Rapid Quantification of Bioactive Lentinan with an Aniline Blue Fluorescent Method

Lentinan is a clinically approved immune modulator and its anticancer and immunomodulatory bioactivity is found to be dependent on its triple helical conformation. Therefore, the development of rapid and convenient method for determination of bioactive lentinan with triple helical conformation holds great promise for the quality control of lentinan healthy products. In this work, an aniline blue fluorescent method was optimized and established to accurately and rapidly detect bioactive lentinan. In the presence of lentinan, the fluorescence intensity of aniline blue with 404 nm excitation and 492 nm emission dramatically enhanced within 15 min in pH 10 glycine-NaOH buffer solution, which allowed the analysis of lentinan in a very simple and fast manner. The method allowed for the sensitive determination of lentinan in the range of 1 to 60 μg/mL with a detection limit of 0.25 μg/mL. Notably, the protocol exhibited excellent selectivity for the determination of triple helical lentinan over other saccharides. The method was successfully applied to the detection of bioactive lentinan in health tonic solution, which demonstrated the method had great potential for quality control of lentinan contained products.

Equilibrium, Kinetic and Thermodynamics Studies of Adsorption of Aniline Blue from Aqueous Media Using Steam-Activated Carbon Prepared from <i>Delonix regia</i>Pod

The adsorption capacity of steam activated carbon prepared from Delonix regia pods SADRC for adsorption of Aniline Blue (AB) from aqueous solution was investigated under various experimental conditions. Batch study was conducted to assess the potential of the activated carbon for the removal of Aniline Blue from aqueous solution. Activated carbon prepared from Delonix regia pods was characterized using Scanning Electron Microscope (SEM) and Fourier Transform Infrared (FTIR) Spectrophotometry before and after adsorption. The FTIR, spectra of SADRC pod before and after Aniline Blue adsorption were compared to study the impact of the Aniline Blue on the activated carbon developed from the Delonix regia pod. The stretching vibration band at 2169.54 cm-1 may be due to strong CN, while the stretching vibration band at around 1580 - 1650 cm-1 may be due to C=C stretching vibration. The bands around 1350 and 426.49 cm-1 are due to C-N and –SO3H group, respectively;this further suggests that some functional groups may be present on the surface of the carbon due to the low temperature of carbonization (300°C) of the adsorbent. Equilibrium isotherm studies were carried out by varying the following four parameters: initial concentration of Aniline Blue dye solution, solution pH and adsorbent dose. The equilibrium data obtained were more fitted to Langmuir than Freundlich isotherm models. The correlation coefficient value (R2) of the pseudo first order kinetics ranged from 0.08 to 0.85 while the R2 of the pseudo second order kinetics ranged from 0.963 to 0.997 at all the temperatures and initial concentrations considered. This suggests that the adsorption kinetics of Aniline Blue onto SADRC can be represented with pseudo second order kinetic model. This study showed that Delonix regia pod could be effectively used as an adsorbent for the removal of Aniline Blue from aqueous solutions over a wide range of concentration and temperature.

Isolation of a Microorganic Strain for the High Volume Degradation of Aniline Blue and Its Application in Natural Sewage Treatment

Aniline blue, one of the triphenylmethane dyes, is the most commonly produced and used of these dyes yet it is also the most dangerous and the most serious cause of pollution amongst them. An exploration of aniline blue degradation is likely to facilitate an understanding of the degradation mechanism for a range of related dyes. In this study, we managed to isolate a particular strain of microorganism, identified to be Lysinibacillus fusiformis N019a, which showed a significant capacity for aniline blue degradation in both laboratory tests and natural sewage treatment. In analysis aided by a UV-Visible spectrophotometer, we found that 96.7% of aniline blue had degraded within 24 hours under laboratory conditions. When treating natural sewage, 80.1% of the aniline blue was removed after just 16 hours. Further analysis has shown that Lysinibacillus fusiformis N019a has a strong resistance to Cu2+, Mn2+, Zn2+, and Pb2+. We also found that the degradation product of aniline blue by Lysinibacillus fusiformis N019a was of reduced toxicity to plants and microbes.

Can the Prediction of Intrauterine Insemination Results by Used Aniline Blue Stain (ABS) and Sperm Chromatin Dispersion (SCD) Levels?

