甲基紫6B共振光散射法测定脱氧核糖核酸

基于甲基紫6B与脱氧核糖核酸在酸性条件下共振光散射的增强效应,建立了测定DNA的共振光散射法。在pH值为2．0～3．0的三羟甲基氨基甲烷一盐酸缓冲溶液中,甲基紫6B与fsDNA、ctDNA分子作用后共振光散射增强,其强度增加值与DNA的浓度呈线性关系,线性范围分别为0～1．0、0～1．5mg／L,相关系数分别为0．9993、0．9997,检出限分别为15．3、12．2ug／L,用于DNA合成样品的测定,测定结果的精密度、准确度较高。     

溴代十六烷基三甲基铵敏化吡罗红B-核酸作用的共振光散射增强法测定核酸

基于表面活性剂溴代十六烷基三甲基铵（CTMAB）对吡罗红B-核酸作用的共振光散射增强效应有敏化作用,建立了一种高灵敏测定核酸的新方法。在pH7．4时,吡罗红B在328nm处的共振光散射的增强与核酸浓度有良好的线性关系。在最佳实验条件下,对小牛胸腺DNA（ct-DNA）、鲱鱼精DNA（fs-DNA）、酵母RNA（yeast—RNA）测定的线性范围分别为0．0～1．2mg／L、0．0～0．8mg／L和0．04～1．4mg／L。检出限分别为6．1μg／L、11．2μg／L和8．6μg／L。该法简便、快捷、重现性好,对合成样品进行了测定,结果令人满意。     

碱性品红共振光散射法测定DNA研究

基于脱氧核糖核酸（DNA）对有机染料碱性品红的共振光散射增强效应，拟订了一种测定DNA的共振光散射法。在pH=6．75～7．25的范围内，碱性品红在594nm处的共振光散射增强与yDNA和ctDNA的浓度呈线性关系，线性范围分别为0．20～1．60μg／mL和0～1．50μg／mL，相关系数分别为0．9997和0．9994，检出限可达26．5μg／L。该方法简便、快速，用于合成样品中DNA的测定，结果满意。     

铬天青S共振光散射法测定脱氧核糖核酸

介绍在阳离子表面活性剂十六烷基三甲基溴化铵(CTMAB)存在下阴离子染料铬天青S(CAS)共振光散射(RLS)法测定脱氧核糖核酸(DNA)的方法。在pH=5．27的六次甲基四铵一盐酸缓冲溶液中，研究了CAS—CT-MAB—yDNA体系的RLS光谱特征、影响因素和最佳反应条件。在最佳条件下，体系的RLS强度增加值△I与yDNA的浓度在50～800μg／L和1000～2000μg／L呈线性关系，其线性回归方程分别为△I=0．48c 2．56和△I=0．14c 17．86，相关系数分别为0．9997和0．9991，检出限为4．3ng／mL。该方法简便、快速，应用于合成样品中yDNA的测定，测定结果的相对标准偏差为2．93％～4．71％，回收率为97．8％～105．2％。     

联吡啶-铜(Ⅱ)络合物荧光增强法测定核酸

利用联吡啶-Cu(Ⅱ)络合物作为荧光探针测定脱氧核糖核酸(DNA),实验发现加入天然和变性DNA均能使联吡啶-Cu(n)络合物(Cu(bpy)2^2 )体系的荧光强度大幅度增强。测定体系的最大激发和发射波长分别为304和334nm。在最佳实验条件下,测定的线性范围分别为：小牛胸腺(CT-DNA)0．09—9mg／L;鱼精DNA(FS—DNA)0．01—12.5mg／L;酵母DNA(yeastDNA)0．30—35mg／L。检出限分别为0.003、0．002和0．02mg／L。实验结果推测该络合物与DNA之间通过静电作用结合。     

邻苯二酚紫-溴化十六烷基三甲铵-核酸共振光散射体系的研究及其分析应用

研究了核酸与阴离子染料邻苯二酚紫(PV)及阳离子表面活性剂溴化十六烷基三甲铵(CTMAB)体系的共振光散射光谱特性。在pH2．35的柠檬酸介质中，核酸(yRNA或fsDNA)与阳离子表面活性剂CTMAB对邻苯二酚紫的共振光散射光谱有协同增强作用，产生最大散射波长为400nm的共振光散射光信号。在最佳实验条件下，共振光散射测定yRNA和fsDNA的线性范围皆为0．02-0.75mg／L，检出限分别为1．4和8．7μg/L。据此建立了一种测定核酸的新方法。     

两性离子表面活性剂作为共振光散射探针测定脱氧核糖核酸

将两性离子表面活性剂用于DNA的共振光散射测定。在pH值为7．96～9．40较宽的范围内,DNA与椰油酰胺丙基-2-羟基-3-磺基丙基甜菜碱(HSB)在392nm处有稳定的共振光散射增强,其强度与DNA浓度呈线性关系,建立了一种测定DNA的新方法。线性范围为0．02—4．25mg／L;检出限达1．5μg／L。该法简便、快速,用于合成样品中DNA的测定,重现性好,结果满意。同时探讨了有关机理。     

