黄嘌呤甲基衍生物的发光特性研究

本文对3种黄嘌呤类化合物(咖啡因，茶碱，可可碱)的液氮低温荧光(LTF)、低温磷光(LTP)、室温流体荧光(RTF)及滤纸表面室温磷光(PS-RTP)发光光谱特性进行了对比研究。研究表明，这3种物质的LTF、LTP、PS—RTP及RTF的最大激发波长λex在270—295nm范围内，最大发射波长λem在385—445nm范围内，且它们的λex和λem不论在什么状态下都非常相近。本文也对这3种物质的荧光量子产率、滤纸基质室温磷光(PS—RTP)寿命、偏振等性质进行了比较研究。实验表明：咖啡因、茶碱和可可碱的PS-RTP的寿命均在0．1s数量级，属于长寿命磷光，且PS--RTP为非完全偏振光。     

黄嘌呤甲基衍生物滤纸基质室温磷光光谱研究

以滤纸为基质,详细讨论了3种黄嘌呤甲基衍生物可可碱、咖啡因、茶碱的固体基质温磷光光谱特性,研究表明：它们的滤纸基质室温磷光（PS－RTP）光谱基本相近,其最大激发波长λem分别为435、432、427nm。40余种无机盐类重原子微扰剂对这3种化合物诱发室温磷光的研究结果为：KI、NaI,Zn(NO3)2,SrCl2对可可碱、咖啡因、茶碱发射PS－RTP都有一定的重原子效应,其中I^-对它们诱发PS－RTP重原子效应最大,酸度实验表明,酸度对可可碱的影响在于咖啡因和茶碱,3种化合物在pH-2-8范围内都有PS－RTP,而且在强酸和强碱笥条件下它们的PS－RTP均发生猝灭,此外,本文也对影响PS－RTP的温度、干燥时间等条件进行了考察。     

光纤与生物分子界面上固定FITC标记抗体的荧光光谱

采用化学方法把FITC标记羊抗人抗体IgG共价固定到三氨基三乙氧基硅烷和戊二醛(APTES-Glu)修饰的石英光纤纤芯表面。通过研究未经过任何修饰的纤芯表面的吸附以及共价固定的FITC标记羊抗人抗体的荧光光谱性质发现:共价结合到纤芯表面的FITC荧光光谱相对于溶液中FITC的荧光峰位红移约9nm,而物理吸附的FITC的荧光峰位移动约4nm,而且两者相对荧光强度相差6倍。而在固定的人血清蛋白进行免疫反应后,FITC的荧光峰位只移动3～4nm,除此之外,研究了抗体共价键固定的稳定性问题。结果表明:共价键结合抗体的数量要大于表面吸附,共价键固定的FITC标记羊抗人抗体与光纤表面间的相互作用较吸附于光纤表面的FITC标记羊抗人抗体间的相互作用更强。这对于建立一种通过荧光光谱识别固体表面与生物分子的吸附还是共价结合的判据提供了物理基础。     

超分子组合流体室温磷光现象(Ⅱ)——醇对环糊精/碘代乙基联苯/溴代环己烷体系室温磷光的影响

在外部重原子微拢剂溴代环己烷存在下，β－ＣＤ／碘代乙基联苯体系可发射一定强度的室温磷光信号，若丁醇作为第四组分存在，则能显著提高体系的室温磷光发射强度。研究了该体系室温磷光发射的适宜条件和多种醇、不同环糊精及其衍生物的影响。认为β－ＣＤ／碘代乙基联苯／溴代环己烷／丁醇体系属于一种超分子组合的发光体系，醇的作用属典型的分子调控作用。β－ＣＤ／碘代乙基联苯／溴代环己烷形成主、客三元包结物，而醇则利用其羟基与ＣＤ端口羟基形成氢键，其烷基键借助于疏水相互作用力，覆盖在ＣＤ上、下端口，对外部氧向ＣＤ腔内的扩散起着隔离作用，减少了三线态氧对发光体激发三线态的猝灭，从而显著提高了体系室温磷光发射强度     

超分子组合流体室温磷光现象（Ⅱ）—醇对环糊精／碘代乙基联 …

在外部重原子微拢剂溴代环己烷存在下，β－ＣＤ／碘代乙基联苯体系可发射一定强度的室温磷光信号，若丁醇作为第四组分存在，则能显著提高体系的室温磷光发射强度。研究了该体系室温磷光发射的适宜条件和多种醇、不同环糊精及其衍生物的影响。认为β－ＣＤ／碘代乙基联苯／溴代环乙烷／丁醇体系属于一种超分子组合的发光体系，醇的作用属典型的分子调控作用。β－ＣＤ／碘代乙基联苯／溴代环乙烷形成主、客三元包结物，可醇则利用其     

