Label-Free Colorimetric Detection of Mercury (II) Ions Based on Gold Nanocatalysis.

Herein, a label-free colorimetric nanosensor for Hg(II) is developed utilizing the hindering effect of Hg(II) on the kinetic aspect of gold nanoparticle (AuNPs) growth on the surface of gold nanostars (AuNSs). H-AuNS probes are synthesized and modified by 2-[4-(2-hydroxyethel) piperazine-1-yl] ethanesulfonic acid (HEPES). After the formulation of the reagents and testing conditions are optimized, HEPES-capped AuNSs (H-AuNSs) demonstrates good selectivity and sensitivity towards Hg(II) determination. A H-AuNS probe, in the presence of HCl/Au(III)/H2O2, is capable of detecting a Hg(II) concentration range of 1.0 nM⁻100 µM, with a detection limit of 0.7 nM, at a signal-to-noise ratio of 3.0, and a visual detection limit of 10 nM with naked eyes. For practicality, the H-AuNS probe is evaluated by measuring Hg(II) in the environmental water matrices (lake water and seawater) by a standard addition and recovery study. The detection limits for environmental samples are found to be higher than the lab samples, but they are still within the maximum allowable Hg concentration in drinking water (10 nM) set by the US Environmental Protection Agency (EPA). To create a unique nanosensor, the competitive interaction between Hg(II) and Pt(IV) toward the H-AuNSs probe is developed into a logic gate, improving the specificity in the detection of Hg(II) ions in water samples.

A dipole-assisted solid-phase extraction microchip combined with inductively coupled plasma-mass spectrometry for online determination of trace heavy metals in natural water.

We employed a polymeric material, poly(methyl methacrylate) (PMMA), for fabricating a microdevice and then implanted the chlorine (Cl)-containing solid-phase extraction (SPE) functionality into the PMMA chip to develop an innovative on-chip dipole-assisted SPE technique. Instead of the ion-ion interactions utilized in on-chip SPE techniques, the dipole-ion interactions between the highly electronegative C-Cl moieties in the channel interior and the positively charged metal ions were employed to facilitate the on-chip SPE procedures. Furthermore, to avoid labor-intensive manual manipulation, a programmable valve manifold was designed as an interface combining the dipole-assisted SPE microchip and inductively coupled plasma-mass spectrometry (ICP-MS) to achieve the fully automated operation. Under the optimized operation conditions for the established system, the detection limits for each analyte ion were obtained based on three times the standard deviation of seven measurements of the blank eluent solution. The limits ranged from 3.48 to 20.68 ng L(-1), suggesting that this technique appears uniquely suited for determining the levels of heavy metal ions in natural water. Indeed, a series of validation procedures demonstrated that the developed method could be satisfactorily applied to the determination of trace heavy metals in natural water. Remarkably, the developed device was durable enough to be reused more than 160 times without any loss in its analytical performance. To the best of our knowledge, this is the first study reporting on the combination of a dipole-assisted SPE microchip and elemental analysis instrument for the online determination of trace heavy metal ions.

Separation of total lipids on human lipoproteins using surfactant-coated multiwalled carbon nanotubes as pseudostationary phase in capillary electrophoresis.

Surfactant-coated multiwalled carbon nanotubes (MWNTs) were used as pseudostationary phase (PSP) in CE to investigate the total lipids of high-density lipoproteins and low-density lipoproteins. To optimize the CE conditions, several experimental factors including carbon nanotube concentration, bile salt concentration, sodium phosphate (PB) concentration, organic modifier concentration and buffer pH value have been examined. In addition, the CE capillary temperature and applied voltage have also been examined. The optimal separation buffer selected was a mixture of 3.2 mg/L MWNT, 50 mM bile salt, 10 mM PB, 20% 1-propanol, pH 9.5. The optimal capillary temperature and applied voltage selected were 50°C and 20 kV, respectively. Phosphatidyl choline (PC) has been used as a model analyte and investigated by the optimal CE method. The linear range for PC was 0.1-3 mg/mL with a correlation coefficient of 0.9934, and the concentration LOD was 0.055 mg/mL. The optimal CE method has been used to characterize the total lipids of high-density lipoprotein and low-density lipoprotein. At absorbance 200 nm, one major peak and two or three minor peaks showed for the total lipids of lipoproteins within 13 minutes. Resolutions of the total lipids were enhanced using surfactant-coated MWNTs as PSPs in the CE separation buffer. However, resolutions of the total lipids were not enhanced using surfactant-coated single-walled carbon nanotubes as PSPs in the CE separation buffer.

