核糖核酸与迈格林华尔色素作用机理及含量测定研究

在弱酸性介质中,小分子迈格林华尔色素与生物大分子核糖核酸发生强烈相互作用,导致分子构象变化,进而引起吸收光谱最大波长和吸收值的变化.研究了缓冲溶液酸度、色素用量、反应时间对反应体系的影响,确定了最佳实验条件,建立了测定核糖核酸的新方法.方法的线性范围为0～8.0 mg/L,相关系数为0.999 6,直接测定样品中的核糖...     

基于阳离子共轭聚合物比色检测HIV逆转录酶的RNase H活性

利用阳离子聚噻吩衍生物与单链DNA和杂合体DNA/RNA通过静电相结合时所产生的紫外吸收变化,建立了一种检测HIV逆转录酶(RT-HIV)的RNase H活性的方法。阳离子聚噻吩衍生物的紫外吸收最大波长位于短波385nm,与单链DNA结合会使聚噻吩衍生物的紫外吸收最大波长红移至525nm;而与杂合体DNA/RNA结合时对其紫外吸收最大波长几乎没有影响,当利用RT-HIV的核糖核酸酶RNase H活性水解掉杂合体中的RNA时,杂合体溶液又会使聚噻吩衍生物的紫外吸收最大波长发生红移。结果表明,紫外吸收最大波长变化明显,甚至直观用肉眼就可以观察到杂合体水解前后溶液颜色的变化。同时还测定了不同时间下RNase H酶水解杂合体中RNA的吸光度变化曲线,计算出了RNase H酶水解的动力学常数和最大初速度。     

铬天青S胶束增溶分光光度法测定高纯氧化铋中微量铝

高纯氧化铋试样用硝酸溶解后 ,加入氢溴酸使铋生成易挥发的溴化铋除去 ,从而消除了铋对测定铝的干扰 ,在pH 6 .7的乙酸 乙酸铵缓冲溶液中 ,在辛基酚聚氧乙烯醚存在条件下 ,铝与铬天青S生成稳定的三元配合物 ,其最大吸收波长为 6 2 0nm ,表观摩尔吸光系数为 2 .30× 10 5L/ (mol·cm) ,铝在 0～ 10 μg/ 5 0mL范围内符合比耳定律 ,适用于高纯氧化铋中的微量铝的测定     

铬天青S-Cu(Ⅱ)配合物探针吸光光度法测定蛋白质的研究

在pH 5.0 B-R缓冲介质中,蛋白质与铬天青S-Cu（Ⅱ）配合物发生结合反应形成超分子复合物,使体系的最大吸收波长红移了30 nm,由此提出了普通光度法测定蛋白质（BSA）的新方法。最佳条件下,该复合物的最大吸收波长位于620 nm处,表观摩尔吸光系数ε=6.13×105L.mol-1.cm-1,蛋白质（BSA）量在10-60μg/mL范围内遵循比耳定律。方法可直接用于蛋清中蛋白质的测定,回收率为100%-105%。     

树形聚酰胺胺与Cu^2＋的络合作用

合成了4．0代聚酰胺胺（PAMAM）树形分子,并合成出端基为羟基的PAMAM树形分子衍生物,用分光光度法研究了4．0代PAMAM树形分子及其衍生物与Cu^2 的络合作用,结果表明当存在树形聚胺胺分子时,Cu2 水溶液的最大吸收波长显著紫移,随n(Cu2 )/n(PAMAM)增加,最大吸收波长红移,PAMAM树形分子与Cu2 的络合作用有多种形式,对端胺基树形分子主要存在Cu-N4和Cu-N2两种配位方式;对端羟基树形分子主要存在Cu-N2的配位方式,随Cu2 的加入,络合形式和各种络合形式的相对比例发生变化,PH对络合形式有较大影响;随代数的增加,杩形分子所能络合Cu2 的最大数目不断增加,但理论与实验值有一定的误差。     

天青Ⅰ分光光度法测定冷却液中的亚硝酸根

研究了亚硝酸根与染料天青Ⅰ在盐酸介质中的显色反应。结果表明,反应产物的最大吸收波长为442nm,据此拟定了快速简便的分光光度测定冷却液中亚硝酸根的新方法。该方法亚硝酸根浓度在0～200ug/50mL范围内服从比尔定律,用于冷却液中亚硝酸根的测定,结果与KMnO_4容量分析法一致。     

微乳液增敏铬天青S分光光度法测定微量铁

研究十六烷基三甲基溴化铵 -正戊醇 -正庚烷 -水 (水包油型 )微乳液介质中Fe (Ⅲ )与铬天青S的显色反应。在pH =6 .0的乙酸 -乙酸铵缓冲溶液中 ,Fe(Ⅲ )与铬天青S形成配合物 ,配合物的最大吸收波长为 6 5 0nm ,表观摩尔吸光系数为 2 .5× 10 5L/ (mol·cm)。该体系较以水为介质的体系有显著的增敏作用。Fe (Ⅲ )含量在 0～ 0 .16 μg/mL范围内符合比耳定律。该法直接用于Fe2 O3 样品中铁的测定 ,测定结果的相对标准偏差为 1.8%～ 2 .2 % ,回收率为 99.2 %～ 10 2 .6 %。     

