Facile synthesis of Vulcan XC-72 nanoparticles-decorated halloysite nanotubes for the highly sensitive electrochemical determination of niclosamide.

We report a scalable and controllable ultrasound-assisted strategy for the preparation of Vulcan XC-72 nanoparticles-decorated halloysite nanotubes (HNTs@VXC-72), which was applied to modify glassy carbon electrode (GCE) for the highly sensitive electrochemical determination of niclosamide (NA). For the HNTs@VXC-72 nanocomposite, VXC-72 nanoparticles with excellent electrical conductivity and good dispersing property contributed to the formation of the interconnected conductive network; HNTs possessed good adsorption performance and promoted the electrochemical redox reaction. The research results showed that the combination of VXC-72 nanoparticles and HNTs produced the effect of synergistic enhancement. The HNTs@VXC-72/GCE sensor could show a relatively low detection limit of 3.28 nM in the great linear NA concentration range of 0.01-1 µM. When used for the NA determination in food samples, the HNTs@VXC-72/GCE sensor exhibited good practical feasibility with low RSD and acceptable recoveries, which provided a promising NA determination approach to ensure food safety.

Development of a micelle-enhanced high-throughput fluorometric method for determination of niclosamide using a microplate reader.

The present paper describes the development and validation of a simple and sensitive micelle-enhanced high-throughput fluorometric method for the determination of niclosamide (NIC) in 96-microwell plates. The proposed method is based on the reduction of the nitro group of niclosamide to an amino group using Zn/HCl to give a highly fluorescent derivative that was developed simultaneously and measured at λem 444 nm after excitation at λex 275 nm. Tween-80 and carboxymethylcellulose (CMC) have been used as fluorescence enhancers and greatly enhanced the fluorescence by factors of 100-150%. The different experimental conditions affecting the fluorescence reaction were carefully investigated and optimized. The proposed method showed good linearity (r2 ≥ 0.9997) over the concentration ranges of 1-5 and 0.5-5 µg/ml with lower detection limits of 0.01 and 0.008 µg/ml and lower quantification limits of 0.04 and 0.03 µg/ml on using Tween-80 and or CMC, respectively. The developed high-throughput method was successfully applied for the determination of niclosamide in both tablets and spiked plasma. The capability of the method for measuring microvolume samples made it convenient for handling a very large number of samples simultaneously. In addition, it is considered an environmentally friendly method with lower consumption of chemicals and solvents.

Residue depletion and risk assessment of niclosamide in three species of freshwater fish.

The residue depletion of niclosamide (NIC) in freshwater fish including blunt snout bream (Megalobrama amblycephala), yellow catfish (Pelteobagrus fulvidraco) and channel catfish (Ietalurus Punetaus) were investigated under laboratory conditions. NIC concentrations in fish were determined by Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) with heated electrospray ionisation. The mean recoveries were in the range of 75.5-98.5%, with relative standard deviations ranging from 1.0 to 9.4%. The depletion results showed that the long half-life of NIC was 8.8-22.1 days. Risk assessment of NIC residue in edible tissues (muscle and skin) of the three species of freshwater fish was performed using the quotient (RQ), because of no maximum residue limit (MRL) has been set for NIC in fish. The results demonstrated that the RQ values were all significantly lower than 1. Therefore, the effect of NIC residues in the three species of freshwater fish at the immersion concentration was negligible to Chinese people.

Indirect photodegradation of the lampricides TFM and niclosamide.

3-Trifluromethyl-4-nitrophenol (TFM) and 2',5-dichloro-4'-nitrosalicylanilide (niclosamide) are lampricides used in tributaries of the Great Lakes to kill the invasive parasitic sea lamprey (Petromyzon marinus). Although the lampricides have been applied since the late 1950s, their photochemical behavior in natural environments is still not well understood. This study examines the indirect photodegradation of these two compounds and the resulting yields of organic and inorganic photoproducts in water samples collected from five tributaries of Lake Michigan. The tributaries were selected to span the length of Lake Michigan and its natural carbonate geologic gradient. In the presence of dissolved organic matter (DOM), the niclosamide photodegradation rate triples, while the rate of TFM photodegradation is unchanged. Additionally, the yield of lampricide organic products is influenced by DOM because many of the organic photoproducts themselves are prone to DOM-mediated indirect photodegradation. The indirect photodegradation of niclosamide is primarily mediated by reaction with singlet oxygen, which accounts for more than 50% of the increased photodegradation rate. Additionally, hydroxyl radicals and carbonate radicals (CO3-˙) influence niclosamide indirect photolysis, and their contribution is dependent on the specific river water chemistry. For example, CO3-˙ contribution to niclosamide photodegradation, while small, is greater in southern tributaries where there is higher carbonate alkalinity.

