Electrochemical Sensors for Nitric Oxide Detection in Biological Applications

Nitric oxide (NO) plays an important role in physiological processes and it has been confirmed some human diseases are related to its biological function. Electrochemical sensors provide an efficient way to explore the NO function in biological processes. This review details different kinds of electrochemical sensors used for NO concentration detection between 2008 and 2013 together with their application in biological samples. Four commonly used electrodes and different assisted analysis membranes used for contributions towards the development of the novel sensors were summarized. Electrochemical sensors employed to measure NO concentration in a single cell, cell culture, tissue homogenate, organ, in vivo, human being, as well as in plant were also detailed. The trends of developing novel NO sensors were outlooked in the last part.     

Analysis of Nitric Oxide from Chemical Donors Using CMOS Platinum Microelectrodes

Electrochemical detection of NO generated from chemical donors is reported. Because NO is an important biological messenger, many donor sources and detection methods have been developed. Few reports have characterized NO donors using electrochemistry despite electrochemical techniques being sensitive and selective. Here, a CMOS platinum microelectrode array is interfaced with a microfluidic device for the electrochemical analysis of NO from (Z)‐1‐[N‐(2‐aminoethyl)‐N‐(2‐ammonioethyl)amino]diazen‐1‐ium‐1,2‐diolate (DETA/NO). The donor parent amine fouls the electrode, resulting in substantial signal loss, but an electrochemical cleaning method was developed that substantially reduces fouling and allows detection of NO between 90 nM and 1 µM.     

Construction and Characterization of a New Flexible and Nonbreakable Nitric Oxide Microsensor

Nitric oxide (NO) is an important molecule in many different physiological phenomena. Investigation of nitric oxide production in vivo requires a specialized sensor capable of real‐time concentration measurement, with a high spatial resolution of NO gas production. In this study, a flexible microsensor is developed specifically for measurement of production of nitric oxide. The new sensor consists of a Pt/Ir working electrode coupled with an integrated Ag/AgCl reference electrode. The sensor is coated with a series of NO‐selective membrane polymers to prevent potential amperometric response due to interfering species. Presented experimental data demonstrates the ability for NO detection between 100 and 400 nM concentrations with a linear response (R²=0.9997). The detection limit of the sensor is 2.14 nM (S/N=3). Various selectivity experiments are indicative of a resistance to interfering species such as dopamine, norepinephrine, L ‐arginine.     

Effects of Siqi Decoction on Concentration of Nitric Oxide and Activity of Nitric Oxide Synthase in Blood Serum of Rats with Myocardial Ischemia

To study the effects of Siqi decoction on rats with Myocardial Ischemia, the model of Myocardium Ischemia was made for the Wistar rats cured with posterior pituitary injection through vein in tail. Siqi decoction, Diaoxinxuekang（DK） and Fufangdanshenpian（FD）, the latter two drugs of which were effective TCM drugs of anti-Myocardial Ischemia at present, were administrated to the rats with Myocardium Ischemia for 5 days to compare the effect of anti-Myocardium Ischemia as reference drugs by measuring the changes of NO concentration and activity of NOS in the rat blood serum with Myocardial Ischemia. There was a remarkable increase in the NO concentration and activity of NOS in serum in Siqi decoction groups compared with those in control group（p〈0.05）. The results of the prevention group in experiment of Siqi decoction are better than those of the cure group. Siqi decoction was really fit for Myocardium Ischemia via increasing NO concentration by stimulating the activity of NOS in serum. The effect of Siqi decoction against Myocardium Ischemia in preventive group is better than the curative that of Siqi decoction in the curative group.     

荧光检测海水介质中的一氧化氮

基于2,3-二氨基萘(DAN)与一氧化氮(NO)在海水介质中生成荧光性2,3-萘酚三唑(NAT)来测定海水中NO.确定了荧光法测定NO的条件(λex=383 nm,λem=410 nm)、温度和时间对DAN捕集NO的影响,以及DAN溶液对捕集NO的影响.测得NO浓度与其荧光强度之间呈线性关系,线性范围1.4～1400 nmol/L, r=0.9985,P<0.0001;检出限1 nmol/L (S/N=3);相对标准偏差1.63%(14 nmol/L NO);不存在介质干扰.同样条件下与NaOH为介质的荧光法相比,本方法将检出限提高了1个数量级.结合吹扫-捕集检测出青岛石老人(36°05′46″ N; 120°29′40″ E)海水中NO浓度为(0.12±0.01) nmol/L,可用于监测硝普钠在海水中释放NO的规律.     

