ACRYLAMIDE QUENCHING OF TRYPTOPHAN PHOTOCHEMISTRY AND PHOTOPHYSICS

Studies of acrylamide quenching of tryptophan (Trp) fluorescence, photochemistry, and photoionization have been conducted. Quenching of Trp fluorescence in aqueous solution by addition of acrylamide in the concentration range 0.0-0.5 M was measured and resulted in a Stern-Volmer quenching constant of KSV = 21 +/- 3 M-1. Photolysis experiments were performed in which Trp was photolyzed at 295 nm in the presence of varying concentrations of acrylamide. The loss of Trp was monitored using reverse-phase high performance liquid chromatography (RP-HPLC) and was observed to follow first order kinetics. Production of N-formylkynurenine (NFK) was observed by RP-HPLC in irradiated Trp samples both in the presence and absence of added acrylamide. In addition, no new photochemical product was detected. This was taken as evidence that acrylamide did not alter the photochemical pathway but just reduced the reaction rate as expected for a physical quenching mechanism. Plotting the reciprocal of photolysis rate constant versus acrylamide concentration produced a Stern-Volmer constant for quenching of Trp photochemistry of KSV = 6 +/- 2 M-1. The KSV values for both fluorescence quenching and photolysis quenching were thus large, implying efficient quenching of both processes by acrylamide. Assuming an excited singlet state lifetime of 2.8 ns, the calculated second-order quenching rate constants for fluorescence and photolysis were kq = 7.5 x 10(9) and 2.1 x 10(9) M-1 s-1 respectively. The possible involvement of photoionization in the photolysis mechanism was investigated by studies of acrylamide quenching of voltage transients produced by xenon flash lamp excitation of Trp at aqueous/teflon or aqueous/mica interfaces.(ABSTRACT TRUNCATED AT 250 WORDS)     

QUENCHING OF TRIPLET STATES OF AROMATIC KETONES BY INDOLE AND 3-METHYL INDOLE IN BENZENE SOLUTION. EVIDENCE FOR HYDROGEN-ATOM ABSTRACTION, FOR QUENCHING DUE TO FAVOURABLE CHARGE TRANSFER INTERACTION AND FOR THE LACK OF ELECTRONIC ENERGY TRANSFER

Abstract— Quenching of the triplet states of the aromatic ketones (KCO), benzophenone, acetophenone and xanthone, by indole and 3-methyl indole gives rise to the neutral radicals resulting from hydrogenatom transfer with variable efficiency (40–100%). Thus in addition to the reaction,
³KCO*+ R H →KCOH + R .
some other quenching path or paths occur. There is no evidence for any triplet energy transfer even when this is energetically favourable, and it is suggested therefore that quenching to give ground state species following favourable charge-transfer interactions accounts for the proportion of quenching without reaction.
The spectra of the indole radicals, R ., were determined and the kinetics of their decay in aerated and deaerated solution were investigated and reaction schemes proposed to explain these observations.     

FLUORESCENCE QUENCHING OF INDOLE BY DIMETHYLFORMAMIDE

Abstract— The effect of dimethylformamide on the lifetime (T) and quantum yield ( F ) of indole has been determined. Negative Stern-Volmer deviations prevail for T and F , but F ⁰/ F T⁰/T at all DMF concentrations. The deviation between F ⁰/ F and T⁰/T is ascribed to changes in F ⁰ and T⁰ induced by the change in solvent composition at the higher quencher concentrations.     

以吖啶染料荧光猝灭为基础的卤素阴离子检测研究

合成了两种吖啶及吖啶橙盐类小分子化合物和带有吖啶盐的三元共聚物.研究了它们在溶液中的荧光被卤素离子猝灭的问题.发现它们能强烈地被碘离子所猝灭而不易被氯离子猝灭,表明这类化合物有可能用于在有氯离子存在条件下对碘、溴离子的检测.对荧光猝灭机理进行的研究发现,卤素离子的猝灭能力并不和它们对发光化合物的系间窜越和三重态的生成等有关,而是和阴离子对极化分子的作用减弱了分子内的电荷转移能力相关.     