Introduction: This study aimed to perform routine seminal fluid analysis, sperm DNA fragmentation, and sperm function tests at the chromatin maturation level and evaluate pregnancy in the patients passing intrauterine insemination before starting Intrauterine Insemination (IUI) method. Materials and Methods: In this prospective study, 111 couples who underwent Intrauterine Insemination (IUI) in unexplained infertility patients were admitted to Al-Farah IVF and assisted reproductive center in Baghdad, Iraq between November 2020 and February 2021 were evaluated. Semen fluid analysis was performed based on (WHO 4th) guiding rules. In addition, Sperm Chromatin Dispersion (halo test) and sperm maturation were performed with Aniline Blue Stain (ABS). Results: Sperm Chromatin Dispersion (SCD) groups were compared in terms of pregnancy outcome;the positive pregnancy rate was found to be above in the normal SCD groups (p = 0.0005). In addition, Aniline Blue Stain (ABS) groups were compared in the terms of pregnancy outcome;the positive pregnancy rate was found to be higher in the normal ABS group (p = 0.017). Conclusion: Our study showed that the use of DNA fragmentation (SCD) and sperm maturation tests (ABS) together with routine semen analysis in intrauterine insemination cases will make a significant contribution to the prediction of Intrauterine Insemination (IUI) increased results. So, these results indicate a defect in the effect of DNA fragmentation on the outcome of intrauterine insemination.

Removal of Aniline Blue from Aqueous Solutions Using Ce_1-xBi_xCrO₃(x = 0, 0.5, 1)

The removal of aniline blue dye from aqueous solutions using the A-site doped perovskite Ce1-xBixCrO3 (x = 0, 0.5, 1) was investigated. The perovskite oxides were synthesised using Sol-Gel method and characterised by conventional powder X-ray diffraction technique. The X-ray diffraction measurements suggested that doping with Bismuth Ion influences both the crystal structure and the particle size of the oxides, and consequently affects the adsorption properties. It was found that both CeCrO3 and Ce0.5Bi0.5CrO3 compounds are orthorhombic and have approximate particle size of 87 and 36 nm respectively, whereas BiCrO3 oxide has rhombohedral space group symmetry and the particle sizes are less than 49 nm. The batch mode study demonstrated that the removal capacities of Aniline Blue at 150 min and pH = 4.3 for Ce0.5Bi0.5CrO3, CeCrO3 and BiCrO3 are 779.67, 705.45 and 440.18 mg/g respectively. The results reflect the influence of the A site doping on the adsorption properties of the oxides. The removal of Aniline Blue was found to be negatively correlated with temperature and pH.

苯胺蓝褪色分光光度法测定卡那霉素

在pH为2 0～3 5的条件下,苯胺蓝(AB)与硫酸卡那霉素(KANA)反应生成复合物,使水溶苯胺蓝溶液褪色,最大褪色波长位于606nm,表观摩尔吸光系数(ε)为2 72×104L·mol-1·cm-1,卡那霉素的浓度在0～1 5×10-5mol/L范围内遵从比耳定律.该法用于实际样品的测定,结果满意.

苯胺蓝分光光度法测定痕量铬(Ⅵ)

在HCl介质中,铬(Ⅵ)与过量I-反应生成黄色I3-,继而与水溶性苯胺蓝阳离子形成离子缔合物,使得苯胺蓝溶液褪色,其在605nm处产生一个强吸收峰。在最佳实验条件下,Cr(Ⅵ)的浓度在4.0×10-7—1.2×10-5mol/L范围内符合比耳定律,表观摩尔吸光系数ε为1.10×106L.mol-1.cm-1。该法用于分析测定沱江水样中Cr(Ⅵ)的含量,结果令人满意。

苯胺蓝增色光度法测定食盐中的碘

在H2SO4介质中，KIO3与苯胺蓝反应增色程度与IO3^-量在一定范围内呈线性关系，从而建立了增色光度法测定碘的新方法。实验结果表明，该体系最大吸收波长为603nm，IO3^-在0．91～4．55μg/L范围内符合比耳定律，表观摩尔吸光系数为5．00×10^5L·mol^-1·cm^-1。本法可直接测定食盐中的碘。

水溶苯胺蓝与蛋白质结合反应的研究及应用

在pH 2.36的Britton-Robinson(B-R)缓冲介质中,水溶苯胺蓝与蛋白质形成显色化合物,其最大吸收波长为688nm,比试剂本身红移88nm,表观摩尔吸光系数达4.25×105L·mol-1·cm-1(牛血清白蛋白)。制定了四种蛋白质的标准工作曲线,用于人血清样品中总蛋白的测定,结果与溴甲酚绿法一致。