Green Synthesis of Carbon Quantum Dots and Carbon Quantum Dot-Gold Nanoparticles for Applications in Bacterial Imaging and Catalytic Reduction of Aromatic Nitro Compounds.

This study delves into the green synthesis and multifaceted applications of three types of carbon quantum dots (CQDs), namely, CQDs-1, CQDs-2, and CQDs-3. These CQDs were innovatively produced through a gentle pyrolysis process from distinct plant-based precursors: genipin with glucose for CQDs-1, genipin with extracted gardenia seeds for CQDs-2, and genipin with whole gardenia seeds for CQDs-3. Advanced analytical techniques, including X-ray photoelectron spectroscopy (XPS) and Fourier-transform infrared spectroscopy (FT-IR), were employed to detail the CQDs' structural and surface characteristics, revealing their unique functional groups and surface chemistries. The study further explores the CQDs' bioimaging potential, where confocal fluorescence microscopy evidenced their swift uptake by Escherichia coli bacteria, indicating their suitability for bacterial imaging. These CQDs were also applied in the synthesis of gold nanoparticles (AuNPs), acting as reducing agents and stabilizers. Among these, CQD3-AuNPs were distinguished by their remarkable stability and catalytic efficiency, achieving a 99.7% reduction of 4-nitrophenol to 4-aminophenol in just 10 min and maintaining near-complete reduction efficiency (99.6%) after 60 days. This performance notably surpasses that of AuNPs synthesized using sodium citrate, underscoring the exceptional capabilities of CQD3-AuNPs. These insights pave the way for leveraging CQDs and CQD-stabilized AuNPs in bacterial imaging and catalysis, presenting valuable directions for future scientific inquiry and practical applications.

Combined Electrochemical Deposition and Photo-Reduction to Fabricate SERS-Active Silver Substrates: Characterization and Application for Malachite Green Detection in Aquaculture Water.

This research introduces a novel approach using silver (Ag) nanostructures generated through electrochemical deposition and photo-reduction of Ag on fluorine-doped tin oxide glass substrates (denoted as X-Ag-AgyFTO, where 'X' and 'y' represent the type of light source and number of deposited cycles, respectively) for surface-enhanced Raman spectroscopy (SERS). This study used malachite green (MG) as a Raman probe to evaluate the enhancement factors (EFs) in SERS-active substrates under varied fabrication conditions. For the substrates produced via electrochemical deposition, we determined a Raman EF of 6.15 × 104 for the Ag2FTO substrate. In photo-reduction, the impact of reductant concentration, light source, and light exposure duration were examined on X-Ag nanoparticle formation to achieve superior Raman EFs. Under optimal conditions (9.0 mM sodium citrate, 460 nm blue-LED at 10 W for 90 min), the combination of blue-LED-reduced Ag (B-Ag) and an Ag2FTO substrate (denoted as B-Ag-Ag2FTO) exhibited the best Raman EF of 2.79 × 105. This substrate enabled MG detection within a linear range of 0.1 to 1.0 µM (R2 = 0.98) and a detection limit of 0.02 µM. Additionally, the spiked recoveries in aquaculture water samples were between 90.0% and 110.0%, with relative standard deviations between 3.9% and 6.3%, indicating the substrate's potential for fungicide detection in aquaculture.

A highly selective and sensitive fluorescence assay for determination of copper(II) and cobalt(II) ions in environmental water and toner samples.

