Binding of malachite green promotes stability and shows preference for a human telomere DNA G-quadruplex

A single-stranded human telomere DNA sequence can fold into an intramolecular G-quadruplex structure, which has been shown to inhibit telomerase activity. Small molecules that selectively target and stabilise the G-quadruplex structure have been proposed as potential anticancer drugs. In this study, we analysed the properties of binding of malachite green, a cationic triphenylmethane dye, to the G-quadruplex of d[(T₂AG₃)₄] by UV spectroscopy of thermal melting analysis, a competitive equilibrium dialysis assay, and absorption and circular dichroism spectroscopies. When binding to malachite green, the quadruplex structure that formed in the presence of K⁺ ions was stabilised with an increase in melting temperatures by 6 °C. Malachite green showed selective binding to the G-quadruplex in the presence of duplex and single-stranded DNAs, owing to which it presents higher potential for anticancer therapy, compared to other triphenylmethane dyes. The induced signals of circular dichroism indicate that the binding mode of malachite green involves intercalation between adjacent guanine tetrads of the G-quadruplex.     

Voltammetric determination of malachite green in fish samples based on the enhancement effect of anionic surfactant

In this work, an electrochemical method for the determination of malachite green was developed on the basis of enhancement effect of an anionic surfactant: sodium dodecyl benzene sulfonate. It is found that the oxidation peak current of malachite green at carbon paste electrode significantly increases in the presence of low concentration of sodium dodecyl benzene sulfonate in pH 6.5 phosphate buffer, suggesting that sodium dodecyl benzene sulfonate shows obvious enhancement effect for the determination of malachite green. The experimental parameters, such as supporting electrolyte, kind of surfactant, concentration of sodium dodecyl benzene sulfonate and accumulation time, were optimized, and then a sensitive and convenient electrochemical method was proposed for the determination of malachite green. The oxidation peak current is proportional to the concentration of malachite green over the range from 8.0 × 10⁻⁹ to 5.0 × 10⁻⁷ mol l⁻¹, and the detection limit is 4.0 × 10⁻⁹ mol l⁻¹ after 5 min of accumulation. Finally, this new method was successfully employed to detect malachite green in fish samples. Published in Russian in Elektrokhimiya, 2008, Vol. 44, No. 8, pp. 1019–1024. The text was submitted by the authors in English.     

Electrochemical determination of malachite green using a multi-wall carbon nanotube modified glassy carbon electrode

A multi-wall carbon nanotube (MWNT) film-modified electrode is described for the determination of malachite green (MG). The electrochemical profile of MG was examined using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), suggesting that the MWNT film facilitates the electron transfer of MG in terms of a potential shift and then significantly enhances the oxidation peak current of MG. The experimental parameters, such as supporting electrolyte, thickness of MWNT film, scan rate and accumulation time, were optimized. Consequently, a sensitive and convenient electrochemical method is proposed for the determination of MG. The oxidation peak current is proportional to the concentration of MG over the range from 5.0 × 10⁻⁸ to 8.0 × 10⁻⁶ mol L⁻¹ obeying the following equation: i_p = 0.09 + 1.19 × 10⁷ C (r = 0.995, i_p in μA, C in mol L⁻¹). The detection limit is 6.0 × 10⁻⁹ mol L⁻¹ (signal to noise = 3) after 5 min of accumulation. Moreover, this method possesses good reproducibility (RSD is 5.6%, n = 8) as well as long-term stability. Finally, the new method was employed to determine MG in fish samples. Correspondence: W. Huang, Department of Chemistry, Hubei Institute for Nationalities, Enshi 445000, P.R. China     

Simple and sensitive fluorometric sensing of malachite green with native double-stranded calf thymus DNA as sensing material

A novel fluorometric sensing of malachite green is proposed in this paper. The native double-stranded calf thymus DNA was used as sensing material. In the presence of native double-stranded calf thymus DNA, malachite green could interact with the DNA, which resulted in a strong fluorescence emission. The fluorescent intensity was linear with malachite green concentration in the range of 4.0 × 10⁻¹⁰ − 1.8 × 10⁻⁷ g ml⁻¹ and the limit of detection was 2.0 × 10⁻¹⁰ g ml⁻¹. Before fluorescence measurement, the only required operation is the mixing of two solutions. So, this method is rather simple and rapid. The method is very safe for the analyst. Furthermore, the mechanism for fluorescence enhancing of native double-stranded calf thymus DNA on MG was proposed based on a series of experiments. The results suggest that the interaction between MG and calf thymus DNA is intercalation in nature.     

