基于液芯波导原理的微流控芯片长光程光度检测系统

提出了一种基于液芯波导(Liquidcorewaveguide,LCW)原理的微流控芯片吸收光度检测系统.通过芯片与外界接口技术实现液芯波导管与芯片的耦合,建立了芯片上长光程(毫米至厘米级)吸收光度检测池.采用邻菲啉-铁(Ⅱ)显色体系验证系统分析性能,以5.5cm外覆TeflonAF液芯波导管作为检测池(检测池体积240nL)时,芯片系统的检测线性范围为0.03～50μmol/L,对邻菲啉-铁(Ⅱ)配合物的检出限为8nmol/L,检测池有效光程达1.7cm,分析精度RSD(n=5)为0.8%.     

基于纳米结构BDD电极的人体尿酸检测

利用纳米结构硼掺杂金刚石(nBDD)电极的优点对人体尿液中的UA含量进行检测,并与常规玻碳(GC)电极做了比较。检测不同尿样的结果可知,nBDD电极检测的回收率为95.3%～98.4%,GC电极79.6%～87.1%;检测同一尿样的重复性实验得出,nBDD电极上峰电流的相对标准偏差(RSD)为8.0%,GC电极上峰电流的RSD为39%。     

信号核酸识体用于药物托普霉素的高灵敏度检测

提出了一种利用RNA核酸识体和光开关化合物[Ru(phen)2dppz\]2+ (RU)高灵敏检测药物分子的荧光分析新方法. 以托普霉素(TOB)为靶分子, 检测限可达到10 nmol/L, 同时该方法有较好的选择性, 能在溶液中实现无需分离的实时检测. 这种无标记检测方法有利于不稳定RNA核酸识体在分析检测中的应用, 简化实验操作, 为其它药物分子的检测提供了新思路.     

芦丁在Au-TiO_2修饰电极上的电化学行为及其测定

利用沉淀沉积法合成了纳米金-二氧化钛(Au-TiO_2)复合纳米材料.构建了Au-TiO_2复合材料修饰的电化学传感器,并对芦丁进行了检测.通过扫描电镜(SEM),X射线衍射(XRD)对Au-TiO_2复合材料进行表征,利用循环伏安(CV)和电流时间曲线(Amperometric i-t Curve)考察了其对芦丁的电催化性能.在最优条件下对芦丁进行检测,检测范围为0.1~10μmol/L,灵敏度为1.47μA·μmol·L~(-1)·cm~(-2),信噪比为3时检测限为0.083μmol/L.     

HA/Ag纳米复合材料的制备与性能

采用一锅热熔胶法制备出多孔花状羟基磷灰石/银(HA/Ag)纳米复合材料。通过检测罗丹明B揭示了HA/Ag作为SERS活性底物的独特拉曼增强效应,其检测极限为10~(-8) mol·L~(-1)。在检测中,改变拉曼激光强度,取点位置等手段能有效提高检测的灵敏度。此外,所制备的多孔纳米复合材料HA/Ag作为对硝基苯酚(4-NP)还原为对氨基苯酚(4-AP)反应的催化剂,可以大大缩短反应时间40 min。     

凹凸棒石修饰丝网印刷碳电极的制备及其在Cd(Ⅱ)检测中应用

以天然廉价的凹凸棒石粘土矿物修饰丝网印刷碳电极为传感电极,采用方波阳极溶出伏安法对Cd(Ⅱ)进行分析检测。研究了凹凸棒石粘土矿物修饰浓度、富集电位、富集时间、溶液pH以及干扰离子对Cd(Ⅱ)检测的影响。在优化实验条件下,传感电极对Cd(Ⅱ)具有敏感的监测特性,峰电流(y)与Cd(Ⅱ)的质量浓度(x)在10～175μg/L范围内具有良好的响应性能,线性方程为y(μA)=0.09056x+0.3350(R²=0.9951),检测限(S/N=3)为1.18μg/L。将该传感器应用于实际河水样品中Cd(Ⅱ)加标分析,回收率为98.6%～103.5%。凹凸棒石粘土矿物修饰电极构建的传感器制备简单,价格低廉,检测效果良好,可用于环境中Cd(Ⅱ)的快速检测。     