In this study, a highly selective and sensitive fluorescence assay has been proposed for the determination of copper(II) and cobalt(II) ions in environmental water and toner samples. In the presence of hydrogen peroxide, copper(II) reacted with a new fluorescence reagent Amplex® UltraRed (AUR), forming a fluorescence product only at pH 7.0, while the fluorescence product of cobalt(II) with AUR formed only at pH 9.0. The fluorescence signal obtained was linear with respect to the copper(II) concentration over the range of 1.6-12.0 µM (R(2) = 0.988) and was linear with respect to the cobalt(II) concentration over the range of 45.0 nM to 1.0 µM (R(2) = 0.992). The limits of detection (at a signal-to-noise ratio of 3) for copper(II) and cobalt(II) were 0.17 µM and 14.0 nM, respectively. Our present approach is simpler, faster, and more cost-effective than other techniques for the detection of copper(II) and cobalt(II) ions in environmental water samples and that of copper(II) ions in toner samples.

Gold nanosponges: green synthesis, characterization, and cytotoxicity.

A simply approach for the synthesis of Au nanostructures in tea infusions at room temperature is developed. By controlling the concentrations of tea infusions, Au nanostructures in various shapes and sizes have been prepared. From 1 x (original concentration) and 0.01 x (100 times diluted) tea infusions, 52.2 +/- 8.1 nm Au nanosponges (T-Au NSs) and 23 +/- 2 nm spherical Au nanoparticles (T-Au NPs) were prepared. The phytochemicals present on the surface of T-Au NSs were proved by surface-assisted laser desorption/ionization mass spectrometry, Fourier transform infrared spectrometry and capillary electrophoresis coupled with UV detection. The energy-dispersive X-ray spectroscopy and powder X-ray diffraction data reveal pure crystalline structures of the T-Au NSs. The dark field scattering images observe that the T-Au NSs have significant affinity toward HeLa cells. The cytotoxicity of T-Au NSs on HeLa cells is through caspase-3 activation.

Green synthesis of gardenia seeds-based carbon dots for bacterial imaging and antioxidant activity in aqueous and oil samples.

In this work, luminescent carbon dots with gardenia seeds as carbon precursors (GCDs) were synthesized using a one-step mild pyrolysis process and were then used as probes for imaging of bacterial (Escherichia coli). The GCDs showed a strong emission at 430 nm when excited at 370 nm. The relative fluorescence quantum yield of GCDs was found to be 1.13% in an aqueous medium. Rapid internalization of the GCDs by bacteria was confirmed by three colors (blue, green, and yellow) images that were obtained using confocal fluorescence microscopy. In addition, GCDs were noted to exhibit potent scavenging activities against DPPH˙, ˙OH, and ˙O2- free radicals. GCDs were also assayed as antioxidants in an oil sample by volumetric determination of the peroxide value. Thus, GCDs exhibited good antioxidant properties both in aqueous and oil media. In addition, a free fatty acid quantification kit in the presence of GCDs showed enhanced fluorescence detection of palmitic acid with a remarkably good limit of detection of 0.08 µM, which is lower than that in the absence of GCDs (0.76 µM). The proposed fluorescence method was then successfully used to determine the concentration of palmitic acid spiked in milk powder samples, with spiked recoveries of 82.6-109.6% and relative standard deviations of 0.9-4.6%.

Nafion/Silver Nanoparticles as an Electrochemically Sensitive Interface for the Detection of Ractopamine in Pork Liver.

