Aptamer-based fluorescent sensor for highly sensitive detection of methamphetamine.

The construction of a fluorescence aptamer sensor was achieved by employing the fundamental principle of fluorescence resonance energy transfer. By employing molecular modeling technologies to identify the binding site, the high-affinity aptamer APT-40nt was derived from the whole sequence and utilized on the graphene oxide (GO) fluorescent platform for the purpose of achieving a highly sensitive detection of methamphetamine (METH). The aptamer tagged with fluorescein (FAM) dye undergoes quenching in the presence of GO due to π-stacking interaction. With the addition of the target, the aptamer that has been tagged was detached from the GO surface, forming a stable complex with METH. This process resulted in fluorescence restoration of the system, and the degree of fluorescence restoration was proportional to METH concentration in the linear range of 1-50 and 50-200 nM. Notably, under optimized conditions, the detection limit of this aptasensor was as low as 0.78 nM, which meets the detection limit requirements of METH detection in saliva and urine in some countries and regions. Moreover, other common illicit drugs and metabolites had minimizing interference with the determination. The established aptasensor, therefore, has been successfully applied to detect METH in saliva and urine samples and exhibited satisfactory recoveries (87%-111%). This aptasensor has the advantages of low detection limit, excellent selectivity, ease of operation, and low cost, providing a promising strategy for on-site detection of METH in saliva and urine.

Ratiometric fluorimetric and colorimetric probe for sensing and imaging pH changes in living cells.

Cell, enzyme, and tissue activity in living organisms are closely related to intracellular pH. Detecting the changes of intracellular pH is important to understanding the physiological and pathological changes in the process of crucial cell metabolism. A pH probe (HTBI) based on hemicyanine was synthesized. The probe solution displayed a marked colour change from yellow to amaranth with the pH increase from neutral to basic; simultaneously, the emission spectra showed a significant red shift. The probe exhibited a ratiometric fluorescence emission (F586nm /F542nm ) characteristic of pKa 8.82. As expected, HTBI exhibited high sensitivity and selectivity for pH, fine photostability, reversibility, and low cytotoxicity. Therefore, it would be a very useful tool for measuring the intracellular pH changes.

A chemiluminescence sensor array for discrimination of seven toxicants.

A chemiluminescence (CL) sensor array was developed based on 11 CL systems cross-combined by three luminescence reagents and four oxidants. Using the CL sensor array, we measured seven toxicants, including morphine, ketamine, diazepam, chlorpromazine, strychnine, paraquat, and fenpropathrin, which represent psychotropic drugs, sedatives and hypnotics, rodenticides, herbicides, and insecticides, respectively. The CL response pattern or 'fingerprints' were obtained for a given compound on the sensor array and then discriminated through principal component analysis. The established sensor array has been applied to real-life samples and the results showed that it possesses excellent discrimination.

Highly Selective Two-Photon Fluorescent Probe for Ratiometric Sensing and Imaging Cysteine in Mitochondria.

A novel ratiometric mitochondrial cysteine (Cys)-selective two-photon fluorescence probe has been developed on the basis of a merocyanine as the fluorophore and an acrylate moiety as the biothiol reaction site. The biocompatible and photostable acrylate-functionalized merocyanine probe shows not only a mitochondria-targeting property but also highly selective detection and monitoring of Cys over other biothiols such as homocysteine (Hcy) and glutathione (GSH) and hydrogen sulfide (H2S) in live cells. In addition, this probe exhibits ratiometric fluorescence emission characteristics (F518/F452), which are linearly proportional to Cys concentrations in the range of 0.5-40 µM. More importantly, the probe and its released fluorophore, merocyanine, exhibit strong two-photon excited fluorescence (TPEF) with two-photon action cross-section (Φσmax) of 65.2 GM at 740 nm and 72.6 GM at 760 nm in aqueous medium, respectively, which is highly desirable for high contrast and brightness ratiometric two-photon fluorescence imaging of the living samples. The probe has been successfully applied to ratiometrically image and detect mitochondrial Cys in live cells and intact tissues down to a depth of 150 µm by two-photon fluorescence microscopy. Thus, this ratiometric two-photon fluorescent probe is practically useful for an investigation of Cys in living biological systems.

