Resonance light scattering and derived techniques in analytical chemistry: past, present, and future

From 1993 to 1995, with a conventional fluorescence spectrophotometer (CFS) (convenient) and working in a synchronous scan model (easy-to-use), Pasternack et al. proposed the resonance light-scattering (RLS) technique, to efficiently characterize self-assemblies or self-aggregations of chromophores with good electronic coupling. Incident wavelengths were specially considered within their absorption envelopes (rather unorthodox), and their amplified signals were observed (good sensitivity and selectivity). Due to these absorbing benefits, RLS technique, as a novel readout method, commenced on its exciting analytical tours soon after Liu et al. and especially Li et al., separately, set out their corresponding pioneering investigations from 1995 to 1997. From then on, it has received an increasing attention by analysts, as a consequence exhibiting more and more fascinating analytical applications. Moreover, various attractive RLS-derived techniques have been developed successively to improve it or to enlarge its possibilities. Later on, Liu et al. and Li et al., Tabak et al., Yguerabide et al., Huang et al., Lakowicz et al. and Fernández Band et al. have made their outstanding contributions. In this review, we concentrate on major achievements of RLS in analytical chemistry for over a decade, involving the developments and analytical applications of RLS derived techniques treated as an impacting progress of RLS technique in analytical chemistry. Finally, an indication of future directions of RLS technique in analytical chemistry is provided.     

Determination of dextrin based on its self-aggregation by resonance light scattering technique

A novel free-probe assay of dextrin was established based on the resonance light scattering (RLS) enhancement in aqueous solution due to the self-aggregation of dextrin. The RLS intensity was well proportional to the concentration of dextrin over the wide range 0.2-14 μg mL⁻¹ and a detection limit 0.02 μg mL⁻¹ was obtained in the optimum conditions. The effect factors such as pH, buffer medium, holding time, ionic strength and temperature were studied in detail. Little or no interference was presented in the detection when adding coexisting substances including various metal ions and some saccharine in the solution. The assay proposed owns the advantages of easy operation, rapidity, sensitivity and practicability. Three synthetic samples and three kinds of medicine samples were analyzed with satisfactory results.     

Use of Gemini surfactant in a one-step ellagic acid assay by resonance light scattering technique

Ellagic acid (EA) reacted with Gemini zwitterionic surfactant, phosphodiesters quaternary ammonium salt (PQAS), and formed fine particles which produced strong enhancement in intensity of resonance light scattering (RLS). The effects of several factors on the RLS signal, such as pH, ionic strength, PQAS concentration and so on, were optimized. The relationship between enhanced RLS intensity and EA concentration was constructed. A novel and rapid method for the determination of EA was built. The linear range of this method was 0.016-4.0 μg mL⁻¹ and the detection limit was 13.9 ng mL⁻¹. Under the optimum conditions, the proposed method was applied to determine EA in body fluids with the results of quantitative recoveries between 98.4-101.4% in human serum samples and 99.1-102% in human urine samples. This method characterized by low limit detection is very sensitive and the cost is low, and constitutes a fast one-step procedure which requires only measuring the RLS intensities. The mechanism of the reaction was also studied. This investigation could contribute to the research on the delivery and release of bioactive molecules by Gemini surfactants.     

On the factors affecting the enhanced resonance light scattering signals of the interactions between proteins and multiply negatively charged chromophores using water blue as an example

Electrostatic interactions of proteins, including bovine serum albumin (BSA), human serum albumin (HSA), γ-globulin (γ-IgG), α-chymotrypsin (Chy), lysozyme (Lys) and cellulase (Cel), with multiply negatively charged chromophores were investigated based on the measurements of the enhanced resonance light scattering (RLS) signals. Using triply negatively charged water blue (WB) as an example, the factors were discussed that affect the enhanced resonance light scattering signals of the interactions between proteins and the negatively charged chromophores. It was found that the enhanced RLS signals with the maximum light scattering peak at 346.0 nm in these interacting systems are strongly dependent on the isoelectric points of proteins and show adverse linear relationships with increasing ionic strength depending on the positive charges of the inorganic metal ions used to control the ionic strength of the medium, sufficiently disclosing that the electrostatic attraction performs an important role in the combination of proteins with WB. Linear responses were discovered between the enhanced RLS signals and the protein molecular weights (M_w), displaying the dimensions of scattered particles formed by proteins and WB make a key contribution to the RLS enhancements. An empirical equation is proposed which possibly displays the factors affecting the enhanced RLS signals of the interactions between proteins with negatively charged chromophores.     