Many countries have allowed farmers to feed ß-adrenergic receptor agonists, such as ractopamine (Rac), to animals to improve the quality of their meat. However, Rac consumption can cause health problems for humans; thus, detecting Rac in meat before its packaging is essential. Consequently, this study developed a simple and sensitive electrochemical sensor by modifying a glassy carbon electrode (GCE) with Nafion/silver nanoparticles (Nafion/AgNPs). When this electrochemical sensor is used to detect Rac, electrostatic interaction occurs between Nafion and Rac, and the AgNPs oxidize Rac; thus, the accumulation and electrochemical sensing of Rac are achieved. Differential pulse voltammetry indicated that the as-prepared Nafion/AgNP-GCE sensor exhibited suitable electrochemical sensing ability under optimum conditions (6.0 µL of 0.10% Nafion/AgNPs in a Britton-Robertson buffer solution with a pH of 1.8, an accumulation potential of -0.2 V, and a Rac accumulation duration of 300 s). Moreover, this sensor has an extremely low limit of detection and high sensitivity (1.60 × 10-3 ppm and 2.14 µA/ppm, respectively) in the Rac concentration range 7.50 × 10-3-1.00 ppm. The as-prepared sensor also exhibits satisfactory reproducibility and storage stability, with the corresponding relative standard deviations (RSDs) being 4.27% (n = 5) and 1.56% (n = 10), respectively. The proposed electrochemical sensor was successfully used to determine the Rac content in pig liver samples, with spiked recoveries of 95.2-101.8% and RSDs of 0.55-4.83% being achieved.

Selective detection of tricyclazole by optical technique using thiomalic acid-modified Au and Ag nanoparticle mixtures.

This study proposes the use of thiomalic acid-modified Au and Ag nanoparticle mixtures (TMA-Au/AgNP mixes) for the selective detection of tricyclazole. Upon the addition of tricyclazole, the color of TMA-Au/AgNP mixes solution changes from orange-red to lavender (red-shift). According to the density-functional theory calculations, tricyclazole-induced aggregation of TMA-Au/AgNP mixes through electron donor-acceptor interactions was proved. The sensitivity and selectivity of the proposed method are affected by the amount of TMA, volume ratio of TMA-AuNPs to TMA-AgNPs, pH value, and buffer concentration. The ratio of absorbance (A654/A520) of TMA-Au/AgNP mixes solution is proportional to the concentration of tricyclazole over the range 0.1-0.5 ppm with a linear correlation (R2 = 0.948). Moreover, the limit of detection was estimated at 0.028 ppm. The practicality of TMA-Au/AgNP mixes was validated for the determination of tricyclazole concentration in real samples (spiked recovery was 97.5%-105.2%), demonstrating its advantages of simplicity, selectivity, and sensitivity.

Aminoglycoside-mimicking carbonized polymer dots for bacteremia treatment.

Bacteremia and associated bacterial sepsis are potentially fatal and occur when the host response to microbial invasion is impaired or compromised. This motivated us to develop carbonized polymer dots (CPDsMan/AA) from a mixture of mannose (Man) and positively charged amino acids [AAs; lysine, arginine (Arg), or histidine] through a one-step mild pyrolysis procedure, which effectively inhibited drug-resistant bacterial strains isolated from septic patients. The as-prepared CPDsMan/AA showed broad-spectrum antibacterial activity, including multidrug-resistant bacteria, even in human plasma. The minimal inhibitory concentration of CPDsMan/Arg is ca. 1.0 µg mL-1, which is comparable to or lower than those of other tested antibiotics (e.g., ampicillin, gentamicin, and vancomycin). In addition to directly disrupting bacterial membranes, the CPDsMan/Arg feature a structure similar to aminoglycoside antibiotics that could bind to 16S rRNA, thereby blocking bacterial protein synthesis. In vitro cytotoxic and hemolytic assays demonstrated the high biocompatibility of the CPDsMan/AA. In addition, in vivo studies on methicillin-resistant Staphylococcus aureus-infected mice treated with the CPDsMan/Arg showed a significant decrease in mortality-even better than that of antibiotics. Overall, the synthesis of the CPDsMan/AA is cost-efficient, straightforward, and effective for treating bacteremia. The polymeric features of the CPDsMan/Arg, including cationic charges and specific groups, can be recognized as a safe and broad-spectrum biocide to lessen our reliance on antibiotics to treat systemic bacterial infections in the future.

