Luminescent and time-resolved determination of gemifloxacin mesylate in pharmaceutical formulations and spiked blood plasma samples using a lanthanide complex as a probe.

A novel luminescence-based analytical methodology was established employing a europium(III) complex with 3-allyl-2-hydroxybenzohydrazide (HAZ) as the coordinating ligand for the quantification of gemifloxacin mesylate (GMF) in pharmaceutical preparations and human plasma samples spiked with the compound. The stoichiometry of the europium complex with HAZ was determined via the Job plot and exhibited a metal-to-ligand ratio of 1 : 2. The analytical procedure relies on a rapid and significant enhancement of luminescence by the Eu(AZ)2 complex when it interacts with gemifloxacin mesylate, which allowed for the rapid detection of 96 samples within approximately 2 minutes. The thermodynamic parameters of the complexation of GMF with Eu(AZ)2 were evaluated and showed that the complexation of GMF was spontaneous with a negative ΔG. The binding constant K was 4.27 × 105 L mol-1 and DFT calculations supported GMF binding and the formation of Eu(AZ)2-GMF without further ligand exchange. The calibration graph for the luminescence quantitation of GMF was linear over a wide concentration range of 0.11-16 µg mL-1 (2.26 × 10-7 to 3.30 × 10-5 mol L-1), with a limit of quantification (LOQ) of 110 ng mL-1 (230 nmol L-1) and a detection limit (LOD) of 40 ng mL-1 (82 nmol L-1). The proposed method showed good accuracy with an average recovery of 99% with relative standard deviations of less than 5% in spiking experiments, even in complex pharmaceutical dosage forms such as tablets and in human blood plasma. Herein, the ability of the suppression of the luminescence background by using the long lag times of the lanthanide probe in a time-resolved detection scheme provided reliable and precise results, which suggests its potential for use in further real or patient samples.

New terbium complex as a luminescent probe for determination of chlorogenic acid in green coffee and roasted coffee infusions.

Green coffee is coming into vogue as a food that contains remarkable contents of antioxidants like chlorogenic acid (ChA) and induces mild stimulation to the consumer. While most methods for determination of ChA require chromatographic separation prior its quantitation, we present the first probe and a simple, sensitive and validated luminescence method for the determination of chlorogenic acid in green and roasted coffee infusion samples that does not require a chromatographic separation. ChA can remarkably quench the luminescence intensity of the Tb3+ complex with 1-(furan-2-ylmethyl)-4-hydroxy-N-(4-methylpyridin-2-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinoline-3-carboxamide (R3) in aqueous solution containing urotropine buffer at a near neutral pH 7.5 at λexc = 315 nm and λem = 545 nm. Under optimal conditions, the quenching of the luminescence intensity is directly proportional to the concentration of ChA in the range of 0.5-30 µg/mL, and the detection limit is 180 ng/mL. From measurements of luminescence decay time, it was determined that both static and dynamic quenching is induced upon coordination of ChA to Tb-R3. The related quenching constants are KS = 5.97∙104 M-1 and KD = 1.05⋅104 M-1. Finally, the method was applied successfully to the determination of ChA in real green and roasted coffee infusion samples and validated by HPLC to yield very closely matching concentrations of both methods. Therefore, this method can serve for a simple quality control of total ChA contents in coffee prior and after roasting.

Critical review of polymer and hydrogel deposition methods for optical and electrochemical bioanalytical sensors correlated to the sensor's applicability in real samples.

