微流控芯片直接化学发光检测方法研究

基于Ru(bipy)₃²⁺的化学发光反应,建立了一种新型微流控芯片化学发光检测方法.以草酸为研究对象,探讨了表面活性剂CTAB的浓度、化学发光反应酸度以及Ru(bipy)₃²⁺浓度等因素对峰形以及检测灵敏度和重现性的影响.草酸的线性范围为1.0×10^-4~5.0×10^-6mol/L,检测限达到2.0×10^-6mol/L.该方法的建立为微流控芯片分离检测某些有机酸、药物、环境物质及核酸等提供了新方法.     

Selective determination of amino acids using flow injection analysis coupled with chemiluminescence detection

The determination of the amino acids proline, histidine, tyrosine, arginine, phenylalanine and tryptophan using flow injection analysis (FIA) with chemiluminescence detection is described. Proline was the only amino acid to exhibit chemiluminescence with the tris(2,2-bipyridyl)ruthenium(III) reaction at pH 10. While, histidine was found to selectively enhance the reaction of luminol with Mn(II) salts in a basic medium. Acidic potassium permanganate chemiluminescence was able to selectively determine tyrosine at pH 6.75. Low pressure separations using a C18 guard column allowed the simultaneous determination of tyrosine and tryptophan or phenylalanine and tryptophan with acidic potassium permanganate and copper(II)-amino acid-hydrogen peroxide chemiluminescence, respectively. Precision for each method was less than 3.9% (R.S.D.) for five replicates of a standard (1×10⁻⁵ M) and the detection limits ranged between 4×10⁻⁹ and 7×10⁻⁶ M. Preliminary investigations revealed that the methodology developed was able to selectively determine the individual amino acids in an equimolar mixture of the 20 naturally occurring amino acids.     

毛细管电泳高频电导法快速分离检测混合氨基酸

氨基酸(Am ino acids,AA s)是组成生物大分子的基本单元,与人的健康状况有极其密切的关系.在医学和生命科学研究中,微量氨基酸的分离检测具有重要意义.     

N-溴代琥珀酰亚胺-二氯荧光素化学发光体系测定废水中5-磺基水杨酸

建立了测定5-磺基水杨酸的流动注射化学发光新方法。 研究了影响化学发光强度的因素,并初步探讨了可能的发光机理。 在最佳化学发光条件下,化学发光强度与5-磺基水杨酸浓度在5.0×10-8~2.0×10-5 mol/L范围内呈良好的线性关系,检出限为1.0×10-8 mol/L,对5.0×10-6 mol/L的5-磺基水杨酸平行测定9次,其相对标准偏差为2.7%。 该法可用于强力霉素废水中5-磺基水杨酸的测定。     

高效液相色谱-激光诱导荧光-增强型电荷耦合器件检测氨基酸的研究

以3-对羟基苯甲酰喹啉-2-甲醛(CBQCA)为检测痕量氨基酸的柱前衍生试剂,利用高效液相色谱-激光诱导荧光-增强型电荷耦合器件检测器(HPLC-LIF-ICCD)系统,考察了氨基酸衍生和HPLC分离过程中多种因素的影响,建立了HPLC-LIF-ICCD分离检测痕量氨基酸的新方法,并应用于鼠脑微透析液的氨基酸检测.当信噪比为3时,对甘氨酸的质量检出限为5fmol.     

反相离子对液相色谱分离-柱后衍生化学发光检测痕量金属离子

报道了反相离子对液相色谱-柱后衍生化学发光检测方法分离测定痕量的钴(Ⅱ)、铁(Ⅱ)、铜(Ⅱ)、铁(Ⅲ)及铬(Ⅲ).利用弱络合性的乳酸作为淋洗剂,加入适量的十二烷基磷酸钠作对离子试剂,研究了分离条件、发光条件及二者的匹配方式等因素对分离检测的影响.测定线性范围分别为(g/mL):Cr(Ⅲ)5.0×10^-9～1.0×10^-5;Fe(Ⅱ)2.0×10^-9～1.0×10^-5;Fe(Ⅲ〕8.0×10^-9～1.0×10^-5;Cu(Ⅱ)8.0×10^-11～1.0×10^-5及Co(Ⅲ)1.0×10^-11～1.0×10^-5.检出限分别为:10pgCr(Ⅲ),0.4pgFe(Ⅱ),20pgFe(Ⅲ),1.0pgCu(Ⅱ)及0.01pgCo(Ⅱ).     

