was based on the competitive reaction of horseradish peroxidase (HRP)-labeled CLB (CLB-HRP) and free CLB with anti-CLB antiserum. The factors affecting the electrophoresis and CL detection were systematically investigated with HRP as a model sample. Under the optimal conditions, the tracer CLB-HRP and the immunoassay complex were separated, and the linear range and the detection limit (S/N = 3) for CLB were 5.0-40 nmol l−1 and 1.2 nmol l−1, respectively. The proposed method has been applied satisfactorily in the analysis of urine sample. 相似文献
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A rapid and simple method using capillary electrophoresis (CE) with chemiluminescence (CL) detection was developed for the determination of levodopa. This method was based on enhance effect of levodopa on the CL reaction between luminol and potassium hexacyanoferrate(III) (K3[Fe(CN)6]) in alkaline aqueous solution. CL detection employed a lab-built reaction flow cell and a photon counter. The optimized conditions for the CL detection were 1.0 × 10−5 M luminol added to the CE running buffer and 5.0 × 10−5 M K3[Fe(CN)6] in 0.6 M NaOH solution introduced postcolumn. Under the optimal conditions, a linear range from 5.0 × 10−8 to 2.5 × 10−6 M (r = 9991), and a detection limit of 2.0 × 10−8 M (signal/noise = 3) for levodopa were achieved. The precision (R.S.D.) on peak area (at 5.0 × 10−7 M of levodopa, n = 11) was 4.1%. The applicability of the method for the analysis of pharmaceutical and human plasma samples was examined. 相似文献
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Indirect detection of paracetamol was accomplished using a capillary electrophoresis-chemiluminescence (CE-CL) detection system, which was based on its inhibitory effect on a luminol-potassium hexacyanoferrate(III) (K3[Fe(CN)6]) CL reaction. Paracetamol migrated in the separation capillary, where it mixed with luminol included in the running buffer. The separation capillary outlet was inserted into the reaction capillary to reach the detection window. A four-way plexiglass joint held the separation capillary and the reaction capillary in place. K3[Fe(CN)6] solution was siphoned into a tee and flowed down to the detection window. CL was observed at the tip of the separation capillary outlet. The CL reaction of K3[Fe(CN)6] oxidized luminol was employed to provide the high and constant background. Since paracetamol inhibits the CL reaction, an inverted paracetamol peak can be detected, and the degree of CL suppression is proportional to the paracetamol concentration. Maximum CL signal was observed with an electrophoretic buffer of 30 mM sodium borate (pH 9.4) containing 0.5 mM luminol and an oxidizer solution of 0.8 mM K3[Fe(CN)6] in 100 mM NaOH solution. Under the optimal conditions, a linear range from 6.6 × 10−10 to 6.6 × 10−8 M (r = 0.9999), and a detection limit of 5.6 × 10−10 M (signal-to-noise ratio = 3) for paracetamol were achieved. The relative standard deviation (R.S.D.) of the peak area for 5.0 × 10−9 M of paracetamol (n = 11) was 2.9%. The applicability of the method for the analysis of pharmaceutical and biological samples was examined. 相似文献
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In this paper we have presented a sensitive and rapid immunoassay (IA) method by capillary electrophoresis with an enhanced chemiluminescence detection system (CE-CL) based on the catalytic effects of horseradish peroxidase (HRP) on the luminol-hydrogen peroxide reaction. The conditions for the CL reaction and electrophoresis were systematically investigated using HRP as a model sample. The linear range from 2.5 x 10(-11) to 1.0 x 10(-9) mol/L (R = 0.999), and the detection limit of 1.0 x 10(-12) mol/L (signal-to-noise ratio = 3) for HRP were achieved using para-iodophenol as CL enhancer. The relative standard deviations of the migration time and peak area for 5.0 x 10(-10) mol/L HRP (n = 7) were 0.26 and 4.8%, respectively, using a CE system with a home-built CL detector. Under the optimal condition, the HRP-labeled CA125 antibody (Ab) and the Ab-antigen complex were well separated within 4 min by CE using a high-pH buffer (pH 10.20). The assay was successfully used for quantification of CA125 in human sera from health controls and patients associated with ovarian cancer, and the recoveries of the standard addition experiments were 93-109%. Our primary results demonstrated that IA based on CE-CL detection is a powerful tool for clinical diagnosis combined with these commercial IA kits. 相似文献
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《Electrophoresis》2017,38(13-14):1780-1787
An enzyme and antibody dual labeled gold nanoparticles enhancing chemiluminescence strategy was developed for highly sensitive CE immunoassay (IA) of prostate‐specific antigen (PSA). In this work, gold nanoparticles were labeled with horseradish peroxidase and antiprostate specific antigen‐antibody, and used as the marker (Ab*). After PSA (antigen, Ag) was added into the system, a noncompetitive immune reaction was happen between Ab* and Ag to form an immune complex (Ag–Ab*). Subsequently, the obtained Ag–Ab* and unreacted Ab* were separated by CE, and the chemiluminescence intensity of Ag‐Ab* was used to estimate PSA concentration. The calibration curve showed a good linearity in the range of 0.25–10 ng/mL. Based on a S/N of 3, the detection limit for PAS was estimated to be 0.092 ng/mL. Proposed CE method was applied for PSA quantification in human serum samples from healthy volunteers and patients with prostate cancer. The obtained results demonstrated that the proposed CE method may serve as an alternative tool for clinical analysis of PSA. 相似文献
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We developed a capillary electrophoresis with chemiluminescence detection system using 2-methyl-6-p-methoxyphenylethynylimidazopyrazinone as a chemiluminescence reagent for determination of antioxidants of superoxide anions. 2-Methyl-6-p-methoxyphenylethynylimidazopyrazinone reacted with superoxide anions generated through the reaction of hypoxanthine and xanthine oxidase, and then emitted chemiluminescence. Suppression of the chemiluminescence in the presence of antioxidants for superoxide anions was introduced as a detection principle for antioxidants into the capillary electrophoresis with chemiluminescence detection system. After optimizing the analytical conditions, various antioxidants, such as superoxide dismutase, nitroblue tetrazolium, ascorbic acid, and catechin, were subjected to the present system. They gave negative peaks due to the quenching effect; the detection limits of superoxide dismutase, nitroblue tetrazolium, ascorbic acid, and catechin were 1, 100, 100, and 10 μM, respectively (S/N = 2). A model sample consisting of superoxide dismutase and nitroblue tetrazolium was satisfactorily separated and detected within ca. 10 min. We also applied the present system to analysis of catechin in green tea as a real sample. 相似文献
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Multichannel homogeneous immunoassay for detection of 2,4,6-trinitrotoluene (TNT) using a microfabricated capillary array electrophoresis chip 总被引:6,自引:0,他引:6
A high-throughput homogeneous immunoassay for the sensitive detection of 2,4,6-trinitrotoluene (TNT) has been developed using radial capillary array electrophoresis microdevices. Samples consisting of equilibrium mixtures of anti-TNT antibody (Ab), fluorescein-labeled TNT, and various concentrations of unlabeled TNT were electrokinetically injected into 48 channels of a radial capillary array electrophoresis microchannel plate. The rapid electrophoretic separation allows us to analyze the equilibrium ratio formed by the competition between the labeled and the unlabeled TNT for Ab binding. The simultaneous parallel TNT separations facilitate determination of a calibration curve for the TNT assay, which has high sensitivity (LOD, 1 ng/mL) and a wide dynamic range (1-300 ng/mL). 相似文献