Environmental monitoring of trace metal pollutants using cellulosic-paper incorporating color change of azo-chromophore.

An essential requirement for colorimetric paper-sensor is to allow the target analytes (heavy metal ions) to access the chromophore while maintaining strong chromophore immobilization on the porous substrate surface. This work evaluates the selection of sensitive chromophores (dithizone, 1-(2-pyridylazo) 2-naphthol and 4-(2-pyridylazo)-resorcinol) and their immobilization strategies on paper sensors. Dithizone (DTz) are capable of producing a significant color transition at unadjusted pH, observed by UV-Vis absorption spectroscopy and visible recognition. After immobilizing DTz on a paper substrate (cellulose acetate/chitosan substrate), the DTz-paper sensor showed a distinctive color change from blue-green to peach-pink upon reaction with Pb2+ ions, and the color intensity was proportional to the metal concentration. Quantitative analysis using RGB (R:Red; G:Green; B:Blue) plots showed that increasing DTz concentration on the CA/CS paper sensor increases the difference in total color intensity (∆IT) and the difference in red code intensity (∆IR). This is due to the formation of more DTz-Pb2+ complexes on the CA/CS paper substrate. The CA/CS paper strips immobilized with 100 ppm DTz showed practical potential for rapid detection of heavy metal ions. The DTz-CA/CS paper sensor showed significant color change when detecting spiked heavy metals ions (0.1 ppm Pb2+, 2.0 ppm Zn2+, and 0.2 ppm Cu2+) in river water samples that prepared at the maximum permissible limit for industrial effluent in Malaysia.

Automatic multicommuted flow-batch setup for photometric determination of mercury in drinking water at ppb level.

Herein, a multicommuted flow-batch setup and a photometric procedure for the determination of mercury at the ppb level in aqueous samples are described. The setup was designed to implement a versatile solvent extraction and pre-concentration strategy by combining flow-batch and multicommuted flow analysis approaches. The photometric method was based on Hg(II) reaction with dithizone in a chloroform medium, which was also used as the extracting organic solvent. The flow analysis system was composed of a homemade syringe pump module, a set of solenoid valves, two Aquarius mini-pumps, and a flow-batch chamber. The homemade photometer was comprised of a light emitting diode (LED), photodiode, and homemade flow cell (50 mm length). The flow system and photometer were controlled using an Arduino Due board, running custom-written software. After optimizing the operational conditions, the effectiveness of the developed system was evaluated for the determination of the mercury concentration in drinking water. For accuracy assessment, samples were analyzed using a spiking methodology and an independent method, yielding a recovery ranging from 92% to 108%. Other important characteristics of the proposed method were found as follows: linear response range, 0.5-10.0 µg L-1 (r = 0.9984); limit of detection 0.38 µg L-1 Hg(II); consumption of dithizone and chloroform, 1.85 µg L-1 and 0.8 mL per analysis, respectively; coefficient of variation, 2% (n = 10); sampling throughput, 20 determinations per h.

Pancreatic human islets and insulin-producing cells derived from embryonic stem cells are rapidly identified by a newly developed Dithizone.

We developed an optimized Dipheylthiocarbazone or Dithizone (DTZ) with improved physical and chemical properties to characterize human islets and insulin-producing cells differentiated from embryonic stem cells. Application of the newly formulated iDTZ (i stands for islet) over a range of temperatures, time intervals and cell and tissue types found it to be robust for identifying these cells. Through high transition zinc binding, the iDTZ compound concentrated in insulin-producing cells and proved effective at delineating zinc levels in vitro.

Surfactant-Assisted Emulsification and Surfactant-Based Dispersive Liquid-Liquid Microextraction Method for Determination of Cu(II) in Food and Water Samples by Flame Atomic Absorption Spectrometry.