Sensors, ranging from in vivo through to single-use systems, employ protective membranes or hydrogels to enhance sample collection or serve as filters, to immobilize or entrap probes or receptors, or to stabilize and enhance a sensor's lifetime. Furthermore, many applications demand specific requirements such as biocompatibility and non-fouling properties for in vivo applications, or fast and inexpensive mass production capabilities for single-use sensors. We critically evaluated how membrane materials and their deposition methods impact optical and electrochemical systems with special focus on analytical figures of merit and potential toward large-scale production. With some chosen examples, we highlight the fact that often a sensor's performance relies heavily on the deposition method, even though other methods or materials could in fact improve the sensor. Over the course of the last 5 years, most sensing applications within healthcare diagnostics included glucose, lactate, uric acid, O2, H+ ions, and many specific metabolites and markers. In the case of food safety and environmental monitoring, the choice of analytes was much more comprehensive regarding a variety of natural and synthetic toxicants like bacteria, pesticides, or pollutants and other relevant substances. We conclude that more attention must be paid toward deposition techniques as these may in the end become a major hurdle in a sensor's likelihood of moving from an academic lab into a real-world product.

Highly sensitive interleukin 6 detection by employing commercially ready liposomes in an LFA format.

Recent years have confirmed the ubiquitous applicability of lateral flow assays (LFA) in point-of-care testing (POCT). To make this technology available for low abundance analytes, strategies towards lower limits of detections (LOD), while maintaining the LFA's ease of use, are still being sought. Here, we demonstrate how liposomes can significantly improve the LOD of traditional gold nanoparticle (AuNP)-based assays while fully supporting a ready-to-use system for commercial application. We fine-tuned liposomes towards photometric and fluorescence performance on the synthesis level and applied them in an established interleukin 6 (IL-6) immunoassay normally using commercial AuNP labels. IL-6's low abundance (< 10 pg mL-1) and increasing relevance as prognostic marker for infections make it an ideal model analyte. It was found that liposomes with a high encapsulant load (150 mmol L-1 sulforhodamine B (SRB)) easily outperform AuNPs in photometric LFAs. Specifically, liposomes with 350 nm in diameter yield a lower LOD even in complex matrices such as human serum below the clinically relevant range (7 pg mL-1) beating AuNP by over an order of magnitude (81 pg mL-1). When dehydrated on the strip, liposomes maintained their signal performance for over a year even when stored at ambient temperature and indicate extraordinary stability of up to 8 years when stored as liquid. Whereas no LOD improvement was obtained by exploiting the liposomes' fluorescence, an extraordinary gain in signal intensity was achieved upon lysis which is a promising feature for high-resolution and low-cost detection devices. Minimizing the procedural steps by inherently fluorescent liposomes, however, is not feasible. Finally, liposomes are ready for commercial applications as they are easy to mass-produce and can simply be substituted for the ubiquitously used AuNPs in the POCT market.

Next generation luminol derivative as powerful benchmark probe for chemiluminescence assays.

Chemiluminescence (CL) provides outstanding analytical performance due to its independence from external light sources, background-free nature and exceptional sensitivity and selectivity. Yet, ultra-sensitive (bio)analysis is impeded by low hydrophilicity, poor quantum yields, fast kinetics or instability of most CL reagents such as luminol, acridinium esters, dioxetanes or peroxyoxalic derivatives. Photophysical studies show that m-carboxy luminol overcomes these limitations as its hydrophilic design provides a 5-fold increase in relative quantum yield resulting in superior performance in H2O2-dependent bioassays with 18-fold higher sensitivity for the quantification of its co-reactant H2O2, and 5-times lower detection limits for the luminophore. Studies with CL enhancers suggest its significance for mechanistic investigations in tandem with peroxidases. Finally, its integration into enzymatic and immunoassay applications demonstrates that m-carboxy luminol will provide signal enhancement, lower detection limits, and increased dynamic ranges for any other luminol-based CL assay, thus comprising the potential to replace luminol as benchmark probe.

Isoquinoline-based Eu(iii) luminescent probes for citrate sensing in complex matrix.

