Association of Dyes Bearing with Opposite Charges and Its Application in the Determination for Micro Amounts of Nucleic Acids

［中文文摘］建立了高灵敏荧光分光光度测定ＤＮＡ 和ＲＮＡ 的新方法． 方法的线性范围分别为０－０４ ～１－２０μｇ／ｍ Ｌ（ 鲑鱼精子ＳＭＤＮＡ 和小牛胸腺ＣＴＤＮＡ） ， ０－１０ ～１－２０μｇ／ｍＬ（ 酵母ＲＮＡ） ， 检测限分别为１７ｎｇ／ｍＬ（ＳＭＤＮＡ） ， ２４ｎｇ／ｍＬ（ＣＴＤＮＡ） ， ９８ｎｇ／ｍＬ（ＲＮＡ） ． 将方法用于实际样品金黄色葡萄球菌中ＤＮＡ 含量的测定， 结果满意．［英文文摘］It has been found that the fluorescence of tetrasulfonated aluminum phthalocyanine was obviously quenched by cationic dyes,but recovered in the presence of nucleic acid. Based on this phenomenon,a novel method for the determination of nucleic acids was investigated.The method presented is senstive and with simplicity in manipulation.Besides,since a fluorescent dye emitting at long wavelength region is employed,this method has the advantage of effectively reducing the interference from background fluorescence and scattering light.国家自然科学基金（29775021

铝酞菁磷酸酯用作碱性磷酸酶红区底物的初步研究

合成了铝酞菁磷酸酯， 对其荧光性质及作为碱性磷酸酶底物的可行性进行了初步考察． 结果表明， 该化合物有望成为一种新型的碱性磷酸酶红区荧光底物

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

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

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

The Determination of Total Protein of Human Serum by Near-IR Fluorescence Enhancement

［中文文摘］建立了以阴离子花菁染料—ＣＴＡＢ体系依近红外荧光增强测定人血清总蛋白的方法． 最大激发及发射波长分别为７６５ｎｍ 及８１２ｎｍ ． 蛋白质测定的线性范围为０－４ ～１２μｇ／ｍＬ．［英文文摘］A fluorometric method has been developed for the determination of total protein in human serum with a new near-IR reagent as a fluorescence probe,based on the fluorescence enhancement of the cyanine-CTAB system in the presence of protein.Maximum fluorescence is produced with maximum excitation and emission wavelengths at 765 and 812nm, respectively. Under optimal conditions,the calibration graphs are linear over the range 0 4～12 0μg/mL for protein. The detection limit is 70ng/mL.The relative standard deviation of six replicate measurement s is 1114 %for BSA of 610μg/ mL level. The results are satisfactory.国家自然科学基金（29775021

A Cationic Cyanine as a Near-IR Fluorescent Probe for Determination of Nucleic Acids

［中文文摘］以阳离子花菁染料为荧光探针， 依近红外荧光猝灭测定核酸． 最大激发及发射波长分别为７６５ 、７９０ｎｍ ． 核酸测定的线性范围为０－０８ ～１－２μｇ／ｍ Ｌ（ＣＴＤＮＡ 及ＳＭ ＤＮＡ） ， ０－１ ～１－６μｇ／ｍＬ（yeａｓｔ ＲＮＡ)．［英文文摘］A method with an cationic near-IR cyanine as a fluorescent probe was developed for the determination of nucleic acids.The near-IR cyanine shows the maximum excitation and emission wavelengths at 765 and 790nm, respectively,in aqueous solution.The method is based on the fluorescence decrease of near-IR cyanine in the presence of nucleic acids.Under optimal conditions,the ratio value of fluorescence intensity in the absence and presence of nucleic acids was proportional to the concentration of nucleic acids over the range of 0108～112μg/ mL for CT DNA or SM DNA , and 011～116μg/ mL for Yeast RNA. The detection limit s were 30 ng/ mL for CT DNA , 25 ng/ mL for SM DNA and 70 ng/ mL for Yeast RNA. The relative standard deviation (n = 6) were 211%for 500 ng/ mL CT DNA level.国家自然科学基金（29775021