Background: Copper (Cu) is an essential metal for humans at certain concentrations. However, it can be toxic at higher concentrations. Therefore, determination of Cu content of foodstuff is important. Objective: The aim of the study was to develop a simple, economical, and environmentally friendly surfactant-mediated extraction method for the determination of Cu using surfactants and flame atomic absorption spectrometry (FAAS). Methods: A nonionic surfactant-assisted emulsification and surfactant-based dispersive liquid-liquid microextraction method was developed for the separation, preconcentration, and determination of Cu by FAAS. Triton X-15 nonionic surfactant, which is insoluble in water, was used as an extractive agent. Triton X-114 (TX-114) nonionic water-soluble surfactant was used as a disperser solvent. Dithizone was used as a complexing agent for complexation of Cu(II) at pH 4. Results: The detection and quantitation limits of the method were determined as 1.61 and 3.82 µg/L, respectively. The preconcentration factor was obtained as 50. Relative SD based on 10 replicates was obtained as 3.7%. Accuracy of the developed method was proved using certified standard reference materials. Cu(II) content of edible mushroom samples was determined between 12 and 19 µg/g. Recoveries were obtained between 96 and 101%. Conclusions: The developed surfactant-assisted emulsification and surfactant-based dispersive liquid-liquid microextraction method has represented the wide linear ranges, low detection limit, and high preconcentration factor for Cu ions. Highlights: TX-114 surfactant was used as both sticking agent and disperser solvent. The method does not require expert personnel and high operational costs. The method is environmentally friendly because mainly surfactants and low-toxicity organic solvents are used in the recommended procedure.

Zinc-Chelating Small Molecules Preferentially Accumulate and Function within Pancreatic ß Cells.

Diabetes is a hyperglycemic condition characterized by pancreatic ß-cell dysfunction and depletion. Whereas methods for monitoring ß-cell function in vivo exist, methods to deliver therapeutics to ß cells are lacking. We leveraged the rare ability of ß cells to concentrate zinc to preferentially trap zinc-binding molecules within ß cells, resulting in ß-cell-targeted compound delivery. We determined that zinc-rich ß cells and islets preferentially accumulated TSQ (6-methoxy-8-p-toluenesulfonamido-quinoline) in a zinc-dependent manner compared with exocrine pancreas. Next, we asked whether appending a zinc-chelating moiety onto a ß-cell replication-inducing compound was sufficient to confer preferential ß-cell accumulation and activity. Indeed, the hybrid compound preferentially accumulated within rodent and human islets in a zinc-dependent manner and increased the selectivity of replication-promoting activity toward ß cells. These data resolve the fundamental question of whether intracellular accumulation of zinc-chelating compounds is influenced by zinc content. Furthermore, application of this principle yielded a proof-of-concept method for ß-cell-targeted drug delivery and bioactivity.

A novel and sensitive carbon paste electrode with clinoptilolite nano-particles containing hexadecyltrimethyl ammonium surfactant and dithizone for the voltammetric determination of Sn(II).

Clinoptilolite nano-particles (CNP) were modified by hexadecyltrimethyl ammonium bromide surfactant (HDTMA) and dithizone (DZ). The modified zeolite was then used for the modification of carbon paste electrode (CPE). The resulted CNP-HDTMA-DZ-CPE was finally used for the voltammetric determination of Sn(II) in aqueous solution. The raw and modified CNP was characterized by SEM-EDX, BET, TG-DTG and FTIR techniques. The best voltammetric response was obtained in 0.1molL-1 NaNO3+HCl at pH 2.2 as supporting electrolyte. The electrode showed a linear response in the concentration range of 1×10-8 to 1×10-2molL-1Sn(II) with LOD about 9×10-9molL-1Sn(II). The electrode showed good selectivity and applicability for the determination of Sn(II) in real samples such as river water, an steel company wastewater, tomato paste and a canned tuna fish.

Single-indicator-based Multidimensional Sensing: Detection and Identification of Heavy Metal Ions and Understanding the Foundations from Experiment to Simulation.

Multidimensional sensing offers advantages in accuracy, diversity and capability for the simultaneous detection and discrimination of multiple analytes, however, the previous reports usually require complicated synthesis/fabrication process and/or need a variety of techniques (or instruments) to acquire signals. Therefore, to take full advantages of this concept, simple designs are highly desirable. Herein, a novel concept is conceived to construct multidimensional sensing platforms based on a single indicator that has capability of showing diverse color/fluorescence responses with the addition of different analytes. Through extracting hidden information from these responses, such as red, green and blue (RGB) alterations, a triple-channel-based multidimensional sensing platform could consequently be fabricated, and the RGB alterations are further applicable to standard statistical methods. As a proof-of-concept study, a triple-channel sensing platform is fabricated solely using dithizone with assistance of cetyltrimethylammonium bromide (CTAB) for hyperchromicity and sensitization, which demonstrates superior capabilities in detection and identification of ten common heavy metal ions at their standard concentrations of wastewater-discharge of China. Moreover, this sensing platform exhibits promising applications in semi-quantitative and even quantitative analysis individuals of these heavy metal ions with high sensitivity as well. Finally, density functional theory calculations are performed to reveal the foundations for this analysis.