A neutral Eu(iii) complex containing the S,S enantiomer of isoQC3A3- ligand (isoQC3A3- = N-isoquinolyl-N,N',N'-trans-l,2-cyclohexylenediaminetriacetate) was synthesized and characterized. The complex was spectroscopically investigated and the results compared with those obtained for the similar bis-anionic ligand bisoQcd2- (bisoQcd2- = N,N'-bis(2-isoquinolinmethyl)-trans-1,2-diaminocyclohexane N,N'-diacetate). Both Eu(iii)-complexes show similar binding constants upon titration with the main analytes contained in interstitial extracellular fluid (i.e. hydrogen carbonate, serum albumin and citrate). However, the analyte affinity is accompanied by different enhancements of the Eu(iii) intrinsic quantum yield (QY). Structures and hydration numbers of the complexes are determined by luminescence decay measurements and DFT calculations. The QYs as well as the binding constants of the individual adducts of the complexes with hydrogen carbonate, bovine serum albumin (BSA) and citrate are determined. The study of the Eu(iii) emission upon the systematic variation of one analyte in a complex mixture has been carried out to predict the performance of the luminescent sensor in conditions close to the real extracellular environment. Both Eu(iii) complexes can detect citrate at extracellular concentrations up to 500 µM, even at millimolar concentrations of the other interfering species. In the case of the Eu(bisoQcd)OTf complex, an increase of 23% of the Eu(iii) luminescence intensity at 615 nm upon addition of 0.3 mM of citrate was recorded. This feature makes the latter complex a viable probe for luminescence analysis of citrate in a complex matrix.

Electrochemical multi-analyte point-of-care perspiration sensors using on-chip three-dimensional graphene electrodes.

Multi-analyte sensing using exclusively laser-induced graphene (LIG)-based planar electrode systems was developed for sweat analysis. LIG provides 3D structures of graphene, can be manufactured easier than any other carbon electrode also on large scale, and in form of electrodes: hence, it is predestinated for affordable, wearable point-of-care sensors. Here, it is demonstrated that LIG facilitates all three electrochemical sensing strategies (voltammetry, potentiometry, impedance) in a multi-analyte system for sweat analysis. A potentiometric potassium-ion-selective electrode in combination with an electrodeposited Ag/AgCl reference electrode (RE) enabled the detection of potassium ions in the entire physiologically relevant range (1 to 500 mM) with a fast response time, unaffected by the presence of main interfering ions and sweat-collecting materials. A kidney-shaped interdigitated LIG electrode enabled the determination of the overall electrolyte concentration by electrochemical impedance spectroscopy at a fixed frequency. Enzyme-based strategies with amperometric detection share a common RE and were realized with Prussian blue as electron mediator and biocompatible chitosan for enzyme immobilization and protection of the electrode. Using glucose and lactate oxidases, lower limits of detection of 13.7 ± 0.5 µM for glucose and 28 ± 3 µM for lactate were obtained, respectively. The sensor showed a good performance at different pH, with sweat-collecting tissues, on a model skin system and furthermore in synthetic sweat as well as in artificial tear fluid. Response time for each analytical cycle totals 75 s, and hence allows a quasi-continuous and simultaneous monitoring of all analytes. This multi-analyte all-LIG system is therefore a practical, versatile, and most simple strategy for point-of-care applications and has the potential to outcompete standard screen-printed electrodes. Graphical abstract.

Optical sensors for determination of biogenic amines in food.

This review presents the state-of-the-art of optical sensors for determination of biogenic amines (BAs) in food by publications covering about the last 10 years. Interest in the development of rapid and preferably on-site methods for quantification of BAs is based on their important role in implementation and regulation of various physiological processes. At the same time, BAs can develop in different kinds of food by fermentation processes or microbial activity or arise due to contamination, which induces toxicological risks and food poisoning and causes serious health issues. Therefore, various optical chemosensor systems have been devised that are easy to assemble and fast responding and low-cost analytical tools. If amenable to on-site analysis, they are an attractive alternative to existing instrumental analytical methods used for BA determination in food. Hence, also portable sensor systems or dipstick sensors are described based on various probes that typically enable signal readouts such as photometry, reflectometry, luminescence, surface-enhanced Raman spectroscopy, or ellipsometry. The quantification of BAs in real food samples and the design of the sensors are highlighted and the analytical figures of merit are compared. Future instrumental trends for BA sensing point to the use of cell phone-based fully automated optical evaluation and devices that could even comprise microfluidic micro total analysis systems.