Nafion修饰铂电极测定天然海水中一氧化氮

用Nafion修饰铂电极测定天然海水介质中的一氧化氮。确定了富集时间、电极修饰时间、修饰膜厚度等最佳实验条件,同时用线性扫描伏安法对海水中的NO进行测定,测得NO的浓度与其氧化峰电流之间有一定线性关系,线性范围1~77μmol/L,r=0.9916;检出限0.4μmol/L;相对标准偏差0.5%,探讨了Nafion对NO的富集机理,并用N-亚硝基-乙腈霉胺分解法(SNAP)对本方法进行了验证。     

羟基取代的多环芳烃与一氧化氮的反应研究

内源性的一氧化氮(NO)是多种生理过程中必不可少的信使分子,它在神经系统递质传导、神经发育、脑血流调节以及免疫调节等过程中具有十分重要的作用。为揭示NO生理功能的化学本质,NO成为化学家近年来研究的重要课题之一。     

Fluorescent Nitric Oxide Photodonors Based on BODIPY and Rhodamine Antennae

Two novel NO photodonors (NOPDs) based on BODIPY and Rhodamine antennae activatable with the highly biocompatible green light are reported. Both NOPDs exhibit considerable fluorescence emission and release NO with remarkable quantum efficiencies. The combination of the photoreleasing and emissive performance for both compounds is superior to those exhibited by other NOPDs based on similar light-harvesting centres, making them very intriguing for image-guided phototherapeutic applications. Preliminary biological data prove their easy visualization in cell environment due to the intense green and orange-red fluorescence and their photodynamic action on cancer cells due to the NO photo-liberated.     

担载型钴镍金属氧化物催化剂上CH4还原NO反应研究

ＮＯｘ是大气的主要污染物之一,近年来,利用甲烷选择还原ＮＯｘ引起了广泛关注,研究发现,Ｃｏ、Ｍｎ和Ｎｉ离子交换的ＺＳＭ－５分子筛具有较高的活性［１～３］．但金属离子交换的分子筛催化剂有热稳定性差、易失活等缺点．最近已有金属氧化物用作甲烷还原ＮＯｘ反应...     

A General Procedure for Synthesis of NG-Alkyl,and NG-Aryl-L-Arginines as Potential Nitric Oxide Synthase inhibitors

A general procedure for the synthesis of N^G-alkyl, and N^G-aryl-L -arginines with relatively high overall yield is reported. The key step involved the coupling of protected L -ornithine 4 with isothiourea 7 to give the fully protected N^G-aryl-L -arginine derivative 8 . Subsequent deprotection of 8 in acidic condition provided the final target compound 9 with an overall yield of more than 80%.     

Calreticulin Transacetylase Mediates the Acetylation of Nitric Oxide Synthase by Polyphenolic Acetate

Our earlier investigations identified acetoxy drug: protein transacetylase (TAase), a unique enzyme in the endoplasmic reticulum (ER) catalyzing the transfer of acetyl groups from polyphenolic acetates (PA) to certain functional proteins. Recently we have established the identity of TAase with ER protein calreticulin (CR) and subsequently transacetylase function of CR was termed calreticulin transacetylase (CRTAase). CRTAase was purified and characterized from human placenta. CRTAase catalyzed the acetylation of a receptor protein nNOS, by a model PA 7, 8-diacetoxy-4-methylcoumarin (DAMC), which was visually confirmed by using antiacetyl lysine. The aim of this report was to provide tacit proof by providing mass spectrometry evidence for CRTAase catalyzed acetylation of purified nNOS by DAMC. For this purpose, purified nNOS was incubated with DAMC and CRTAase, the modified nNOS was analyzed by nanoscale LC-MS/MS, which recorded 11 distinct peptides with a significant score as acetylated on lysine residues. The distribution was in order: lysines-24, -33, -38, -131, and -229 of the PDZ domain, Lys-245 of the oxygenase domain, Lys-754 and -856 of FMN binding domain, Lys-989 of connecting domain and Lys-1300, -1321, and -1371 of the NADPH-binding domain were acetylated. The results documented in this paper highlighted for the first time modification of nNOS by way of acetylation. Our earlier work recorded the profound activation of platelet NADPH cytochrome P-450 reductase and the acetylation of the reductase protein by DAMC, which also remarkably enhanced intracellular levels of nitric oxide. The results reported here coupled with the aforementioned previous observations strongly implicate the possible role of the acetylation of the reductase domain of nitric oxide synthase (NOS) in the NOS activation. In addition, the acetylation of nNOS can be expected to potentiate the interaction with CR, eventually leading to the augmented catalytic activity of NOS and expression of the related biological effects.     

