Identification of major degradation products of Clorsulon drug substance including its degradation pathways by high resolution mass spectrometry and NMR.

Clorsulon is an effective anthelmintic drug substance extensively used in the treatment of parasitic infestations in both cattle and sheep. An in-depth investigation of Clorsulon's degradation products (DPs) was carried out through forced degradation study to comprehend its degradation path. A total of eight degradation products were separated under various stress degradation conditions. Structural elucidation of these DPs was conducted using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS), and their fragmentation patterns were compared to that of the parent compound. Adequate amount of DP-4 was isolated and purified by semi-preparative high-performance liquid chromatography (HPLC) methods. Subsequently, it was examined in detail using both 1D and 2D NMR (nuclear magnetic resonance spectroscopy). Most probable mechanistic pathways for the formation of degradation products under various stress degradation conditions were proposed to better understand the degradation profile. Based on the results of the stress study, Clorsulon drug substance was found to be unstable under photolytic and oxidative conditions. Understanding Clorsulon's degradation pathway is essential for determining shelf-life prediction of the finished product, safety and efficacy assurance, and guiding the development of stable, high-quality formulations.

Determination of Clorsulon and its related substances in commercial bulk drug substance batches by a rapid ion-pair UPLC method.

Clorsulon is a potent anthelmintic agent and is widely used for the treatment and control of parasites including adult liver flukes in cattle and sheep. A rapid ion-paired reversed phase ultraperformance liquid chromatography (IP-UPLC) method has been developed and validated for determination of Clorsulon and its related substances in bulk drug substance batches of Clorsulon with a short octadecyl column. Analytes were eluted by a gradient elution on a Acquity UPLC® BEH C18 column (50 mm × 2.1 mm i.d., 1.7 µm particle size). Column temperature was maintained at 55 °C and all analytes were monitored by UV detection at 268 nm. Mobile phase-A constitutes 3 mM tetrabutylammonium hydroxide in H2O and mobile phase-B constitutes acetonitrile/methanol (50/50, v/v), respectively. All analytes of interest were adequately separated within 5 min. The analytes were quantitated against an external reference standard of Clorsulon. The chemical structures of degradation products of Clorsulon were proposed based on two-dimensional UPLC-MS data. The IP-UPLC method has been successfully validated and demonstrated to be accurate, sensitive, robust, and specific.

A durable non-contact reciprocated triboelectric nanogenerator for low-frequency vibration energy harvesting.

Harvesting vibration energy using a triboelectric nanogenerator (TENG) is a promising approach in solving the power supply restriction of the Internet of Things. Currently, the low durability due to friction surface wearing is the primary limitation of TENGs, which restricts their applicability and practicability. This study introduces a non-contact-type TENG aimed at significantly enhancing its durability by increasing its anti-wear capability. The configuration of the proposed TENG includes permanent magnets and rolling-balls. The reciprocating motion of functional friction surfaces, facilitated by the permanent magnets, enhances the efficiency of harvesting low-frequency vibration energy. The embedded rolling-balls are utilized to separate two functional friction surfaces, which minimizes the friction surface wearing between different dielectric materials. The electrical output characteristics of this non-contact TENG under variable load resistances are explored according to sinusoidal excitation based on either variable frequencies or accelerations. The results demonstrate that the proposed nanogenerator can generate a short-circuit current of 2118.2 nA and achieve a peak power density of 9.891 mW/m2. The electrical responses of this non-contact TENG remain stable over 120 000 continuous working cycles, lasting for more than 200 min. Furthermore, the nanogenerator can identify and harvest energy from running or jumping motions performed by individuals in different postures and at various speeds or heights. With its exceptional durability and stability, this non-contact nanogenerator offers a novel approach to low-frequency vibration energy harvesting, paving the way for practical applications in the field.

Development and Validation of a Stability-Indicating Reversed-Phase HPLC Method for Assay and Estimation of Related Substances of Ivermectin in an Oral Paste.