6-巯基嘌呤滤纸基质室温磷光光谱研究

本文建立了 6 -巯基嘌呤 (6 - MP)的滤纸基质室温磷光 (PS- RTP)检测法 ,实验条件已经最佳化。研究表明 ,6 - MP的 PS- RTP光谱的最大激发波长 λex与最大发射波长 λem分别为 312 nm和 4 5 5 nm。30多种无机盐类重原子微扰剂对 6 - MP的 RTP影响的研究结果表明 ,Cd盐可增强 RTP,其中 Cd(OAc) 2 重原子效应最为显著。酸度条件对 6 - MP的 PS- RTP有较大影响 ,在 p H为 7— 8时 RTP发射较强。此外 ,本文也对影响 6 - MP的 PS- RTP的固体基质类型、重原子浓度、干燥条件等进行了详细的研究。实验表明 ,方法的线性范围为 4 .2 6— 10 6 6 ng/spot,检出限为 3.31ng/spot,相关系数为 0 .997,相对标准偏差为 4 .0 7% ,回收率为 99.6 %— 10 0 .5 %。该方法简便、快速、灵敏 ,应用于商业药片的分析 ,结果令人满意。     

基于Mn掺杂的ZnS量子点/CTAB纳米复合材料米托蒽醌的检测EI北大核心CSCD

以3-巯基丙酸为稳定剂,采用水相合成法合成Mn掺杂的ZnS量子点,该量子点在室温条件下能够发射较强的磷光信号。十六烷基三甲基溴化铵（CTAB）作为一种阳离子表面活性剂能够与该量子点发生静电作用,最终与量子点聚合形成Mn掺杂的ZnS量子点/CTAB纳米复合材料,使量子点的室温磷光（RTP）强度明显增强。加入米托蒽醌（MXT）后,MXT能够与CTAB通过疏水作用和结构作用结合成为更加稳定的混合物,最终导致CTAB从量子点的表面脱离,进而使该量子点的室温磷光强度降低。结果表明该纳米复合材料能够大大提高量子点对MXT的检测性能,可由此建立高效、灵敏的检测MXT的室温磷光传感器。在最优条件下,该传感器对MXT的检出限为0.23nmol/L,线性范围为0~200nmol/L,相关系数R为0.99,且尿液和血清实际样品的检测回收率为98.6%~102.5%。该量子点磷光分析方法简便快速、灵敏度高、选择性好,能够用于体液中MXT含量的分析与检测。     

蛋白质的室温磷光法研究

提出了牛血清蛋白(BSA)和人血清蛋白(HSA)的室温磷光法,蛋白质的磷光主要来自于包埋于其中的色氨酸残基,并对影响其室温磷光强度的的各种因素:如重原子浓度,酸度,除氧剂浓度等进行了详细研究,建立了测定痕量HSA和BSA的室温磷光法.该法是以KI为重原子微扰剂,在水溶液中用Na2SO3化学除氧后,以287nm为激发波长,BSA和HSA在443nm左右的磷光具有较好的重复性和稳定性.在此条件下它们的线性范围分别为:1×10-6-8.6×10-5mol/L和3×10-6-13×10-5mol/L,检出限分别为:2.20×10-7mol/L和4.10×10-7mol/L.同时,还对它们的光谱性质如荧光、磷光寿命,偏振等进行了测量.结果显示BSA中色氨酸残基有两种荧光衰减形式,HSA则只有一种衰减形式,它们的磷光属于长寿命磷光,都能引起光的偏振.     

荧光光谱成像在生物芯片蛋白量化分析中的应用研究

采用荧光光谱成像并结合椭圆偏振技术研究了３-氨基３-乙氧基硅烷(APTES)修饰及其与戊二醛(APTES-Glu)共同修饰的两种不同表面上固定的羊抗人抗体活性和数量及其荧光免疫结合。研究结果表明：应用荧光光谱成像在APTES-Glu表面上检测到的FITC标记人血清蛋白分子的数量为APTES表面结合的2.8倍,而应用椭圆偏振技术在前者表面上检测到的FITC标记人血清蛋白分子的数量为后者表面上的2.2倍。这个结果说明：在荧光免疫检测中,荧光光谱成像完全可用于分析不同表面固定蛋白的免疫活性和半定量的检测。     

2种卤代荧光素滤纸基质室温磷光特性及其与DNA作用的研究

对四溴荧光素(TBF)、四氯四溴荧光素(TTF)两种卤代荧光素滤纸基质室温磷光光谱(PSRTP)特性及其与DNA的作用进行了研究.结果表明:TBF和TTF的PS-RTP的最大λex/λem为526/652nm和557/699nm;酸度实验表明:两种物质在碱性范围内有强发射.小牛胸腺(DNA)的存在会使TBF和TTF的PS-RTP强度发生变化;偏振实验表明:TBF、TTF与DNA的作用方式有嵌插作用;TBF和TTF的磷光寿命分别为136.4ms和131.0ms属长寿命磷光.     

