Study on Fluorometric Determination of Hydrogen Peroxide Catalyzed by Manganese(Ⅱ)- Tetrasulfonatophthalocyanine with L-Tyrosine as a Substrate

以四磺基锰酞菁为过氧化物模拟酶， 研究其对H2 O2 氧化酪氨酸反应的催化作用， 建立了测定雨水中H2O2 的方法， 结果满意.A sensitive spectrofluorimetric method for the determination of trace H 2 O 2 has been proposed based on the catalytic effect of manganese(Ⅱ)-tetrasulfonatophthalocya- nine(MnTSPc) on the reaction of L-tyrosine and hydrogen peroxide.The maximum emission wavelength of the product is located at 410nm with excitation at 330 nm.Under the optimal conditions,the calibration graph is linear in the range of 0～2 0μmol/L H 2 O 2 with detection limit of 9 7nmol/L.The relative standard deviation of five replicate measurements is 3 6% for a 0 5μmol/L H 2 O 2 .This method has been applied to determine H 2 O 2 in rain water with satisfactory results.国家自然科学基金!（29775021

Determination of DNA using Rivanol as the Fluorescent Probe in the Presence of SDS

建立了在SdS存在下， 以利凡诺（rVn） 为荧光探针测定dnA 的新方法. 在适量SdS存在下， rVn 的荧光被强烈地猝灭直至最低值. 但于上述rVn- SdS体系中加入dnA 后， 体系的荧光强度增强， 且荧光光谱发生移动. 在一定条件下， 荧光强度增强值与加入dnA 的量在一定浓度范围内呈现良好的线性关系.The quantitative method for DNA using rivanol(RVN) as the fluorescent probe in the presence of SDS was proposed.Under proper conditions,addition of DNA to a mixture of RVN and SDS resulted in enhanced fluorescence and spectral shifts of RVN-SDS system.The calibration graph was linear in the range of 0～10 μg/mL,the limit of detection was 62 ng/mL CT DNA

A Rapid Method for the Determination of Molar Ratio of Fluorochrome to Protein Techniques by Fluorescence Anisotropy Detection

免疫荧光技术是免疫学检测的重要手段之一，该技术在病原微生物的早期诊断、自身免疫研究、抗原或抗体的免疫组化定位等方面都得到了广泛应用［1］.荧光色素对抗体（或抗原）标记比的测定是免疫荧光技术的重要部分，这是因为标记比合适与否直接关系到检测结果的优劣甚至...The Determination of molar ratio of fluorochrome to protein is an important part in fluorescent antibody techniques.The conventional method is time consuming and with troublesome manipulations.A rapid homogeneous method based on the anisotropy change of the fluorochrome after reacting with protein was presented here.In our experiments, fluorescein isothiocyanate(FITC) and bovine serum albumin(BSA) were chosen to form the FITC/BSA conjugate.A series of solutions containing the two components were prepared and allow to react according to the standard procedure.Fluorescence anisotropy of the mixtures were detected, a diagram of r lg c is then obtained, where c is the concentration of protein.Because FITC in the mixture exists in both free form(F) and binding form(B), the fluorescence anisotropy observed is given by r=f F r F+ f B r B, where r F and r B refer to the anisotropy of free and bound FITC, respectively; and f F and f B represent the fraction of the free and bound forms, respectively, f F+ f B=1.A formula as f B= (r-r F)/( r B- r F) is achieved.Here r B correspods to the value of r in the upper platform region of the r lg c curve, where FITC is bound to protein completely, while r F can be obtained by the determination of anisotropy of FITC in the absence of BSA.So, for each of the mixtures, the binding fraction of FITC can then be calculated.Correspondingly, the content of bound form of FITC that was used to calculate the molar ratio of FITC to BSA can be gained.Since the method avoids the tiresome separation procedure, it bears the merit of time saving and can be used to estimate the molar ratio of fluorochrome to protein rapidly.The determination results of samples by this method were compared with that got from a spectrophotometric analysis.The results were in good agreement.国家自然科学基

Study on Fluorimetry for Determination of Phenylalanine Using Enzyme Substituted by the UV-Light

在碱性介质中， 光子可以取代苯丙氨酸氧化酶使苯丙氨酸分解并生成H2 O2 ， 然后用氯化血红素作为模拟酶检测H2O2 ， 从而建立间接测定苯丙氨酸的新方法.In alkaline medium,the photon can substitute phenylalanine oxydase,and cause phenylalanine to decompose and produce H 2 O 2 ,which are detected by using HPA to be coupled and produce strong fluorescent substance in present of mimetic peroxydase-hemin.Thus,a new fluorimetry for the determination of phenylalanine was developed.福建省自然科学基金!（B96003

Mimetic-Enzyme Fluoroimmunoassay for HBsAg Based on Thermal Phase Separating Technique

以聚n- 异丙基丙烯酰胺（PnIP） 作为免疫反应载体， 以铁酞菁作为HrP的新型模拟酶来标记抗人HbSAg 抗体， 建立了基于热相分离技术的夹心型荧光免疫分析HbSAg 的方法. HbSAg浓度在20 ~5000ng/Ml范围与体系的相对荧光强度呈良好线性关系. 检测限8ng/Ml.An anti-HBsAg antibody was conjugated to Poly-N-Isopropylacrylamide,and a novel polymer-mimetic enzyme fluoroimmunoassay method for the determination of HBsAg with sandwich format was proposed by using iron-tetrasulfonatophthalocyanine (FeTSPc) as a labelling reagent.The calibration graph for HBsAg was linear over the range of 20～5 000 ng/mL with a detection limit of 8 ng/mL.福建省自然科学基金!（9810004）;国家教育部博士点基