海带中昆布素的荧光定量检测及提取工艺优化

针对昆布素检测技术的难点，利用昆布素与苯胺蓝能特异性结合的特点，采用荧光分光光度计建立了昆布素含量的荧光快速检测方法。实验结果表明：NaOH浓度对昆布素立体结构影响较大，在昆布素解旋过程中加入70μL 3mol／L的NaOH能使昆布素-苯胺蓝荧光复合物具有最大的荧光信号；通过三维荧光扫描，确定昆布素．苯胺蓝荧光复合物的最佳激发和发射波长分别为398nm和502nm，在此条件下昆布素-苯胺蓝荧光复合物浓度与其荧光值正相关（R^2=0．9994）；方法学分析表明：该测定方法具有较高的精密度（RSD为2．52％）和稳定性（RSD为2．09％），检测范围为20～80mg／L。基于所建立的昆布素含量快速检测方法，本文设计了有关提取温度、提取时间、pH以及乙醇浓度四个因素三个水平的正交试验对海带中昆布素的提取条件作了初步优化，结果表明：海带中昆布素的最佳提取条件是：置于稀盐酸（pH为1）溶液中于25℃下提取4h，90％乙醇沉淀。

白腐真菌XX-3对苯胺蓝染料的脱色条件优化

采用静置开放式培养法研究碳源、氮源、盐度、金属离子对白腐真菌降解苯胺蓝的影响。结果表明,菌株脱色最适降解条件为丙二酸2.0 g/L,氯化铵4.0 g/L,Cd2+0.10 mmol/L,盐浓度≤4.0 g/L,在上述培养条件下,对浓度为100 mg/L不灭菌的苯胺蓝溶液静止培养5 d,降解率达80%以上。因此,该菌对处理以苯胺蓝为主要成分的印染废水具有较好的应用潜力。

一株苯胺蓝降解菌的分离鉴定及其降解特性

从河水中驯化、筛选、分离得到一株对染料苯胺蓝有降解能力的菌株WZR-B,该菌能以苯胺蓝为唯一碳源、能源生长.通过对菌株WZR-B的形态特征、生理生化,以及16Sr DNA序列分析,初步鉴定为铜绿假单孢菌(Pseudomonas aeruginosa).研究苯胺蓝的质量浓度、pH值和温度等因素对该菌株生长,以及对降解苯胺蓝的影响.结果表明,菌株生长和脱色的最适pH值为6.5,最适合温度为30℃,苯胺蓝的最适脱色质量浓度为200 mg.L-1.此外,总有机碳(TOC)残留量测定结果表明,该菌株对苯胺蓝的脱色率可达83%,TOC去除率为80%以上,不仅能对苯胺蓝脱色,而且还能进行矿化降解.

贮前苯胺蓝染色预测苹果冷藏末期表皮病害

为了实现对苹果贮藏末期表皮病害的预测,以红富士苹果(Malus domestica Borkh cv.Fuji)为试材,通过苯胺兰染色的方法,研究了苹果采后苯胺兰染色与果实贮藏末期几种表皮病害的关系。结果表明:富士苹果采后苯胺兰染色指数与贮藏末期的好果率呈明显负相关,相关系数为-0.947;果实苯胺兰染色的级别与二氧化碳伤害指数和腐烂指数密切相关,其决定系数分别为0.9779和0.8394,但苯胺兰染色的级别与贮藏末期果实的虎皮病和皮孔斑点病之间无明显的相关性。因此,苹果采后苯胺兰染色可用于预测果实贮藏末期的好果率。

水溶苯胺蓝吸光光度法测定人血清蛋白

用吸光光度法研究了水溶苯胺蓝(AnB)与牛血清白蛋白(BSA)在Britton-Robinson广泛pH缓冲介质中的结合反应,在最佳试验条件下,以试剂空白为参比,AnB-BSA复合物在600nm波长处的吸光度,与BSA的浓度呈良好的线性关系,BSA的线性范围为1.6×10-8～2.4×107mol.L-1,摩尔吸光系数ε=1.34×106L·mol-1·cm-1,Sandell灵敏度为0.051μg·cm-2,BSA的检出限为1.0×10-8mol·L-1,方法具有灵敏度较高、选择性好等特点,用于人血清样品中蛋白质的测定,与溴甲酚绿方法结果一致。

利用苯胺蓝染色检测苹果冷藏表皮病害的研究

为阐明苯胺蓝染色检测苹果冷藏表皮病害的机理,以富士苹果(Malus domestica Borkh cv.Fuji)为试材,通过苯胺蓝染色和扫描电镜的方法,研究了苹果采收后苯胺蓝染色和0℃冷藏210 d后几种表皮病害的表皮结构。结果表明,(1)苹果表皮苯胺蓝染色部位存在皮孔发育不完善、表皮裂缝愈合不完善等质量缺陷,且该缺陷可以通过苯胺蓝染色的方法显现出来;(2)苹果贮藏210 d后表皮病害部位表皮及其蜡质层存在皮孔和表皮裂缝愈合不完善的质量缺陷;(3)苹果苯胺蓝处理后,发生染色部位往往与冷藏210 d后表皮病害发生部位相吻合。因此,苯胺蓝染色可用于检测果实贮藏病害的结构机理是:染色部位与病害部位存在共同结构基础,即发育不完善的表皮质量;该质量缺陷可用苯胺蓝染色的方法在果实入贮前使其显现出来。