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.     

A novel histidine assay using tetraphenylporphyrin manganese (III) chloride as a molecular recognition probe by resonance light scattering technique

A novel histidine-selective method has been developed for the determination of histidine in aqueous solutions by resonance light scattering (RLS) technique. At pH 8.0, the weak RLS intensity of tetraphenylporphyrin manganese (III) chloride [MnTPPCl] was greatly enhanced by the addition of histidine with the maximum peak located at 483 nm. Under the optimum conditions, it was found that the enhanced RLS intensity was in proportion to the concentration of histidine in the range 7.8 × 10⁻⁷-2.4 × 10⁻⁵ mol l⁻¹. Low detection limit of 9.2 × 10^-8 mol l⁻¹ has been achieved. The histidine concentrations in synthetic samples and real samples were determined with satisfactory results. The sensitivity and selectivity of this method are high enough to permit the determination of trace amounts of histidine without any significant interference from high levels of other components such as common anions and especially, other amino acids.     

甲醇-水、乙醇-水体系聚集态的共振散射光谱

自从Ｐａｓｔｅｒｎａｃｋ[1,2]使用普通荧光分光光度计建立共振光散射(ＲＬＳ)技术以来,人们用该技术建立了十分灵敏的蛋白质和核酸的分析方法[3,5]。同时该技术在研究化合物在溶液中的聚集态方面也有了较为广泛的应用,一般认为分子聚集体的形成是引起ＲＬＳ增强的主要原因[6]。已知甲醇、乙醇与水互溶形成均匀的溶液体系,那么在这些溶液体系中是否也存在某种形式的分子聚集体呢?为此本文研究了不同浓度的甲醇水溶液和乙醇水溶液的ＲＬＳ光谱。1　实验部分1 1　试剂和仪器甲醇(上海建鑫化工试剂厂,分析纯,含量%≥99 5);乙醇(安徽特酒总…     

蛋白质-四羧基铜酞菁体系的共振散射行为及其分析应用

在pH 3.0的Clark-Lub's 缓冲溶液中,蛋白质与四羧基铜酞菁(CuPc(COOH)4)作用,使体系的共振光散射增强,在λ=389 nm处光散射强度最大.增强作用的强弱与蛋白质的含量成正比,据此建立了共振光散射测定蛋白质的新方法.此方法对牛血清白蛋白(BSA)、人血清白蛋白(HSA)、免疫球蛋白G(IgG)、鱼精蛋白(Protamine)、血清总蛋白(TP)的测定范围分别为0.05～1.60 mg/L、 0.025～1.60 mg/L、 0～1.45 mg/L、 0.1～1.50 mg/L、 0～1.60 mg/L,相应检出限分别为1.12×10-2 mg/L、 1.11×10-2 mg/L、 1.95×10-2 mg/L、 3.61×10-3 mg/L、 4.34×10-3 mg/L.方法应用于实际人血清样品中总蛋白的测定,结果与考马斯亮蓝法基本一致.     

Determination of Nucleic Acids Based on their Resonance Light Scattering Enhancement Effect on Metalloporphyrin Derivatives

A new method for the determination of nucleic acids at nanogram per mL level is proposed based on the enhanced resonance light scattering (RLS) signal resulting from the interaction of metalloporphyrins with nucleic acids. Under optimum conditions, the weak RLS signal of metalloporphyrin is enhanced by nucleic acids, and the enhanced RLS intensity is proportional to the concentration of nucleic acids. The detection limits of calf thymus DNA were 3.5ngmL^–1, 2.9ngmL^–1 and 1.0ngmL^–1 for three metalloporphyrins, respectively. Synthetic samples were determined with satisfactory results.     