Ivermectin is a potent semi-synthetic antiparasitic drug used in veterinary medicine. A reversed-phase high performance liquid chromatography (HPLC) method has been developed and validated for the identification and assay of Ivermectin, including the identification and estimation of its related impurities in an oral paste. Analytes were separated using a gradient elution at a flow rate of 1.5 mL/min on a Zorbax Extend-C18 column (150 mm × 4.6 mm i.d., 3.5-µm particle size) maintained at 30 °C. The mobile phase was composed of water as mobile-phase A and acetonitrile/methanol (85/15, v/v) as mobile-phase B. Ultraviolet detection at 245 nm was employed to monitor the analytes. Limit of quantitation (LOQ) and limit of detection (LOD) of the method are 0.6 and 0.2 µg/mL, respectively. The validation results demonstrated excellent linearity of the method in the range of 0.1-150% of the analytical concentration (0.6 mg/mL) of the method. The stability-indicating capability of the method has been demonstrated by adequately separating the degradation products from the stress degraded samples of the oral paste as per method validation requirements prescribed in the current International Council for Harmonisation guidelines.

Simultaneous Determination of Esafoxolaner, Eprinomectin, Praziquantel, BHT and Their Related Substances in a Topical Finished Product by a Single Reversed-Phase High-Performance Liquid Chromatography Method.

The topical product with three active pharmaceutical ingredients (APIs), namely, esafoxolaner, eprinomectin and praziquantel has demonstrated its efficacy in the treatment of cats with mixed infections with ectoparasites and nematodes and cestodes. A reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed and validated for assay and determination of related substances peaks of three APIs including the assay of antioxidant butylated hydroxytoluene (BHT) in the finished product. Analytes were separated on a short Zorbax SB-C18 column (50 × 4.6 mm I.D., 5 µm particle size, pore size: 80 Å) with gradient elution at 40 °C column temperature. Analytes were detected at 245 nm for praziquantel, esafoxolaner, eprinomectin and their degradation products. BHT and eprinomectin degradation product 8a-oxo-B1a were detected at 280 nm. All analytes of interest were adequately separated within 40 min. The assay for praziquantel, esafoxolaner, eprinomectin and BHT was conducted against their corresponding external reference standards. The related substances peaks of each API were determined by peak area and relative response factor against total peak area of their corresponding API peak in sample solution. This method has been demonstrated to be accurate, robust, specific and stability indicating.

Development and Validation of a Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) Method for Identification, Assay and Estimation of Related Substances of Ivermectin in Bulk Drug Batches of Ivermectin Drug Substance.

A reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed and validated for the identification and assay of Ivermectin, including the identification and estimation of its process-related impurities and degradation products in bulk drug substance of Ivermectin. Analytes were separated on a HALO C18 column (100 mm × 4.6 mm I.D., 2.7 µm particle size) maintained at 40 °C (column temperature) with gradient elution. All analytes of interests were adequately separated within 25 min. All degradation products, process-related impurities and assay were monitored by ultraviolet detection at 254 nm. The new HPLC method described here successfully separated an isomer peak of the active pharmaceutical ingredient (API) from the major API peak. This newly separated isomer peak is around 1.2 to 1.5% (peak area) in typical API samples, and coelutes with the major API peak by all current HPLC methods. Quantitation limit of the HPLC method is 0.1% of target analytical concentration (~1.0 µg/mL). This method has been demonstrated to be accurate, robust, significantly higher degree of selectivity compared to the HPLC methods of Ivermectin drug substance reported in the literature and in the compendial HPLC methods prescribed in the current USA and European Pharmacopeia.

Development and Validation of a Stability-Indicating Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) Method for Identification, Assay, and Estimation of Related Substances of Ivermectin Drug Substance.

BACKGROUND: Ivermectin is a potent semi-synthetic antiparasitic drug used in veterinary medicine. It is widely used for the treatment of parasites. OBJECTIVE: This study aimed to develop a stability-indicating reversed-phase HPLC (RP-HPLC) method for assay and identification of ivermectin including identification and estimation of its related substances in bulk drug substance batches of ivermectin. METHOD: Ivermectin and its related substances were separated on an Ascentis Express C18 column (100 mm × 4.6 mm id, 2.7 µm particle size) maintained at 45°C (column temperature) on an HPLC system with gradient elution. The mobile phase was composed of water - acetonitrile (ACN; 50 + 50, v/v) as mobile phase A, and isopropanol - ACN (15 + 85, v/v) as mobile phase B. Analytes were detected with a detection wavelength of 252 nm and quantitated against an external reference standard of ivermectin with a quantitation limit of 0.1% of the target (analytical) concentration. RESULTS: The HPLC method was able to separate all analytes of interest by gradient elution within 25 min. The method was validated according to the guidelines described in the International Conference on Harmonization guideline Q2(R1). CONCLUSIONS: The HPLC method for assay of ivermectin and estimation of its related substances was successfully developed, validated, and demonstrated to be accurate, robust, specific, and stability indicating. HIGHLIGHTS: The performance of the HPLC method is significantly faster and possesses a higher degree of selectivity. Implementation of this method for routine analysis in QC laboratories would save significant time, resources and solvents.