Ratiometric emission fluorescent pH probe for imaging of living cells in extreme acidity.

A novel ratiometric emission fluorescent probe, 1,1-dimethyl-2-[2-(quinolin-4-yl)vinyl]-1H-benzo[e]indole (QVBI), is facilely synthesized via ethylene bridging of benzoindole and quinoline. The probe exhibits ratiometric fluorescence emission (F(522nm)/F(630nm)) characteristics with pKa 3.27 and linear response to extreme-acidity range of 3.8-2.0. Also, its high fluorescence quantum yield (Φ = 0.89) and large Stokes shift (110 nm) are favorable. Moreover, QVBI possesses highly selective response to H(+) over metal ions and some bioactive molecules, good photostability, and excellent reversibility. The probe has excellent cell membrane permeability and is further applied successfully to monitor pH fluctuations in live cells and imaging extreme acidity in Escherichia coli cells without influence of autofluorescence and native cellular species in biological systems.

Discrimination of vinegars using a catalytic nanomaterials-based chemiluminescence sensor array.

Quality control of foods is important for both industrial and personal concerns. In the past decade, a variety of sensor techniques have been developed and various applications realized for the analysis of foods in both the liquid and gas phases. In this paper, we report a chemiluminescence (CL) sensor array based on nine catalytic nanomaterials for the discrimination of eight vinegars. CL response patterns can be obtained as 'fingerprints' for a given compound on the sensor array and then discriminated using linear discriminant analysis (LDA). The experiments demonstrate that the sensor array has excellent differentiability and reversibility.

Enzyme-aided amplification strategy for sensitive detection of methamphetamine based on fluorescence aptamer sensor.

Methamphetamine (METH) abuse poses a serious risk to human health and social stability. It is critical to develop sensitive and selective methods for detecting METH. Here, we develop a fluorescence aptamer sensor to detect METH based on DNA exonuclease III (Exo III), graphene oxide (GO), and FAM-labeled aptamer. First, the sensor used GO's strong binding capacity to adsorb and quench the fluorescence of the aptamer attached to GO surface. When METH was added to the system, the formation of stable complex for aptamer and METH dissociated from the surface of GO, leading to a fluorescence restoration. Then, the fluorescence signal was further amplified by using Exo III to liberate target METH for cyclic hybridization. And the gel electrophoresis experiment further verified the reliability of this strategy. This aptamer sensor exhibited a low detection limit (0.52 nM) and excellent selectivity under optimal conditions. Notably, this sensor has been successfully validated in the detection of METH in urine and saliva samples, exhibited commendable recovery (94.00-104.65%). Its benefits include facile, sensitive, and rapid. Expected to be used in practical METH detection.

A chemiluminescence sensor array based on nanomaterials for discrimination of teas.

In recent years, electronic tongue and nose devices have been developed that consist of an array of cross-responsive sensors. In this study, we report a chemiluminescence (CL) sensor array based on oxidation at twelve different catalytic nanomaterial locations for the discrimination of eight teas. CL response patterns or "fingerprints" were obtained for a given compound on the sensor array and then discriminated through linear discriminant analysis. The experiments demonstrate that the sensor array had excellent differentiability and reversibility.

Forensic toxicokinetics of difenidol hydrochloride in vivo verified by practical lethal cases.