孔雀石绿的磁性免疫层析快速检测方法研究

开发了一种适用于现场快速检测孔雀石绿(MG)的免疫层析试纸条,在超顺磁性纳米微球上偶联MG单克隆抗体作为检测探针,分别将孔雀石绿完全抗原(MG-B SA)和羊抗鼠IgG喷涂于NC膜的T线和C线.结果 发现,T线最佳喷涂量为0.25 mg/mL,抗体最佳偶联量为20 μg,构建的试纸条可在25 min内实现养殖用水及鱼肉...     

免标记比色核酸适配体传感器同步快速检测孔雀石绿和无色孔雀石绿

研究以双特异性核酸适配体A3作为传感探针、纳米金(AuNPs)为指示剂、NaCl溶液为聚集诱导剂,构建了一种新型的免标记AuNPs比色生物传感器,可实现水产品中孔雀石绿(MG)和无色孔雀石绿(LMG)的同步、快速、可视化检测。该方法的检测原理是核酸适配体A3对MG和LMG有双特异性识别能力,可作为MG和LMG理想的识别受体。它可通过静电作用吸附到AuNPs表面,保护AuNPs并抑制高盐溶液诱导的聚集,AuNPs溶液颜色不变,即为红色;当加入靶标MG或LMG后,该核酸适配体能够与靶标特异性结合,并从AuNPs表面上解离,AuNPs失去保护作用而在高盐溶液诱导下发生聚集,溶液颜色由红变蓝。根据颜色变化,可通过肉眼定性或通过光谱仪定量分析MG和LMG的残留量。该方法首先将50μL的核酸适配体A3(终浓度150 nmol/L)与150μL的AuNPs(终浓度1.25 nmol/L)混合,室温孵育6 min。随后加入50μL待测液,室温孵育30 min。最后加入50μL NaCl(终浓度150 mmol/L), 4 min后观察溶液颜色变化,并分别测定MG和LMG在520 nm和650 nm下的...     

Surfactant-sensitized malachite green method for trace determination of orthophosphate in aqueous solution

A surfactant-sensitized spectrophotometric method for determination of trace orthophosphate has been developed using anion surfactant (Ultrawet 60 L) with molybdate and malachite green in low acidic medium (pH_T 1.0). The method detection limit (3 × standard deviation of blank, n = 10) was 8 nM and the calibration curve was linear over a range of 10-400 nM (r² = 0.997). The molar absorptivity was 1.26 × 10⁵ L mol⁻¹ cm⁻¹ at 600 nm with the background correction at 530 nm. The precision of method was 3.4% at 50 nM and 2.4% at 100 nM orthophosphate (n = 10). The hydrolysis of eight organic phosphorus and polyphosphate compounds was less than 2% of the total phosphorus present (5-10 μM). This method showed less arsenate interference than previous methods, with only 3% even in the presence of orthophosphate in the samples. No interference of silicate up to 40 μM was observed. Background anions (in an order of SO₄²⁻ > NO₃⁻ > Cl⁻) have greater effects than cations (Ca²⁺ > Mg²⁺ > Na⁺) on the reagent blank and the molar absorptivity of the color product.     

Elimination of zinc interference in the voltammetric determination of cobalt by using iminodiacetic acid

The voltammetric determination of cobalt carried out by adsorbing cobalt dimethylglyoxime onto a mercury electrode and subsequently measuring the current for the reduction of the adsorbed species is subject to interference by zinc. If this interference is neglected and the determination carried out by standard additions large errors may result. The interference can be eliminated by adding iminodiacetic acid.     

A new reagent system for the highly sensitive spectrophotometric determination of selenium

Highly sensitive and simple spectrophotometric determination of selenium is described for the determination of selenium(IV) using a new reagent leuco malachite green. The method is based on the reaction of selenium(IV) with potassium iodide in an acidic condition to liberate iodine, the liberated iodine oxidizes leuco malachite green to malachite green dye. The green coloration was developed in an acetate buffer (pH 4.2–4.9) on heating in a water bath (∼ 40 °C). The formed dye exhibits an absorption maximum at 615 nm. The method obeys Beer’s law over a concentration range of 0.04–0.4 μg mL⁻¹ selenium. The molar absorptivity and Sandell’s sensitivity of the color system were found to be 1.67 × 10⁵ L mol⁻¹ cm-¹ and 0.5 ng cm⁻², respectively. The optimum reaction conditions and other analytical parameters have been evaluated. The proposed procedure has been successfully applied to the determination of selenium in real samples of water, soil, plant material, human hair, and cosmetic samples. The results were compared to those obtained with the reference method. Statistical analysis of the results confirms the precision and accuracy of the proposed method. In addition, the developed method is cost-effective and involves easily accessible instrumentation technique which can be used by ordinary research laboratories.     