孔雀绿分子印迹96孔检测板的制备及应用研究

为更加快速、灵敏、简单地检测水产品中的孔雀绿(MG)残留,通过原位聚合法,以乙腈/甲苯溶液(V/V=3∶1)为溶剂、甲基丙烯酸(MAA)为功能单体、二甲基丙烯酸乙二醇酯(EGDMA)为交联剂(模板分子与之比例为1∶4∶20),制备了分子印迹聚合物(MIP)。通过包被MIP到96孔板上研发了以化学发光免疫分析法(CLIA)检测的MG-MIP-96孔检测板,其标准曲线为y=-29197.49x+1945998.45,R2=0.9416,线性范围为30~66.64 ng/m L,检测限为3.355ng/m L。批内变异系数范围为7.38%~18.07%,批内回收率范围为103.41%~144.45%;批间变异系数范围为12.45%~17.52%,批间回收率范围为100.07%~143.20%。以孔雀石绿结构类似物副品红(PA)和结晶紫(CV)进行特异性检测的交叉率分别为27.97%、30.97%。说明此新型检测方法具有很大的研究价值和实际应用价值。     

芯片毛细管电泳电化学发光法快速测定盐酸普鲁卡因的含量

建立了芯片毛细管电泳电化学发光法快速测定盐酸普鲁卡因含量的新方法。采用三联吡啶钌(Ru(bpy)2+3)为电化学发光试剂,三电极体系(直径300μm的铂圆盘电极为工作电极,集成在铂圆盘工作电极外的钛管为对电极,Ag/AgCl丝为参比电极)进行检测。分别考察了运行缓冲溶液pH值、检测缓冲溶液pH值、检测电位以及分离电压对分离和检测性能的影响。在优化条件下,即运行缓冲溶液为10mmol/L磷酸盐溶液(pH4.0),检测池缓冲溶液为含5mmol/LRu(bpy)2+3的50mmol/L磷酸盐缓冲溶液(pH7.0),检测电位为1.25V,分离电压为300V/cm时,盐酸普鲁卡因可在40s内实现较好的分离与检测,其线性范围为10~2000μg/mL(r2=0.9991),检出限(S/N=3)为3.0μg/mL,加标回收率为97%~99%,相对标准偏差为1.8%~2.2%。该方法简便、快速、准确,可用于盐酸普鲁卡因注射液的质量控制。     

反相高效液相色谱法测定硝酸咪康唑及其相关物质的含量

在LUNAC18色谱柱(250×4.6mmI.D,5μm)上,研究了硝酸咪康唑(Mi conazoleNitrate)及其相关物质(Alpha (2,4 Dichlorophenyl) 2 1H Imidazole 1 Ethanol)的反相高效液相色谱分离检测的最适宜条件,采用乙腈∶缓冲溶液=70∶30(V/V)为流动相,流速为1.0mL/min,检测波长为235nm,经实际样品测定,结果满意。这为跟踪产品的合成工艺,提高产品的质量提供了一个快速有效的检测方法。     

制备金纳米棒标记的免疫层析试纸条用于临床血清样本中心肌肌钙蛋白Ⅰ的快速检测

心肌肌钙蛋白Ⅰ(Cardiac troponin Ⅰ, cTnⅠ)是心肌损伤的生物标志物之一,快速检测其血清水平对急性心肌梗死(Acute myocardial infarction, AMI)的临床诊断至关重要。本研究以鼠抗cTnⅠ单克隆抗体(4T21cc-19C7cc, dAb)修饰的金纳米棒(Gold nanorod, GNR)为标记探针,采用双抗体夹心法制备侧流免疫层析试纸条(Lateral flow immunochromatographic test strip, LFITS),用于快速检测临床血清样本中的cTnⅠ。此GNR标记的LFITS(GNR-LFITS)具有可定量检测、灵敏度高和特异性强的优点,检测血清中cTnⅠ的线性范围为5～100 ng/mL,检出限(Limit of detection, LOD)为1.2 ng/mL,与其它蛋白的交叉反应值均小于5%。GNR-LFITS具有较高的实际应用能力,对临床血清样本中cTnⅠ的检测结果与商用酶联免疫(Enzyme-linked immunosorbent assay, ELISA)检测试剂盒具有良好的相关性(R     

