流动注射碱性降解荧光法快速测定微量金霉素

提出了流动注射碱性降解荧光法快速测定微量金霉素的新方法.金霉素在流动管路中与0.25 mol/L NaOH溶液快速反应,生成强荧光降解物.在流通池中用荧光法进行检测.反应产物荧光强度在10~(-8)～10~(-5)mol/L范围与金霉素含量成正比.共存四环素和脱水金霉素杂质不干扰金霉素测定.本法既可用于金霉素产品含量测定,又可用于四环素产品中杂质金霉素的测定,还成功地应用于尿液和血清中微量金霉素的测定.     

荧光光度法测定血清中碱性磷酸酶

合成了一种新的底物水杨酸磷酸酯 (SP),用于荧光光度法测定血清中碱性磷酸酶(ALP)活力.在37.0 ℃的Tris-HCl缓冲液(pH 9.0)条件下, 碱性磷酸酶作用于荧光底物SP,水解生成强荧光产物水杨酸(SA),生成的SA的量与参与反应的ALP 的活力成正比.据此建立了荧光光度法测定血清中碱性磷酸酶活力的新方法.测定碱性磷酸酶的线性范围是0.03～6.00 U/L,检测限为7.04 mU/L.本方法适用于血清中碱性磷酸酶的测定.测定结果与临床上常用的以对硝基苯磷酸酯为底物的分光光度法相比,无显著性差异.     

痕量溴的阻抑动力学荧光法测定

基于在磷酸介质中溴对溴酸钾氧化丁基罗丹明B反应的抑制作用，建立了痕量溴的阻抑动力学荧光分析的新方法；应用单纯形最优化法确定了 实验条件，方法的检出限0.075μg/L，线性范围0.40-6.40μg/L；将该法应用于地下水、人发中溴的分析，获得满意结果，并对反应机理进行了初步探讨。     

阻抑动力学荧光法测定痕量苯胺

苯胺及其衍生物有剧毒且是致癌性物质。环境中苯胺的含量已成为各国环保、卫生部门重要的检测指标之一。测定苯胺的方法有色谱法、电化学法、分光光度法及荧光法等。动力学分析法灵敏度高、简便快速,已经有测定苯胺的报道,但动力学荧光法测定苯胺的报道极少。有人基于苯胺对溴酸钾氧化荧光素的抑制作用,建立了抑制—褪色分光光度     

血清碱性磷酸酶对人体缺锌、补锌的估价作用

概述了碱性磷酸酶（AKP）的基本结构、分型及其血清AKP变化的病理、生理因素，并回顾与总结了多年来国内外学者对血清AKP活力与锌(Zn)含量之间关系的基础与临床研究状况。     

蛋白质测定的流动注射荧光法

根据荧光素的单体－二聚体平衡转化的原理，提出了一种检测蛋白质的方法；荧光素在表面活性剂作用下形成二聚体后荧光强度明显降低，加入蛋白质后二聚体解聚使荧光强度增加；结合流动注射法测定牛血清蛋白（BSA），该法简便，灵敏度高，对血清中蛋白质进行测定，结果令人满意。     

镁—8羟基喹啉—5—磺酸荧光法测定血清镁浓度

血清中的镁离子与8-羟基喹啉-5-磺酸的络合物在碱性条件下可产生黄绿色荧光(Ex370 nm,E_M500 nm),当血清镁浓度在0.04～2.5μg/ml范围内与荧光强度直线相关(r=0.9996);平均回收率为99.765=3.51(SD)％,相对标准偏差为3.52％(n=21)。方法简便灵敏,为监测血清镁浓度及镁离子药物动力学研究提供了可行方法。     

阻抑动力学荧光法测定痕量间苯二酚

基于在ＨＣｌＯ４介质中,间苯二酚对Ｆｅ（Ⅲ）催化Ｈ２Ｏ２氧化罗丹明６Ｇ使其荧光猝灭的抑制作用,建立了动力学荧光法测定痕量间苯二酚的新方法。线性范围为０．００８～０．０９６ｍｇ／Ｌ间苯二酚,方法检测限为５．９ｕｇ／Ｌ。用于测定合成样中间苯二酚的含量,结果令人满意。     