A Promising CdSe@CdS-Quantum Dots-Cysteine for the Determination of Trace IgE by Solid Substrate Room Temperature Phosphorescence Immunoassay

The labelling reagent CdSe@CdS-QDs-Cys (QDs-Cys) with the grain diameter of 4.5 nm was synthesized by modifying CdSe@CdS quantum dots (QDs) with cysteine (Cys). At the same time, QDs-Cys-Ab_IgE, a phosphorescent quantum dot probe, was developed based on the labelling reaction between -COOH of QDs-Cys and -NH₂ of goat anti human IgE antibody (Ab_IgE). This probe with excellent biocompatibility and high specificity could not only emit strong and stable room temperature phosphorescence (RTP), but also could carry out specific immunoassay (IA) with immunoglobulin E (IgE), causing the RTP of the system to sharply enhance. Thus, a new solid substrate room temperature phosphorescence immunoassay (SSRTPIA) for the determination of IgE was established. The limit of quantification (LOQ) of the method was 0.12 fg spot⁻¹, corresponding concentration was 3.0 × 10⁻¹³ g mL⁻¹ and sampling quantity was 0.40 μL spot⁻¹. This highly selective, sensitive and accurate SSRTPIA has been applied to determine IgE in biological samples and diagnose diseases, and the results agreed well with those obtained by enzyme-link immunoassay (ELISA). Meanwhile, the mechanisms of QDs-Cys labelling Ab_IgE and the determination of IgE by SSRTPIA were also discussed.     

菲的无保护性介质流体室温磷光性质研究

在无任何保护性介质存在下,以Na2SO3作化学除氧剂,KI为重原子微扰剂,菲即能产生强而稳定的流体室温磷光发射.激光和发射波长λex/λem为283/482,504nm,不同有机溶剂存在对其磷光发射的性质有不同影响.1%乙腈存在时,菲浓度在8.0×10-7～6.0×10-6mol*L-1和6.0×10-6～4.0×10-5mol*L-1范围内分别与磷光发射强度呈良好的线性关系,检出限为2.6×10-8mol*L-1.     

苯并咪唑的三维荧光光谱与三维室温磷光光谱

测量了浓度为1×10^-4mol/L苯并咪唑水溶液的三维荧光光谱,三维室温磷光光谱和紫外/可见吸收光谱,还测量了苯并咪唑固体紫外/可见吸收光谱对化合物的荧光和室温磷光进行了分析、比较,发现苯并咪唑在290nm、580nm和870nm区域均有强而丰富的荧光谱线,而室温磷光谱线(RTP)单一地出现在290nm区域,且强度很小;同时还讨论了苯并咪唑的升频转换荧光现象.     

磷光寿命法研究无保护流体室温磷光的重原子效应和发光动力学参数

以两种卤代萘为模型化合物，基于磷光寿命的定义τ＝τ^-，τ0＝1／kp和其与各速率常数的关系，导出了一种类似于Stern-Volmer方程的线性方程：τ0／τ＝（kp_kic)/kp=1 kic/kp=1 Ksv.c。通过测定不同重原子微扰剂浓度时的磷光寿命，探讨了从两种途径计算流体室温磷光发射相关动力学参数的可行性和方法，通过这些参数对比讨论了KI和TINO3两种重原子微扰剂对这两种卤代萘无保护流体室温磷光发射的重原子效应的差异。     

Determination of Bioactive Matter by Affinity Adsorption Solid Substrate–Room Temperature Phosphorimetry Based on Lectin Labeled with Self-ordered Ring of Eosin Y

A new phosphorescent labeling reagent named self-ordered ring (ESOR) of eosin Y (E) was developed. And the application of the determination of bioactive matter by affinity adsorption solid substrate–room temperature phosphorimetry (AA-SS-RTP) based on ESOR labeling lectin was studied. Results showed that pink and homogeneous ESOR could be formed by E on polyamide membrane (PAM) in the presence of cetyltrimethylammonium bromide (CTAB) and ammonia water. ESOR could emit strong and stable room temperature phosphorescence (RTP) signal of E in the presence of heavy atom perturber. Specific affinity adsorption (AA) reactions could be carried out between the products of concanavalin agglutinin (Con A), triticum vulgaris lectin (WGA) labeled with ESOR and alpha-fetoprotein variant (AFP-V), alkaline phosphatase (ALP), glucose (G), respectively. Not only did the products of the affinity adsorption reactions preserve good RTP characteristic of E, but also the ΔI _p (ΔI _p = I _p2 − I _p1, I _p1 is the RTP intensity of blank reagent, I _p2 is the RTP intensity of sample) of these products was proportional to the content of AFP-V, ALP and G, respectively. According to the facts above, a new method of AA-SS-RTP for the determination of AFP-V, ALP and G was established, based on ESOR labeling lectin. Detection limits (LD) of this method were 0.040 fg spot⁻¹ for AFP-V, 0.045 fg spot⁻¹ for ALP and 0.090 fg spot⁻¹ for G. And it has been successfully applied to the determination of AFP-V in human serum as well as the survey and forecast of human diseases. This method had high sensitivity, good repeatability, long RTP lifetime and little background interference with at the long wavelength area. Meanwhile, the mechanism for the determination of trace AFP-V by AA-SS-RTP based on Con A labeled with ESOR was also discussed.     