A voltammetric method for diuron determination in river water

A sensitive, simple, fast, and inexpensive method for determining diuron in water using square wave voltammetry and a glassy carbon electrode is presented. This method was developed to evaluate diuron contamination in river waters close to sugarcane cultivation, located in the metropolitan region of João Pessoa, in the state of Paraíba, Brazil. An analytical curve with a linear response ranging from 38.5–115.0 nmol L⁻¹ (r² = 0.993), a LOD equal 0.2 nmol L⁻¹, and recovery rates from 88.0 to 108.0 %. The proposed method does not require any previous chemical treatment of the sample, thus contributing to green analytical chemistry.     

磁性碳纳米管对水中孔雀石绿的吸附性能研究

（1）以磁性碳纳米管吸附处理孔雀石绿溶液,考察了吸附剂用量、孔雀石绿浓度以及温度等对吸附平衡的影响。结果表明,该吸附不受溶液pH值的影响,符合准二级动力学方程（R2＞0.999）。不同温度下,该吸附过程满足Langmuir方程（R2>0.97）和Freundlich方程（R2>0.98）,qmax、KF随温度升高而变大。D-R等温方程拟合结果表明该吸附的机理是以化学吸附为主。热力学参数计算结果表明,该吸附是一个熵增的自发吸热过程,温度越高越利于吸附的进行。     

Development of a PRiME cartridge purification method for rapid determination of malachite green and leucomalachite green in Chinese softshell turtle

A high‐throughput PRiME (process, robustness, improvements, matrix effects, ease of use) sample purification procedure was developed to simplify the multiple steps of traditional SPE in extracting the malachite green and leucomalachite green in Chinese softshell turtle (Pelodiscus sinensis). The sample loading volume, extracting solvent type, and pH value of the employed PRiME hydrophilic‐lipophilic balance cartridge for sample purification were optimized to be 3 mL, acetonitrile, and pH 5, respectively. In comparison with traditional SPE, the PRiME process is cost‐effective, solvent‐saving, and simple to operate, which only consists of a passing through step without traditional sorbent conditioning and impurity washing. Afterward, eluate was analyzed by ultra‐performance liquid chromatography‐tandem mass spectrometry, and the proposed method was validated for linearity (R² > 0.9992), intraday precision (2.44–3.22%), interday precision (3.28–6.58%), sensitivity (LOD ≤ 0.18 μg/kg and, LOQ ≤ 0.60 μg/kg), and recovery (88.7–94.1%, RSD < 6.79%). The results indicated that the PRiME technique can simplify the sample preparation procedure by avoiding the tedious steps, such as conditioning, washing, etc. It would be of significant interest for environmental and food safety applications in the market of Chinese softshell turtle and related products.     

A New Spectrophotometric Method for the Determination of Ascorbic Acid Using Leuco Malachite Green

A new simple and sensitive and selective spectrophotometric method has been developed for the determination of ascorbic acid (AA) at trace level using a new reagent, leuco malachite green (LMG). AAreacts with potassium iodide‐iodate solution under acidic conditions to liberate iodine and the liberated iodine selectively oxidizes LMG to MG dye. The colour of the dye was measured at 620 nm. Beer's law is obeyed over the concentration range of 0.8–8 iμg μAA per 25 mL of final solution (0.032–0.32 ppm). The apparent molar absorptivity and Sandell's sensitivity of the method were found to be 2.98 × 10⁵ l mol⁻¹ cm⁻¹, 0.0042 μg cm⁻², and respectively. Statistical treatment of the experimental results indicates that the method is precise and accurate. The method is free from interference of common ions and many of the ingredients commonly found in pharmaceuticals. The reliability of the method was established by parallel determination against Leucocrystal violet (LCV) method. The method described was satisfactorily applied for the determination of AA in fruit juices, pharmaceuticals and biological samples.     

Resonance Rayleigh scattering method for the recognition and determination of double-stranded DNA using amikacin

In weakly acidic buffer medium, the interaction of amikacin with calf thymus DNA, yeast RNA and denatured DNA has been investigated by using resonance Rayleigh scattering (RRS) technique. The result shows that calf thymus DNA is capable of enhancing the RRS intensity of the amikacin, while yeast RNA and denatured DNA have very little enhancement effect. Based on the characteristics, a sensitive assay for detecting double-stranded DNA in the presence of denatured DNA and yeast RNA has been developed. The enhancement of the RRS signal is directly proportional to the concentration of double-stranded DNA in the range 0.02-12.0 μg ml⁻¹ for calf thymus DNA and its detection limit (3σ) is 2.5 ng ml⁻¹. The method shows a wide linear range and high sensitivity, and almost no interference can be observed from RNA, denatured DNA, amino acid and most of the metal ions. The trace amounts of nucleic acid in synthetic samples and practical samples are determined with satisfactory results. Therefore, the proposed method is promising for as an effect means for recognition in vivo and determination in situ of double-stranded DNA.     

