基于双层酶信号放大及纳米功能界面的微囊藻毒素电化学免疫传感器

设计了一种基于纳米复合膜双层酶信号放大的竞争型电化学免疫传感器,用于检测痕量微囊藻毒素-LR(Microcystin-(leucine-arginine),MCLR)。在多壁碳纳米管修饰玻碳电极上电沉积金纳米粒子形成纳米复合膜,作为MCLR抗体(anti-MCLR)和辣根过氧化物酶(HRP)的固定化载体;引入HRP作为封闭剂或者通过抗体捕获HRP标记的MCLR(HRP-MCLR),起封闭活性位点及协同催化的作用。利用MCLR与HRPMCLR对纳米复合膜所固载抗体活性位点的竞争结合模式,采用微分脉冲伏安法(DPV)测定电极界面上HRP酶催化过氧化氢(H_2O_2)氧化对苯二酚(HQ)产生的还原电流,实现MCLR的定量检测。此传感器具有良好的特异性、稳定性与灵敏度,线性检测范围为0.1~100.0 μg/L,检出限为0.038 μg/L(S/N=3),对实际水样的加标回收率为72.9%~117.3%。     

基于Fe3O4@Au磁性纳米粒子修饰丝网印刷电极的微囊藻毒素免疫传感器研究

将核壳型Fe3O4@Au磁性纳米粒子修饰在丝网印刷工作电极表面,再通过纳米金和微囊藻毒素-(亮氨酸-精氨酸)抗体(anti-MCLR)之间的吸附作用,将抗体固定于电极表面,以牛血清白蛋白(BSA)封闭非特异性吸附位点,制得了检测MCLR的电流型免疫传感器。该传感器基于直接竞争的免疫分析模式,以辣根过氧化物酶偶联的微囊藻毒素(MCLR-HRP)为标记物,用差分脉冲伏安法检测微囊藻毒素,在优化的实验条件下,此免疫传感器响应的峰电流值与微囊藻毒素浓度在0.79～12.9μg/L范围内呈良好的线性关系,检测限为0.38μg/L。对实际水样进行了微囊藻毒素的加标回收实验,回收率在95%～107%之间。此免疫传感器具有测定速度快、灵敏高、携带方便等优点。     

Ti/RuO2-CoO电极制备及其氨氮降解性能

采用热氧化法制备Ti/RuO_2-CoO电极,通过SEM和XRD对电极涂层的表面形貌和晶体结构进行表征;并利用最佳条件制备的电极开展电化学氧化氨氮模拟废水研究。实验研究了电流密度、初始pH值、Cl~-浓度和NH_3-N初始浓度对NH_3-N降解效果的影响。研究结果表明:当n(Ru)/n(Co)=7∶3时Ti/RuO_2-CoO电极对NH_3-N的去除效果较好。随着电流密度的增加,NH_3-N去除率随之升高;初始pH为碱性条件时,NH_3-N去除效果较好;NH_3-N去除率随Cl~-浓度的增加呈现先增大后减小,当Cl~-浓度为3 000 mg/L时,氨氮去除率达到97.95%。当电流密度为70 mA/cm~2、Cl~-浓度为3 000 mg/L、初始pH=9时,经过180 min电解后,氨氮初始浓度从500 mg/L降低至10.25 mg/L,且电化学氧化氨氮的产物主要为氮气和硝酸盐氮。     

三维电极系统电化学氧化三唑酮

以三唑酮(TDF)为目标污染物,以钛基钌铱电极和不锈钢板作阳极、阴极,颗粒活性炭(GAC)为粒子电极构建三维电极系统,通过搅拌实现粒子电极呈流化状态。研究电流密度、TDF初始浓度、GAC投加量、初始pH对三维电极系统的TDF去除效率的影响。研究结果表明:增大电流密度有助于提高TDF去除效率;TDF去除率随初始pH的增大呈现逐渐增大的趋势,碱性条件有助于TDF的去除;当GAC投加量为1～4 g/L时,增大GAC投加量有助于TDF的降解;增大TDF初始浓度会导致去除率的降低。当TDF初始浓度为200μg/L、电流密度为8m A/cm2、初始pH=11、GAC投加量为4 g/L时,电化学氧化10 min时TDF去除率达到99.95%。三维电极系统的传质效率高于二维电极系统,三维电极电化学氧化系统是一种适用于降解TDF的高级氧化技术。     