Fluorescence detection of lead(II) ions through their induced catalytic activity of DNAzymes.

We have developed a fluorescence approach for the highly selective and sensitive detection of Pb(2+) ions using AGRO100, a G-quadruplex DNAzyme. The sensing strategy is based on Pb(2+) ions inducing increased DNAzyme activity of AGRO100 in the presence of hemin, which acts as a cofactor to catalyze H(2)O(2)-mediated oxidation of Amplex UltraRed (AUR). A test of eight aptamers of various sequences for the detection of Pb(2+) ions revealed that AGRO100 performed the best in terms of sensitivity. The AGRO100-AUR probe exhibited high selectivity (>100-fold) toward Pb(2+) ions over other tested metal ions. The fluorescence intensity (excitation/emission maxima, ca. 561/592 nm) of the AUR product was proportional to the concentration of Pb(2+) ions over the range 0-1000 nM, with a linear correlation (R(2) = 0.98). For 5 mM Tris-acetate (pH 7.4) solutions in the presence and absence of 100 mM NaCl, the AGRO100-AUR probe provided limits of detection (signal-to-noise ratio = 3) for Pb(2+) ions of 1.0 and 0.4 nM, respectively. We validated the practicality of the use of the AGRO100-AUR probe for the determination of the concentrations of Pb(2+) ions in soil samples. This approach allows the determination of the concentrations of Pb(2+) ions with simplicity, selectivity, and sensitivity.

Detection of mercury and phenylmercury ions using DNA-based fluorescent probe.

A DNA probe labeled with a 4-([4-(dimethylamino)phenyl]azo)benzoic acid (DABCYL) quencher and a carboxyfluorescein (FAM) donor at its 5'- and 3'-termini can be used for the detection of Hg(2+) ions and phenylmercury ions (PhHg(+)). This DNA probe possesses a random coil structure that changes into a hairpin-like structure upon binding Hg(2+) and PhHg(+) ions. As a result, the fluorescence of the FAM unit decreased through quenching between the donor and the quencher. In the presence of ethylenediaminetetraacetic acid (EDTA), the DNA probe allowed the selective detection of PhHg(+) ions at concentrations as low as 70.0 nM, mainly as a result of T-Hg(2+)-T coordination and π-π stacking between the Ph unit and DNA bases. A linear correlation existed between the fluorescence intensity and the concentration of PhHg(+) ions over the range from 0.10 to 1.0 µM (R(2) = 0.99). After acid hydrolysis and neutralization of the samples, all of the mercury species are converted to Hg(2+) ions, allowing us to use the DNA-based probe to determine the concentrations of total mercury species at the nM level. The practicality of this probe has been validated by the analyses of pond water and fish samples, showing its advantages of sensitivity, selectivity, and simplicity.

Gold nanoparticle probes for the detection of mercury, lead and copper ions.

Monitoring the levels of potentially toxic metal (PTM) ions (e.g., Hg(2+), Pb(2+), Cu(2+)) in aquatic ecosystems is important because these ions can have severe effects on human health and the environment. Gold (Au) nanomaterials are attractive sensing materials because of their unique size- and shape-dependent optical properties. This review focuses on optical assays for Hg(2+), Pb(2+), and Cu(2+) ions using functionalized Au nanomaterials. The syntheses of functionalized Au nanomaterials are discussed. We briefly review sensing approaches based on changes in absorbance resulting from metal ion-induced aggregation of Au nanoparticles (NPs) or direct deposition of metal ions onto Au NPs. The super-quenching properties of Au NPs allow them to be employed in 'turn on' and 'turn off' fluorescence approaches for the sensitive and selective detection of Hg(2+), Pb(2+), and Cu(2+) ions. We highlight approaches based on fluorescence quenching through analyte-induced aggregation or the formation of metallophilic complexes of Au nanodots (NDs). We discuss the roles of several factors affecting the selectivity and sensitivity of the nanosensors toward the analytes: the size of the Au nanomaterial, the length and sequence of the DNA or the nature of the thiol, the surface density of the recognition ligand, and the ionic strength and pH of the buffer solution. In addition, we emphasize the potential of using new nanomaterials (e.g., fluorescent silver nanoclusters) for the detection of PTM ions.