Cationic liposomes for generic signal amplification strategies in bioassays.

Liposomes have been widely applied in bioanalytical assays. Most liposomes used bare negative charges to prevent non-specific binding and increase colloidal stability. Here, in contrast, highly stable, positively charged liposomes entrapping the fluorescent dye sulforhodamine B (SRB) were developed to serve as a secondary, non-specific label' and signal amplification tool in bioanalytical systems by exploiting their electrostatic interaction with negatively charged vesicles, surfaces, and microorganisms. The cationic liposomes were optimized for long-term stability (> 5 months) and high dye entrapment yield. Their capability as secondary, non-specific labels was first successfully proven through electrostatic interactions of cationic and anionic liposomes using dynamic light scattering, and then in a bioassay with fluorescence detection leading to an enhancement factor of 8.5 without any additional surface blocking steps. Moreover, the cationic liposomes bound efficiently to anionic magnetic beads were stable throughout magnetic separation procedures and could hence serve directly as labels in magnetic separation and purification strategies. Finally, the electrostatic interaction was exploited for the direct, simple, non-specific labeling of gram-negative bacteria. Isolated Escherichia coli cells were chosen as models and direct detection was demonstrated via fluorescent and chemiluminescent liposomes. Thus, these cationic liposomes can be used as generic labels for the development of ultrasensitive bioassays based on electrostatic interaction without the need for additional expensive recognition units like antibodies, where desired specificity is already afforded through other strategies. Graphical abstract.

Magnetosomes for bioassays by merging fluorescent liposomes and magnetic nanoparticles: encapsulation and bilayer insertion strategies.

Magnetized liposome (magnetosomes) labels can overcome diffusion limitations in bioassays through fast and easy magnetic attraction. Our aim therefore was to advance the understanding of factors influencing their synthesis focusing on encapsulation strategies and synthesis parameters. Magnetosome synthesis is governed by the surface chemistry and the size of the magnetic nanoparticles used. We therefore studied the two possible magnetic labelling strategies, which are the incorporation of small, hydrophobic magnetic nanoparticles (MNPs) into the bilayer core (b-liposomes) and the entrapment of larger hydrophilic MNPs into the liposomes' inner cavity (i-liposomes). Furthermore, they were optimized and compared for application in a DNA bioassay. The major obstacles observed for each of these strategies were on the one hand the need for highly concentrated hydrophilic MNPs, which is limited by their colloidal stability and costs, and on the other hand the balancing of magnetic strength vs. size for the hydrophobic MNPs. In the end, both strategies yielded magnetosomes with good performance, which improved the limit of detection of a non-magnetic DNA hybridization assay by a factor of 3-8-fold. Here, i-liposomes with a magnetization yield of 5% could be further improved through a simple magnetic pre-concentration step and provided in the end an 8-fold improvement of the limit of detection compared with non-magnetic conditions. In the case of b-liposomes, Janus-like particles were generated during the synthesis and yielded a fraction of 15% magnetosomes directly. Surprisingly, further magnetic pre-concentration did not improve their bioassay performance. It is thus assumed that magnetosomes pull normal liposomes through the magnetic field towards the surface and the presence of more magnetosomes is not needed. The overall stability of magnetosomes during storage and magnetic action, their superior bioassay performance, and their adaptability towards size and surface chemistry of MNPs makes them highly valuable signal enhancers in bioanalysis and potential tools for bioseparations. Graphical abstract.

Shedding Light on the Diversity of Surfactant Interactions with Luminol Electrochemiluminescence for Bioanalysis.