Effect of Lithium and Aluminum on the Mechanical Properties, In Vivo and In Vitro Degradation, and Toxicity of Multiphase Ultrahigh Ductility Mg-Li-Al-Zn Quaternary Alloys for Vascular Stent Application.

Magnesium alloys are the most widely studied biodegradable metals for biodegradable vascular stent application. Two major issues with current magnesium alloy based stents are their low ductility and fast corrosion rates. Several studies have validated that introduction of Li into the magnesium alloys will significantly improve the ductility while alloying with Al will improve the corrosion resistance and strength. In the present study, we studied the effects of alloying different amounts of Li and Al on the Mg-Li-Al-Zn (LAZ) quaternary alloy system. Rods were made from four different LAZ alloys, namely, LAZ611, LAZ631, LAZ911, and LAZ931 following melting, casting, and then extrusion. Systematic assessment of mechanical properties, in vitro corrosion, cytotoxicity, and in vivo degradation including local and systemic toxicity conducted demonstrated the beneficial effects of Li and Al on the mechanical properties. Our results specifically suggest that alloying with Li significantly improved the ductility while Al enhanced the strength of the LAZ alloys. Four of the LAZ alloys exhibited different corrosion rates in Hank's balanced salt solution depending on the chemical composition. Indirect in vitro cytotoxicity tests also showed lower cytotoxicity for the alloys exhibiting higher corrosion resistance. In vivo corrosion rates in the mouse subcutaneous model showed different corrosion rates compared to the in vitro tests. Nevertheless, all of the four LAZ alloys displayed no local and systemic toxicity based on the histology analysis. This research study, therefore, demonstrated the benefits of using Li and Al as alloying elements in LAZ alloys and the potential use of LAZ alloys for vascular stent application.

In vivo quantification of hydrogen gas concentration in bone marrow surrounding magnesium fracture fixation hardware using an electrochemical hydrogen gas sensor.

Magnesium (Mg) medical devices are currently being marketed for orthopedic applications and have a complex degradation process which includes the evolution of hydrogen gas (H2). The effect of H2 exposure on relevant cell types has not been studied; and the concentration surrounding degrading Mg devices has not been quantified to enable such mechanistic studies. A simple and effective method to measure the concentration of H2 in varying microenvironments surrounding Mg implants is the first step to understanding the biological impact of H2 on these cells. Here, the in vivo measurement of H2 surrounding fracture fixation devices implanted in vivo is demonstrated. An electrochemical H2 microsensor detected increased levels of H2 at three anatomical sites with a response time of about 30 s. The sensor showed the H2 concentration in the bone marrow at 1 week post-implantation (1460 ±â€¯320 µM) to be much higher than measured in the subcutaneous tissue (550 ±â€¯210 µM) and at the skin surface (120 ±â€¯50 µM). Additionally, the H2 concentrations measured in the bone marrow exceeded the concentration in a H2 saturated water solution (∼800 µM). These results suggest that H2 emanating from Mg implants in bone during degradation pass through the bone marrow and become at least partially trapped because of slow permeation through the bone. This study is the first to identify H2 concentrations in the bone marrow environment and will enable in vitro experiments to be executed at clinically relevant H2 concentrations to explore possible biological effects of H2 exposure. STATEMENT OF SIGNIFICANCE: An electrochemical H2 sensor was used to monitor the degradation of a Mg fracture fixation system in a lapine ulna fracture model. Interestingly, the H2 concentration in the bone marrow is 82% higher than H2 saturated water solution. This suggests H2 generated in situ is trapped in the bone marrow and bone is less permeable than the surrounding tissues. The detectable H2 at the rabbit skin also demonstrates a H2 sensor's ability to monitor the degradation process under thin layers of tissue. H2 sensing shows promise as a tool for monitoring the degradation of Mg alloy in vivo and creating in vitro test beds to more mechanistically evaluate the effects of varying H2 concentrations on cell types relevant to osteogenesis.

In Vitro and in Vivo Evaluation of Multiphase Ultrahigh Ductility Mg-Li-Zn Alloys for Cardiovascular Stent Application.