Synthesis of a NearIR Cyanine and Its Application in Determination of Biomolecules

合成了一种新型阴离子型近红外花菁染料，通过核磁共振及质谱验证了结构，并研究了其水溶液的光谱性质.发现在酸性条件下，蛋白的加入使该花菁染料在776nM处的吸收发生减色作用，因而可以作为测定血清总蛋白质的近红外生色探针.另外，还研究了其在阳离子表面活性剂作用下形成的弱荧光现场二聚体做为核酸探针的可能性.A novel water soluble nearIR cyanine dye(HMC) was synthesized and its structure was characterized by NMR and MS.The absorption and fluorescence spectral characteristics of HMC in aqueous solution were also studied.It was found that in acidic solution of HMC the presence of protein could cause a hypochromic effect at 776 nm, that was used as a probe for the determination of total protein in human serum.The low fluorescence of in situ HMC dimer caused by cationic surfactant CTAB has been tested as a probe for nucleic acid determination.国家自然科学基

Determination of Nucleic Acids by a Near-IR Spectrophotometric Method

［中文文摘］以阳离子型近红外花菁染料做为生色探针 ,研究了该染料与核酸的结合反应 .在 p H6 .0的条件下 ,该染料在 771nm处有最大吸收峰 ,在核酸存在下 ,其吸收峰显著下降 ,而在 6 4 0 nm处出现新的吸收峰 .据此 ,以该近红外花菁染料为生色探针 ,在 771nm处依其吸光度的下降可定量测定核酸 .SM DNA的线性响应范围为 6 0～ 2 0 0 0 ng/ m L ,yeast RNA的线性响应范围为 10 0～ 2 0 0 0 ng/m L;检测限分别为 30 ng/ m L及 50 ng/ m L;六次平行测定 DNA(4 50 ng/ m L)的相对标准偏差为3.4 % ,测定 RNA (4 50 ng/ m L )的相对标准偏差为 3.7% ,方法具有较高的灵敏度 .用于合成试样分析, 结果满意.［英文文摘］A cationic near IR cyanine was used as a chromic probe and its binding reaction with nucleic acids was studied. In the medium of pH 6.0, the near IR cyanine showd a decrease in absorption at its maximal absorption peak (771nm) on binding to nucleic acids, and a new absorption peak appeared at 640 nm. Based on the fact, a novel near IR spectrophotometric method for the determination of nucleic acids was developed. The calibration graphs are linear over the range 60~2000 ng/mL for Salmon Sperm DNA，100～ 2 000 ng/mL for Yeast RNA , respectively. The detection lim its is 30 ng/mL for Salmon Sperm DNA , 50 ng/mL for Yeast RNA, respectively.国家自然科学基金资助项目(29775021

Determination of Human Serum Albumin by Aluminium Tetrasulponated Phthalocyanine

以红区荧光染料四磺基铝酞菁(AlS_4PC)为探针建立了荧光猝灭法测定白蛋白的新方法.考察了各种影响因素,在最佳实验条件下,该方法的线性区间0.10~4.5Mg/l,检测限为40μg/l.将该方法用于人血清样品中白蛋白的直接测定并与临床方法(溴甲酚绿法)比较,二者符合良好.Based on the fluorescence quenching of aluminium tetrasulphonated phthalocyanine (AlS4Pc), a red-region fluorescent dye, reacted with albumin, a new method was developed for the determination of human serum albumin(HSA).Under the experimental conditions, the linear range ofthis assay was 0.10-4.5 mg/Land the detection limit was 40μg/L.The method has been applied to theanalysis of human serum samples and the results were in agreement well with those achieved by bromocresol green method, which indicated that the method was not only sensitive but also reliable