A field analysis of lampricide photodegradation in Great Lakes tributaries.

The lampricides 3-trifluoromethyl-4-nitrophenol (TFM) and 2',5-dichloro-4'-nitrosalicylanilide (niclosamide) are added to Great Lakes tributaries to target the sea lamprey, an invasive parasitic fish. This study examines the photochemical behavior of the lampricides in Carpenter Creek, Sullivan Creek, and the Manistique River. The observed loss of TFM in Carpenter and Sullivan Creeks (i.e., 34 and 19%) was similar to the loss of bromide in parallel time of passage studies (i.e., 30 and 29%), demonstrating that TFM photodegradation was minimal in both tributaries during the lampricide application. Furthermore, the absence of inorganic and organic photoproducts in the Manistique River demonstrates that TFM and niclosamide photodegradation was minimal in this large tributary, despite its long residence time (i.e., 3.3 days). Kinetic modeling was used to identify environmental variables primarily responsible for the limited photodegradation of TFM in the field compared to estimates from laboratory data. This analysis demonstrates that the lack of TFM photodegradation was attributable to the short residence times in Carpenter and Sullivan Creeks, while depth, time of year, time of day, and cloud cover influenced photochemical fate in the Manistique River. The modeling approach was extended to assess how many of the 140 United States tributaries treated with lampricides in 2015 and 2016 were amenable to TFM photolysis. While >50% removal of TFM due to photolysis could occur in 13 long and shallow tributaries, in most systems lampricides will reach the Great Lakes untransformed.

Evaluation of quantitative assays for the identification of direct signal transducer and activator of transcription 3 (STAT3) inhibitors.

In many forms of cancer the signal transducer and activator of transcription 3 (STAT3) transcription factor remains constitutively active, driving cancer survival and progression. The critical role of STAT3 in tumorigenesis has prompted a campaign of drug discovery programs to identify small molecules that disrupt the function of STAT3, with more recent efforts focusing on direct STAT3 inhibition. There are two target binding sites for direct STAT3 inhibitors: the SH2 dimerization domain and the DNA-binding domain. An in vitro fluorescence polarization assay, using recombinant STAT3 protein, has successfully identified compounds that target the SH2 domain; however, no assay has been reported to identify inhibitors that bind the DNA-binding domain. The lack of such a quantitative assay has limited the identification and development of STAT3 DNA-binding domain inhibitors. Here, we report a modified DNA-binding ELISA to incorporate recombinant STAT3 protein to evaluate small molecules that prevent STAT3-DNA binding. The concomitant use of the ELISA and fluorescence polarization assay enables the classification of direct STAT3 inhibitors by their site of action. Our data provide further support that niclosamide inhibits STAT3 through interaction with the DNA-binding domain. Furthermore, the ELISA can support medicinal chemistry efforts by identifying DNA-binding domain inhibitors and allowing the determination of an IC50 value, supporting the ranking of inhibitors and development of structure-activity relationships. Therefore, we propose a tandem evaluation approach to identify small molecules that target the SH2 domain or the DNA-binding domain of STAT3, which allows for quantitative evaluation of candidate STAT3 inhibitors.

[Determination of niclosamide ethanolamine residue in rice and paddy field by high performance liquid chromatography].

A high performance liquid chromatographic (HPLC) method was established for the analysis of niclosamide ethanolamine residue in rice and paddy field. The paddy water and plant were extracted with alkaline ethylacetate, while the paddy soil, rice husk and unpolished rice were first extracted with alkaline ethanol, and then with ethylacetate. The extracts were then cleaned-up by a Florisil column and detected by high performance liquid chromatography with an ultraviolet detector (UVD) on a Welchrom C18 column (250 mm x 4.6 mm, 5 microm). The calibration curve showed good linearity from 0.01 mg/L to 10.00 mg/L with the correlation coefficient more than 0. 999 8. The average recoveries of this method were from 93.47% to 100.9% with the relative standard deviations of 1.46% - 5.82% at the spiked levels of 0.01 - 5.00 mg/kg. This method is fast, simple, sensitive, reproducible and practical for the determination of niclosamide ethanolamine residue in paddy fields, and can meet the requirement of the determination of pesticide residues.

Residues of the lampricides 3-trifluoromethyl-4-nitrophenol and niclosamide in muscle tissue of rainbow trout.