Magnesium alloys have been extensively studied as a novel biodegradable metallic material for cardiovascular stent application. However, the ductility limitation of magnesium alloy has been a key issue for biodegradable stents applications. In this study, two different multiphase ultrahigh ductility Mg-Li-Zn alloys, LZ61 and LZ91, are fabricated in the form of extruded rods and evaluated both in vitro and in vivo. The microstructure, mechanical properties and in vitro degradation are evaluated as well as in vitro cytotoxicity. The in vivo degradation, tissue response, and systematic toxicity are evaluated in a mouse subcutaneous model. Measurements show that LZ61 and LZ91 exhibit more than 40% elongation at fracture without significantly compromising the strength. Both in vitro and in vivo degradation showed low degradation rates for LZ61 but high degradation rate for the LZ91 alloy. Excellent biocompatibility is observed both in vivo and in vitro for LZ61 and LZ91. In summary, this study successfully demonstrates that the ultraductility multiphase Mg-Li-Zn alloy has the potential to be used for stent applications. Compared to LZ91, the LZ61 alloy shows better balance of mechanical properties, corrosion resistance, and biocompatibility, indicating its promise for cardiovascular stent applications.

Carbon Nanotube Thread Electrochemical Cell: Detection of Heavy Metals.

In this work, all three electrodes in an electrochemical cell were fabricated based on carbon nanotube (CNT) thread. CNT thread partially insulated with a thin polystyrene coating to define the microelectrode area was used as the working electrode; bare CNT thread was used as the auxiliary electrode; and a micro quasi-reference electrode was fabricated by electroplating CNT thread with Ag and then anodizing it in chloride solution to form a layer of AgCl. The Ag|AgCl coated CNT thread electrode provided a stable potential comparable to the conventional liquid-junction type Ag|AgCl reference electrode. The CNT thread auxiliary electrode provided a stable current, which is comparable to a Pt wire auxiliary electrode. This all-CNT thread three electrode cell has been evaluated as a microsensor for the simultaneous determination of trace levels of heavy metal ions by anodic stripping voltammetry (ASV). Hg2+, Cu2+, and Pb2+ were used as a representative system for this study. The calculated detection limits (based on the 3σ method) with a 120 s deposition time are 1.05, 0.53, and 0.57 nM for Hg2+, Cu2+, and Pb2+, respectively. These electrodes significantly reduce the dimensions of the conventional three electrode electrochemical cell to the microscale.

In vivo characterization of magnesium alloy biodegradation using electrochemical H2 monitoring, ICP-MS, and XPS.

The effect of widely different corrosion rates of Mg alloys on four parameters of interest for in vivo characterization was evaluated: (1) the effectiveness of transdermal H2 measurements with an electrochemical sensor for noninvasively monitoring biodegradation compared to the standard techniques of in vivo X-ray imaging and weight loss measurement of explanted samples, (2) the chemical compositions of the corrosion layers of the explanted samples by XPS, (3) the effect on animal organs by histology, and (4) the accumulation of corrosion by-products in multiple organs by ICP-MS. The in vivo biodegradation of three magnesium alloys chosen for their widely varying corrosion rates - ZJ41 (fast), WKX41 (intermediate) and AZ31 (slow) - were evaluated in a subcutaneous implant mouse model. Measuring H2 with an electrochemical H2 sensor is a simple and effective method to monitor the biodegradation process in vivo by sensing H2 transdermally above magnesium alloys implanted subcutaneously in mice. The correlation of H2 levels and biodegradation rate measured by weight loss shows that this non-invasive method is fast, reliable and accurate. Analysis of the insoluble biodegradation products on the explanted alloys by XPS showed all of them to consist primarily of Mg(OH)2, MgO, MgCO3 and Mg3(PO4)2 with ZJ41 also having ZnO. The accumulation of magnesium and zinc were measured in 9 different organs by ICP-MS. Histological and ICP-MS studies reveal that there is no significant accumulation of magnesium in these organs for all three alloys; however, zinc accumulation in intestine, kidney and lung for the faster biodegrading alloy ZJ41 was observed. Although zinc accumulates in these three organs, no toxicity response was observed in the histological study. ICP-MS also shows higher levels of magnesium and zinc in the skull than in the other organs. STATEMENT OF SIGNIFICANCE: Biodegradable devices based on magnesium and its alloys are promising because they gradually dissolve and thereby avoid the need for subsequent removal by surgery if complications arise. In vivo biodegradation rate is one of the crucial parameters for the development of these alloys. Promising alloys are first evaluated in vivo by being implanted subcutaneously in mice for 1month. Here, we evaluated several magnesium alloys with widely varying corrosion rates in vivo using multiple characterization techniques. Since the alloys biodegrade by reacting with water forming H2 gas, we used a recently demonstrated, simple, fast and noninvasive method to monitor the biodegradation process by just pressing the tip of a H2 sensor against the skin above the implant. The analysis of 9 organs (intestine, kidney, spleen, lung, heart, liver, skin, brain and skull) for accumulation of Mg and Zn revealed no significant accumulation of magnesium in these organs. Zinc accumulation in intestine, kidney and lung was observed for the faster corroding implant ZJ41. The surfaces of explanted alloys were analyzed to determine the composition of the insoluble biodegradation products. The results suggest that these tested alloys are potential candidates for biodegradable implant applications.