A gas chromatography-mass spectrometry (GC-MS) method for the determination of difenidol hydrochloride in biological specimens has been developed. The method exhibited excellent recovery (> 90%) and precision (RSD < 10%), and the LOD was 0.05 µg/mL or µg/g, which met the requirements of bioanalytical method. Through the animal model of the forensic toxicokinetics, the dynamic distribution, postmortem redistribution (PMR) and stability in specimen preservation process of difenidol in animals were studied. The experimental results showed that after intragastric administration, the difenidol's concentrations in the heart-blood and various organs increased over time except stomach, and then decreased gradually after reaching the peaks of concentration. The toxicological kinetics equation and toxicokinetic parameters were established by processing the data of the mean drug concentration of difenidol changing with time. In PMR experiment, the concentrations of difenidol in some organs closer to the gastrointestinal tract (heart-blood, heart, liver, lung, kidney, and spleen) changed significantly at different time points. But the concentration of difenidol in brain tissues which were far away from the gastrointestinal tract and muscles with larger overall mass was relatively stable. PMR of difenidol was therefore confirmed. Thus, the effect of PMR on the concentration of difenidol in the specimens should be considered in cases involving difenidol poisoning or death. Furthermore, the stability of difenidol in heart-blood samples from poisoned rats was investigated at various time points and under different preservation conditions (20 °C, 4 °C, - 20 °C and 20 °C (1% NaF)) for a period of two months. Difenidol was stable and did not decompose in the preserved blood. Therefore, this study provided the experimental basis for the forensic identification of the cases of difenidol hydrochloride poisoning (death). PMR has been verified by practical lethal cases.

A comparative study of postmortem distribution and postmortem diffusion of tramadol in rabbits.

In recent years, the cases of tramadol intoxication have become more frequent in many countries. However, most of the previous studies have been based on cases of tramadol intoxication, and the detailed information on the differences between postmortem distribution and diffusion of tramadol remains unclear. To investigate this issue systematically, we established a postmortem distribution model and two postmortem diffusion models. Then, gas chromatography-mass spectrometry (GC/MS) was used to measure the concentrations of tramadol in various biological specimens of fluids and tissues. In postmortem distribution, the results showed an uneven distribution of tramadol in various biological specimens, and the concentrations of tramadol in urine were significantly higher than those in other fluids. In postmortem diffusion, the results showed a dosage-dependent increase of tramadol concentration in most specimens; at all time points from 0.25 to 6 h after postmortem administration, the concentrations of tramadol in fluids were not significantly different from those in tissues, and the concentrations of tramadol in urine were lower than those in both tissues and other fluids in most time points. We recommend a quantitative examination of the specimens of both fluids and tissues to provide more evidence for the forensic identification, and the realization that there is a correlation between the concentrations of fluids and tissues is important for determining antemortem and postmortem administration of tramadol. This information can serve as ancillary data in inferring the contribution of a drug to death in cases of suspected tramadol poisoning.

A chemiluminescence sensor array for discriminating natural sugars and artificial sweeteners.

In this paper, we report a chemiluminescence (CL) sensor array based on catalytic nanomaterials for the discrimination of ten sweeteners, including five natural sugars and five artificial sweeteners. The CL response patterns ("fingerprints") can be obtained for a given compound on the nanomaterial array and then identified through linear discriminant analysis (LDA). Moreover, each pure sweetener was quantified based on the emission intensities of selected sensor elements. The linear ranges for these sweeteners lie within 0.05-100 mM, but vary with the type of sweetener. The applicability of this array to real-life samples was demonstrated by applying it to various beverages, and the results showed that the sensor array possesses excellent discrimination power and reversibility.

A label-free Exonuclease I-assisted fluorescence aptasensor for highly selective and sensitive detection of silver ions.

Based on the specific interaction of Ag+ and cytosine-cytosine (C-C) base mismatch and using berberine (Ber) as the fluorescent probe and Exonuclease I (Exo I) as the background fluorescence reducing tool, a label-free Exo I-assisted fluorescence aptamer sensing platform was established for the detection of silver ions with high sensitivity and selectivity. Exo I reduced the fluorescence background of the Ber/Ag+-aptamer complex to a level similar to that of Ber itself in the absence of Ag+. After introducing Ag+ into the sensing system, it induces the aptamer rich in base C to form C-Ag+-C i-motif structure which are resistant to degradation mediated by Exo I. The concentration of Ber, Ag+-aptamer, Exo I and the temperature and reaction time for Exo I were all optimized. Under the optimal experimental conditions, the detection limit of Ag+ was 4.4 nM and the linear range was from 0.0059 µM to 235.48 µM with a coefficient of determination (R2) > 0.99. Moreover, the proposed strategy had been successfully applied to the detection of Ag+ in tap water and human serum with a good recovery ranging from 88.4% to 106.9%.