Rainbow trout (Oncorhyncus mykiss) were exposed to the (14)C-labeled lampricide 3-trifluoromethyl-4-nitrophenol (TFM) (2.1 mg/L) or niclosamide (0.055 mg/L) in an aerated static water bath for 24 h. Fish were sacrificed immediately after exposure. Subsamples of skin-on muscle tissue were analyzed for residues of the lampricides. The primary residues in muscle tissue from fish exposed to TFM were parent TFM (1.08 +/- 0.82 nmol/g) and TFM-glucuronide (0.44 +/- 0.24 nmol/g). Muscle tissue from fish exposed to niclosamide contained niclosamide (1.42 +/- 0.51 nmol/g), niclosamide-glucuronide (0.0644 +/- 0.0276 nmol/g), and a metabolite not previously reported, niclosamide sulfate ester (1.12 +/- 0.33 nmol/g).

Detection of the damage caused to DNA by niclosamide using an electrochemical DNA-biosensor.

Niclosamide is the only commercially available molluscicide recommended by the WHO for large-scale use in schistosomiasis control programs. The electrochemical reduction and oxidation mechanism of niclosamide was studied using cyclic, differential and square wave voltammetry, at a glassy carbon electrode. An indirect procedure for in situ quantification of niclosamide using batch injection analysis with electrochemical detection, possible to be used for in situ determinations in river streams and effluents, was developed. It enabled a detection limit of 8 x 10(-7) M. The investigation of the niclosamide-DNA interaction using an electrochemical DNA-biosensor showed for the first time clear evidence of interaction with DNA and suggested that niclosamide toxicity can be caused by this interaction, after reductive activation.

Rapid loss of lampricide from catfish and rainbow trout following routine treatment.

Rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) were exposed to 3-trifluoromethyl-4-nitrophenol (TFM) and Bayluscide (niclosamide) during a sea lamprey control treatment of the Ford River, located in the upper peninsula of Michigan. Caged fish were exposed to a nominal concentration of 0.02 mg/L of niclosamide for a period of approximately 12 h. Samples of fillet tissue were collected from each fish species before treatment and at 6, 12, 18, 24, 48, 96, and 192 h following the arrival of the block of chemical at the exposure site. The fish were dissected, homogenized, extracted, and analyzed by high-performance liquid chromatography. The major residues found in the fillet tissues were TFM and niclosamide. Niclosamide concentrations were highest 12 h after arrival of the chemical block for rainbow trout (0.0395 +/- 0.0251 microg/g) and 18 h after arrival of the chemical block for channel catfish (0.0465 +/- 0.0212 microg/g). Residues decreased rapidly after the block of lampricide had passed and were below the detection limits in fillets of rainbow trout within 24 h and channel catfish within 96 h after the arrival of the lampricide.

Effect of soil moisture and sample depth on pesticide photolysis.

The effects of soil depth and moisture on pesticide photolysis were studied. Moist soil at depths of 3, 2.5, 2, 1.5, 1, and 0.5 mm were each dosed at 2.5 microg/g with (14)C-niclosamide and photolyzed under a xenon lamp at constant temperature. Samples were removed after 20, 40, 110, and 153 h of continuous irradiation. The decrease in percent of niclosamide and the appearance of degradates were followed by analyzing the soil extracts by HPLC. A corresponding set of experiments used air-dried soil. An experiment was also performed using initially moist soil which was permitted to dry during photolysis but returned to moist conditions at each sampling. Qualitative and quantitative differences were found in the rate and route of degradation of niclosamide under these conditions. These differences have resulted from a combination of reduced photochemical activity and microbial population in dry soil. The half-lives of niclosamide in the dry soils were 2 to 5 times longer than those in the moisture-maintained soil. There was also a noticeable difference in the half-lives in soil of different depths. Moisture-maintained soil showed a uniform linear increase in half-life from 95 to 195 h as soil depth increased from 0.5 mm to 3.0 mm. With air-dried soil the half-lives were greatly dependent on soil depth, showing a much broader range of 199 h at 0.5-mm to 1064 h in 3.0-mm soil. An experimental design is described which maintains soil temperature and moisture to preset conditions.

Voltammetric determination of niclosamide at a glassy carbon electrode.

A very sensitive and selective procedure was developed for the determination of niclosamide based on square-wave voltammetry at a glassy carbon electrode. Cyclic voltammetry was used to investigate the electrochemical reduction of niclosamide at a glassy carbon electrode. Niclosamide was first irreversibly reduced from NO2 to NHOH at -0.659 V in aqueous buffer solution of pH 8.5. Reversible and well defined peaks at -0.164 V and -0.195 V (vs. Ag/AgCl) were obtained that are responsible for two electron peaks between NHOH and NO. Following optimisation of the voltammetric parameters, pH and reproducibility, a linear calibration curve over the range 5 x 10(-8)-1 x 10(-6) mol dm(-3) was achieved. The detection limit was found to be 2.05 x 10(-8) mol dm(-3) niclosamide. For eight successive determinations of 5 x 10(-7) mol dm(-3) niclosamide, a relative standard deviation of 2.4% was obtained. This voltammetric method was applied to the direct determination of niclosamide in tablets. The results of the analysis suggest that the proposed method has promise for the routine determination of niclosamide in the products examined.