Luminol is a major probe for chemiluminescence (CL) and electrochemiluminescence (ECL) detection technologies in (bio)analysis. Surfactants are added to ECL assay cocktails to enhance signals or are present, owing to given bioassay protocols, yet little is known regarding their effects on luminol ECL. In-depth understanding is provided here through a broad study with bioanalytically relevant surfactants (cationic, anionic, and nonionic), four common electrode materials, and two luminol derivatives. Naturally, in ECL, surface effects are dominant; however, bulk solution, diffusion, and luminescence-stabilization processes also contribute significantly to the overall reaction. It was found that in contrast to CL the effect surfactants have on luminol ECL cannot be linked to general surfactant characteristics such as ionic nature, hydrophilic lipophilic balance (HLB) value, and critical micellar concentration (CMC). Instead, surfactants act in an all-encompassing mechanism, including surface electrochemistry, their solution and interfacial phases, and the chemical luminescence pathway. This leads to dramatic differences in signals obtained, ranging from 5-fold increases to total quenching. Within this complexity, we defined six guiding principles that are extrapolated from the underlying mechanisms and selection guides for surfactant, electrode, and environmental condition combinations. Those will now assist in developing highly sensitive luminol-ECL-based bioassays, because the surfactant selection can be based not only on properties needed for the assay protocol but also on identifying the optimal electrode-surfactant pair to maximize detection efficiency.

Tethering functionality to lipid interfaces by a fast, simple and controllable post synthesis method.

HYPOTHESIS: Liposomes require careful control of the surface design to ensure colloidal stability in complex matrices and target-specific binding to desired receptor units. Ideally, surface functionalization should be smart and controllable in terms of composition which is seldomly achieved by conventional methods. Therefore, a new strategy (insertion method) was developed and compared to the standard method (modification post-synthesis) using the model receptor biotin. EXPERIMENTS: Dipalmitoylphosphatidylethanolamine-biotin (DPPE-biotin) was used in both procedures, lipopeptide-biotin and cholesterol-biotin were tested additionally for insertion into the intact lipid bilayer. The insertion method was optimized regarding incubation time, temperature and vesicle stability. The biotinylated vesicles of both functionalization methods were characterized with respect to their size, ζ-potential and binding functionality. FINDINGS: Standard incorporation resulted in large variations in insertion-efficiency, high batch-to-batch differences, and an incorporation limit of 4 mol%. Best results were obtained through effortless insertion of the lipopeptide-biotin at room temperature. The concentration-controlled functionalization of liposomes (up to 10 mol%) could easily be monitored by the ζ-potential, resulted in reliable, quantitative binding to streptavidin and did not affect the analytical properties of the nanomaterial. This offers the possibility for a general modification strategy for lipid-based nanomaterials ideal for assay optimizations or multi-analyte detection.

Nanocontainers for Analytical Applications.

Nanocontainers such as mesoporous silica particles and polymersomes are versatile structures containing holes or pores which are used for the entrapment of small molecules and the introduction of specific functionalities. They are widely applied in drug delivery, biomedicine, bioreactors, and analytical applications. In the last case, nanocontainers usually serve as amplification systems. They are hence synthesized to entrap signaling molecules and to bear functional moieties at the outer surface, which in turn enable specific analyte recognition and control of the nanocontainer pore permeability. This Review outlines the most important nanocontainer materials and discusses their synthesis, surface chemistry modifications, and strategies for molecule entrapment. Their advantages, challenges, and limitations are critically discussed in view of other common signal amplification strategies for different assay formats and various detection methods.

A luminescent europium complex for wide-range pH sensors and sensor microtiterplates.

In this work, we show a new concept of a luminescent wide-range pH indicator based on a europium complex and its implementation in sensors and sensor microtiterplates. For this purpose, we complexed europium with the ligands gallic acid (Gall) and pyridine dicarboxylic acid (PDA) that carry groups with wide ranging pKa values and the antenna ligand theonyl trifluoroacetone (TTA). This yields an indicator that overcomes the typical drawbacks of many optical pH indicator dyes which mostly cover three pH units, only. The spectral characterization of the new europium complex Eu3+-TTA-Gall-PDA in a molar ratio of 1 : 3 : 1 : 1 in aqueous buffer delivers a luminescence emission that is dependent on pH over up to 8 orders of magnitude from pH 2-10 in aqueous solution. It was therefore embedded into sensor membranes made of cellulose acetate (CA) which respond reversibly from pH 2-7 over 5 orders of magnitude within several minutes. Moreover, the first high-throughput pH sensor microtiterplate based on this indicator showed a dynamic range from pH 2-8 over 6 orders of magnitude and is suitable for up to 24 h of continuous use.