Dynamic Distribution and Postmortem Redistribution of Tramadol in Poisoned Rats.

In the past dozen years, the cases of tramadol intoxication have become frequent in many countries. Most previous studies focused on tramadol's pharmacology, such as pharmacokinetics, pharmacodynamics and pharmacogenetics. However, the dynamic distribution and postmortem redistribution (PMR) of tramadol remain unclear. Our study aimed to investigate these two issues systematically in various specimens of 216 poisoned male rats. A validated gas chromatography-mass spectrometry method was used in this study to measure the concentrations of tramadol. In the first part, 66 tramadol poisoned rats were sacrificed at 11 different time points and their organs were collected separately for the study of tramadol's dynamic distribution, which made it feasible to investigate its PMR later on. The results of this part showed that tramadol's concentrations varied according to the organ and time, and peaked 2 h after intragastric administration in the specimens of liver, kidney, spleen, lung, brain and heart-blood (except stomach and heart). Based on the results of the first part, the concentration of tramadol peaked 2 h in most tissues. Therefore, this time point was used for the study of tramadol's PMR. In the second part, the remaining 150 rats were sacrificed 2 h after intragastric administration of tramadol, and the carcasses were stored under three different conditions (-20, 4 and 20°C). The autopsy was carried out at eight different time points and their organs were collected separately. The results of this part showed that under storage temperatures of -20 and 4°C, the concentrations of tramadol in individual organs showed no significant changes at different time points whereas under a storage temperature of 20°C, the concentrations in certain organs (liver, kidney, spleen, lung, brain and heart-blood) increased significantly at the last few time points. PMR of tramadol was therefore confirmed. The process of PMR of tramadol could be slowed or stopped at lower storage temperatures (-20 or 4°C), which is significant in cases of suspected tramadol poisoning.

Facile and green synthesis of fluorescent carbon dots with tunable emission for sensors and cells imaging.

Most carbon dots (CDs) conventional fabrication approaches produce single colored fluorescent materials, different methods are required to synthesize distinct carbon dots for specific optical applications. Herein, using one-pot hydrothermal treatment of Syringa obtata Lindl, a facile, low-cost and green assay is achieved in the controllable synthesis of blue and green fluorescent carbon dots. The fluorescent emission of CDs can be well-tuned by adding sodium hydroxide in the precursor solution. Blue fluorescent CDs are applied to Fe3+ sensing with a low detection limit of 0.11 µM of linear range from 0.5 to 80 µM, and then further extended to analysis river water samples. Green fluorescent CDs can be applied to pH detection, which show a remarkable linear enhancement in the green fluorescence emission region when the pH is increased from 1.98 to 8.95. Eventually, the detection of Fe3+ and pH are applied for the living cells fluorescent images in MCF-7 cells are achieved successfully, indicating as-synthesized CDs potential toward diverse application as promising candidate.

Chemiluminescence sensor for the determination of perphenazine based on a molecular imprinted polymer.

A luminol-potassium ferricyanide-perphenazine CL reaction with high sensitivity for the determination of perphenazine was found. A perphenazine molecular imprinted polymer (MIP) was synthesized. Using the perphenazine MIP as a recognition material, a novel molecular imprinting-chemiluminescence (MI-CL) sensor for the determination of perphenazine was made based on the luminol-potassium ferricyanide-perphenazine CL reaction. The sensor displayed good selectivity and high sensitivity. The linear-response range of the sensor was 5.0 x 10(-8) - 1.0 x 10(-5) g/ml with a linear correlation coefficient of 0.9961. The detection limit was 2 x 10(-8) g/ml perphenazine and the relative standard deviation for 1.0 x 10(-6) g/ml perphenazine solution was 3.7% (n = 11). The sensor was applied to the determination of perphenazine in urine samples with satisfactory results.