利用H2O2-KI-鲁米诺体系化学发光法测定痕量钼

钼(Ⅵ)可以催化H₂O₂氧化I^-的反应,生成的I₂在碱性水溶液中能氧化鲁米诺而产生化学发光^[1],本文将这两个反应结合起来,建立了铝的化学发光分析法,方法有较高的灵敏度,检出限为6×10^-10g/cm³Mo,测定的相对标准偏差小于6%,工作曲线的线性范围是1×10^-9～1×10^-7g/cm³Mo,用EDTA作掩蔽剂,排除了大多数金属离子的干扰,方法有较好的选择性,应用此法对一些粮食样品中的痕量钼进行了测定,结果比较满意。     

LONG WAVELENGTH UV RADIATION AFFECTS CHEMILUMINESCENCE OF HUMAN POLYMORPHONUCLEAR LEUCOCYTES

Abstract— Luminol-enhanced chemiluminescence in suspensions of human polymorphonuclear leucocytes stimulated with the chemotactic oligopeptide formyl-methionyl-leucyl-phenylalanine was increased by exposure of the cells to long wavelength UV radiation (mainly320–400 nm). Leucocytes treated with 0.3 × 10⁴ J m^-2 of UV responded with doubled peak values, and 4 × 10⁴ J m^-2 lead to a five-fold increase in peak chemiluminescence, as compared with non-exposed cells. Supernatants isolated from irradiated leucocytes contained increased amounts of both myeloperoxidase and lactate dehydrogenase and showed higher chemiluminescence values, when compared with supernatants from sham-irradiated cells. The results suggest that leucocyte degranulation contributes to the inflammatory properties of long wavelength UV.     

对氨基苯磺酸氧化化学发光反应的研究及应用——测定痕量钴

研究了对氮基苯磺酸在金属离子钻(Ⅱ)作为催化剂存在下,在碱性溶液中被过氧化氢氧化的化学发光行为,测定了发光光谱,最大发射波长为445nm.基于钻(Ⅱ)的催化作用建立了分析采量钻的化学发光新方法,其线性范围为5.0×10^-11～5.0×10^-7g/mL,检出限为0.01ng/mLGo²⁺,具有较好的选择性,通过对化学发光反应产物及有关反应试剂的紫外、红外和荧光光谱的测定研究,提出了氧化发光反应的机理,还对发光猝灭作用进行了讨论。     

亚铁氰化钾-NO2--鲁米诺化学发光体系测定痕量亚硝酸盐的研究

本文在调查和了解天津市独立科技信息服务机构现状的基础上，研究和探讨了其调整原则和调整目标，确立了调整的步骤和要点。最后提出天津市独立科技信息服务机构的七点调整对策。     

毛细管电泳-间接激光光热干涉检测方法研究

本文提出毛细管电泳-间接激光光热干涉检测新方法。采用吸收系数大、吸收波长与泵浦激光(He-Ne)波长匹配性能较好的亚甲蓝溶液为背景电解质溶液,加入乙醇减少了毛细管壁对亚甲蓝的吸附作用。并将间接光热干涉检测法用于氨基酸毛细管电泳分离检测,对赖氨酸检测限达5×10^-6mol/L(S/N=2).     

A New Method for Brucine Analysis Based on Its Chemiluminescence Reaction with Potassium Permanganat

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Electrochemical Determination of Reduced Glutathione at Multiwalled Carbon Nanotubes/Poly(bromocresol green)Modified Glassy Carbon Electrode

A multiwalled carbon nanotubes/poly(bromocresol green) modfied glassy carbon electrode (MWNTs-PBG/GCE) was used to investigate the electrochemical behavior of reduced glutathione(GSH). GSH showed an irreversible oxidation process on MWNTs-PBG/GCE with an oxidation peak at 0.77 V(vs. SCE) in a phosphate buffer solution(pH=4.0). The kinetic parameters of the electrochemical behavior of GSH on MWNTs-PBG/GCE were calculated. Under the optimal conditions and with the help of amperometric method, a linear relationship was obtained between the oxidation peak current and GSH concentration in the range from 2×10–7 mol/L to 5×10–6 mol/L with the detection limit as 1×10–8 mol/L(signal-to-noise ratio of 3). The current reached the steady-state current within about 5 s. The modified electrode surface had very good reproducibility and stability.     

A capillary electrophoresis detection scheme for underivatized amino acids based on luminol-BrO- chemiluminescence system

A novel chemiluminescence (CL) detection scheme has been developed for detecting underivatized amino acids following capillary electrophoresis (CE) separation. This detection was based on the inhibitory effect of amino acids on the CL reaction between luminol and BrO⁻ in alkaline aqueous solution. Detection of amino acids was accomplished with a borate-based background electrolyte at pH 9.2. The luminol was used as a component of the separation carrier electrolyte. Parameters affecting CE-CL separation and detection, such as the pH value, the concentration of electrolyte and CL reagent on the resolution were optimized. The relative standard deviation for the analysis of amino acids was less than 1.5% for the migration time and 4% for the peak height. The mass limits of detection were from 7 to 144 fmol for the 7 amino acids. This method has been applied of 7 amino acids in amino acid injection.     