植物小孢子母细胞减数分裂过程中胼胝质染色的新方法

利用改良苯酚品红-苯胺蓝压片法,观察小孢子母细胞减数分裂过程中胼胝质的动态变化。使用该方法简便、快速且省时,获得的照片颜色鲜艳,细胞质呈红色,染色体为深红色,胼胝质呈黄绿色荧光,对比明显,有三维效果。单用改良苯酚品红染液对新鲜材料进行压片,在蓝光激发下,细胞质与染色体呈红色荧光,染色体清晰。实验结果表明,改良苯酚品红染液可作为荧光染料代替DAPI及H33258等昂贵的核染料,从而降低实验成本。

分光光度法测定血清中的总蛋白含量

在 p H=2 .36的 Britton- Robinson缓冲介质中 ,紫色的水溶苯胺蓝与蛋白质形成蓝紫色的结合物 ,其最大吸收波长 λmax为 688nm,比水溶苯胺蓝试剂本身红移 88nm,表观摩尔吸光系数达4.2 5× 1 0 5L/( mol· cm) (与牛血清白蛋白结合 )。制定了 4种蛋白质的标准曲线 ,用于人血清样品中总蛋白的测定 。

荧光法研究苯胺蓝与人血清白蛋白的相互作用

用荧光光谱、同步荧光光谱研究了苯胺蓝和人血清白蛋白的相互作用。研究表明,苯胺蓝对人血清白蛋白的荧光发射有明显的猝灭作用,根据不同温度下的猝灭数据,由Stern-Volmer方程推断苯胺蓝对人血清白蛋白的猝灭属于静态猝灭。计算得到了结合常数KA、结合位点数n,同时计算得到的热力学常数表明苯胺蓝和人血清白蛋白作用力类型为静电作用和疏水作用结合。同时用同步荧光光谱探讨了苯胺蓝对人血清白蛋白构象的影响。

中心组合设计优化S-Cr-TiO_2催化剂光降解工艺条件

在确定的温度和光照条件下,采用中心组合设计法研究了固相反应法合成的硫与铬共掺杂二氧化钛(S-Cr-TiO)2光催化剂与水溶苯胺蓝(ANB)可见光降解过程中的三个作用因子:催化剂的浓度、水溶苯胺蓝水溶液的pH值及其初始浓度之间的相互作用关系,得到了影响因子与降解速率之间的回归方程及因素之间相互作用的响应面图。结果表明,S-Cr-TiO2光催化剂对水溶苯胺蓝的可见光降解过程遵循一级反应动力学规律。当催化剂浓度为2.05g/L,水溶苯胺蓝的浓度为42.57mg/L、pH=6.63时,光降解速率常数为0.01432 min-1,比未优化工艺条件时所得光降解反应速率常数0.01348min-1提高了6.2%。中心组合设计对筛选光降解工艺条件,提高光降解效率是十分有效的。

窄带成像技术联合苯胺蓝染色在鼻咽癌内镜诊断中的价值

目的探讨窄带成像技术(NBI)联合苯胺蓝染色在鼻咽癌内镜筛查及诊断中的价值。方法选择2017年10月至2021年10月在海军军医大学(第二军医大学)附属公利医院和银川市第一人民医院就诊且怀疑有鼻咽部占位性病变的68例患者,使用具有NBI模式的电子鼻咽镜进行鼻咽部检查,取疑似病变表面的渗液进行苯胺蓝染色,并对疑似病变进行病理活检。以病理检查结果作为诊断金标准,分别计算白光模式、NBI模式、苯胺蓝染色及NBI联合苯胺蓝染色4种检查手段的灵敏度、特异度、阳性预测值、阴性预测值和准确度及Kappa系数,并使用χ^(2)检验比较各检查方法的诊断阳性率。结果NBI与苯胺蓝染色联合诊断鼻咽癌的阳性率高于白光内镜、NBI、苯胺蓝染色单独诊断[97.83%(45/46)vs 56.52%(26/46)、84.78%(39/46)、80.43%(37/46)],差异均有统计学意义(P均<0.001)。NBI联合苯胺蓝染色诊断鼻咽癌的灵敏度、特异度、阳性预测值、阴性预测值和准确度分别为97.83%、86.36%、93.75%、95.00%和94.12%。NBI联合苯胺蓝染色对鼻咽癌的诊断结果与病理检查结果有较好的一致性(Kappa系数为0.862)。结论相较于白光模式及单一NBI模式和苯胺蓝染色,NBI联合苯胺蓝染色对鼻咽癌有较高的灵敏度和特异度。