过氧化氢氧化罗丹明6G催化荧光法测定铁

基于在高氯酸溶液中痕量铁对过氧化氢氧化罗丹明 6G具有催化作用 ,建立了催化荧光法测定痕量铁的新方法。方法的检出限为 1.4× 10 - 8g·ml- 1,线性范围为 0 .0 2 0 .6 μg·ml- 1。该方法用于人发、面粉中痕量铁的测定 ,结果满意     

A New Voltammetric Enzyme Immunoassay System for the Detection of Alkaline Phosphatase

Enzymes are widely used as labels in immunoassay and especially useful at low concentrations of the analyte in the sample. Most routine detection methods are based on the formation of color or fluorescent products, which are determined by spectropho-tometry or fluorometry, respectively. Electrochemical detection has been applied to the enzyme immunoassay with the potentials of combining the low detection limits and good selectivity of electroanalytical chemistry with the specificity of immun…     

以对硝基磷酸苯酯为底物电化学分析法检测碱性磷酸酯酶

研究了对硝基磷酸苯酯（ＰＮＰＰ）为底物伏安法测定碱性磷酸酯酶（ＡＬＰ）的方法。ＰＮＰＰ在ＡＬＰ的催化作用下水解生成对硝基苯酚（ＰＮＰ）。ＰＮＰ在玻碳电极上＋１．０２Ｖ（ｖｓＡｇ／ＡｇＣｌ）左右产生氧化峰,借助此氧化电流可以测定ＡＬＰ,并进而可用于以ＡＬＰ为标记物的酶免疫分析。用微分脉冲伏安法对酶催化反应条件和酶催化反应产物的测定条件进行了详细的研究,测定ＡＬＰ的线性范围是４．０×１０２～１．０×１０６ｍＵ／Ｌ;检测限为２．８ ×１０２ｍＵ／Ｌ。     

Electrochemical Determination of Alkaline Phosphatase in Human Serum by Differential Pulse Voltammetry

Differential pulse voltammetry(DPV) was applied to the determination of alkaline phosphatase(ALP) activity in human serum with phenyl phosphate as the substrate. Phenyl phosphate can enzymatically be hydrolyzed to produce phenol which is quantified by DPV at 660 mV(vs. Ag/AgCl) in the concentration rangeof 2.0--100 μmol/L. The standard curve for ALP is linear over the range from 0.06 to 1000U/L with a relative standard deviation of 3.0%. The conditions for the enzymatic reaction and voltammetric detection were optimized and the kinetic constants were also examined. The human serum samples were tested by this method and the results were in good agreement with those obtained by the routine p-nitrophenyl phosphate spectrophotometric method.     

Indirect Measurement of Yoctomole Alkaline Phosphatase by Capillary Electrophoresis with Electrochemical Detection

A method for indirectly detecting yoctomole (ymol) alkaline phosphatase was developed by capillary electrophoresis with electrochemical detection. In this method, disodium phenyl phosphate was used as the enzyme substrate and the product (phenol) of its hydrolysis reaction catalyzed by alkaline phosphatase was detected at the carbon fiber electrode. The optimum conditions of detection are 1.0×10^?2 mol/L Na₂B₄O₇ (pH 9.8) for the running buffer; 1.00×10^?3 mol/L disodium phenyl phosphate for the enzyme substrate; 20.0 kV for the separation voltage; 5 kV and 10 s for the injection voltage and injection time; 1.05 V (vs. saturated calomel electrode) for the detection potential and 10 min for the incubation time, respectively. In order to enhance the ratio of signal to noise, the shape and size of the working electrode, the shape of detection end of the capillary, and the capillary/electrode alignment method were studied in detail. When a single carbon fiber microcylinder electrode of 6 μm, a capillary of 10 μm ID with the etched detection end and the in‐capillary alignment were used, a ymol mass limit of detection for alkaline phosphatase was achieved.     