基于HDT自组装金纳米粒子修饰电极的微囊藻毒素-LR电化学免疫传感器研究

构建了一种新型的基于金纳米粒子(Au NPs)修饰金电极的微囊藻毒素-亮氨酸-精氨酸(MCLR)电化学免疫传感器。采用柠檬酸钠还原法制备了Au NPs溶胶,分别用透射电子显微镜和紫外-可见吸收光谱对其进行表征。将Au NPs组装到1,6-己二硫醇(HDT)自组装单分子层修饰的金电极表面,再将MCLR抗体(anti-MCLR)固定于该修饰电极上,利用扫描探针显微镜法、循环伏安法和电化学交流阻抗法(EIS)表征了自制化学修饰电极表面的形貌特征和电化学免疫传感器的电化学特征。通过辣根过氧化物酶标记的MCLR(MCLR-HRP)与MCLR竞争结合抗体,建立了检测MCLR的差分脉冲伏安法(DPV)。在最佳实验条件下,用DPV对MCLR检测的线性范围为0.01~25μg/L,检出限为0.005μg/L。对构建的免疫传感器的重现性、稳定性和选择性进行了考察。该方法对实际水样中MCLR的加标回收率为100%~102%,测定结果与高效液相色谱法的测定结果一致。     

Ti/α-PbO2/β-PbO2电极电化学降解2-氯酚

采用电化学沉积法在Ti基底上制备了复合电极Ti/α-PbO2/β-PbO2,扫描电镜结果表明电极呈现由β-PbO2小晶体组成的菜花状微观形貌.所制电极在电化学降解环境污染物2-氯酚时表现出较高的电催化效率、较好的电极稳定性和较长的电极寿命.用正交实验优化了电化学降解2-氯酚的实验条件.在最优的实验条件(2-氯酚初始浓度50 mg/L,电解质0.1 mol/L Na2SO4,温度35oC,阳极电流密度20 mA/cm2)下电化学降解180 min后,2-氯酚的去除率达100%.动力学结果表明, Ti/α-PbO2/β-PbO2电极上2-氯酚的电化学氧化符合准一级动力学过程.     

基于石墨烯和金纳米笼修饰的无标记型微囊藻毒素免疫传感器的研制

利用石墨烯及中空结构的金纳米笼构建了无标记型电化学免疫传感器,并用于微囊藻毒素的检测。利用多元醇还原法合成制备了导电性好、催化性强、生物相容性好的金纳米笼;再利用高分散的石墨烯将其固定于玻碳电极表面,进一步吸附固定微囊藻毒素抗体。在无微囊藻毒素存在时,电化学探针[Fe(CN)6]3!/4!在传感器界面上能获得较高的电流响应信号。当培育了微囊藻毒素后,抗体与微囊藻毒素形成免疫结合物,增加了电极表面的电荷密度和传质阻力,阻碍[Fe(CN)6]3!/4!扩散到电极表面,导致[Fe(CN)6]3!/4!的电流响应信号明显降低,电流减小的程度间接地与微囊藻毒素的浓度成比例,可实现对微囊藻毒素的检测。实验考察了抗原培育时间,抗体浓度等条件对该传感器响应性能的影响。结果表明,此传感器对微囊藻毒素的线性响应范围为0.05~1000μg/L,检出限为0.017μg/L,优于文献报道。此传感器操作简单,并且具有良好的稳定性,将其用于实际水样中微囊藻毒素的检测,平均加标回收率为94.1%。     

La-Ti/SnO2-Sb/RuO2-Co电极制备及其处理垃圾渗滤液研究

以纯钛为基体材料,并以热氧化的方式制备La-Ti/SnO_2-Sb/RuO_2-Co电极,采用X-射线衍射仪(XRD)、扫描电子显微镜(SEM)及X-射线能量色散谱(EDS)对电极涂层表面形貌和晶体结构进行表征。采用制备的La-Ti/SnO_2-Sb/RuO_2-Co电极作为阳极,不锈钢为阴极,构建电化学反应器,对老龄垃圾渗滤液进行降解。结果表明在电流密度为65 mA/cm~2、Cl-浓度为4 500 mg/L、初始pH=9和反应时间为6 h的最佳反应条件下,La-Ti/SnO_2-Sb/RuO_2-Co电极对NH_3-N和COD的去除率分别达到86.5%、61%。     