Determination of proteins with Alizarin Red S by Rayleigh light scattering technique

A new protein determination method by enhanced Rayleigh light scattering (RLS) technique has been developed. In acid condition (pH=3.60), RLS of 1,2-dihydroxyanthraquinone-3-sulfonate (Alizarin Red S) can be greatly enhanced by addition of proteins, resulting in two characteristic peaks, 360 and 505 nm, respectively. The new protein assay is based on the RLS enhancement and spectrum change. The optimum condition for the reaction was investigated. The linear range is 0.20-24.9 μg ml⁻¹ for BSA and 0.20-15.5 μg ml⁻¹ for HSA. The detection limits (S/N=3) are 9.59 ng ml⁻¹ for BSA and 9.51 ng ml⁻¹ for HSA. The results of determination for human serum samples were comparable to those obtained by Bradford method. The binding stoichiometry was determined.     

蛋白质-SDS-罗丹明B体系的共振光散射光谱及其分析应用

研究了阴离子表面活性剂十二烷基硫酸钠(SDS),阳离子染料罗丹明B,与蛋白质相互作用的共振光散射(RLS)光谱及用于蛋白质的测定.实验表明,在pH 4.35的酸性介质中,SDS的共振光散射强度较小,它与蛋白质结合后,共振光散射强度能得到增强,但加入阳离子染料罗丹明B后,共振光散射强度显著增强.在λ=332.0 nm处,ΔIRLS最大,并且增强的共振光散射信号与蛋白质的浓度成正比.据此建立了一种测定蛋白质的新方法,该方法灵敏度高,对HSA的检出限达到1.9 ng/mL,线性范围为0.01～5.0 μg/mL.用于人血清样品中蛋白质的测定,回收率为94.0%～105.5%.     

Rapid and sensitive determination of proteins by enhanced resonance light scattering spectroscopy of sodium lauroyl glutamate

A rapid and sensitive method for the determination of proteins is proposed based on the measurements of the enhanced resonance light scattering (RLS) spectroscopy of sodium lauroyl glutamate (SLG). Under the optimum conditions, the interaction between SLG and proteins occurred rapidly, resulting in greatly enhanced RLS intensity with the maximum peak located at 394 nm. It was found that the enhanced RLS intensities were in proportion to the concentrations of proteins in the range of 0.01-3.1 μg ml⁻¹ depending on the kind of proteins. The detection limits were below 6 ng ml⁻¹. Compared with some other methods for the determination of proteins, this method shows high sensitivity, low detection limit and simplicity. This is an inexpensive, simple and fast one-step procedure which requires only measuring the RLS intensities. Human serum samples were determined with satisfactory results.     

钙试剂-蛋白质体系的共振光散射光谱研究

基于蛋白质对钙试剂共振光散射的增强作用,拟定了一种测定蛋白质的共振光散射法。在pH 4.00的水溶液中,钙试剂在595 nm处的共振光散射增强与蛋白质浓度呈线性关系。此方法对人血清白蛋白、牛血清白蛋白的检出限分别可达0.068和0.085μg/mL。同时得到钙试剂与人血清白蛋白和牛血清白蛋白的结合常数(K)以及结合位点数(n)分别为8.15×104mol/L,0.18和7.67×104mol/L,1.5。     

Determination of deoxyribonucleic acids by a resonance light scattering technique and its application

For the first time, acetamiprid has been used to determine nucleic acid (DNA) using the resonance light scattering (RLS). The RLS of acetamiprid was greatly enhanced by DNA in the range of pH 1.6-1.8. A RLS peak at 313 nm was found, and the enhanced intensity of RLS at this wavelength was proportional to the concentration of DNA. The linear range of the calibration curve was 0-11.0 microg ml(-1) with the detection limit of 20 ng ml(-1). The nucleic acids in synthetic sample and in rice seedling extraction were determined satisfactorily. The interaction mechanism of acetamiprid and DNA is discussed. Mechanism studies show that the enhanced RLS is due to the aggregation of acetamiprid in the presence of DNA.     