Resonance Light-Scattering Spectra of Bovine Serum Albumin and Its Analytical Application

研究了牛血清白蛋白水溶液本身的共振光散射光谱信号， 基于其光谱强度与牛血清白蛋白的浓度呈线性相关的现象， 初步建立了一种步骤简单的测定方法.The resonance light-scattering spectra of BSA have been studied using an ordinary spectrofluorometer.The signal intensity of BSA is enhanced with the increase of concentration of BSA,and the enhancement is linear.Based on this principle,a new method for the determination of BSA was proposed.The calibration curve of this method is essentially linear over the range of 0-200 μg/mL

Study on the Mimetic Peroxidase Activity of Iron-Tetrasulfonatophthalocyanine Modified by Temperature-Sensitive Polymer

合成了带活性末端的热敏高分子并用其修饰四磺基铁酞菁. 研究了修饰后的铁酞菁模拟过氧化物酶的催化活性. 结果表明， 修饰后铁酞菁的模拟酶活性明显高于游离铁酞菁.Iron-tetrasulfonatophthalocyanine was covalently conjugated to a temperature- seneitive polymer, oligo(N-isopropylacrylamide) containing a terminal carboxlyic group,and the mimetic peroxidase activity of this conjugate was studied.The results show that the catalytic activity of the modified mimetic enzyme is higher than that of the free mimetic enzyme.福建省自然科学基金!（B9810004）;国家教育部博士点基

Studies on Tetra-substituted Amino Aluminum Phthalocyanine as a New Red-region Substrate for Mimetic Peroxidase

Tetra substituted amino aluminum phthalocyanine (TAAlPc) has been synthesized and used for the first time as a new red region fluorescent substrate for the determination of hydrogen peroxide catalyzed by peroxidase or mimetic peroxidase. Under optimum conditions, the calibration graph has a linear range of 0.0-3.0×10 -7 mol/L H 2O 2 with a detection limit of 3.7×10 -9 mol/L. The feasibility of TAAlPc as a new promising red region substrate in practical application has been proven in the determination of H2O2 in rainwater. The proposed method can largely minimize the nterference that results from background fluorescence or scattering light and has a high analytical sensitivity.国家自然科学基金(批准号:29775021

Application of a Red-Region Fluorescence Dye,Nile Blue, in Nucleic Acids Assay

基于dnA或rnA对耐尔蓝荧光的猝灭效应， 建立一种新的dnA 或rnA 的测定方法. 在最优条件下， 测定小牛胸腺dnA、鲑鱼精子dnA、鲱鱼精子dnA 和酵母rnA 的线性范围分别为： 3-0 ~2-0 x103ng/Ml、9-0 ~2-0 x103ng/Ml、5-4 ~5-0 x103ng/Ml 和27 ~10 x103ng/Ml.检测限分别是3-0ng/Ml、9-0ng/Ml、5-4ng/Ml和27ng/Ml， 相对标准差（n ＝ 6） 是2-1 % . 本文还讨论了干扰物质的影响.A novel fluorimetric method was proposed for the rapid determination of DNA and RNA based on their quenching effect on the cationic red-region fluorescent dye,nile blue.Under optimum conditions,the calibration curves for the determination of Calf thymus DNA(CT DNA),Salmon DNA(SM DNA),Herring sperm (HS DNA) and yeast RNA were linear over the range of 3 0-2 0×10 3 ng/mL,9 0-2 0×10 3 ng/mL,5 4-5 0×10 3 ng/mL and 27-10×10 3 ng/mL,respectively.The detection limits were 3 ng/mL for CT DNA,9 0 ng/mL for SM DNA,5 4 ng/mL for HS DNA and 27 ng/mL for RNA,respectively.The relative standard deviation(n=6) was within 2 1% in the middle of linear range.Interference concerning some interesting coexisting substances with the determination of DNA was also examined.国家自然科学基金!（29775021

A rapid method for the determination of molar ratio of fluorochrome to protein techniques by fluorescence anisotropy detection

The Determination of molar ratio of fluorochrome to protein is an important part in fluorescent antibody techniques. The conventional method is time consuming and with troublesome manipulations. A rapid homogeneous method based on the anisotropy change of the fluorochrome after reacting with protein was presented here. In our experiments, fluorescein isothiocyanate (FITC) and bovine serum albumin(BSA) were chosen to form the FITC/BSA conjugate. A series of solutions containing the two components were prepared and allow to react according to the standard procedure. Fluorescence anisotropy of the mixtures were detected, a diagram of r-lgc is then obtained, where c is the concentration of protein. Because FITC in the mixture exists in both free form(F) and binding form(B), the fluorescence anisotropy observed is given by r = f(F)r(F) + f(B)r(B), where r(F) and r(B) refer to the anisotropy of free and bound FITC, respectively; and f(F) and f(B) represent the fraction of the free and bound forms, respectively, f(F) + f(B) = 1. A formula as f(B) = (r-r(F))/(r(B)-r(F)) is achieved. Here r(B) correspods to the value of r in the upper platform region of the r-lgc curve, where FITC is bound to protein completely, while r(F) can be obtained by the determination of anisotropy of FITC in the absence of BSA. So, for each of the mixtures, the binding fraction of FITC can then be calculated. Correspondingly, the content of bound form of FITC that was used to calculate the molar ratio of FITC to BSA can be gained. Since the method avoids the tiresome separation procedure, it bears the merit of time saving and can be used to estimate the molar ratio of fluorochrome to protein rapidly. The determination results of samples by this method were compared with that got from a spectrophotometric analysis. The results were in good agreement