吖啶橙共振光散射法测定痕量脱氧核糖核酸

研究了三环杂芳香类染料吖啶橙(AO)与DNA作用的共振光散射光谱,在pH11．5～12．5的范围内,加入DNA导致吖啶橙共振光散射增强,在339nm处,存在一共振光散射增强峰,其强度与DNA浓度呈线性关系,据此建立了一种测定DNA的共振光散射新方法?对于ctDNA,方法的线性范围为14．3～1000μg／L,检出限为2．86μg／L,RSD为3．6％;对于fsDNA,方法的线性范围为24．0～1250μg／L,检出限为4．78μg／L,RSD为6．0％。已用于合成样品中DNA的测定。     

荧光素共振光散射法测定脱氧核糖核酸

研究了咕吨类染料荧光素Fluorescein(FL)与DNA作用的共振光散射光谱。加入DNA后,在pH6～8的范围内,荧光素在DNA分子表面发生长距离自组装,在400nm处产生了增强的共振光散射峰,其发光强度与DNA的浓度呈线性关系,线性范围为0．04～3．1mg／L;检出限为16μg／L。考察了影响因素和最佳反应条件,建立了用RLS光谱测定ng级DNA的新方法。     

十六烷基三甲基溴化铵增敏砂罗铬花青R共振光散射法检测DNA

在pH=3．92的三羟甲基氨基甲烷-HCl缓冲溶液中,阳离子表面活性剂十六烷基三甲基溴化铵对砂罗铬花青R与脱氧核糖核酸(DNA)的共振光散射(RLS)有协同增强作用。考察了影响因素,研究了在优化条件下RLS强度与DNA浓度之间的关系,鱼精DNA和小牛胸腺DNA的线性范围均为0．05—3．00mg／L,检出限分别31．03、35．98μg/L,回收率为97．9％～99．6％,测定结果的相对标准偏差为0．5％-2．1％。     

meso—四（对—羟基苯基）卟啉与脱氧核糖核酸的作用及其分析应用

报道了非离子型卟啉ｍｅｓｏ－四卟啉与脱氧核糖核酸的作用。在ＰＨ４．９－５．４范围内，ＤＮＡ能与聚合态的ＴＨＰ作用，产生不规则的电子吸收光谱。但如果体系中含有３０％（Ｖ／Ｖ）乙醇，ＤＮＡ与ＴＨＰＰ作用产生４５９．０ｎｍ和７００．０ｎｍ两个新峰。     

溴代十六烷基三甲基铵敏化亮绿-脱氧核糖核酸作用的共振光散射增强研究

首次报道在表面活性剂存在下阳离子染料与脱氧核糖核酸(DNA)作用所导致的共振光散射增强(RLSE)。在pH 10.4 ～11.8 和离子强度低于0.050 mol/L的条件下,亮绿(BG)与DNA作用产生398.0 和 467.2 nm的特征RLSE信号,而溴代十六烷基三甲基铵(CTMAB)进一步敏化该RLSE信号。受CTMAB敏化的RLSE信号与0～1.2 mg/L的ctDNA和fsDNA呈正比,检测限低于5 mg/L。方法成功应用于合成样中DNA的测定。     

Determination of nucleic acids with tetra-(N-hexadecylpyridiniumyl) porphyrin sensitized by cetyltrimethylammonium bromide (CTMAB) using a Rayleigh light-scattering technique

Using a common spectrofluorometer to measure the intensity of Rayleigh light-scattering (RLS), a method for determination of nucleic acids has been developed. At pH 10.24 and ionic strength 0.01 mol l-1 (NaCl), the Rayleigh light-scattering of the tetra-(N-hexadecylpyridiniumyl) porphyrin (TC16PyP) is greatly enhanced by nucleic acids in the presence of cetyltrimethylammonium bromide (CTMAB), with the scattering peak located at 311.8 nm. The enhanced RLS intensity is in proportion to the concentration of calf thymus DNA (ctDNA) in the range 0.2-6.0 microg ml-1 and to that of fish sperm DNA (fsDNA) in the range 0.05-3.0microg ml-1. The limits of detection are 0.016 microg ml-1 for calf thymus DNA and 0.023 microg ml-1 for fish sperm DNA when the concentration of TPP was chosen 2.0 x 10(-6) mol l-1. Four synthetic samples were determined satisfactorily.     

结晶紫共振光散射法测定脱氧核糖核酸

研究了三苯甲烷类碱性染料结晶此与DNA作用的共振光散射光谱,在pH9.2-10.5的范围内,加入DNA导致结晶紫共振光散射增强,在512nm处,存在一共振光散射增强峰,其强度与DNA的浓度呈线性关系,据此建立了一种测定DNA的菜振光散射法,方法的线性范围为0－900ng/mL,检出限为5.02mg/mL.已用于混合样品中的DNA的测定.     

酚藏花红与DNA作用的共振光散射特征及微量DNA的光散射测定

研究了酚藏花红与脱氧核糖核酸在近中性条件下作用中的共振光散射特征，并以此建立起纳克级核酸的分析方法。考察了影响因素和最佳反映条件，建立了用ＲＬＳ光谱测定纳克级ＤＮＡ的新方法。     

Determination of DNA with cetyltrimethylammonium bromide by the measurement of Resonance Light Scattering.