Insulin-Producing Cells Differentiated from Human Bone Marrow Mesenchymal Stem Cells In Vitro Ameliorate Streptozotocin-Induced Diabetic Hyperglycemia.

BACKGROUND: The two major obstacles in the successful transplantation of islets for diabetes treatment are inadequate supply of insulin-producing tissue and immune rejection. Induction of the differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs) into insulin-producing cells (IPCs) for autologous transplantation may alleviate those limitations. METHODS: hMSCs were isolated and induced to differentiate into IPCs through a three-stage differentiation protocol in a defined media with high glucose, nicotinamide, and exendin-4. The physiological characteristics and functions of IPCs were then evaluated. Next, about 3 × 10(6) differentiated cells were transplanted into the renal sub-capsular space of streptozotocin (STZ)-induced diabetic nude mice. Graft survival and function were assessed by immunohistochemistry, TUNEL staining and measurements of blood glucose levels in the mice. RESULTS: The differentiated IPCs were characterized by Dithizone (DTZ) positive staining, expression of pancreatic ß-cell markers, and human insulin secretion in response to glucose stimulation. Moreover, 43% of the IPCs showed L-type Ca2+ channel activity and similar changes in intracellular Ca2+ in response to glucose stimulation as that seen in pancreatic ß-cells in the process of glucose-stimulated insulin secretion. Transplantation of functional IPCs into the renal subcapsular space of STZ-induced diabetic nude mice ameliorated the hyperglycemia. Immunofluorescence staining revealed that transplanted IPCs sustainably expressed insulin, c-peptide, and PDX-1 without apparent apoptosis in vivo. CONCLUSIONS: IPCs derived from hMSCs in vitro can ameliorate STZ-induced diabetic hyperglycemia, which indicates that these hMSCs may be a promising approach to overcome the limitations of islet transplantation.

Dithizone-modified graphene oxide nano-sheet as a sorbent for pre-concentration and determination of cadmium and lead ions in food.

Graphene oxide nano-sheet was modified with dithizone as a novel sorbent for selective pre-concentration and determination of Cd(II) and Pb(II) in food. The sorbent was characterised by various analytical methods and the effective parameters for Cd(II) and Pb(II) adsorption were optimised during this work. The high adsorption capacity and selectivity of this sorbent makes the method capable of fast determinations of the Cd(II) and Pb(II) content in complicated matrices even at µg l(-1) levels using commonly available instrumentation. The precision of this method was < 1.9% from 10 duplicate determinations and its accuracy verified using standard reference materials. Finally, this method was applied to the determination of Cd(II) and Pb(II) ions in common food samples and satisfactory results were obtained.

Zinc-Dithizone Complex Engineered Upconverting Nanosensors for the Detection of Hypochlorite in Living Cells.

Current chemo/biosensors for hypochlorous acid or hypochlorite detections are usually limited to the submicromolar level because of their insufficient sensitivity, which is a problem because the concentrations in biological matrices is generally on the nanomolar scale or even lower. Developing a probe with a high enough sensitivity remains a challenge. Using the minimal background fluorescence of upconversion nanocrystals to our advantage, we herein report on an energy-transfer mechanism-based upconversion luminescent nanosensor for the sensitive and selective detection of hypochlorite in aqueous solution. In this nanosensor water-dispersible upconversion nanoparticles act as the energy donor and a novel hypochlorite-responsive coordination complex Zn(DZ)3 is employed as the energy acceptor. The quenched upconversion luminescence, induced by the Zn(DZ)3 complex, can be efficiently recovered after addition of hypochlorite through the selective oxidative breakage of the Zn-S-C bonds in the Zn(DZ)3 complex, which was verified by mass spectrometry. The detection limit for hypochlorite of this sensing system is as low as 3 nM. Furthermore, this newly coordination-complex engineered upconversion nanosensor is successfully applied to image different amounts of exogenous hypochlorite in living HeLa cells.