DISTANCE-DEPENDENT FLUORESCENCE QUENCHING OF TRYPTOPHAN BY ACRYLAMIDE

Abstract We used GHz frequency-domain fluorometry to investigate the time-dependent intensity decays of N -acetyl -L-trytophanamide (NATA) when collisionally quenched by acrylamide in propylene glycol at 20°C. The intensity decays of NATA became increasingly heterogeneous in the presence of acrylamide. The NATA intensity decays were not consistent with the Collins-Kimball radiation boundary condition (RBC) model for quenching. The steady-state Stern-Volmer plots show significant upward curvature. At low temperature in vitrified propylene glycol (-60%), where translational diffusion cannot occur during the lifetime of the excited state, quenching of NATA by acrylamide was observed. The Smoluchowski and RBC quenching models do not predict any quenching in the absence of translational diffusion. Hence, these frequency-domain and steady-state data indicate a through-space quenching interaction between NATA and acrylamide. The rate for quenching of NAT A by acrylamide appears to depend exponentially on the fluorophore-quencher separation distance. Comparison of the time-resolved and steady-state data provides a sensitive method to determine the distance dependence of the fluorophore-quencher interaction. The distance-dependent rate of quenching also explains the upward curvature of the Stern-Volmer plot, which is often observed for quenching by acrylamide. These results suggest that the distance-dependent quenching rates need to be considered in the interpretation of quenching data of proteins by acrylamide.     

GEOMETRIC REQUIREMENTS FOR THE INTRAMOLECULAR QUENCHING OF INDOLE FLUORESCENCE BY AMIDE AND CARBOXYLIC ACID GROUPS IN RIGID MOLECULES WITH THE [2,2,1]BICYCLOHEPTENYL SKELETON

Abstract— The geometric requirements for intramolecular fluorescence quenching of indole by carboxylic acid and amide functions have been determined in rigid norbornyl ([2.2,1]bicycloheptenyl) skeleton: trans and cis 3-(3'-indolyl) norbornene 2-dimethylcarboxamides or carboxylic acids. The reference compound was the 3-(3'-indolyl) N-dimethylpropionamide or -propanoic acid. The quenching occurs mainly in protic solvents. In the cis compounds, the quantum yields are strongly decreased when compared to the trans and open chain compounds. As determined by the pH dependence of fluorescence of the acids, it is in the acid form—COOH that the fluorescence is quenched; in the ionized species -CO⁻₂, only small quenching is observed. The K_s of Stern-Volmer equations are very small for the cis bicyclic acid and amide, the indole nucleus being shielded on one side only. The results support the hypothesis that small rearrangement of the peptide bond in protein conformation changes may be detected by fluorescence.     

DELAYED FLUORESCENCE AND SOME PROPERTIES OF THE CHLOROPHYLL TRIPLETS

–P-Type delayed fluorescence and sensitised P-Type delayed fluorescence have been observed from solutions of chlorophylls a and b in ethanol. E-Type delayed fluorescence has been observed from solutions in propylene glycol. The measurements in propylene glycol have yielded approximate values for the triplet energies of chlorophylls a and b. Measurements of the emissions from the ethanolic solutions have been used to calculate approximate values of the triplet formation efficiencies in this solvent. The sums of the fluorescence and triplet formation efficiencies in ethanol fall far short of unity and the triplet formation efficiencies therefore need to be confirmed by an independent method before they can be accepted with confidence.     