Electrochemiluminescence Bioassays with a Water-Soluble Luminol Derivative Can Outperform Fluorescence Assays.

The most efficient and commonly used electrochemiluminescence (ECL) emitters are luminol, [Ru(bpy)3 ]2+ , and derivatives thereof. Luminol stands out due to its low excitation potential, but applications are limited by its insolubility under physiological conditions. The water-soluble m-carboxy luminol was synthesized in 15 % yield and exhibited high solubility under physiological conditions and afforded a four-fold ECL signal increase (vs. luminol). Entrapment in DNA-tagged liposomes enabled a DNA assay with a detection limit of 3.2 pmol L-1 , which is 150 times lower than the corresponding fluorescence approach. This remarkable sensitivity gain and the low excitation potential establish m-carboxy luminol as a superior ECL probe with direct relevance to chemiluminescence and enzymatic bioanalytical approaches.

Functional electrospun nanofibers for multimodal sensitive detection of biogenic amines in food via a simple dipstick assay.

Electrospun nanofibers (ENFs) are promising materials for rapid diagnostic tests like lateral flow assays and dipsticks because they offer an immense surface area while excluding minimal volume, a variety of functional surface groups, and can entrap functional additives within their interior. Here, we show that ENFs on sample pads are superior in comparison to standard polymer membranes for the optical detection of biogenic amines (BAs) in food using a dipstick format. Specifically, cellulose acetate (CA) fibers doped with 2 mg/mL of the chromogenic and fluorogenic amine-reactive chameleon dye Py-1 were electrospun into uniform anionic mats. Those extract cationic BAs from real samples and Py-1 transduces BA concentrations into a change of color, reflectance, and fluorescence. Dropping a BA sample onto the nanofiber mat converts the weakly fluorescent pyrylium dye Py-1 into a strongly red emitting pyridinium dye. For the first time, a simple UV lamp excites fluorescence and a digital camera acts as detector. The intensity ratio of the red to the blue channel of the digital image is dependent on the concentration of most relevant BAs indicating food spoilage from 10 to 250 µM. This matches the permitted limits for BAs in foods and no false positive signals arise from secondary and tertiary amines. BA detection in seafood samples was also demonstrated successfully. The nanofiber mat dipsticks were up to sixfold more sensitive than those using a polymer membrane with the same dye embedded. Hence, nanofiber-based tests are not only superior to polymer-based dipstick assays, but will also improve the performance of established tests related to food safety, medical diagnostics, and environmental testing. Graphical Absract ᅟ.

Signal enhancement and low oxidation potentials for miniaturized ECL biosensors via N-butyldiethanolamine.