β修正光度法测定食品中的微量锰

在pH=8．6的硼砂-磷酸二氢钾缓冲溶液中，锰(Ⅱ)-铬黑T-吐温80显色体系的最大吸收波长为550nm，利用此显色体系，用卢修正光度法测定食品中的微量锰，Mn(Ⅱ)含量在0．45～35μg／(25mL)内服从比耳定律，线性方程为A。=0．0281X-0．0026，相关系数r=0．9997，用该方法测定调味料八角和奶粉中的微量锰。测定结果的相对标准偏差为0．52％～2．86％，回收率为96．21％～102．2％。     

铝与络黑T显色反应的研究和应用

研究了络黑T(EBT)与铝发生显色反应的适宜条件。在0.15mol·L-1NaOAc溶液中,发生配位反应生成紫色的二元配合物,配位比为Al∶EBT=1∶3。最大吸收波长为536nm,ε值为1.9×104L·mol-1·cm-1,线性范围为0～30μg/50ml。方法可用于一些胃药中铝的测定,测定结果与络天青S法十分吻合。     

分光光度法快速测定低合金钢中的铌

用盐酸—过氧化氢溶解试样，在2．9mol／L盐酸溶液介质中，铌与氯磺酚—S形成稳定的蓝紫色络合物，其最大吸收波长约为650nm，用分光光度法可以测定低合金钢中的铌。用该法对含铌标准样品进行测定，测得值与标准值基本一致，测定结果的相对标准偏差为2．1％～6．7％。     

TiO_2-SiO_2纳米粒子的合成及二级散射光谱法测定核糖核酸

采用溶胶-凝胶法制备了TiO2-SiO2纳米粒子.通过X-射线衍射谱、透射电子显微镜等对纳米粒子进行了表征.研究了TiO2-SiO2纳米粒子与核糖核酸(RNA)的相互作用,建立了基于纳米粒子与RNA吸附反应,二级散射光谱法测定痕量RNA的新方法.方法的线性范围是0.005～5 mg/L; 检出限为1.46 μg/L.方法用于RNA合成样品的测定,回收率为99.4%～103.9%;相对标准偏差为019%～0.24%.     

Specific Occurence of Selenium in Certain Enzymes and Amino Acid Transfer Ribonucleic Acids

Abstract

Some enzymes known to contain selenium are enumerated. In four of them which catalyze coupled oxidation-reduction reactions the selenium occurs exclusively in the form of selenocysteine residues. Their structure and function are described in detail. Two other bacterial enzymes which contain selenium in the form of a labile, readily dissociable component are also described.

Results on the isolation, identification and structure determination of selenium-containing amino acid transfer ribonucleic acids are presented. These seleno-t RNA's are shown to contain either 5-methyl-aminomethyl-2-selenouridine or other 2-selenouridine derivatives. The role of selenium in a glutamate iso-accepter species is discussed.     

液相色谱法检测水产品中亚甲蓝及其代谢物残留

建立了液相色谱同时检测水产品中亚甲基蓝及其代谢物天青A、天青B、天青C残留的方法并用于样品检测.试样中残留的药物以含离子对试剂对甲苯磺酸的乙酸铵缓冲液-乙腈提取,二氯甲烷反萃,MCAX固相萃取小柱净化,液相色谱测定.对提取、净化及仪器分离分析条件进行了探讨,对亚甲基蓝在分析过程的稳定性进行了研究.水产品中4种化合物在0.005～0.50 mg/kg范围内线性关系良好,相关系数为0 9946～0.9978.在0.005～0.5 mg/kg浓度范围内,平均加标回收率为75.9%～93.2%; 相对标准偏差为3 3%～11.9%; AZA、AZB、AZC、MB的检出限分别为1.27、0.98、1.48和1.33μg/kg.方法稳定可靠,可满足水产品中亚甲基蓝及其代谢物的检测需要.     

Simultaneous determination of malachite green, crystal violet, methylene blue and the metabolite residues in aquatic products by ultra-performance liquid chromatography with electrospray ionization tandem mass spectrometry

