核黄素在不同极性溶剂中的光谱特性研究

实验测得核黄素在水、二甲基亚砜(DMSO)和三氯甲烷三种不同极性溶剂中的稳态吸收光谱、荧光光谱和时间分辨荧光光谱,研究了溶剂对核黄素光谱性质的影响。实验结果表明,在不同极性的溶剂中,核黄素的吸收峰位置几乎不变,而荧光光谱峰值随着溶剂极性的增大而出现红移。这是由于溶质分子的电子激发及溶剂化效应引起的电子重新分布导致它在极性溶剂中第一激发单重态能级的变化。在时间分辨荧光光谱实验中,核黄素在水溶液中荧光寿命也高于在其他两种溶剂中,荧光寿命的延长可归因于核黄素与氢键对体溶剂之间的分子间氢键相互作用。应用Gaussian09软件,采用密度泛函理论和含时密度泛函理论,结合基于密度的溶剂化模型,对不同极性溶剂中的核黄素分子进行基态和激发态优化和计算。通过前线分子轨道分析,核黄素的受激跃迁属于苯环和含氮杂环上的π电子向苯环及C N,C O共轭双键的反键轨道π*的跃迁。分子偶极矩的计算结果表明,核黄素分子的第一激发态偶极矩大于基态偶极矩,偶极矩的增大,导致溶质与溶剂分子之间的相互作用的增大。而溶剂分子与溶质分子基态和激发态的相互作用程度不同,使得吸收峰和荧光峰出现不同变化情况。极性越大的溶剂越有利于对激发态的稳定作用,使激发态能量降低,相应的发射波长发生红移。最后,通过分子表面静电势和弱相互作用分析,在水溶剂中考虑氢键作用对核黄素分子光谱的影响。多聚体结构的理论吸收和发射峰值更接近实验结果,说明多聚体结构合理。水分子二聚体与核黄素形成的环状结构,有利于提高核黄素分子的刚性,有利于荧光的发射,减少非辐射跃迁的几率,荧光寿命延长。     

荧光光谱法研究核黄素与鲱鱼精DNA的相互作用

在B-R缓冲溶液中, 以吖啶橙(AO)作荧光探针研究了核黄素(RF)与DNA的相互作用. 在pH值为7.25时, 荧光光谱研究表明, RF与DNA发生作用生成了复合物, 其结合比为n_RF∶n_DNA＝5∶1, 28 ℃时RF与DNA的结合常数K＝2.53×10⁶ L/mol; 并运用双倒数法求得RF与DNA相互作用的 为－6.06×10⁴ J/mol, 为－78.70 J/(mol•K), 28 ℃时的 为－3.69×10⁴ J/mol. 同时通过粘度法、盐效应和Scatchard法等研究了RF与DNA的作用方式, 确定了在该实验条件下RF与鲱鱼精DNA之间主要为沟面和静电作用方式.     

核黄素的燐光分析

核黄素无磷光,加碱在沸水中作用３０ｍｉｎ后产生强的磷光，λｅｘ３９０，λｅｍ５２ ０ｎｍ寿命０．７３ｓ。研究了１６个化合物对核黄素磷光的影响，结果表明，只有硫胺、叶酸、色氨酸、Ｂa＾２＋、Ｉ＾－在浓度比核黄素大１０～１００倍以上才有影响。建立了核黄素的磷光分析法，用于制剂分析及血浆中核黄素的测定，方法简单快速。     

维生素B2片中核黄素含量的快速测定

利用维生素B2光解分解为光黄素这一反应,以光黄素荧光法快速测定了维生素B2片中核黄素的含量。改进方法简便、快速、稳定性高,相对标准偏差为1.60%—2.30%,加标平均回收率为99.2%,准确度及精密度均能满足分析要求。     

葫芦脲[7]与核黄素包结物的可能结构和光谱性质

利用密度泛函理论对葫芦脲[7]和核黄素包结物的可能结构进行了理论计算.从5种可能结构分析主客体在包结物中的作用得出:核黄素作为客体分子与主体葫芦脲[7]分子通过不同官能团作用可形成稳定性不同的包结物,其中以核糖醇的羟基插入到葫芦脲[7]分子的腔体与其端羰基氧原子形成较多的氢键最稳定.在理论上证明了此包结反应为一放热反应的实验事实,并进一步说明了包结物分子的相对稳定性受主客体分子的几何畸变能和主客体之间作用能的影响,说明了成键能和稳定性之间的区别.在此基础上利用时间相关的密度泛函理论进行了不同构型包结物的激发态计算并对光谱性质进行了理论预测.比较了包结物中核黄素分子的跃迁轨道的组成变化,分析得出:葫芦脲[7]的包结作用改变了核黄素分子的电子跃迁性质,使其吸收光谱发生红移.并预测出其荧光猝灭的微观机理,此理论计算结果很好地解释了相应的实验现象.     

