微板式磁化学发光酶免疫分析法对人绒毛膜促性腺激素(HCG)的灵敏快速测定

以磁性酶免疫测量分析为模型,建立了微板式磁化学发光酶免疫分析法.利用3-(2′-螺旋金刚烷)-4-甲氧基-4-(3″-磷酰氧基)苯-1,2-二氧杂环丁烷(AMPPD)-碱性磷酸酶(ALP)化学发光体系对人绒毛膜促性腺激素(HCG)进行测定,测量的灵敏度较分光光度法提高了13倍.测定的线性范围为0.15～500 mIU/mL;批内变异系数(C.V.%)和批间变异系数(C.V.%) 均在18%之内;回收率在80%～116%之间;利用本法对血清样品进行了测定,并与其它化学发光免疫分析方法进行了比较,其相关系数为0.965.首次将酶免疫分析方法用于唾液中人绒毛膜促性腺激素(HCG)的测定,为建立非侵入式样品测定方法打下了基础.     

流动注射化学发光法测定人绒毛膜促性腺激素

采用反相流动注射法,系统研究了碱性介质和阳离子表面活性剂十六烷基三甲基溴化铵(CTMAB)存在下,荧光素-人绒毛膜促性腺激素(HCG)复合物的化学发光行为和化学发光条件,提出了反相流动注射化学发光法测定HCG含量的新方法。该法测定HCG的线性范围为0.8～200mIU/mL,检出限(3σ/k)为0.01mIU/mL;对浓度为10mIU/mL的HCG平行测定11次,其相对标准偏差为0.8%。该法可用于人工混合样品和孕妇尿液中HCG含量的测定。     

电位溶出法同时测定水质和生物样品中的铜、铅、镉、锌、铁、锰

本文提出了在乙二胺和乙二胺-盐酸体系中以计时电位溶出法测定Fe、Mn及以电位溶出法测定Cu、Pb、Cd、Zn的新方法。各元素的浓度在Fe 0.1～2600 ng/ml、Mn0.05～3200 ng/ml、Cu0.2～2800 ng/ml、Pb0.1～3600ng/ml、Cd 0.05～1200 ng/ml、Zn0.2～4200ng/ml时,浓度与溶出峰高呈良好的线性关系。本方法应用于水质及生物样品中上述各元素的测定,结果良好。     

光化学-荧光分析法研究——Ⅲ.片剂及注射液中安定的测定

本文提出了测定安定的光化学—荧光分析新方法。安定在含有十二烷基硫酸钠的稀硫酸介质中,经紫外光照射后,产物能发射荧光(=368nm,=476nm)。安定浓度在0～2.5μg/ml范围内,荧光强度与浓度成良好的线性关系。方法的相对标准偏差为1.5％,检出限为0.044μg/ml,可用于片剂及注射液中安定含量的测定。     

地高辛等强心甙的单扫示波极谱测定

本文报道在0.01M ICH_2COONH_4-0.2M NH_4Cl底液中,用单扫示波极谱测定地高辛、洋地黄毒甙和毛花强心丙等心血管药物。它们的还原峰电位顺次为-1.54,-1.53和-1.60V(vs.SCE)。峰电流与地高辛浓度在2.0—60ng/ml,洋地黄毒甙在5.0—400ng/ml,毛花强心丙在15—125ng/ml范围均有良好的线性关系。对25ng/ml地高辛测定12次的变动系数为1.74％。     

催化光度法测定痕量铜的研究 Cu(Ⅱ)-次甲基蓝-H_2O_2体系

在氨水介质中,痕量Cu(Ⅱ)对H_2O_2氧化次甲基蓝的反应有强烈的催化作用。由此建立的痕量CU(Ⅱ)的催化光度分析法的检测限为1.5×10~(-10)g/ml。测定范围为0～0.7μg/25ml。测得催化及非催化反应的表观活化能分别为9.15kJ/mol及14.13kJ/mol。样品分析结果良好。     

鲁米诺-氰化物化学发光反应的研究——环境水样中氰化物的测定

本文提出了利用鲁米诺-氰化物化学发光体系测定氰化物的新方法。方法的检出限是1.2ng/ml,校正曲线在2～100ng/ml CN~-范围内具有良好的线性。相对标准偏差小于4％。此方法已用于自来水和工业废水中氰化物的测定。文中对鲁米诺与氰化物化学发光反应的可能机理作了初步探讨。     

蒙脱石修饰碳糊电极测定氨基酸的研究：Ⅰ.色氨酸的测定

本文用含10％蒙脱石的修饰碳糊电极研究了测定色氨酸的方法。发现在0.001mol/L KCl-HCl(pH=2.0)溶液中富集、介质交换后用微分脉冲伏安法测定,于+0.75V出峰,在20.～10.0μg/ml范围内有线性。相对标准偏差为2.60％。检测限为70ng/ml,该电极具有很强的抗干扰能力。用此方法直接测定氨基酸药物样品中的色氨酸,结果良好。     