化学发光淬灭测定磺胺甲基异噁唑(SMZ)的研究

复方新诺明系磺胺甲基异噁唑(Sulfamethoxazole,简称SMZ)和甲氧苄氨嘧啶(Trime-thoprim,简称TMP)的复方制剂,用于抗脑膜炎球菌、肺炎球菌、溶血性链球菌及革蓝氏阴性杆菌,为临床广泛使用的磺胺类抗菌药物之一。药典规定用永停滴定法,重氮化外指示剂法进行含量测定。文献报道也有用三波长分光光度法、双波长法、吸收度比值法,核磁共振法等。本文根据SMZ的结构     

脱落酸对伊文思蓝的荧光猝灭机理研究及其含量测定

利用荧光光谱研究了脱落酸(Abscisic acid)与伊文思蓝(Evans blue)的相互作用。用266 nm激发光激发时,伊文思蓝的最大发射波长为396 nm。伊文思蓝与脱落酸相互作用,使伊文思蓝发生荧光猝灭,根据Stern-Volmer方程研究了荧光猝灭的类型及机理,实验证明脱落酸与伊文思蓝发生的静态猝灭,即脱落酸和伊文思蓝形成了一种稳定的复合物。伊文思蓝的相对荧光强度与脱落酸的浓度线性相关,线性方程为F0/F=0.78153+3.9153×104c,线性相关系数为0.9976。通过实验求算了脱落酸与伊文思蓝的结合点常数KABA-EB=1.838×105L/mol和结合点数n=1.171,由此建立了测定植物中脱落酸的新方法。     

AN INDIRECT METHOD FOR DETERMINATION OF PARAMETERS IN THE FORECAST MODEL

An indirect method for determining some parameters in a forecast model has been developed, by which the program of the original model with little modification can be used as a subroutine of the inversion system and the optimum estimate of the parameters can be obtained by calling the subroutine time after time. It is specially useful for the sophisticated numerical weather prediction model. It may also be utilized to improve the forecast timely in case of obvious differences between the recent observations and the forecasts. The effect of the method is verified by numerical simulation tests with simple models.     

芦丁对罗丹明6G的荧光猝灭机理研究及其含量测定

根据Stem - Volmer方程和罗丹明6G吸收光谱的变化研究了芦丁对其荧光猝灭的类型及机理,结果表明,芦丁与罗丹明6G作用,形成一种稳定的复合物,而发生荧光静态猝灭,求得复合物的结合常数K=5.55×105L/mol,结合点数n=1.26.芦丁含量在0.92～137.8μg/mL范围内与F0/F成正比,检出限为0....     

STUDIES OF THE EFFICIENCY and MECHANISM OF FLUORESCENCE QUENCHING REACTIONS USING ACRYLAMIDE and SUCCINIMIDE AS QUENCHERS

Abstract Here we report data for the quenching by acrylamide and succinimide of the fluorescence of a number of simple aromatic fluorophores in aqueous solution. Acrylamide is an efficient quencher of the fluorescence of most of these aromatic fluorophores, but succinimide is less efficient for all fluorophores and shows a very crude dependence on the ionization potential of the fluorophore. When the solvent is ethanol, the quenching efficiency by acrylamide and succinimide is found to decrease for the fluorophores, indole, naphthalene, and carbazole. These studies are consistent with an electron transfer quenching mechanism for the two quenchers. Quenching parameters (by acrylamide, succinimide, and iodide) are also reported for a number of fluorescent probes commonly used in biochemical studies. In general, the efficiency of acrylamide and succinimide quenching of these probes is low in aqueous solution.     

气相苯在TiO2光催化剂上吸附常数和光催化反应常数测定

采用溶胶-凝胶法制备TiO2光催化剂以及掺杂Fe3 和Ce3 的TiO2光催化剂,进行间歇式光催化降解气相苯动力学实验,基于光催化Langmuir-Hinshelwood反应动力学模型(L-H模型),测定气相苯在3种光催化剂上的降解动力学常数和吸附平衡常数.根据光催化降解气相苯实验动力学曲线和L-H模型,估算出TiO2、Fe3 /TiO2和Ce3 /TiO2光催化剂光催化降解苯的反应速率常数k和Langmuir吸附常数K分别为0.5247g/m3·min、1.523g/m3·min、1.010g/m3·min和8.605×10-2m3/g、2.390×10-2m3/g、3.928×10-2m3/g.掺杂Fe3 和Ce3 可明显提高光催化剂光催化降解苯的反应速率常数k,其中Fe3 /TiO2,的反应速率常数k最大.     