Determination of niclosamide residues in rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) fillet tissue by high-performance liquid chromatography.

Bayluscide [the ethanolamine salt of niclosamide (NIC)] is a registered piscicide used in combination with 3-(trifluoromethyl)-4-nitrophenol (TFM) to control sea lamprey populations in streams tributary to the Great Lakes. A high-performance liquid chromatography (HPLC) method was developed for the determination of NIC residues in muscle fillet tissues of fish exposed to NIC and TFM during sea lamprey control treatments. NIC was extracted from fortified channel catfish and rainbow trout fillet tissue with a series of acetone extractions and cleaned up on C(18) solid-phase extraction cartridges. NIC concentrations were determined by HPLC with detection at 360 and 335 nm for rainbow trout and catfish, respectively. Recovery of NIC from rainbow trout (n = 7) fortified at 0.04 microg/g was 77 +/- 6.5% and from channel catfish (n = 7) fortified at 0.02 microg/g was 113 +/- 11%. NIC detection limit was 0.0107 microg/g for rainbow trout and 0.0063 microg/g for catfish. Percent recovery of incurred radioactive residues by this method from catfish exposed to [(14)C]NIC was 89.3 +/- 4.1%. Percent recoveries of NIC from fortified storage stability tissue samples for rainbow trout (n = 3) analyzed at 5 and 7.5 month periods were 78 +/- 5.1 and 68 +/- 2.4%, respectively. Percent recoveries of NIC from fortified storage stability tissue samples for channel catfish (n = 3) analyzed at 5 and 7.5 month periods were 88 +/- 13 and 76 +/- 21%, respectively.

Spectrophotometric determination on niclosamide in chewable tablets

Amostras de comprimidos mastigáveis contendo niclosamida foram analisadas por método espectrofotométrico no ultravioleta padronizado a 333 nm. A reta de calibraçäo com coeficiente de correlaçäo r= 0,9884 passa pela origem. A lei de Lambert-Beer é obedecida no intervalo de concentraçäo 6,4 a 20,8 g/mL. O coeficiente de variaçäo foi de 0,27 por cento e 0,51 por cento para amostras comerciais e simuladas, respectivamente. Os excipientes näo interferem na determinaçäo

Extractive alkylation of 5,2'-dichloro-4'-nitrosalicylanilide (niclosamide) for gas-liquid chromatographic analysis.

Niclosamide (5,2'-dichloro-4'-nitrosalicylanilide), formulated as the ethanolamine salt, is one of the most effective and widely used molluscicides for the control of snail vectors of schistosomiasis, a parasitic disease afflicting over 200 million people in more than 70 countries. This report details the development of a sensitive, specific analytical method for the evaluation of niclosamide formulations and assessment of the impact of residues on the environment. Efficient (greater than 85%) phase-transfer, N,O-dimethylation of niclosamide and the synthesized 5-deschloro analog internal standard, followed by gas-liquid chromatographic separation and electron-capture detection, permits the determination of as little as 10 ppb analyte in fortified, stagnant water. Tetrabutylammonium chloride was the phase-transfer agent, while methyl iodide was the methylating agent of choice among four investigated.

Gas-liquid chromatographics determination of Bayer 73 in fish, aquatic invertebrates, mud, and water.

A gas-liquid chromatographic (GLC) method is described for determining residues of Bayer 73 (2-aminoethanol salt of 2',5-dichloro-4'-nitrosalicylanilide) in fish muscle, aquatic invertebrates, mud, and water by analyzing for 2-chloro-4-nitroaniline (CNA), a hydrolysis product of Bayer 73. Bayer 73 residues are extracted from fish muscle tissue, invertebrates, and mud with acetone-formic acid (98+2), and partitioned from water samples with chloroform. After sample cleanup by solvent and acid-base partitioning, the concentrated extract is hydrolyzed with 2N NaOH and H2O2 for 10 min at 95 degrees C. The CNA is then partitioned into hexane-ethyl ether (7+3) and determined by electron capture GLC. Average recoveries were 88% for fish, 82% for invertebrates, 82% for mud, and 98% for water at 3 or more fortification levels.