微生物燃料电池处理餐饮废水及同步发电性能研究

针对食堂餐饮废水,建立微生物燃料电池实验系统,研究微生物燃料电池废水处理与同步发电性能。首先使用Fe(NO3)3溶液作为阴极电解液进行实验,证明餐饮废水生物降解及产电的可行性;分别采用NaCl溶液和K3[Fe(CN)6]溶液作为阴极电解液进行对比实验,研究不同运行环境下微生物燃料电池的发电性能和污水净化效果。结果表明,采用NaCl溶液和K3[Fe(CN)6]溶液作为阴极电解液时的COD去除率分别是30%和22%左右,平均电流密度分别为5.6和5.2mA/m2。在污水稀释比为2∶1、NaCl电解液浓度为0.4mol/L的情况下,微生物燃料电池系统的发电性能和净水效果达到最佳状态,稳态电流密度为8.8mA/m2,COD去除率为33.3%。     

电生成Fenton试剂及其对染料降解脱色的研究

以可溶性铁为阳极,多孔石墨电极为阴极,Na2SO4为支持电解质.通电后,铁从阳极溶解生成二价亚铁离子Fe2+,氧在阴极上还原为过氧化氢H2O2,于电解现场发生Fenton反应,产生羟基自由基即Fenton试剂.在低电流密度(10mA/cm2)下,可有效地抑制阴、阳两极副反应的发生,所产生的羟基自由基浓度足以有效地进行染料废水的降解与脱色,脱色率达100%,CODcr去除率达80%.     

Resonance Raman spectra of cytochrome C and oxyhemoglobin using pulsed laser excitation and optical multichannel detection

The resonance Raman spectra of dilute cytochrome c and oxyhemoglobin solutions obtained with high peak power (>1 MW) laser excitation at 532.0 nm and optical multichannel analysis (OMA) are presented. The frequencies and intensities of bands in resonance Raman spectra obtained under these conditions are directly comparable to spectra derived from 0.15 W peak power excitation using cw lasers. No anomalous spectral features are observed. These pulsed-laser/OMA spectra are used to comment on the applicability of such techniques for time-resolved resonance Raman spectroscopy of biopolymers.     

Characteristics of the Raman spectra of single-walled carbon nanotube bundles under electrochemical potential control

Resonant Raman scattering spectra of single-walled carbon nanotube–sodium dodecyl sulfate (SWNT–SDS) bundles adsorbed on Au electrodes have been investigated in aqueous electrolytes. Raman intensities of the radial breathing mode (RBM) with 785-nm laser excitation were monitored at different electrode potentials between −0.5 and +0.8 V relative to the SCE. Six resolved RBM peaks assignable to different diameter tubes all decreased in intensity when the electrode was positively biased, because of depletion of valence-band electrons associated with resonant excitation. The attenuation occurred at more positive potentials for narrower-diameter tubes with higher RBM frequencies consistent with their larger bandgaps. The results suggest the Fermi level is equilibrated in bundled SWNTs in contrast with the large Fermi-level shifts reported for isolated SWNTs.     

Characterization and surface-enhanced Raman spectral probing of silver hydrosols prepared by two-wavelength laser ablation and fragmentation

A four step Ag foil laser ablation-Ag nanoparticle fragmentation procedure in ultrapure water was carried out both under argon and in air. Pulses of a high power Nd/YAG laser were used for laser ablation (1064 nm) and for the three step Ag hydrosol treatment in the absence of Ag foil in the sequence 1064-532-1064 nm. Transmission electron microscopy (TEM) and surface plasmon (SP) extinction spectra provide evidence of Ag nanoparticle fragmentation in the second and third step of the procedure carried out under argon. While polydispersity of Ag hydrosol increases in the second step, both the polydispersity and the mean size of the nanoparticles are reduced in the third step. Qualitative and quantitative surface-enhanced Raman scattering (SERS)/surface-enhanced resonance Raman scattering (SERRS) spectral probing of systems with Ag hydrosols and the selected adsorbates at 514.5 nm excitation shows that Ag hydrosols obtained in the second step of the preparation procedure carried out in air are the most suitable substrates for SERS/SERRS experiments performed at this excitation wavelength.     