Screening of cadmium and lead in potentially contaminated waters using a spectrophotometric sequential injection lab-on-valve methodology.

The present work describes the development of a µSI-LOV method for the simultaneous screening of cadmium and lead in potentially contaminated water samples. To attain the biparametric determination, dithizone was chosen as the spectrophotometric reagent as it forms a colored complex with both metal ions, at different pH conditions. The cadmium determination was attained in strong alkaline conditions (pH≈12); the lead determination was calculated by the difference with the determination of both metals in mild alkaline conditions (pH≈8). The colored complex was measured at 550 nm and the method presented a LOD of 34 µg L(-1) for cadmium and 56 µg L(-1) for lead, with a sample consumption of 20 µL per assay and a determination rate of 55 h(-1). The results obtained were in agreement with those obtained by FAAS. The developed method was efficiently applied to the screening of cadmium and lead in marine port waters.

Interaction evaluation of silver and dithizone complexes using DFT calculations and NMR analysis.

Silver has distinct antibacterial properties and has been used as a component of commercial products with many applications. An increasing number of commercial products cause risks of silver effects for human and environment such as the symptoms of Argyria and the release of silver to the environment. Therefore, the detection of silver in the aquatic environment is important. The colorimetric chemosensor is designed by the basic of ligand interactions with metal ion, leading to the change of signals for the naked-eyes which is very useful method to this application. Dithizone ligand is considered as one of the effective chelating reagents for metal ions due to its high selectivity and sensitivity of a photochromic reaction for silver as well as the linear backbone of dithizone affords the rotation of various isomeric forms. The present study is focused on the conformation and interaction of dithizone with silver using density functional theory (DFT). The interaction parameters were determined in term of binding energy of complexes and the geometry optimization, frequency of the structures and calculation of binding energies using density functional approaches B3LYP and the 6-31G(d,p) basis set. Moreover, the interaction of silver-dithizone complexes was supported by UV-Vis spectroscopy, FT-IR spectrum that were simulated by using B3LYP/6-31G(d,p) and (1)H NMR spectra calculation using B3LYP/6-311+G(2d,p) method compared with the experimental data. The results showed the ion exchange interaction between hydrogen of dithizone and silver atom with minimized binding energies of silver-dithizone interaction. Therefore, the results can be the useful information for determination of complex interaction using the analysis of computer simulations.

A highly selective voltammetric sensor for nanomolar detection of mercury ions using a carbon ionic liquid paste electrode impregnated with novel ion imprinted polymeric nanobeads.

This work reports the preparation of a voltammetric sensor for selective recognition and sensitive determination of mercury ions using a carbon ionic liquid paste electrode (CILE) impregnated with novel Hg(2+)-ion imprinted polymeric nanobeads (IIP) based on dithizone, as a suitable ligand for complex formation with Hg(2+) ions. The differential pulse anodic stripping voltammetric technique was employed to investigate the performance of the prepared IIP-CILE for determination of hazardous mercury ions. The designed modified electrode revealed linear responses in the ranges of 0.5nM-10nM and 0.08µM-2µM with a limit of detection of 0.1nM (S/N=3). It was found that the peak currents of the modified electrode for Hg(2+) ions were at a maximum value in phosphate buffer of pH4.5. The optimized preconcentration potential and accumulation time were to be -0.9V and 35s, respectively. The applicability of the proposed sensor to mercury determination in waste water samples is reported.

Modification of an Iranian clinoptilolite nano-particles by hexadecyltrimethyl ammonium cationic surfactant and dithizone for removal of Pb(II) from aqueous solution.