流动注射-分光光度-火焰光度串联技术同时快速测定磷、钾、钠的研究

本文利用流动注射-分光光度-火焰光度串联技术,使磷生成磷钼兰以721型分光光度计检测,钾、钠以6400型火焰光度计检测,从而建立了磷、钾和钠三元素同时快速测定的新方法,该方法操作简单、方便、快速,每小时可进行60次测定,磷、钾、钠兰元素的线性范围分别为1.0～4.0μg/ml、3.0～60μg/ml和6.0～40μg/ml。利用该方法测定了植物样品中的磷、钾和钠,获得了良好的结果。     

停流流动注射-动力学方法多组份同时测定的研究——铁(Ⅱ)、钛(Ⅲ)和钒(Ⅳ)的同时测定

依据诱导反应原理,提出了同时测定多组份的动力学方法,并以停流流动注射分析研究了Fe(Ⅱ)、Ti(Ⅲ)及V(Ⅳ)诱导的Cr(Ⅵ)-I~-反应体系,建立了铁、钛和钒同时测定的条件。对铁、钛和钒测定的线性范围分别为0～2.2μg/ml,0～3.1μg/ml及0～1.8μg/ml,检测限分别为0.012,0.020和0.018μg/ml。     

The study of a chemiluminescence immunoassay using the peroxyoxalate chemiluminescent reaction and its application

The paper presented a novel chemiluminescence (CL) immunoassay method, which combines the advantages of traditional enzyme-linked immunosorbent assays (ELISA) and bis (2,4,6-trichlorophenyl) oxalate (TCPO)-hydrogen peroxide CL detection system. A fluorescent product 2,3-diaminophenazine (DAPN) was produced by reaction between o-phenylenediamine (OPDA, 1,2-diaminobenzene) and H₂O₂ catalyzed by horseradish peroxidase (HRP). DAPN was excited by the reactive intermediate of TCPO-H₂O₂ chemiluminescent reaction, and led to CL. The dependence of the CL intensity on the concentrations of antigen was studied. As analytical application, the proposed method was used for determination of recombinant human interleukin 6 (rHu IL-6) and β-human chorionic gonadotropin (β-HCG). Under the selected experimental conditions, a linear relationship was obtained between the CL intensity and the concentration of rHu IL-6 in the range of 4.0-625.0 pg/ml, and β-HCG in the range of 12.5-400.0 mIU/ml. The detection limits were 0.5 pg/ml for rHu IL-6 and 3 mIU/ml for β-HCG with relative standard deviation of 2.3 for 78.0 pg/ml rHu IL-6, and 3.9 for 50.0 mIU/ml β-HCG. This method has been applied to the determination of rHu IL-6 in human serum and β-HCG in urine with satisfactory results.     

Level of Human Chorionic Gonadotrop in-Like Substance in Serum of Normal Subjects

Abstract

The level of human chorionic gonadotropin (hCG)-like substance in normal serum was measured using 50 μl of serum specimen by a highly specific and sensitive sandwich enzyme immunoassay for hCG which did not significantly cross-react with hLH. The level was calculated using a standard hCG and expressed in mIU/ml as that of hCG. In normal male (20–59 years of age) and normal, non-pregnant female (20–42 years of age) adults, the level was below 0.6 mIU/ml with mean levels of 0.16 mIU/ml (n=209) and 0.14 mIU/ml (n=36), respectively. In older subjects, however, the level tended to rise. In normal male adults, aged 60–91 years, the level was mostly below 0.8 mIU/ml (mean=0.56 mIU/ml, n=99), but levels of 15 specimens out of 99 were 1.1–5.5 mIU/ml. In normal, non-pregnant female adults, aged 45–88 years, the level was widely distributed between 0.1 and 11.7 mIU/ml (mean=1.6 mIU/ml, n=141). In these older female adults, levels of hCG-like substance were not well correlated to either levels of hLH and hFSH, while levels of hLH and hFSH were well correlated to each other.     

Estimation of plasma catecholamines by high-performance liquid chromatography with electrochemical detection in patients with subarachnoid haemorrhage

Plasma catecholamine levels were determined by high-performance liquid chromatography with electrochemical detection following alumina extraction. Mean individual recoveries of 50.5, 49.9 and 48% were obtained for norepinephrine, epinephrine and dopamine, respectively, and the limits of detection for each catecholamine were 0.15, 0.34 and 0.6 pmol/ml. Total analysis time for each plasma sample was approximately 1 h. Catecholamine levels were measured in plasma from control subjects and the ranges obtained were: norepinephrine, 0.33-5.98 pmol/ml; epinephrine, 0-4.77 pmol/ml; dopamine, 0-0.8 pmol/ml. When patients with subarachnoid haemorrhage were investigated, the ranges were found to be: norepinephrine, 0.23-7.27 pmol/ml; epinephrine, 0-4.91 pmol/ml; dopamine, 0-0.23 pmol/ml.     