还原剂与鲁米诺化学发光反应的研究(Ⅱ)——钒(Ⅱ)-鲁米诺化学发光反应

Fe(Ⅱ)和Ti(Ⅲ)与鲁米诺的化学发光反应已有报道,我们发现,Cr(Ⅱ)、Mo(Ⅲ)、W(Ⅲ)、U(Ⅲ)、CN^-、SO₃^2-、抗坏血酸等一大类还原剂均可与鲁米诺溶液作用产生化学发光,本文利用Jones还原柱产生V(Ⅱ),首次研究了V(Ⅱ)与鲁米诺的化学发光反应,在此基础上建立了钒的流动注射化学发光分析法,方法的检出限是8×10^-11g/mL钒,线性范围是4×10^-10～1×10^-5g/mL钒。测定的相对标准偏差小于2％,考察了20余种常见离子对测定的干扰情况,方法已用于水样中痕量钒的测定,初步探讨了发光反应的机理。     

Chemiluminescence Detection of Amino Acids,Peptides, and Proteins Using Tris-2,2′-Bipyridine Ruthenium (III)

Abstract

The feasibility of using the tris-2-2′-bipyridine ruthenium (III) (Ru(bpy)₃ ^{3 +}) chemiluminescent (CL) reaction for the detection of amino acids, peptides, and proteins has been studied.

Detection limits of the amino acids as determined by flow injection analysis (FIA) ranged from 20 pmol of proline to 50 nmol of asparagine. In general, amino acids containing secondary amine groups yielded the strongest responses. A reaction mechanism for Ru (bpy)₃ ^{3 +} chemiluminescence of aliphatic amines has been proposed. Studies of peptide molecules and poly-prolines showed that the peptide bond barely contributes to the detection signals. The separation of hydroxyproline and proline in synthetic collagen by HPLC with Ru (bpy)₃ ^{3 +} chemiluminescence detection has been shown to be possible.     

HPLC分离化学发光检测药物中的马钱子碱、士的宁和麻黄类生物碱

在KMnO₄-H⁺-还原剂体系中研究了马钱子碱、士的宁和麻黄类生物碱的化学发光(CL)行为,建立了其快速灵敏的流动注射化学发光分析方法,将此法与HPLC联用,实现了上述物质的选择性测定,该方法的检出限为1.0×10^-10g/mL马钱子碱、1.0×10^-8g/mL士的宁、1.0×10^-5g/mL麻黄碱和伪麻黄碱,测定的相对标准偏差小于3%,HPLC流动相的脉动及其中的溶解氧不影响测定的精密度。此法已用于一些药物中马钱子碱等生物碱的测定,回收率为94%～101%.     

二维阀切换离子色谱法测定海带中游离氨基酸

建立一种测定海带中游离氨基酸的阀切换高效阴离子交换色谱耦合脉冲安培检测器法。采用一根阳离子交换柱对氨基酸进行富集,而后经阀切换至氨基酸分析柱Amino Pac~PA-10(250 mm×2 mm)上分离并进入安培检测器检测。在最佳分离条件下,20种氨基酸的质量浓度在0.1~20.0 mg/L范围内与其色谱峰面积线性关系良好,线性相关系数r~20.99,20种氨基酸的检出限为0.01 mg/L,加标回收率为83.12%~117.34%,测定结果的相对标准偏为1.02%~13.05%(n=8)。该方法样品前处理简单,无基底杂质干扰,适用于海带样品中游离氨基酸的测定。     

高效液相色谱化学发光检测法测定氨基酸

本文根据铜与氨基酸组成的１：１不饱和络合物对Ｌｕｍｉｎｏｌ－Ｈ２Ｏ２化学发光体系的催化活性，设计了一种新型氨基酸高效液相色谱化学发光检测器，即在色谱柱后安装一个氢氧化铜柱，从柱后流出的氨基酸通过氢氧化铜柱时，产生配合溶解形成氨基酸－铜（１：１）络合物，进行化学发光检测，１４种常见氨基酸的检测限均在ｐｍｏｌ级，已用于血清、尿和啤酒中氨基酸的测定。     

Determination of fumaric and maleic acids with stacking analytes by transient moving chemical reaction boundary method in capillary electrophoresis

The paper presents an on-line transient moving chemical reaction boundary (MCRB) method for simply but efficiently stacking analytes in capillary electrophoresis (CE). The CE technique was developed for a rapid determination of fumaric and maleic acid. Based on the theory of MCRB, Effects of several important factors such as the pH and concentration of running buffer and the conditions of stacking analytes were investigated to acquire the optimum conditions. The optimized separations were carried out in a 20 mmol/L sulphate neutralized with ethylenediamine to pH 6.0 electrolytes using a capillary coated with poly (diallyldimethylammonium chloride) and direct UV detection at 214 nm. The optimized preconcentrations were carried out in 50 mmol/L borax (pH 9.0). The calibration curves were linear in the concentration range of 1.0 × 10⁻⁷–1.0 × 10⁻⁴ mol/L and 5.0 × 10⁻⁷–1.0 × 10⁻⁴ mol/L for fumaric and maleic acid with correlation coefficients higher than 0.9991. The detection limits were 5.34 × 10⁻⁸ mol/L for fumaric acid and 1.92 × 10⁻⁷ mol/L for maleic acid. This method was applied for determination of fumaric acid in apple juice and of fumaric and maleic acid in dl-malic, the recovery tests established for real samples were within the range 95–105%. This work provided a valid and simple approach to detect fumaric and maleic acid.