基于金粒子修饰二硫化钼纳米复合材料的电化学发光免疫传感器的构建及其应用研究

生物标志物的快速、精准检测对控制和预防疾病或病毒的暴发具有重要意义.该研究将三维花状Au@(MoS2/GO/o-MWNTs)纳米复合物(AMGMs)作为一种理想的基底,利用"三明治"免疫夹心组装法构筑一种高灵敏的电化学发光免疫传感器,并用于前列腺特异性抗原(PSA)的高灵敏检测.AMGMs不仅具有良好的导电性和生物相容...     

Electrochemiluminescence immunosensor for α-fetoprotein using Ru(bpy)3(2+)-encapsulated liposome as labels

In this work, an electrochemiluminescence (ECL) immunosensor for ultrasensitive detection of α-fetoprotein (AFP) was fabricated using Ru(bpy)(3)(2+)-encapsulated liposome as the label and electrodeposited gold nanoparticles (GNPs) as the immobilizing support. Great signal amplification was achieved since liposome could encapsulate large amount of reporter molecules and GNPs could provide large active surface. Under optimized conditions, with sandwich type format, a linear range of AFP from 0.005 to 0.2 pg/mL and an extremely low detection limit of 0.001 pg/mL was obtained, much lower than that in previous reports. The proposed ECL immnuosensor showed high sensitivity, specificity, and good stability, which may open a new door to ultrasensitive detection of proteins in clinical analysis.     

显微荧光光谱原位测定红树植物根表面微区中的蒽

自行搭建显微荧光光谱分析( Microscopic fluorescence spectrometric analysis, MFSA)系统,并将其用于吸附于红树植物秋茄( Kandelia obovata, K. obovata)和白骨壤( Avicennia marina, A. marina)根表面微区中蒽( Anthrancene, Ant)的原位测定。在同步扫描模式下,以波长差为60 nm,吸附于K. obovata和A. marina根表面微区中Ant(5.3 pg/μm2)的荧光信号有最大信噪比(5.5和6.8)。所建方法的线性范围分别为5.3 ~63.2 pg/μm2和10.5~52.6 pg/μm2,检出限分别为1.1和5.5 pg/μm2,相对标准偏差均小于12.5%(n=9),加标回收率分别为98.1%~117.0%和81.2%~110.9%。结果表明,搭建的MFSA系统具备原位获取植物根表面微区Ant荧光光谱定量信息和荧光图像信息的能力。     

Disposable electrochemical immunosensor by using carbon sphere/gold nanoparticle composites as labels for signal amplification

This work designed a simple, sensitive, and low-cost immunosensor for the detection of protein marker by using a carbon sphere/gold nanoparticle (CNS/AuNP) composite as an electrochemical label. The nanoscale carbon spheres, prepared with a hydrothermal method by using glucose as raw material, were used to load AuNPs for labeling antibody by electrostatic interaction, which provided a feasible pathway for electron transfer due to the remarkable conductivity. The disposable immunosensor was constructed by coating a polyethylene glycol (PEG) film on a screen-printed carbon-working electrode and then immobilizing capture antibody on the film. With a sandwich-type immunoassay format, the analyte and then the CNS/AuNP-labeled antibody were successively bound to the immunosensor. The bound AuNPs were finally electro-oxidized in 0.1 M HCl to produce AuCl(4)(-) for differential pulse voltammetric (DPV) detection. The high-loading capability of AuNPs on CNS for the sandwich-type immunorecognition led to obvious signal amplification. By using human immunoglobulin?G (IgG) as model target, the DPV signal of AuNPs after electro-oxidized at optimal potential of +1.40?V for 40?s showed a wide linear dependence on the logarithm of target concentration ranging from 10?pg mL(-1) to 10?ng mL(-1). The detection limit was around 9?pg mL(-1). The immunosensor showed excellent analytical performance with cost effectivity, good fabrication reproducibility, and acceptable precision and accuracy, providing significant potential application in clinical analysis.     