Simple, rapid and green one-step strategy to synthesis of graphene/carbon nanotubes/chitosan hybrid as solid-phase extraction for square-wave voltammetric detection of methyl parathion.

Simple, rapid, green and one-step electrodeposition strategy was first proposed to synthesis of graphene/carbon nanotubes/chitosan (GR/CNTs/CS) hybrid. The one-step electrodeposition approach for the construction of GR-based hybrid is green environmentally, which would not involve the chemical reduction of graphene oxide (GO) and therefore result in no further contamination. The whole procedure is simple and needs only several minutes. Combining the advantages of GR (large surface area, high conductivity and good adsorption ability), CNTs (high surface area, high enrichment capability and good adsorption ability) and CS (good adsorption and excellent film-forming ability), the obtained GR/CNTs/CS composite could be highly efficient to capture organophosphate pesticides (OPs) and used as solid phase extraction (SPE). The GR/CNTs/CS sensor is used for enzymeless detection of OPs, using methyl parathion (MP) as a model analyte. Significant redox response of MP on GR/CNTs/CS sensor is proved. The linear range is wide from 2.0ngmL(-1) to 500ngmL(-1), with a detection limit of 0.5ngmL(-1). Detection limit of the proposed sensor is much lower than those enzyme-based sensors and many other enzymeless sensors. Moreover, the proposed sensor exhibits high reproducibility, long-time storage stability and satisfactory anti-interference ability. This work provides a green and one-step route for the preparation of GR-based hybrid, and also offers a new promising protocol for OPs analysis.

Sensitive determination of phenothiazines in pharmaceutical preparation and biological fluid by flow injection chemiluminescence method using luminol-KMnO(4) system.

A flow injection chemiluminescence method was described for the determination of four phenothiazine drugs, namely, chlorpromazine hydrochloride, perphenazine hydrochloride, fluphenazine hydrochloride and thioridazine hydrochloride. Strong Chemiluminescence (CL) signal was produced when above-mentioned drug was injected into the mixed stream of luminol with KMnO(4). The linear ranges of the method were 0.0020-1.0mug/mL chlorpromazine hydrochloride, 0.0040-3.0mug/mL perphenazine hydrochloride, 0.0020-5.0mug/mL fluphenazine hydrochloride and 0.0050-1.0mug/mL thioridazine hydrochloride. The detection limits were 0.4ng/mL chlorpromazine hydrochloride, 0.7ng/mL perphenazine hydrochloride, 2ng/mL fluphenazine hydrochloride and 0.7ng/mL thioridazine hydrochloride. The proposed method was applied to the determination of chlorpromazine hydrochloride in injections and in mental patient's urine samples and the satisfactory results were achieved. The possible CL reaction mechanism was also discussed briefly.

Investigating the post-chemiluminescence behavior of phenothiazine medications in the luminol-potassium ferricyanide system: molecular imprinting-post-chemiluminescence method for the determination of chlorpromazine hydrochloride.

A new post-chemiluminescence (PCL) phenomenon was observed when phenothiazine medications were injected into the reaction mixture after the chemiluminescence (CL) reaction of luminol and potassium ferricyanide had finished. A possible reaction mechanism was proposed based on studies of the kinetic characteristics of the CL, CL spectra, fluorescence spectra, and on other experiments. The feasibility of determining various phenothiazine medications by utilizing these PCL reactions was examined. A molecular imprinting-post-chemiluminescence (MI-PCL) method was established for the determination of chlorpromazine hydrochloride using a chlorpromazine hydrochloride-imprinted polymer (MIP) as the recognition material. The method displayed high selectivity and high sensitivity. The linear range of the method was 1.0 x 10(-8) approximately 1.0 x 10(-6), with a linear correlation coefficient of 0.9985. The detection limit was 3 x 10(-9) g/ml chlorpromazine hydrochloride, and the relative standard deviation for a 1.0 x 10(-7) g/ml chlorpromazine hydrochloride solution was 4.0% (n = 11). The method has been applied to the determination of chlorpromazine hydrochloride in urine and animal drinking water with satisfactory results.