Quantitative analysis of methyl green using surface-enhanced resonance Raman scattering

Surface-enhanced resonance Raman scattering (SERRS) spectra of aqueous solutions of the triphenylmethane dye methyl green have been obtained for the first time by use of citrate-reduced silver colloids and a laser excitation wavelength of 632.8 nm. Given the highly fluorescent nature of the analyte, which precluded collection of normal Raman spectra of the dye in solution and powdered state, it was highly encouraging that SERRS spectra showed no fluorescence due to quenching by the silver sol. The pH conditions for SERRS were optimised over the pH range 0.5–10 and the biggest enhancement for SERRS of this charged dye was found to be at pH 2.02, thus this condition was used for quantitative analysis. SERRS was found to be highly sensitive and enabled quantitative determination of methyl green over the range 10⁻⁹ to 10⁻⁷ mol dm⁻³. Good fits to correlation coefficients were obtained over this range using the areas under the vibrational bands at 1615 and 737 cm⁻¹. Finally, a limit of detection of 83 ppb was calculated, demonstrating the sensitivity of the technique.     

Library of FT-Raman spectra of pigments, minerals, pigment media and varnishes, and supplement to existing library of Raman spectra of pigments with visible excitation.

Sixty pigments, minerals and media have been analysed by Fourier-transform Raman (FT-Raman) microscopy in order to assemble a database of reference FT-Raman spectra for scientists working at the Arts-Science interface. An earlier library of Raman spectra compiled using visible excitation has been extended by the addition of 22 further reference spectra obtained with 780.0, 647.1, 632.8 and/or 514.5 nm excitation. The relative merits of 1064 nm and visible excitation are discussed.     

四苯基卟啉及其金属配合物在溴化银胶体上的表面增强喇曼光谱研究

研究了四苯基卟啉(H2TPP)及其金属配合物(AgTPP和MgTPP)在AgBr胶体上的表面增强喇曼光谱(SERS).SERS光谱表明,吸附在AgBr胶体粒子表面的MgTPP和H2TPP分子分别发生银离子交换和银配位反应生成AgTPP,这种表面反应可能与激光照射有关.AgTPP分子在胶体粒子表面的吸附导致卟啉大环的非平面化,使vs振动(M-N键伸缩振动)向高波数方向移动近10 cm-1.632.8 nm激发下的表面喇曼谱以化学增强为主,而488.0 nm激发下表面喇曼谱除化学增强效应外,还存在共振增强效应.     

Enhancement at the junction of silver nanorods

The enhancement of surface enhanced Raman scattering (SERS) at the junction of linearly joined silver nanorods (31 nm in diameter) deposited in the pores of anodic aluminum oxide templates was studied systematically by excitation with a 632.8 nm laser line. The single and joined silver nanorod arrays showed a similar extinction spectrum when their length was the same. Maximum enhancement was observed from the junction system of two nanorods of the same size with a total length of 62 nm. This length also corresponded to the optimum length of single nanorods for SERS by excitation with a 632.8 nm laser line. The enhancement at the junction was approximately 40 times higher than that of the 31 nm single nanorod, while it was 4 times higher than that of the 62 nm single nanorod. The enhancement factor at the junction after oxide removal was approximately 3.9 x 10 (9).     

Raman scattering of 4-aminobenzenethiol sandwiched between Ag nanoparticle and macroscopically smooth Au substrate: effects of size of Ag nanoparticles and the excitation wavelength