A study on LH- and FSH-RIA kits by immunoradiometric assay

An evaluation of LH- and FSH-RIA BEAD kits based on IRMA was carried out. The results obtained with the methods characterized by the use of monoclonal antibodies, i.e., one is linked to solid phase, and another is isotopically labeled, were compared with those determined by the Daiichi LH- and FSH-kits. Intra- and inter-assay precision, recovery, linearity, and specificity of both methods were favorable without exceptions. The cross reactivity of the LH kit to 5,000 mIU/ml hCG revealed within the range of less than 3 mIU/ml. Significant correlations were observed between the results derived from conventional Daiichi LH- and FSH-kits. The results from the conventional kits exhibited 30 to 40% of those from the Daiichi kits, considered to be mainly due to the difference in standard calibrations used. Among the individuals within the normal menstrual cycle, the serum LH- and FSH-levels determined by the present kits gave a typical pattern with a peak in the preovulatory phase. On the other hand, the LH- and FSH-values of individuals in normal pregnancy revealed strikingly decreased in comparison with those of non-pregnant women.     

Solid phase inclusive immunoradiometric assay for human chorionic gonadotropin using monoclonal antibodies

Summary A simple, one step, inclusive immunoradiometric assay for human chorionic gonadotropin (hCG) employing monoclonal antibodies is described. Commercially available monoclonal antibodies from various commercial sources were screened. Identified “detection” antibody was radiolabeled with ¹²⁵I and the selected “capture” antibody was chemically coupled to magnetizable cellulose to form a solid phase. In the procedure developed, standard/sample, radiolabeled antibody and capture antibody were incubated together for 3 hours at room temperature with shaking. After incubation, the bound complex was quantitated for the associated radioactivity. The analytical sensitivity observed was 1.0 mIU/ml with a wide concentration range up to 1000 mIU/ml of hCG. “High dose hook” of the developed assay was observed beyond 2000 mIU/ml. Results showed that the developed assay had a good precision: intra-assay CV less than 8%, inter-assay CV less than 10% and good analytical recovery of 97-109%. The clinical samples analyzed by the developed procedure showed a good correlation with that of the commercial kit (r = 0.92; y = 0.99x+0.51).     

流动注射-阴离子交换预浓集-电感耦合等离子体发射光谱法检测痕量元素——砷、硒、钼、硫、铬

本文研究了阴离子交换树脂D296对待测元素As、Se、Mo、S、Cr的预浓集性能,并与改进的流动注射(FI)(双流路)相结合,建立了一套新型、高效的FI-阴离子交换-ICP-AES分析体系。从而极大地提高了分析的频率和灵敏度,分析速度可达每小时14个样品。As、Se、Mo、S、Cr各元素的检测限分别为13.0 ng/ml、25.2 ng/ml、2.04 ng/ml、6.19 ng/ml、3.64ng/ml。利用此体系检测了标准土壤、头发、牛肝粉等,结果与标准值均很接近。     

Reversed-phase high-performance liquid chromatographic separation and electrochemical detection of retinol and its isomers

Baseline separation of the isomers of retinol using reversed-phase high-performance liquid chromatography (HPLC) in less than 30 min is presented. A new approach to the detection of retinol using electrochemical detection is developed. The oxidative electrochemistry of retinol is studied at a glassy-carbon electrode using coulometry, ultraviolet-visible spectrophotometry and HPLC. Amperometric detection in HPLC for retinol provided a linear response from 0 to 1.5 micrograms/ml and a detection limit of 4.1 ng/ml. Electrochemical detection was compared to ultraviolet-visible absorbance detection for the determination of retinol in human serum extracts. Good agreement is found for the results obtained with the two detectors.     

MCM-41 mesoporous material modified carbon paste electrode for the determination of cardiac troponin I by anodic stripping voltammetry

An anodic stripping voltammetric method for the determination of cardiac troponin I (cTnI) at a MCM-41 mesoporous material modified carbon paste electrode (MCM-MCPE) was investigated. The test was based on the dual monoclonal antibody “sandwich” principle using colloidal gold as a labeled substrate. Four main steps were carried out to obtain the analytical signal, i.e. electrode preparation, immunoreaction, silver enhancement, and anodic stripping voltammetric detection. The anodic stripping peak current increased linearly with the concentration of cTnI over the range of 0.8-5.0 ng/ml. A detection limit of 0.5 ng/ml was obtained. The established method was applied to detect cTnI in acute myocardial infarction (AMI) samples using routine enzyme-linked immunoadsorbent assay (ELISA) for comparison analysis, and good results were obtained.     

Alumina sol-gel/sonogel-carbon electrode based on acetylcholinesterase for detection of organophosphorus pesticides

Two new amperometric biosensors based on immobilization of acetylcholinesterase on a sonogel-carbon electrode for detection of organophosphorous compounds are proposed. The electrodes were prepared applying high-energy ultrasounds directly to the precursors. The first biosensor was obtained by simple entrapping acetylcholinesterase in Al₂O₃ sol-gel matrix on the sonogel-carbon. The second biosensor was produced in a sandwich configuration. Its preparation involved adsorption of the enzyme and modification via a polymeric membrane such as polyethylene glycol and the ion-exchanger Nafion. The optimal enzyme loading was found to be 0.7 mIU. Both biosensors showed optimal activity in 0.2 M phosphate buffer, pH 7.0, at an operating potential of 210 mV. The detection limit achieved for chlorpyriphos-ethyl-oxon was 2.5 × 10⁻¹⁰ M at a 10-min incubation time.