Visual H2 sensor for monitoring biodegradation of magnesium implants in vivo.

A visual sensor for H2 was used to transdermally monitor H2 that originated from biodegrading magnesium (Mg) alloys implanted subcutaneously in mice. The visual sensor consisted of a thin film of H2-sensitive material (MoO3 and Pd catalyst) coated on a flexible plastic sheet that was pressed against the mouse skin directly above the implant. Although the H2 levels permeating through the skin during the degradation process were very low, the sensor changed color to give a three dimensional (3D) visualization of H2 permeation. The correlation between the visual sensor response and measurements made with an electrochemical H2 microsensor on several magnesium alloys demonstrates that the visual sensor has the capability to monitor in real-time the dissolution rate of implants in vivo. This detection method is noninvasive, easy to implement, effective and potentially low cost compared to electrochemical detection. STATEMENT OF SIGNIFICANCE: Biodegradable Mg implants offer advantages over permanent implants such as stainless steel that are used for broken bone repair. Mg alloys gradually dissolve, avoiding the need for removal by a later surgery if complications arise. Here we report a visual H2 sensor that can be used in the research laboratory to monitor the corrosion process in vivo during animal testing of different Mg alloys. The sensor consists of a plastic sheet with a thin coating that changes color in the presence of H2 gas. The sensor is easily used by taping it on the skin over the Mg implant. The color change gives a map of the H2 level permeating from the degrading Mg through the skin above it. This low cost, simple method of monitoring the dissolution of biodegradable implants would greatly facilitate the development of the biodegradable materials, especially in animal studies where in vivo biodegradation is tested.

In vivo monitoring the biodegradation of magnesium alloys with an electrochemical H2 sensor.

UNLABELLED: Monitoring the biodegradation process of magnesium and its alloys in vivo is challenging. Currently, this process is monitored by micro-CT and X-ray imaging in vivo, which require large and costly instrumentation. Here we report a simple and effective methodology to monitor the biodegradation process in vivo by sensing H2 transdermally above a magnesium sample implanted subcutaneously in a mouse. An electrochemical H2 microsensor was used to measure the biodegradation product H2 at the surface of the skin for two magnesium alloys (ZK40 and AZ31) and one high purity magnesium single crystal (Mg8H). The sensor was able to easily detect low levels of H2 (30-400µM) permeating through the skin with a response time of about 30s. H2 levels were correlated with the biodegradation rate as determined from weight loss measurements of the implants. This new method is noninvasive, fast and requires no major equipment. STATEMENT OF SIGNIFICANCE: Biomedical devices such as plates and screws used for broken bone repair are being developed out of biodegradable magnesium alloys that gradually dissolve when no longer needed. This avoids subsequent removal by surgery, which may be necessary if complications arise. A rapid, non-invasive means for monitoring the biodegradation process in vivo is needed for animal testing and point of care (POC) evaluation of patients. Here we report a novel, simple, fast, and noninvasive method to monitor the biodegradation of magnesium in vivo by measuring the biodegradation product H2 with an electrochemical H2 sensor. Since H2 rapidly permeates through biological tissue, measurements are made by simply pressing the sensor tip against the skin above the implant; the response is within 30s.

Electrospun carbon nanofiber modified electrodes for stripping voltammetry.