Lighting‐Up of the Dye Malachite Green with Mercury(II)–DNA and Its Application for Fluorescence Turn‐Off Detection of Cysteine and Glutathione

This work describes a novel strategy for the highly sensitive and selective detection of cysteine (Cys) and glutathione (GSH) based on the Hg²⁺–AGRO100–malachite green (MG) complex system. The dye MG, which has a very low quantum yield in aqueous solution by itself, can bind with the thymine‐rich DNA AGRO100 in the presence of Hg²⁺ ions to generate a striking fluorescence intensity enhancement of 1000‐fold. As sulfur‐containing amino acids, Cys and GSH effectively sequester Hg²⁺ ions from the Hg²⁺–AGRO100–MG complex structure to switch the ‘lit‐up’ chemosensor to the ‘off’ state (about a 50‐fold fluorescence intensity decrease), thus providing a facile, but effective, method to probe for Cys/GSH. The fluorescence titration, UV absorption, CD, and Raman spectra provide some insight into the structural and chemical basis for the enhancement effect. The formation of the Hg²⁺–AGRO100–MG complex significantly affects the electronic structure and conformation of the MG molecule by leading to an extended π system, which is the likely origin of the observed striking fluorescence intensity enhancement. Notably, the proposed sensing platform exhibits exquisite selectivity and sensitivity toward Cys/GSH with limits of detection of 5 nM for Cys and 10 nM for GSH, respectively. Furthermore, the straightforward assay design avoids labeling of the probe, uses only commercially available materials, and still displays comparable sensitivity and excellent selectivity.     

A simple visual method for DNA detection based on the formation of gold nanoparticles

A simple visual method for DNA detection during the formation of gold nanoparticles (AuNPs) was developed based on different electrostatic properties of single strand DNA (ssDNA) and double strand DNA (dsDNA). Since the ssDNA is easy to bind to AuNPs due to its exposed bases which could prevent salt-induced aggregation of AuNPs. The dsDNA always present negative charge because its negatively charged phosphate backbone is exposed. In this case, the dsDNA could disturb the adsorption between dsDNA and AuNPs and result in non-aggregation of AuNPs. After hybridization, chloroauric acid and ascorbic acid were added to the mixture solution, and the solution changed to red immediately and turned to purple in 10 min in the present of target DNA. TEM results confirmed that the change of color stemed from aggregation of AuNPs. In order to obtain accurate results by naked eye, the DNA detection assay should be conducted under pH 7.0.     

A simple method for the determination of malachite green and leucomalachite green residues in fish by a modified QuEChERS extraction and LC/MS/MS

A simple method using LC/MS/MS was developed and validated to determine residues of malachite green (MG) and leucomalachite green (LMG) in fish fillet. A modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) technique was used to perform the sample preparation. The optimal extraction and cleanup conditions were established using an experimental design. The validation parameters obtained to determine both MG and LMG complied with the requirements established by regulatory agencies for the presence of such substances in fish, which establish that the method must attain, at least, a minimum required performance limit of 2 ng/g. The accuracy values ranged between 95 and 107%, and the precision values were lower than 11.2%. The method was used in the analysis of tilapia samples (n = 20) commercialized in Campinas, SP, Brazil. None of the samples presented detectable levels of MG or LMG residues.     

A simple and green analytical method for determination of iron based on micro flow analysis

A simple, inexpensive and reagent-less colorimetric micro flow analysis (μFA) system was implemented in a polymethyl methacrylate (PMMA) micro fluidic manifold. A T-shaped micro channel on a PMMA chip was fabricated by laser ablation and topped with molded polydimethylsiloxane (PDMS). The fabricated μFA system was integrated with the optical components as detector and applied to the determination of iron in water samples. It is based on the measurement of Fe(III)-nitroso-R salt complex at 720 nm formed by the reaction between Fe(III) and nitroso-R salt in an acetate buffer solution pH 5. The proposed μFA consumed very small amount of reagent and sample, it released waste of less than 2.0 mL h⁻¹. The relative standard deviation (R.S.D.) was less than 2% (n = 11) with the recovery of 98.7 ± 0.12 (n = 5). The linear range for the determination of iron in water samples was over the range of 0.05-4.0 μg mL⁻¹ with a correlation coefficient (r²) of 0.9994. The limit of detection (3σ) and limit of quantitation (10σ) were 0.021 μg mL⁻¹ and 0.081 μg mL⁻¹, respectively with a sample throughput of 40 h⁻¹.     

A simple visual method for DNA detection based on the formation of gold nanoparticles

A simple visual method for DNA detection during the formation of gold nanoparticles (AuNPs) was developed based on different electrostatic properties of single strand DNA (ssDNA) and double strand DNA (dsDNA). It could identify target DNA in 10 min.