一种检测NO的修饰电极

报道了[Fe(Sal₂en)]²⁺的咪唑加合物对NO电氧化的影响.实验表明,它将[Fe(Sal₂en)]²⁺对NO的电催化氧化活性提高T₂倍.过量咪唑会降低[Fe(Sal₂en)]²⁺对NO的催化活性.证实了NO的电催化氧化形成NO[Fe(Sal₂en)]²⁺中间体的推断.用制得的修饰电极测定NO,当c(NO)在3×10^-6～2×10^-9mol/L时,有良好线性关系,相对标准偏差小于0.5％,线性相关系数为0.9994,检测限为2×10^-9mol/L.满足生理分析的要求.     

乙烯基炔与一氧化氮反应机理的理论探讨

利用半经验AM1方法，研究了乙烯基乙炔与一氧化氮反应生成丙炔腈（反应1）和丙烯腈（反应2）的两条竞争反应途径．采用Berny梯度法优化得到了反应势能面上的所有驻点，并逐一进行了振动分析确认．结果表明，反应互较反应Ⅱ易引发（即；NO比较容易与乙烯基乙炔进行分步的1，4-加成，形成六元环过渡态及中间体，最终消除CH2O和H自由基后生成丙炔腈），而反应Ⅱ较难引发（NO与乙烯基乙炔较难进行1，2-加成，形成四元环过渡态及中间体），但一经引发，就能较容易地进行分步裂解，生成丙烯腈和CHO自由基．     

一氧化氮标准储备水溶液浓度的测定及稳定性分析

用紫外-可见分光光度法,分别使用酸性和中性两种G riess-Saltzm an(GS)试剂对NO标准储备液的浓度及稳定性进行了分析。结果表明NO在酸性GS试剂中其检测灵敏度远大于中性GS试剂中的灵敏度(近19倍),但在酸性GS试剂中NO2-对NO的检测有干扰,其灵敏度约为NO的1/3;而在中性GS试剂中,当NO2-浓度小于10μmol/L时,NO2-对NO的检测没有影响。实验室制备的一氧化氮饱和溶液的浓度在10℃时为2.7 mmol/L,20℃时为2.0 mmol/L,其稳定期均为3 d。     

高等植物叶绿体中光诱导生成NO的ESR证据

NO分子在生物细胞体系中是一个十分重要的信使分子,它具有调节细胞凋亡、松弛平滑肌、抑制血小板聚集与黏附、抑制病原体和肿瘤生长等多种生理功能^[1,2],因而备受生物学家的关注.众所周知,动物细胞中NO主要来源于一氧化氮合酶(NOS)的表达^[3];然而,直到最近才有研究证明植物细胞中存在参与调节植物生长和激素信号转导的NOS基因^[4].一般认为植物细胞中NO主要来源于硝酸盐还原酶(NR)^[5].我们曾在高等植物光系统中研究发现了与电子传递相关的活性氧生成机制^[6～8],但其间是否也会通过光诱导电子传递产生NO尚不清楚.Kozlov等^[9]曾报道在小鼠线粒体内存在与亚硝酸盐还原酶功能类似的还原型烟酰胺腺嘌呤二核苷酸(NADH)诱导电子传递生成NO的反应,本文采用电子自旋共振(ESR)方法验证了高等植物叶绿体内光诱导电子传递可产生NO的设想.     

关于一氧化氮生成反应的讨论

In the course of frontier orbital theory, the teacher and students discussed the conditions for the nitric oxide formation reaction, by using this theory. In an extracurricular practice, students testified the reaction mechanism obtained from the course by molecular orbital calculations. The course practice improved students' understanding of these theories and their abilities to apply them to exploration of chemical problems.     

基于表面活性剂单分子层修饰碳糊电极的一氧化氮电化学传感器及其应用

报道了一种表面活性剂单分子层修饰碳糊电极,并用于NO的高灵敏电化学检测。研究表明,表面活性剂通过烷基链在电极表面形成的疏水性单分子层微环境对NO的电化学响应具有较好的促进作用。其中,阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)对NO的电化学氧化表现出最强的催化活性和增敏作用。在Nafion膜覆盖的CTAB修饰碳糊电极上,NO的安培响应与其浓度在3.6×10-8～1.8×10-5mol/L范围内呈良好的线性关系,检出限为1.8×10-8mol/L。该电极作为低成本、高灵敏的NO电化学传感器,被成功应用于大鼠肺组织细胞中NO释放的实时监测。     

检测生理内源性一氧化氮分子探针的研究及应用进展

本文介绍和评述了荧光光度法、化学发光法、分光度度法、电化学和电子自旋共振光谱法中的分子探针及在检测生理内源性一氧化氮的应用及研究进展。引用文献42篇。