Simultaneous determination of acetylsalicylic acid and salicylic acid in human plasma by isocratic high‐pressure liquid chromatography with post‐column hydrolysis and fluorescence detection

A selective, sensitive and rapid high‐performance liquid chromatography method with post‐column hydrolysis and fluorescence detection was developed for the simultaneous quantification of acetylsalicylic acid and its metabolite salicylic acid in human plasma. Following the addition of 2‐hydroxy‐3‐methoxybenzoic acid as internal standard and simple protein precipitation with acetonitrile, the analytes were separated on a ProntoSIL 120 C₁₈ ace‐EPS column (150 × 2 mm, 3 µm) protected by a C₈ guard column (5 µm). The mobile phase, 10 mm formic acid in water (pH 2.9) and acetonitrile (70:30, v/v), was used at a flow rate of 0.35 mL/min. After on‐line post‐column hydrolysis of acetylsalicylic acid (ASA) to salicylic acid (SA) by addition of alkaline solution, the analytes were measured at 290 nm (λ_ex) and 400 nm (λ_em). The method was linear in the concentration ranges between 0.05 and 20 ng/μL for both ASA and SA with a lower limit of quantification of 25 pg/μL for SA and 50 pg/μL for ASA. The limit of detection was 15 pg/μL for SA and 32.5 pg/μL for ASA. The analysis of ASA and SA can be carried out within 8 min; therefore this method is suitable for measuring plasma concentrations of salicylates in clinical routine. Copyright © 2012 John Wiley & Sons, Ltd.     

液相色谱-串联质谱法检测贝类产品中的原多甲藻酸贝类毒素

建立了一种贝类组织中原多甲藻酸(azaspiracid, AZA)贝类毒素主要成分AZA1的高效液相色谱-串联质谱检测方法。本方法采用甲醇-水(80:20, v/v)溶液对贝类组织中AZA1进行提取,并用MAX阴离子交换固相萃取(SPE)柱富集净化,使用Atlantis dC18(150 mm×4.6 mm, 5.0 μm)色谱柱分离,以含有50 mmol/L甲酸和2 mmol/L甲酸铵的乙腈-水溶液(80:20, v/v)为流动相进行等度洗脱,质谱采用选择反应监测(SRM)模式。AZA1在5 min内获得完全分离,且在48.85～2 442 ng/L范围内线性良好,相关系数为0.998 1。该方法检出限(S/N=3)为11.00 pg/g,添加水平为36.64、73.27、146.54 pg/g时的平均回收率为75.8%～82.5%(n=6),相对标准偏差小于10%。利用该方法对采自大连、青岛、广州水产品市场上的112个贝类样品进行了分析,发现采自大连和广州的部分贝类样品中含有AZA1。结果表明,该方法具有简单、快速、灵敏度高等特点,能充分满足贝类中AZA1检测的要求。     

Amplified inhibition of the electrochemical signal of ferrocene by enzyme-functionalized graphene oxide nanoprobe for ultrasensitive immunoassay