This work describes solid-phase extraction-ultra-performance liquid chromatography with electrospray ionization tandem spectrometry for determination of malachite green and metabolite leucomalachite green, crystal violet and metabolite leucocrystal violet, methylene blue and metabolites including azure A, azure B and azure C in aquatic products. Samples were extracted with acetonitrile and ammonium acetate buffer and purified by liquid extraction with dichloromethane, and then on MCAX solid-phase extraction cartridges. Then the extract was evaporated at 45°C by nitrogen blow. The residue was dissolved and separated by an Acquity BEH C18 column. The mobile phase was acetonitrile (A) and 5 mmol/L of ammonium acetate containing 0.1% formic acid (B). Analytes were confirmed and quantified using a tandem mass spectrometry system in multiple reaction mode with triple quadrupole analyzer using positive polarity mode. The limits of detection of malachite green, leucomalachite green, crystal violet and leucocrystal violet were 0.15 μg/kg, the limits of quantification were 0.50 μg/kg, and the average recoveries were more than 75% with spiked residues from 0.5 to 10 μg/kg. The relative standard deviations were less than 13%. The limits of detection of methylene blue, azure A, azure B and azure C were 0.3 μg/kg, the limits of quantification were 1.0 μg/kg, the average recoveries were more than 70% with spiked residues from 1.0 to 10 μg/kg and the relative standard deviations were less than 15%. The method has the merits of simplicity, sensitivity and rapidity, and can be used for simultaneous determination of the analytes in aquatic products.     

Use of polyelectrolytes for purification of crude ribonucleic acid

The effect of polymeric reagents on a ribonucleic acid suspension was studied. The polyelectrolytic complex based on polyacrylic acid proved to be an effective flocculant for colloidal impurities in crude ribonucleic acid. The efficiency of ribonucleic acid purification was evaluated.     

Determination of five nucleosides by LC‐MS/MS and the application of the method to quantify N6‐methyladenosine level in liver messenger ribonucleic acid of an acetaminophen‐induced hepatotoxicity mouse model

Ribonucleic acid N⁶‐methyladenosine methylation plays an important role in a variety of biological processes and diseases. Acetaminophen‐induced hepatotoxicity is one of the major challenges faced by clinicians. To date, the link between N⁶‐methyladenosine and acetaminophen‐induced hepatotoxicity has not been studied. In this study, a simple ultra high performance liquid chromatography with tandem mass spectrometry method was developed for the simultaneous determination of five nucleosides (adenosine, uridine, cytidine, guanosine, and N⁶‐methyladenosine) in messenger ribonucleic acid. After enzymatic digestion of messenger ribonucleic acid, the nucleosides sample was separated on an Acquity UPLC column with gradient elution using methanol and 0.02% formic acid water, and detected by a Qtrap 4500 mass spectrometer with an electrospray ionization mode. The method was validated over the concentration ranges of 4–800 ng/mL for adenosine, uridine, cytidine, and guanosine and 0.1–20 ng/mL for N⁶‐methyladenosine. It was successfully applied to the determination of N⁶‐methyladenosine levels in liver messenger ribonucleic acid in an acetaminophen‐induced hepatotoxicity mouse model and a control group. This study offers a method for the determination of nucleoside contents in epigenetic studies and constitutes the first step toward the investigation of ribonucleic acid methylation in acetaminophen‐induced hepatotoxicity, which will facilitate the elucidation of its mechanism.     

A thermodynamic investigation on the binding of phenothiazinium dyes azure A and azure B to double stranded RNA polynucleotides

The thermodynamics of the reactions of the two phenothiazinium dyes azure A and azure B with the three double stranded ribonucleic acids, poly(A).poly(U), poly(C).poly(G), poly(I).poly(C) were investigated using DSC and ITC. The bound dyes stabilized the RNAs against thermal strand separation. The binding of azure A to the RNAs was predominantly enthalpy dominated while the binding of azure B was favoured by both negative enthalpy and favourable entropy changes. Although electrostatic interaction had a significant role in the binding, non-polyelectrolytic forces dominated the binding process. The negative values of heat capacity changes for the binding suggested a substantial hydrophobic contribution to the binding process. The overall binding affinity of both the dyes to the RNAs varied in the order, poly(A).poly(U) > poly(C).poly(G) > poly(I).poly(C).     

中药对超氧阴离子自由基清除率的测定

邻苯三酚在碱性溶液中的自氧化反应产生超氧阴离子自由基(O2-.),可迅速氧化天青Ⅰ褪色。中药提取物可以抑制O2-.氧化天青Ⅰ褪色,根据该原理建立了一种测定中药对O2-.清除率的新方法。测定体系最佳实验条件为pH 9.0,天青Ⅰ用量为5.0 mL,邻苯三酚用量为0.8 mL,反应时间9 min。测得六种中药的提取液清除O2-.的效果,结果表明,清除能力顺序为:车前子>枸杞>川芎>当归>茯苓>射干。     

Single-step separation of major and rare ribonucleosides and deoxyribonucleosides by high-performance liquid cation-exchange chromatography for the determination of the purity of nucleic acid preparations

A method is described for the separation of 13 major and minor ribo- and deoxyribonucleosides in a single chromatographic run using high-performance liquid chromatography on strongly acidic cation-exchange columns. The method proved useful for the routine determination of small amounts of ribonucleic acid impurities in deoxyribonucleic acid preparations and vice versa. About 3% or even less of nucleic acid contamination in a given sample can be easily detected and quantitatively determined under the conditions used.