Au@Ag Dendritic Nanoforests for Surface-Enhanced Raman Scattering Sensing.

The effects of Au cores in Ag shells in enhancing surface-enhanced Raman scattering (SERS) were evaluated with samples of various Au/Ag ratios. High-density Ag shell/Au core dendritic nanoforests (Au@Ag-DNFs) on silicon (Au@Ag-DNFs/Si) were synthesized using the fluoride-assisted Galvanic replacement reaction method. The synthesized Au@Ag-DNFs/Si samples were characterized using scanning electron microscopy, energy-dispersive X-ray spectroscopy, reflection spectroscopy, X-ray diffraction, and Raman spectroscopy. The ultraviolet-visible extinction spectrum exhibited increased extinction induced by the addition of Ag when creating the metal DNFs layer. The pure Ag DNFs exhibited high optical extinction of visible light, but low SERS response compared with Au@Ag DNFs. The Au core (with high refractive index real part) in Au@Ag DNFs maintained a long-leaf structure that focused the illumination light, resulting in the apparent SERS enhancement of the Ag coverage.

Dendritic Forest-Like Ag Nanostructures Prepared Using Fluoride-Assisted Galvanic Replacement Reaction for SERS Applications.

Dendritic forest-like Ag nanostructures were deposited on a silicon wafer through fluoride-assisted galvanic replacement reaction (FAGRR) in aqueous AgNO3 and buffered oxide etchant. The prepared nanostructures were characterized using scanning electron microscopy, energy-dispersive X-ray spectroscopy, inductively coupled plasma-optical emission spectroscopy, a surface profiler (alpha step), and X-ray diffraction. Additionally, the dendritic forest-like Ag nanostructures were characterized using surface-enhanced Raman scattering (SERS) when a 4-mercaptobenzoic acid (4-MBA) monolayer was adsorbed on the Ag surface. The Ag nanostructures exhibited intense SERS signal from 4-MBA because of their rough surface, and this intense signal led to an intense local electromagnetic field upon electromagnetic excitation. The enhancement factor for 4-MBA molecules adsorbed on the Ag nanostructures was calculated to be 9.18 × 108. Furthermore, common Raman reporters such as rhodamine 6G, 4-aminothiolphenol, 5,5'-dithiobis-2-nitrobenzoic acid, and carboxyfluorescein (FAM) were characterized on these dendritic forest-like Ag nanostructures, leading to the development of an ultrasensitive SERS-based DNA sensor with a limit of detection of 33.5 nM of 15-mer oligonucleotide.

Detection of proteins and protein-ligand complexes using HgTe nanostructure matrixes in surface-assisted laser desorption/ionization mass spectrometry.

We have analyzed peptides, proteins, and protein-drug complexes through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using HgTe nanostructures as matrixes. We investigated the effects of several parameters, including the concentration of the HgTe nanostructures, the pH of the buffer, and the concentration of salt, on the performance of this system. When HgTe nanostructures are used as matrixes, [M + H](+) ions were the dominant signals. Relative to other commonly used nanomaterials, HgTe nanostructures provided lower background signals from metal clusters, fewer fragment ions, less interference from alkali-adducted analyte ions, and a higher mass range (up to 150,000 Da). The present approach provides limits of detection for angiotensin I and bovine serum albumin of 200 pM and 14 nM, respectively, with great reproducibility (RSD: <25%). We validated the applicability of this method through the detections of (i) the recombinant proteins that were transformed in E. coli, (ii) the specific complex between bovine serum albumin and l-tryptophan, and (iii) a carbonic anhydrase-acetazolamide complex. Our results suggest that this novel and simple SALDI-MS approach using HgTe nanostructures as matrixes might open several new ways for proteomics and the analysis of drug-protein complexes.