A nanogap formed by a metal nanoparticle and a flat metal substrate is one kind of "hot site" for surface-enhanced Raman scattering (SERS). Accordingly, although no Raman signal is observable when 4-aminobenzenethiol (4-ABT), for instance, is self-assembled on a flat Au substrate, a distinct spectrum is obtained when Ag or Au nanoparticles are adsorbed on the pendent amine groups of 4-ABT. This is definitely due to the electromagnetic coupling between the localized surface plasmon of Ag or Au nanoparticle with the surface plasmon polariton of the planar Au substrate, allowing an intense electric field to be induced in the gap even by visible light. To appreciate the Raman scattering enhancement and also to seek the optimal condition for SERS at the nanogap, we have thoroughly examined the size effect of Ag nanoparticles, along with the excitation wavelength dependence, by assembling 4-ABT between planar Au and a variable-size Ag nanoparticle (from 20- to 80-nm in diameter). Regarding the size dependence, a higher Raman signal was observed when larger Ag nanoparticles were attached onto 4-ABT, irrespective of the excitation wavelength. Regarding the excitation wavelength, the highest Raman signal was measured at 568 nm excitation, slightly larger than that at 632.8 nm excitation. The Raman signal measured at 514.5 and 488 nm excitation was an order of magnitude weaker than that at 568 nm excitation, in agreement with the finite-difference time domain simulation. It is noteworthy that placing an Au nanoparticle on 4-ABT, instead of an Ag nanoparticle, the enhancement at the 568 nm excitation was several tens of times weaker than that at the 632.8 nm excitation, suggesting the importance of the localized surface plasmon resonance of the Ag nanoparticles for an effective coupling with the surface plasmon polariton of the planar Au substrate to induce a very intense electric field at the nanogap.     

Semi-quantitative trace analysis of nuclear fast red by surface enhanced resonance Raman scattering

The dye nuclear fast red has been detected and determined semi-quantitatively by means of surface enhanced resonance Raman scattering (SERRS) and surface enhanced Raman scattering (SERS), using laser exciting wavelengths of 514.5 and 632.8 nm, respectively, by employing a citrate-reduced silver colloid. A good linear correlation is observed for the dependence of the intensities of the SERRS bands at 989 cm⁻¹ (R=0.9897) and 1278 cm⁻¹ (R=0.9872) on dye concentration over the range 10⁻⁹ to 10⁻⁷ M, when using an exciting wavelength of 514.5 nm. At dye concentrations above 10⁻⁷ M, the concentration dependence of the SERRS signals is non-linear. This is almost certainly due to the coverage of the colloidal silver particles being in excess of a full monolayer of the dye. A linear correlation is also observed for the dependence of the intensities of the SERS bands at 989 cm⁻¹ (R=0.9739) and 1278 cm⁻¹ (R=0.9838) on the dye concentration over the range 10⁻⁸ to 10⁻⁶ M when using an exciting wavelength of 632.8 nm. Strong fluorescence prevented collection of resonance Raman scattering (RRS) spectra from powdered samples or aqueous solutions of the dye using an exciting wavelength of 514.5 nm, but weak bands were observed in the spectra obtained from both powdered and aqueous samples of the dye using an exciting wavelength of 632.8 nm. A study of the pH dependence of SERRS/SERS and UV–VIS absorption spectra revealed the presence of different ionisation states of the dye. The limits of detection for nuclear fast red by SERRS (514.5 nm), SERS (632.8 nm) and visible spectroscopy (535 nm) are 9, 89 and 1000 ng ml⁻¹, respectively.     

Time-resolved spatially offset Raman spectroscopy for depth analysis of diffusely scattering layers

The objective of this study is to use time-resolved (TR) Raman spectroscopy, spatially offset Raman spectroscopy (SORS), and a combination of these approaches to obtain high quality Raman spectra from materials hidden underneath an opaque layer. Both TR Raman and SORS are advanced techniques that allow for an increased relative selectivity of photons from deeper layers within a sample. Time-resolved detection reduces fluorescence background, and the selectivity for the second layer is improved. By combining this with spatially offset excitation we additionally increased selectivity for deeper layers. Test samples were opaque white polymer blocks of several mm thicknesses. Excitation was carried out with a frequency-doubled Ti:sapphire laser at 460 nm, 3 ps pulse width and 76 MHz repetition rate. Detection was either with a continuous-wave CCD camera or in time-resolved mode using an intensified CCD camera with a 250 ps gate width. The Raman photons were collected in backscatter mode, with or without lateral offset. By measuring the delay of the Raman signal from the second layer (polyethylene terephthalate/PET/Arnite), the net photon migration speeds through Teflon, polythene, Delrin and Nylon were determined. Raman spectra could be obtained from a second layer of PET through Teflon layers up to 7 mm of thickness. The ability to obtain chemical information through layers of diffusely scattering materials has powerful potential for biomedical applications.