吲哚-3-丁酸无保护流体室温燐光和胶束增稳室温燐光性质的比较

仔细研究了吲哚-3-丁酸(IBA)的无保护流体室温燐光(NP—RTP)及以高分子分散剂聚乙二醇-200,聚乙二醇-400和非离子表面活性剂Tween-20,Tween-40,Tween-80,Tween-85,Brij35和乳化剂OP为介质的流体室温燐光性质。表面活性剂和高分子分散剂能抑制IBA燐光猝灭,使其具有更低的检出限,同时也使燐光强度-激发光照射时间曲线发生改变,但不影响IBA燐光光谱特性。无论是否存在表面活性剂或高分子分散剂,TINO3都不能诱导IBA产生燐光,KI却能诱导其产生强烈燐光。用于强化水样和土壤样品中IBA的测定,回收率95．2％～104％,相对标准偏差2．4％～4．0％。     

Room‐Temperature Phosphorescence in Metal‐Free Organic Materials

Purely organic materials with room‐temperature phosphorescence (RTP) have attracted a growing interest for their potential applications in biological imaging, digital encryption, optoelectronic devices, and so on. To date, many strategies have succeeded in designing efficient organic RTP materials by overcoming the spin‐forbidden transition between singlet and triplet states. However, the underlying mechanisms of RTP still remain ambiguous. Such spin prohibition in phosphorescence are clarified, herein, from the perspective of perturbation theory, helping to understand the intrinsic relationship among various phosphorescence parameters, like phosphorescence efficiency, lifetime, intersystem crossing rate, as well as radiative and nonradiative rates. Taking into consideration the recent progress in organic RTP materials, these factors are further illustrated by a selection of the most relevant molecules. In addition, some novel RTP phenomena are also reviewed, thus providing an excellent guideline to constructing efficient RTP materials.     

Highly Sensitive Detection of Residual Chlorpromazine Hydrochloride with Solid Substrate Room Temperature Phosphorimetry

Under the condition of 60?°C and 20?min at pH 6.12, chlorpromazine hydrochloride (CPZ) could react with fluorescein isothiocyanate (FITC) to produce FITC-CPZ, which increased the π-electron density (δ) of carbon atom in FITC conjugated system and the room temperature phosphorescence (RTP) intensity of FITC. Thus, a new solid substrate room temperature phosphorimetry (SSRTP) for the determination of residual CPZ was established. The regression equation of working curve was ΔI (p)?=?4.254?+?7.906 m(CPZ) (ag spot(-1)) with the correlation coefficient (r) of 0.9990 in the range of 0.036-9.6 ag spot(-1) (corresponding concentration: 0.090-24?fg?ml(-1), sample volume: 0.40?μl spot(-1)), and the detection limit (LD) was 0.018 ag spot(-1) (corresponding concentration: 4.5?×?10(-17)?g?ml(-1)). This method with wide linear range and high sensitivity was not only used to diagnose human disease based on the correlation between the residual quantity and lethal dose of CPZ in human serum, but also used to determine residual CPZ in biological samples with the results consisting with those obtained by gas chromatography (GC), showing good accuracy. The constituent of FITC-CPZ was analyzed by GC-MS (mass spectrometry) and the reaction mechanism of SSRTP for the determination of trace CPZ was also discussed.     

Second-Order Data Obtained by Time-Resolved Room Temperature Phosphorescence. A New Approach for PARAFAC Multicomponent Analysis

A second-order multivariate calibration approach, based on a combination of PARAFAC with time-resolved room temperature phosphorescence (RTP), has been applied to resolve a binary mixture of Phenanthrene and 1,10-Phenanthroline, as model compounds. The RTP signals were obtained in aqueous β-cyclodextrin solutions, in the presence of several heavy atom containing compounds. No deoxygenation was necessary to obtain the phosphorescence signals, which adds simplicity to the method. The resolution of the model compounds was possible in base to the differences in the delay-time of the RTP signals of the investigated analytes, opening a new approach for second-order data generation and subsequent second order multivariate calibration.