Determination of Riboflavin in Cereal Grains by Capillary Electrophoresis with Laser-Induced Fluorescence Detection with On-Line Concentration

A very simple, sensitive, and reliable method for the routine determination of riboflavin in cereal grains by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was established. Two on-line concentration techniques, namely, stacking in reverse migrating micelles (SRMM) and sweeping, were examined to enhance the detection signal. The optimum separation buffer consisted of 20 mM phosphoric acid and 140 mM sodium dodecyl sulfate (SDS), and a hydrodynamic injection of 30 s at 0.5 psi was confirmed for the introduction of a sample. In addition, it was found that salt in sample matrix influenced the performance of SRMM, showing the standard addition method was required for the quantitative analysis. The linearity of the method was good with a range of 0.5–500 ng/mL (r = 0.9990). The limit of detection (LOD) was 0.29 ng/mL at a signal-to-noise ratio of 3. This procedure presented was successfully applied to determine riboflavin in 18 samples of 9 types of cereal grains. The recovery rates varied from 94% to 98%, and the relative standard deviation (RSDs) was less than 4.1%.     

A Sensitive Photoinduced Chemiluminescence Method for the Determination of Riboflavin with Flow Injection Analysis

ABSTRACT

A sensitive chemiluminescence (CL) induced by light for the determination of riboflavin with flow injection analysis is developed. Riboflavin shows a strong enhancement effect on the CL reaction of luminol oxidized by periodate after the photochemical reaction of riboflavin in alkaline solution, and the enhancing effect of riboflavin disappears under dark conditions. Oxygen can further improve the enhancing effect of the riboflavin in this CL system, and the effects were also investigated. And based on this phenomenon, a very sensitive method of various surfactants, β-cyclodextrin and organic solvents on this CL system for the determination of riboflavin was described. The range of the linear response for riboflavin using this method is ca. 2.4×10^?8 ~ 1.0 × 10^?6 mol/L, and the detection limit (S/N = 3) is l.0×10^?8 mol/L. The applicability of the method was demonstrated by the determination of riboflavin in pharmaceutical preparation.     

胃癌患者胃液的荧光研究

利用外加荧光物质在肿瘤部位的特征吸附,或通过恶性肿瘤的自体荧光进行临床诊断及治疗均有报道,但对外加或自体荧光物质的认识大都集中于卟啉类分子。我们在利用胃液对胃癌特征荧光物质的研究中,首次提出了黄素蛋白类为胃癌胃液的特征荧光物质及存在形式,为胃癌的临床诊断和治疗及癌变机理的阐明提供了新的依据。     

The effect of riboflavin-photoinduced degradation of alginate matrices on the diffusion of poly(oxyethylene) probes in the polymer network

The effect of irradiation, in the wavelength range of 310-800 nm, on the tracer diffusion of poly(oxyethylene) (POE) of different molecular weights embedded in various alginate matrices with the photosensitizer riboflavin (RF) present, has been investigated with the aid of pulsed field gradient spin-echo NMR (PGSE-NMR). Both alginate solutions of different polymer concentration were studied as well as corresponding acid gels of alginate produced by introduction of different amount of glucono-δ-lactone (GDL). In 2 wt.% alginate solutions, the values of the tracer diffusion coefficient suggest a strong obstruction effect as the probe molecular weight increases. Faster probe diffusion was observed for the irradiated samples, which indicates a photochemical scission of the polymer chains and the formation of a fragmented polymer network that facilitates the migration of the tracer chains. A semidilute alginate/RF solution was transformed into a gel by adding sufficient amount of GDL. GDL lowers the pH of the solution under the pK_a of alginate, favouring intermolecular associations and the evolution of a less homogeneous network with more open structure. Therefore, the POE chains were shown to diffuse faster in the acid gel matrix than in the corresponding more homogeneous alginate solutions. The photochemically induced cleavage again promoted faster migration of the probe chains in the irradiated samples. The probe diffusion of the eight-arm star-shaped POE sample revealed an augmented obstruction effect with increasing alginate concentration and higher values of the diffusion coefficient were found in gels. The evolution of a tighter network inhibits the diffusion of the probe molecules. At lower alginate concentrations the values of the tracer diffusion coefficients are higher for the irradiated samples than for the non-exposed systems.