Exploring the interactions between gold nanoparticles and analytes through surface-assisted laser desorption/ionization mass spectrometry.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is applied to provide strong evidence for the chemical reactions of functionalized gold nanoparticles (Au NPs) with analytes--Hg(2+) ions induced MPA-Au NPs aggregation in the presence of 2,6-pyridinedicarboxylic acid (PDCA) and H(2)O(2) induced fluorescence quenching of 11-MUA-Au NDs. PDCA-Hg(2+)-MPA coordination is responsible for Au NPs aggregation, while the formation of 11-MUA disulfide compounds that release into the bulk solution is responsible for H(2)O(2)-induced fluorescence quenching. In addition to providing information about the chemical structures, SALDI-MS is also selective and sensitive for the detection of Hg(2+) ions and H(2)O(2). The limits of detection (LODs) for Hg(2+) ions and H(2)O(2) by SALDI-MS were 300 nM and 250 microM, respectively. The spot-to-spot variations in the two studies were both less than 18% (50 sample spots). Our results reveal that SALDI-MS can be used to study analyte-induced changes in the surface properties of nanoparticles.

Fluorescence detection of single nucleotide polymorphisms using a universal molecular beacon.

We present a simple and novel assay-employing a universal molecular beacon (MB) in the presence of Hg(2+)-for the detection of single nucleotide polymorphisms (SNPs) based on Hg(2+)-DNA complexes inducing a conformational change in the MB. The MB (T(7)-MB) contains a 19-mer loop and a stem of a pair of seven thymidine (T) bases, a carboxyfluorescein (FAM) unit at the 5'-end, and a 4-([4-(dimethylamino)phenyl]azo)benzoic acid (DABCYL) unit at the 3'-end. Upon formation of Hg(2+)-T(7)-MB complexes through T-Hg(2+)-T bonding, the conformation of T(7)-MB changes from a random coil to a folded structure, leading to a decreased distance between the FAM and DABCYL units and, hence, increased efficiency of fluorescence resonance energy transfer (FRET) between the FAM and DABCYL units, resulting in decreased fluorescence intensity of the MB. In the presence of complementary DNA, double-stranded DNA complexes form (instead of the Hg(2+)-T(7)-MB complexes), with FRET between the FAM and DABCYL units occurring to a lesser extent than in the folded structure. Under the optimal conditions (20 nM T(7)-MB, 20 mM NaCl, 1.0 muM Hg(2+), 5.0 mM phosphate buffer solution, pH 7.4), the linear plot of the fluorescence intensity against the concentration of perfectly matched DNA was linear over the range 2-30 nM (R(2) = 0.991), with a limit of detection of 0.5 nM at a signal-to-noise ratio of 3. This new probe provides higher selectivity toward DNA than that exhibited by conventional MBs.

Accurate quantitation of glutathione in cell lysates through surface-assisted laser desorption/ionization mass spectrometry using gold nanoparticles.

We developed a method for the determination of three aminothiols--cysteine, glutathione (GSH), and homocysteine--using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The analytes were first captured using the unmodified 14-nm gold nanoparticles; N-2-mercaptopropionylglycine-modified gold nanoparticles serving as internal standard were sequentially added, and then the sample was analyzed using SALDI-MS. This approach provided good quantitative linearity of the three analytes (R(2) = approximately 0.99), with good reproducibility (relative standard deviations: <10%), in the analyses of GSH in the lysates of human red blood cells and MCF-7 cancer breast cells in the presence and absence of the anti-inflammatory drug sulfasalazine. The internal-standard SALDI-MS approach provides simplicity, accuracy, and precision to the determination of GSH in cells under drug invasion, to open an avenue for SALDI-MS to be used for the precise quantitative determination of a variety of analytes. From the clinical editor: This paper reports the development of a surface assisted laser desorption/ionization mass spectrometry method to precisely determine aminothiols-cysteine (Cys), glutathione (GSH), and homocysteine (HCys).