FLUORESCENCE QUENCHING FOR FLAVINS INTERACTING WITH EGG WHITE RIBOFLAVIN-BINDING PROTEIN

Abstract— The effect of flavin structure variation upon the binding process of flavin to hen egg white riboflavin was studied using fluorescence methods for formylmethylflavin (FMF), riboflavin (RF) and flavomononucleotide (FMN).
Measurements of flavin fluorescence intensities (steady state and phase-sensitive) and lifetimes were performed in a variety of RBP concentrations and temperatures (4 to 40°C). No fluorescence of flavoproteins was detected, while the fluorescence of flavins was found to be quenched by RBP. The overall quenching process is dominated by the static quenching (about 88%) due to the flavoprotein complex formation in the ground state, presumably a charge transfer complex.     

磺化竹红菌素对蛋白质荧光猝灭机理的研究

本文详细研究了磺化竹红菌素对多种蛋白质在溶液状态下的荧光猝灭过程。结果表明,磺化竹红菌素对多种蛋白质荧光猝灭服从Stern-Volmer曲线,实验观察了温度、粘度、pH值和盐酸胍对荧光猝灭过程的影响。由于磺化竹红菌素是一两性分子,对于不同蛋白具有不同猝灭过程;磺化竹红菌素对蛋白质的荧光猝灭常数K_q在10¹³mol/L·s^-1左右,这说明,磺化竹红菌素是一种比其它蛋白质荧光猝灭剂更加有效的荧光猝灭剂。     

常山碱的比色测定法

根据常山碱和异常山碱分子中的喹唑酮能碱水解而形成邻氨基苯甲酸的特点,制订常山植株中碱类含量的比色方法。邻氨基苯甲酸可与经重氮化的N-1-萘基-乙二胺偶合后,于530毫微米处读光密度。此法的灵敏度以光密度值/微克异常山碱/毫升比色液表示为0.111。用此法检查通常用的氯仿萃取法,得知其所回收的常山碱类尚不及实际含量之半。     

荧光染料吖啶红作测定蛋白质的生物探针——蛋白质的染料分析法新进展

简述染料结合分析法的发展过程,吖啶红染料自取和体系测定蛋白质的最佳试验条件和测定结果。阳离子荧光染料吖淀红及其自聚的二聚体的可平衡转化,形成自聚平衡体系,在表面活性剂十二烷基磺酸钠的存在下,蛋白质的定量加入可调节这一平衡,从而引起体系 荧光有规律的变化。方法线性范围宽、灵敏度高、反应速度快、准确度高 、抗干扰 能力强,是染料分析法定量测定蛋白质的最新方法之一。     

4-乙烯基吡啶与丙烯酰胺共聚合的研究

以过硫酸钾为引发剂 ,采用溶液自由基共聚合方法 ,实现了丙烯酰胺 (AM)与 4 乙烯基吡啶 (4 VP)的共聚合 .通过详细研究共溶剂体系、单体总浓度、反应温度、反应时间及引发剂量对共聚合过程中转化率和分子量的影响 ,从而确定了适宜的共溶剂体系和最佳的工艺条件 .用紫外分光光度法测得了共聚物的组成 .用Kelen Tudos方法 ,求得 4 乙烯基吡啶 (4 VP)和丙烯酰胺 (AM)单体的竞聚率 ,r4 VP =0 6 4 4 ,rAM =0 371.最后通过FTIR和1 3C NMR表征共聚物的结构并验证了共聚物的组成 .     

A QUENCHING FLUORESCENCE IMMUNOASSAY METHOD FOR DETERMINATION OF TRACE ALBUMIN USING UNLABELED TERBIUM CHELATE

A fluoroimmunoassay method using unlabeled Terbium chelate isdescribed.The principle is similar to that of fluoroimmunoassay method usinglanthanide chelate as labels.The procedure is simpte because labeling processis unnecessary.The recovery of HSA and albumin in urine is 107% and95% respectively.The standard deviation is tess than 10%.