Natural clinoptilolite tuff was mechanically converted to micro (MCP) and nano (NCP) particles. The MCP and NCP powders were respectively modified with hexadecyltrimethyl ammonium bromide (HDTMA) and dithizone (DTZ). The raw and modified samples were characterized by X-ray diffraction (XRD), Fourier transformation infra red (FT-IR), scanning electron microscope (SEM), transmission electron microscope (TEM) and thermogravimetry (TG) and used for the removal of Pb(II) from aqueous solution. The results confirm that both ion exchange and complexation processes are responsible for removal of Pb(II) cations in the modified samples, while Pb(II) cations were only removed via an ion exchange process by the raw clinoptilolite. In this sorbent, the anionic removal property of surfactant modified zeolites (SMZs) changed to cationic removal property by an additional modification step. The best removal efficiency was observed by NCP-HDTMA-DTZ at the following experimental conditions: C(Pb(II)): 800 mg L(-1), HDTMA dosage: 0.2 mol L(-1), DTZ dosage: 5 mmol L(-1), contact time of DTZ with NCP-HDTMA: 1800 min and contact time of the sorbent with Pb(II): 360 min. The NCP-HDTMA-DTZ sorbent showed good efficiency for the removal of lead in the presence of different multivalent cations. Adsorption isotherms of Pb(II) ions obey the Langmuir equation that indicate the monolayer sorption of Pb(II). The adsorption kinetics based on the pseudo-second-order rate equation indicates that the rate limiting step involving a chemical reaction. The negative ΔH and ΔG indicate an exothermic and spontaneous process.

Highly selective and sensitive optical sensor for determination of Pb2+ and Hg2+ ions based on the covalent immobilization of dithizone on agarose membrane.

A highly sensitive and selective optical membrane for determination of Hg(2+) and Pb(2+) was prepared by covalent immobilization of dithizone on agarose membrane. In addition to its high stability, reproducibility and relatively long lifetime, the proposed optical sensor revealed good selectivity for target ions over a large number of alkali, alkaline earth, transition, and heavy metal ions. The proposed optical membrane displays linear responses from 1.1×10(-8) to 2.0×10(-6) mol L(-1) and 1.2×10(-8) to 2.4×10(-6) mol L(-1) for Hg(2+) and Pb(2+), respectively. The limits of detection (LOD) were 2.0×10(-9) mol L(-1) and 4.0×10(-9) mol L(-1) for Hg(2+) and Pb(2), respectively. The prepared optical membrane was successfully applied to the determination of Hg(2+) and Pb(2+) in industrial wastes, spiked tap water and natural waters without any preconcentration step.

Dithizone as novel and efficient chromogenic probe for cyanide detection in aqueous media through nucleophilic addition into diazenylthione moiety.

A new selective chemodosimeter probe was developed by the introduction of dithizone (DTZ) as a simple and available dye for detection of cyanide in aqueous media which enables recognition of cyanide over other competing anions such as acetate, dihydrogen phosphate, fluoride and benzoate through covalent bonding. The sensing properties of DTZ were investigated in DMSO/H2O (1:9) and have demonstrated a very high selectivity toward the cyanide anions. A reasonable recognition mechanism was suggested using UV-Vis, (1)H NMR and FTIR spectroscopy techniques. Time dependent density function theory (TDDFT) computations of UV-Vis excitation for DTZ2-CN adduct agreed well with our experimental findings. The detection limit of the new chromogenic probe was measured to be 0.48 µmol L(-1) which is much lower than most recently reported chromogenic probes for cyanide determination. The analytical utility of the method for the analysis of cyanide ions in electroplating wastewater (EPWW), human serum, tap and mineral water samples was demonstrated and the results were compared successfully with the conventional reference method. The short time response and the detection by the naked eye make the method available for the detection and quantitative determination of cyanide in a variety of real samples.

Cloud point extraction of Cu(II) using a mixture of Triton X-100 and dithizone with a salting-out effect and its application to visual determination.

A method for the separation and concentration of trace copper(II) ion (Cu(II)) via cloud point extraction (CPE) using a nonionic surfactant with a salting-out effect was developed and applied as a technique for the visual determination of Cu(II). Triton X-100 (TX-100), which has a cloud point at 64-67 °C in aqueous solutions, was used as the nonionic surfactant for the CPE of Cu(II). Although CPE with TX-100 requires heating of the solution to separate the surfactant-rich phase from the aqueous phase, the new method achieves phase separation at 15-30 °C owing to the addition of a large amount of salt to the solution, which lowers the cloud point. The compound 1,5-diphenylthiocarbazone (dithizone) was selected as the chelating agent for complexation and transfer of Cu(II) to the surfactant-rich phase. The extractability of Cu(II) (initial concentration: 10 µM) was 96.6±2.1% when Na2SO4 was added to a 20% TX-100/4 µM dithizone solution (pH 2). Using this method, the visual determination of Cu(II) was possible for concentrations ranging from 0.01 to 10 µM. In addition, the extraction system was successfully applied to the visual determination of Cu(II) in a river water sample.