A High-Sensitivity Assay of Nucleic Acids with Tetraphenyl Porphyrin Cobalt Chlorine by Resonance Light Scattering Technique

Abstract

A novel probe, tetraphenyl porphyrin cobalt chlorine (CoTPPCl), is first applied to determine nucleic acids at the nanogram level based on the measurement of resonance light scattering (RLS) signals, which result from the interaction of CoTPPCl with nucleic acids. Under pH 6.37 conditions, the reaction between CoTPPCl and nucleic acid enhances the weak resonance light scattering (RLS) signal of CoTPPCl, and the enhanced light scattering intensity is proportional to the concentration of nucleic acid. The method is sensitive (3.45 ng/mL for ctDNA), simple (one step and a common fluorimeter), and tolerant of the metal ions and other coexistent substances. The mode of the combination between CoTPPCl and nucleic acids and the reasons for RLS enhancement are clearly clarified. Synthetic samples were determined with satisfactory results.     

Immunoassay by detecting enhanced resonance light scattering signals of immunocomplex using a common spectrofluorometer

A sensitive, highly specific immunoassay method has been developed by measuring the enhanced resonance light scattering (RLS) signals of immunoreactions with simultaneously scanning both the excitation and the emission monochromators of a common spectrofluorometer. For a given content of antibody (Ab), the RLS signals of an immunoreaction follow Gaussian distribution with antigen (Ag) concentration. The central position of the Gaussian curve represents the concentration of given Ab, and the half bandwidth has proved to be a characteristic constant of a given Ab-Ag immunoreaction. With the RLS signals, the limit of detection for human immunoglobulin G (HIgG) in serum samples could reach 10 ng ml⁻¹, and the concentration of HIgG in blood serum samples could be detected with the recovery of 90.2-107.7% and R.S.D. of 0.8-2.7%. The results of determination for three human serum samples are identical to those obtained by immunoturbidimetry.     

Resonance Light Scattering Imaging Determination of Heparin

Light scattering submicroscopic particles such as metallic particles and particles of other composition have high-producing power and can be used as fluorescent analogs and tracers labels in clinical and biological applications1-3. Their light scattering …     

溴化十六烷基三甲铵存在下固红VR盐与fsDNA作用的共振光散射增强研究及其分析应用

研究了在阳离子表面活性剂溴化十六烷基三甲铵(CTMAB)存在下, 阴离子染料固红VR盐(FVR)和鱼精脱氧核糖核酸(fsDNA)作用的共振光散射(RLS)光谱特性、影响因素和最佳反应条件. 在pH 5.72和离子强度低于0.01 mol/L的条件下, fsDNA和CTMAB对FVR的共振光散射光谱有协同增强作用, 产生最大散射波长为361 nm的共振光散射增强(RLSE)信号. 在优化实验条件下, 测定fsDNA的线性范围为0.01～2.0 mg/L, 检出限可达2.5 μg/L. 方法能用于合成样中DNA的测定.     

花菁-脱氧核糖核酸的共振光散射光谱及其分析应用

研究了一种苯并噻唑阳离子花菁与脱氧核糖核酸(DNA)作用的共振光散射光谱,在pH 6.0的六次甲基四胺-HCl缓冲介质中,痕量DNA的加入使花菁在590nm的共振光散射强度显著增强。在最佳实验条件下,增强的共振光散射强度与DNA浓度具有良好的线性关系,据此建立了一种测定DNA的共振光散射光谱法。方法的线性范围为:小牛胸腺DNA(CT DNA),0～20μg/mL,鱼精子DNA(FS DNA),0～15μg/mL;检出限分别为0.005μg/mL和0.008μg/mL。该方法已用于合成样品中DNA的测定。     

藉L-半胱氨酸包覆的ZnS纳米粒子的共振光散射定量分析蛋白质

合成和表征了功能性L-半胱氨酸包覆的ZnS纳米粒子。在pH5．12的NaAc-HAc溶液介质中,L-半胱氨酸包覆ZnS纳米粒子于波长308．0nm处出现共振光散射峰。一定量蛋白质的加入能明显增强体系的共振光散射,且峰强度增加值与蛋白质浓度间存在良好线性关系,据此建立了一种灵敏的测定微量蛋白质的方法。用L-半胱氨酸包覆ZnS纳米粒子作为探针,不仅克服了有机染料可能出现的光漂白等缺点,而且本身不具毒性。该法用于人血清试样中总蛋白的测定,其结果与临床数据一致。