We present studies on ruthenium-based electrochemiluminescence (ECL) focusing on conditions supporting signal enhancement and low oxidation potentials. Low oxidation potentials (LOPs) are especially attractive for miniaturized ECL biosensors, as microfabricated electrodes tend to detach from their support when used with high currents and operated at high potentials. Furthermore, high potentials or current densities can lead to damage of typical biosensor surface coatings and biological probes. The possibility of generating LOP ECL signals at a potential below 900 mV was therefore studied for Ru(bpy)32+ with two typical coreactants, i.e. 2-(dibutylamino)ethanol (DBAE) and tripropylamine (TPA), as well as with the tertiary amine N-butyldiethanolamine (NBEA). Furthermore, the effect of buffer components and pH values on ECL signal generation was investigated. We could show a significant LOP ECL signal for NBEA. We found that Tris buffer, with its ability to form complexes with transition metal ions, has a positive influence on this ECL signal in terms of signal strength and LOP capabilities. Specifically, at basic pH values significant increases in ECL signals were observed at 900 mV and at 1.2 V. In fact, the ECL signal at 1.2 V was three times higher than the signal observed in phosphate buffer at a pH of 7, and it was thirty times higher than the ECL signal for TPA under these conditions. The LOP signal for NBEA in Tris buffer at pH 8.5 was similar to the signal obtained for TPA in phosphate buffer at pH 8.5 but three times higher than for TPA at pH 7.0. Interestingly, the coreactant DBAE was neither significantly influenced by the buffer system or pH nor did it present a valuable LOP ECL signal. Finally, it was found that high peak currents in cyclic voltammograms are not the indicators for high ECL signals, which should be obvious because the ECL mechanism requires more complex electron transfers. Overall, the standard TPA ECL at 1.2 V in phosphate buffer at pH 7.0 can successfully be replaced by NBEA ECL at 900 mV in Tris at pH 8.5 providing significantly higher signals accompanied by more gentle electrochemical conditions.

Improving ruthenium-based ECL through nonionic surfactants and tertiary amines.

The influence of surfactants and coreactants on Ru(bpy)32+ electrogenerated chemiluminescence (ECL) was investigated comparatively. Specifically, the influence that the two tertiary amines, N-butyldiethanolamine (NBEA) and 2-(dibutylamino)ethanol (DBAE) have on the ECL reaction, alone and in the presence of the two surfactants Triton™ X-100 and Zonyl® FSN, was investigated, in comparison with that of the well-studied and established coreactant tripropylamine (TPA). Experiments were conducted on poly(methyl methacrylate) substrates coated with plasma-evaporated gold as used in many miniaturized systems. Upon optimization and study of the various combinations, the combination of NBEA/Zonyl FSN provided superior ECL signal characteristics. A limit of detection (LOD) of 2.2 nM Ru(bpy)32+ was obtained. Compared with the LOD of 0.59 µM for the commonly used TPA/Triton™ X-100 system, the resulting LOD is enhanced by a factor of 250. In addition, significantly more stable signals lead to an increase in sensitivity by a factor of 50. This makes the NBEA/Zonyl® FSN system an attractive ECL strategy, especially for miniaturized analytical systems. Furthermore, it became clear that previously postulated effects of surfactants on the enhancement of coreactant-based ECL do not translate to other surfactants and coreactants. We could demonstrate that more complicated mechanisms are at play as the ECL intensity for Ru(bpy)32+/NBEA was the highest in the presence of Zonyl® FSN, whereas the ECL signal decreased significantly upon introduction of the surfactant with DBAE as coreactant.

Design, selective alkylation and X-ray crystal structure determination of dihydro-indolyl-1,2,4-triazole-3-thione and its 3-benzylsulfanyl analogue as potent anticancer agents.

Three sets of substituted indolyl-triazoles were synthesized by the alkylation of 1,2-dihydro-5-(1H-indol-2-yl)-1,2,4-triazole-3-thione with different alkyl halides. The use of pyridine restricted the alkylation to sulfur. Whereas, upon using K2CO3, the alkylation exceeded sulfur to one of the remaining triazole nitrogens. The assignment of which nitrogen is alkylated besides sulfur is made for the first time using X-ray analysis of single crystals and 2D NMR which indicated that S-, 2-N-isomers will be preferably formed over the S-, 1-N-isomers. The antiproliferative activity on HEPG-2 and MCF-7 cancer cell lines was tested. The results showed that compound 2a is the most active with an IC50 3.58 µg/mL and 4.53 µg/mL for HEPG-2 and MCF-7 respectively and compound 7 is the least active with an IC50 > 100 µg/mL compared to the standard drug doxorubicin (IC50 4.0 µg/mL). The interaction of the synthesized compounds with tyrosine kinases, namely, Akt, PI3, and EGFR was also studied using molecular docking simulation to predict their mode of action which will drive future work directions.