Nanocomposite-based rapid, visual, and selective luminescence turn-on assay for Hg2+ sensing in aqueous media.

Composite nanospheres containing dithizone, luminescent LaVO4:Eu(3+) nanoparticles (NPs), and amphiphilic polymer have been composed for the rapid, selective, and visual luminescence turn-on detection of mercury ions (Hg(2+)) in water. Due to the absorption of dithizone, the strong red luminescence of LaVO4:Eu(3+) NPs encapsulated in nanospheres was quenched noticeably. As a result, these as-prepared nanocomposites (NCs) demonstrate very weak red luminescence. However, in the presence of Hg(2+), the red luminescence of nanocomposites was turned on dramatically, which can be attributed to the strong binding of mercury (II) ions by dithizone and forming a complex without absorption in the red emission range. Meanwhile, other cations have no influence on the detection of Hg(2+), suggesting a good selectivity for Hg(2+) sensing. Due to the high photostability and chemical stability of the nanocomposites, operation simplicity, low cost, and good selectivity, this newly developed method is highly desirable for field assay of Hg(2+) in aqueous media ranging from 40.0 nM to 4.0 µM with a limit of detection of 32.0 nM and a good linearity (r=0.9980). Therefore, a facile, rapid, selective, and visual luminescence turn-on technology has been successfully developed for Hg(2+) detection.

A novel development of dithizone as a dual-analyte colorimetric chemosensor: detection and determination of cyanide and cobalt (II) ions in dimethyl sulfoxide/water media with biological applications.

The behavior of dithizone (DTZ), an easily available dye has been studied for the first time in chromogenic sensing of CN(-) as an anionic species and for Co(2+) as a cationic species in DMSO/H2O media. So employing DTZ an efficient colorimetric chemosensor was afforded with a chromogenic selectivity for Co(2+) over other cations with detection limit of 0.04 µmol L(-1). The complex of Co(2+) with DTZ also displayed ability to detect up to 0.43 µmol L(-1) CN(-) (K(+) salts) among other competing anions through a fast response time of less than 30s which is much lower than most recently reported chromogenic probes. The linear dynamic ranges for the determination of Co(2+) and CN(-) were 0.3-4.4 and 3.3-58.6 µmol L(-1) respectively. This method could have potential application in a variety of cases requiring rapid and accurate analysis of Co(2+) and CN(-) for human serum and water samples.

Colorimetric response of dithizone product and hexadecyl trimethyl ammonium bromide modified gold nanoparticle dispersion to 10 types of heavy metal ions: understanding the involved molecules from experiment to simulation.

A new kind of analytical reagent, hexadecyl trimethyl ammonium bromide (CTAB), and dithizone product-modified gold nanoparticle dispersion, is developed for colorimetric response to 10 types of heavy metal ions (M(n+)), including Cr(VI), Cr(3+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+), and Pb(2+). The color change of the modified gold nanoparticle dispersion is instantaneous and distinct for Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+), and Pb(2+). The color change results from the multiple reasons, such as electronic transitions, cation-π interactions, formation of coordination bonds, and M(n+)-induced aggregation of gold nanoparticles (AuNPs). The different combining capacity of heavy metal ions to modifiers results in the different broadening and red-shifting of the plasmon peak of modified AuNPs. In addition, Cr(VI), Cu(2+), Co(2+), Ni(2+), and Mn(2+) cause the new UV-vis absorption peaks in the region of 360-460 nm. The interactions between the modifiers and AuNPs, and between the modifiers and M(n+), are investigated by using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The results confirm that AuNPs are modified by CTAB and dithizone products through electrostatic interactions and Au-S bonds, respectively, and the M(n+)-N bonds form between M(n+) and dithizone products. Furthermore, the experimental and density functional theory calculated IR spectra prove that dithizone reacts with NaOH to produce C6H5O(-) and [SCH2N4](2-). The validation of this method is carried out by analysis of heavy metal ions in tap water.