A nanoprobe-induced signal inhibition mechanism was designed for ultrasensitive electrochemical immunoassay at a chitosan-ferrocene (CS-Fc) based immunosensor. The nanoprobe was prepared by covalently loading signal antibody and high-content horseradish peroxidase (HRP) on the graphene oxide (GO) nanocarrier. The immunosensor was prepared through the stepwise assembly of gold nanoparticles (Au NPs) and capture antibody at a CS-Fc modified electrode. After sandwich immunoreaction, the GO-HRP nanoprobes were quantitatively captured onto the immunosensor surface and thus induced the production of a layer of insoluble film through the enzymatically catalytic reaction of the HRP labels. Both the dielectric immunocomplex formed on the immunosensor surface and the enzymatic precipitate with low electroconductivity led to the electrochemical signal decease of the Fc indicator, which was greatly amplified by the multi-enzyme signal amplification of the nanoprobe. Based on this amplified signal inhibition mechanism, a new ultrasensitive electrochemical immunoassay method was developed. Using carcinoembryonic antigen as a model analyte, this method showed a wide linear range over 5 orders of magnitude with a detection limit down to 0.54 pg/mL. Besides, the immunosensor showed good specificity, acceptable reproducibility and stability as well as satisfactory reliability for the serum sample analysis.     

基于硫化镍的C-反应蛋白免疫传感器的研制

本文合成了负载金纳米颗粒(Au NPs)的NiS纳米材料,通过壳聚糖(CHIT)将其固定在玻碳电极表面作为电化学生物传感器的固定基质。将C-反应蛋白(CRP)抗体固定到修饰过的玻碳电极表面,利用二茂铁甲酸标记CRP抗体,构建夹心型CRP生物传感器。采用差分脉冲伏安法(DPV)检测标记物二茂铁甲酸在0.3V左右的特征峰信号,该电流与培育的CRP抗原量成正比,从而实现对CRP的定量检测。传感器检测CRP的线性范围为0.01~500ng/mL,线性相关系数为0.9939,检测限为3.3pg/mL。     

大流量采样-GC-NCI-MS测定环境空气中的多溴联苯醚

建立了大流量采样-气相色谱负化学电离质谱法测定环境空气中痕量多溴联苯醚的方法.用PS-1型大流量空气采样器采集环境空气样品,样品经提取、纯化后采用气相色谱负化学电离质谱法测定环境空气中多溴联苯醚.方法的线性范围在5～10000 pg/m3之间,检出限1～50 pg/m3.用于检测2006年5月在广州市采集的环境空气样品,多溴联苯醚组分含量在5.4～4989 pg/m3范围.该方法适合用于监测环境空气中的痕量PBDEs.     

Determination of human IgG by solid substrate room temperature phosphorescence immunoassay based on an antibody labeled with nanoparticles containing Rhodamine 6G luminescent molecules

Luminescent silicon dioxide nanoparticles (R-SiO2) with size of 50 nm containing Rhodamine 6G (R) were synthesized by sol-gel method. In the presence of Pb(Ac)2 as a heavy atom perturber, the particle can emit intense and stable room temperature phosphorescence signal of R, respectively, on polyamide membrane, with the lambda(ex)(max)/lambda(em)(max) = 470/635 nm for R. Our research indicates that the specific immune reaction between goat-anti-human IgG antibody labeled with R-SiO2 and human IgG can be carried on polyamide membrane quantitatively, and the phosphorescence intensity was enhanced after the immunoreactions. Thus, a new method of solid substrate room temperature phosphorescence immunoassay (SS-RTP-IA) for the determination of human IgG was established basing on antibody labeled with the nanoparticles containing binary luminescent molecules. The linear range of this method is 0.0624-20.0 pg spot(-1) of human IgG (corresponding concentration, 0.156-50.0 ng mL(-1); sample volume, 0.40 microL spot(-1)). The regression equations of working curves are delta I(p) = 88.16. + 16.79 m(IgG) (pg spot(-1)) (485/646 nm, r = 0.9997). Detection limits calculated by 3Sb/k are 0.017 pg spot(-1). For samples containing 0.156 and 50.0 ng mL(-1) of IgG, we measured repeatedly for 11 times, RSDs are 3.9 and 2.8%, respectively. This method is sensitive, accurate and of high precision.