基于离子液体修饰碳纳米管电极的碱性磷酸酶电化学检测

利用自制的离子液体修饰碳纳米管电极(CNTs-ILE),在对CNTs-ILE的特性及对碱性磷酸酶(AP)酶促反应的底物磷酸对硝基苯酯(PNPP)和产物对硝基苯酚(PNP)进行电化学研究的基础上,采用安培法对酶促反应进程实现了持续动态的检测.实验表明,CNTs-ILE的循环伏安特性优于传统玻碳电极,且它在检测PNP时呈现出良好的抗钝化特性.当AP浓度范围在1.0～100 U/L时,电流大小与AP的浓度呈良好的线性关系.本方法可快速检测AP浓度,检测时间在20 min内.CNTs-ILE具有良好的抗钝化、操作简单及制作成本低等特点,在碱性磷酸酶的检测领域中有望得到进一步发展.在传统ELISA基础上,利用CNTs-ILE对具体样本最终的AP酶促反应产物进行电化学检测,通过峰值电流大小确定抗原浓度,为检测以AP为标记酶的抗原和抗体提供了理论依据.     

Umbelliferone Phosphate as a Substrate for Acid and Alkaline Phosphatase

Abstract

The results of a complete study of 8 substrates for acid and alkaline phosphatase indicated 7-hydroxycoumarin (umbelliferone) phosphate to be the best substrate for the analysis of these enzymes. Using this ester, from 10^?6 to 2 × 10^?2 units per ml. of alkaline phosphatase and 10^?5 to 0.06 units per ml. of acid phosphatase can be determined with an accuracy and precision of about 1.5%. Samples of serum as small as 1 μ;1. can be assayed. Analysis is performed by a direct initial reaction rate method in 2–3 minutes.     

荧光素二磷酸酯的合成和荧光分光光度法分析碱性、酸性磷酸酯酶

目前荧光光度法测定碱性和酸性磷酸酯酶的底物常用伞形酮磷酸酯和萘酚As-BI(6-溴-3-羟基-2-萘基邻茴香胺)的磷酸二氢酯.它们具有很高的灵敏度,在荧光免疫分析中被广泛应用. 本文用荧光素和三氯氧磷在吡啶介质中合成了荧光素二磷酸酯,再用氢氧化钠处理为四钠盐.该醇易溶于水,其水溶液无荧光.它在碱性或酸性磷酸酯酶的催化下水解,产生具有很强荧光的荧光素,且荧光强度与酶量成正比,因此可进行碱性或酸性磷酸酯酶的测定.     

An Automated Spectrophotometric Flow Injection Assay of Alkaline Phosphatase

Abstract

Assay of alkaline phosphatase using an automated flow injection manifold is described. The detector used is a spectrophotometer. The measurement is based on the rate of formation of p-nitrophenol at 405 nm, from the substrate p-nitropheny1 phosphate. The linear range, precision and accuracy of the technique for the assay of the enzyme is given. The advantages of the approach to assay of plasma enzymes are also discussed.     

阵列纸芯片比色法检测碱性磷酸酶

采用5-溴-4-氯-3-吲哚磷酸盐(BCIP)/氯化硝基四氮唑蓝(NBT)显色体系,构建了阵列纸芯片比色检测碱性磷酸酶(ALP)的方法.首先,借助烘干处理方式在光刻法制备的阵列纸芯片微孔中固定显色试剂,然后加入ALP进行显色反应,最后,采用凝胶成像仪和普通照相机成像,读取显色强度(灰度值)进行比色检测.详细考察了显色条件对检测结果的影响,探讨了人血清白蛋白对ALP检测的增色效应,在最佳实验条件下,ALP检测的线性范围为1.5～20 U/L,检出限(3 σ)为0.78 U/L(n=18),比文献报道中纸芯片上检测ALP方法的检出限低约两个数量级.本方法成功用于实际血清样品检测,测定结果与临床值一致.在此基础上,构建了双色阵列纸芯片,通过颜色的变化实现了ALP的可视化半定量检测.