A simple assay of DNA was developed based on the measurements of enhanced signals of Resonance Light Scattering (RLS) of cetyltrimethylammonium bromide (CTMAB) by DNA. The enhanced RLS signals, measured by simultaneously scanning the excitation and emission monochromators of a common spectrofluorometer with lambda ex = lambda em, was optimized for the DNA assay with CTMAB. On the conditions of pH 2.21 and ionic strength 0.002, the enhanced RLS intensity at 470.0 nm, delta I, was found to be proportional to the concentration of DNA in the range 0-2.5 micrograms/ml if 1.5 x 10(-5) M CTMAB was used. Limits of determination for calf thymus DNA and fish sperm DNA were 4.9 ng/ml and 9.2 ng/ml, respectively. Synthetic samples were determined with the recovery ratio ranging from 93.2% to 105.1%, and the RSD is lower than 2.7%.     

A simple and sensitive assay of nucleic acids based on the enhanced resonance light scattering of zwitterionics

A new assay of nucleic acids at nanogram level was established based on the enhanced resonance light scattering (RLS) signals of two zwitterionics cocamidopropyl hydroxysultaine (HSB) and lauryl betaine (BS-12). Under optimum conditions, the weak RLS signal of HSB is enhanced by nucleic acids, and the enhanced RLS intensity is proportional to the concentration of nucleic acids in the range of 0.02–7.3 mg l⁻¹ for calf thymus DNA and 0.01–8.6 mg l⁻¹ for fish sperm DNA. The detection limits were 1.5 ng ml⁻¹ for calf thymus DNA and 1.9 ng ml⁻¹ for fish sperm DNA. Plasmid DNA extracted from K-12-HB101 colt was determined with satisfactory results.     

Cetyltrimethylammonium bromide sensitized resonance light-scattering of nucleic acid--Pyronine B and its analytical application

This is the first report on the determination of nucleic acids with Pyronine B (PB) sensitized by cetyltrimethylammonium bromide (CTMAB) with resonance light-scattering (RLS) technique. Under the experimental conditions (1 x 10(-5) mol l(-1) PB, 1 x 10(-5) mol l(-1) CTMAB, pH 7.4, at room temperature, ionic strength 0.02 mol l(-1) NaCl), the interaction of PB with DNA sensitized by CTMAB results in enhanced RLS signals at 328 and 377 nm in the enhanced regions. It was found that the enhanced RLS intensity at 328 nm was proportional to the concentration of DNA in the suitable ranges. The linear range of this assay is 0.0-1.2 microg ml(-1) for calf thymus, 0.0-0.8 microg ml(-1) for fish sperm DNA (fsDNA), and 0.04-1.4 microg ml(-1) for yeast RNA, respectively. The detection limits (3 sigma) are 6.1 ng ml(-1) for calf thymus DNA (ctDNA), 11.2 ng ml(-1) for fish sperm DNA, and 8.6 ng ml(-1) for yeast RNA, respectively. Six synthetic samples were determined satisfactorily. This method is simple, rapid and the dye is inexpensive and stable.     

Interactions of Nile Blue Sulphate with Nucleic Acids as Studied by Resonance Light-Scattering Measurements and Determination of Nucleic Acids at Nanogram Levels

ABSTRACT

The interactions of nile blue sulphate (NBS) with nucleic acids, including calf thymus DNA, fish sperm DNA and yeast RNA, were characterized with resonance light-scattering (RLS) measurements by using a common spectrofluorometer. Accordingly a method for the determination of nucleic acids at nanogram levels was established. At pH's of 7.20～7.60 and ionic strengths lower than 0.012, the interactions of NBS with nucleic acids result in three characteristic RLS peaks at 293.4 nm, 349.4 nm and 560.4 nm. Mechanism study shows that these peaks are ascribed to the long range assembly of NBS on the molecular surface of nucleic acids, which depends on pH, ionic strength and the stranded structure of nucleic acids. A Scatchard plot was constructed by using the RLS data, yielding the assembly number and assembly constant being 6.4 and 7.13x10⁶ mol^?1 1 for NBS assembly on the molecular surface of calf thymus DNA. The same parameters are 6.6 and 4.58x10⁶ mol^?1 1 for the assembly on that of fish sperm DNA, 3.9 and 1.67x10⁶ mol^?1 1 on that of yeast RNA, respectively. Linear relationships were found between the enhanced RLS intensity at 293.4 nm and nucleic acid concentration. If 1.2x10^?5 mol I^?1 NBS was employed, 0～0.80 μg ml^?1 calf thymus DNA and fish sperm DNA, 0.20～0.60 μg ml^?1 yeast RNA can be determined with the determination limits being 3.2 ng ml^?1 for calf thymus DNA, 11.5 ng ml^?1 for fish sperm DNA and 38.3 ng ml^?1 for yeast RNA, respectively. Four synthetic samples were determined with satisfaction.