Electrospun polyacrylonitrile (PAN) based carbon nanofibers (CNFs) have attracted intense attention due to their easy processing, high carbon yield, and robust mechanical properties. In this work, a CNF modified glassy carbon (GC) electrode that was coated with Nafion polymer was evaluated as a new electrode material for the simultaneous determination of trace levels of heavy metal ions by anodic stripping voltammetry (ASV). Pb(2+) and Cd(2+) were used as a representative system for this initial study. Well-defined stripping voltammograms were obtained when Pb(2+) and Cd(2+) were determined individually and then simultaneously in a mixture. Compared to a bare GC electrode, the CNF/Nafion modified GC (CNF/Nafion/GC) electrode improved the sensitivity for lead detection by 8-fold. The interface properties of the CNF/Nafion/GC were characterized by electrochemical impedance spectroscopy (EIS), which showed the importance of the ratio of CNF/Nafion on electrode performance. Under optimized conditions, the detection limits are 0.9 and 1.5 nM for Pb(2+) and Cd(2+), respectively.

Optically transparent carbon nanotube film electrode for thin layer spectroelectrochemistry.

Carbon nanotube (CNT) film was evaluated as an optically transparent electrode (OTE) for thin layer spectroelectrochemistry. Chemically inert CNT arrays were synthesized by chemical vapor deposition (CVD) using thin films of Fe and Co as catalysts. Vertically aligned CNT arrays were drawn onto a quartz slide to form CNT films that constituted the OTE. Adequate conductivity and transparency make this material a good OTE for spectroelectrochemistry. These properties could be varied by the number of layers of CNTs used to form the OTE. Detection in the UV/near UV region down to 200 nm can be achieved using these transparent CNT films on quartz. The OTE was characterized by transmission electron microscopy, scanning electron microscopy, Raman spectroscopy, UV-visible spectroscopy, cyclic voltammetry, electrochemical impedance spectroscopy, and thin layer spectroelectrochemistry. Ferricyanide, tris(2,2'-bipyridine) ruthenium(II) chloride, and cytochrome c were used as representative redox probes for thin layer spectroelectrochemistry using the CNT film OTE, and the results correlated well with their known properties. Direct electron transfer of cytochrome c was achieved on the CNT film electrode.

Platinum(II) Monomer and Dimer Complexes with a Bis(oxazolinyl)phenyl Pincer Ligand.

A series of platinum(II) complexes with the formulas Pt(phebox)(L)(+) (phebox(-) = 1,3-bis(4,4'-dimethyl-2'-oxazolinyl)phenyl anion; L = pyridine (py), 4-phenylpyridine, quinoline, acridine) and Pt2(phebox)2(µ-L')(2+) (L' = pyrazine, 4,4'-bipyridine, 1,2-bis(4-pyridyl)ethane) was prepared. Crystallographic data establish that the metal center is bonded to the tridentate phebox(-) and monodentate pyridyl ligands. The five-membered oxazoline rings favor a CH2-CMe2 twist conformation. Pt(phebox)Cl and Pt(phebox)(py)(+) undergo a ligand-based chemically reversible redox reaction, whereas the electrochemistry of the other complexes is chemically and electrochemically less reversible. In contrast to complexes with the 1,3-bis(piperdylmethyl)phenyl anion ligand (pip2NCN(-)) or related pincer ligands, each of the phebox(-) complexes described here exhibits intense emission in room-temperature methylene chloride solution, which is assigned as originating from a lowest, predominantly phebox(-) ligand-centered excited state. In acetonitrile, the complexes undergo solvolysis resulting in displacement of the pyridyl ligands. The accumulated data demonstrate that subtle variations in the nature of the NCN and ancillary ligands of platinum(II) complexes provide access to at least five orbitally distinct emissive excited states.

Target validation and structure-activity analysis of a series of novel PCNA inhibitors.

Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair. Tumor cells express high levels of PCNA, identifying it as a potentially ideal target for cancer therapy. Previously, we identified nine compounds termed PCNA inhibitors (PCNA-Is) that bind directly to PCNA, stabilize PCNA trimer structure, reduce chromatin-associated PCNA, and selectively inhibit tumor cell growth. Of these compounds, PCNA-I1 was most potent. The purpose of this study is to further establish targeting of PCNA by PCNA-I1 and to identify PCNA-I1 analogs with superior potencies. We found that PCNA-I1 does not affect the level of chromatin-associated PCNA harboring point mutations at the predicted binding site of PCNA-I1. Forty-six PCNA-I1 analogs with structures of 1-hydrazonomethyl-2-hydroxy (scaffold A), 2-hydrazonomethyl-1-hydroxy (scaffold B), 2-hydrazonomethyl-3-hydroxy (scaffold C), and 4-pyridyl hydrazine (scaffold D) were analyzed for their effects on cell growth in four tumor cell lines and PCNA trimer stabilization. Compounds in scaffold group A and group B showed the highest trimer stabilization and the most potent cell growth inhibitory activities with a significant potency advantage observed in the Z isomers of scaffold A. The absence of trimer stabilization and growth inhibitory effects in compounds of scaffold group D confirms the essentiality of the hydroxynaphthyl substructure. Compounds structure-activity relationship (SAR)-6 and SAR-24 were analyzed for their effects on and found to reduce chromatin-associated PCNA in tumor cells. This study led to the identification of SAR-24, a compound with superior potencies and potentially improved solubility, which will be used for future development of PCNA-targeting cancer therapies.

In-house and synchrotron X-ray diffraction studies of 2-phenyl-1,10-phenanthroline, protonated salts, complexes with gold(III) and copper(II), and an orthometallation product with palladium(II).

Different salts of the 2-phenyl-1,10-phenanthrolin-1-ium cation, (pnpH)(+), are obtained by reacting 2-phenyl-1,10-phenanthroline (pnp), C18H12N2, (I), with a variety of anions, such as hexafluoridophosphate, C18H13N2(+)·PF6(-), (II), trifluoromethanesulfonate, C18H13N2(+)·CF3SO3(-), (III), tetrachloridoaurate, (C18H13N2)[AuCl4], (IV), and bromide (as the dihydrate), C18H13N2(+)·Br(-)·2H2O, (V). Compound (I) crystallizes with Z' = 2, with both independent molecules adopting a coplanar conformation. In (II)-(IV), a hydrogen bond exists between the cation and anion, while one of the lattice water molecules serves as a hydrogen-bonded bridge between the cation and anion in (V). Reaction of (I) with HAuCl4 gives the salt complex (IV); however, reaction with KAuCl4 produces the monodentate complex trichlorido(2-phenyl-1,10-phenanthroline-κN(10))gold(III), [AuCl3(C18H12N2)], (VI). Dichlorido(2-phenyl-1,10-phenanthroline-κ(2)N,N')copper(II), [CuCl2(C18H12N2)], (VII), results from the reaction of CuCl2·2H2O and (I), in which the Cu(II) center adopts a tetrahedrally distorted square-planar geometry. The pendent phenyl ring twists to a bisecting position relative to the phenanthroline plane. The square-planar Pd(II) complex, bromido[2-(phenanthrolin-2-yl)phenyl-κ(3)C(1),N,N']palladium(II), [PdBr(C18H11N2)], (VIII), is obtained from the reaction of (I) with [PdCl2(cycloocta-1,5-diene)], followed by addition of bromine. A coplanar geometry for the pendent ring is adopted as a result of the tridentate bonding motif.

Carbon nanotube-loaded Nafion film electrochemical sensor for metal ions: europium.

A Nafion film loaded with novel catalyst-free multiwalled carbon nanotubes (MWCNTs) was used to modify a glassy carbon (GC) electrode to detect trace concentrations of metal ions, with europium ion (Eu(3+)) as a model. The interaction between the sidewalls of MWCNTs and the hydrophobic backbone of Nafion allows the MWCNTs to be dispersed in Nafion, which was then coated as a thin film on the GC electrode surface. The electrochemical response to Eu(3+) was found to be ∼10 times improved by MWCNT concentrations between 0.5 and 2 mg/mL, which effectively expanded the electrode surface into the Nafion film and thereby reduced the diffusion distance of Eu(3+) to the electrode surface. At low MWCNT concentrations of 0.25 and 0.5 mg/mL, no significant improvement in signal was obtained compared with Nafion alone. Scanning electron microscopy and electrochemical impedance spectroscopy were used to characterize the structure of the MWCNT-Nafion film, followed by electrochemical characterization with Eu(3+) via cyclic voltammetry and preconcentration voltammetry. Under the optimized conditions, a linear range of 1-100 nM with a calculated detection limit of 0.37 nM (signal/noise = 3) was obtained for determination of Eu(3+) by Osteryoung square-wave voltammetry after a preconcentration time of 480 s.