Determination of mercury(II) by invertase enzyme inhibition coupled with batch injection analysis

The determination of mercury(II) ions at the trace level by inhibition of the invertase enzyme-catalysed hydrolysis of sucrose into glucose and fructose coupled to electrochemical batch injection analysis was investigated using two approaches. In the first, the glucose produced was detected by injection of 100 microliters samples into the batch injection cell containing a platinum electrode modified by immobilised glucose oxidase. In the second, the glucose and fructose present in injected samples were oxidised directly at a copper-modified glassy carbon electrode. The experimental parameters were optimised and the degree of enzyme inhibition by mercury(II) ions under both conditions was measured. Mercury concentrations in the ng ml-1 range were determined by these two techniques with low sample and reagent consumption. Comparison is made between the two methods and perspectives as a screening test for field application are indicated.     

Determination of mercury(II), methylmercury and ethylmercury in the ng/ml range with an electrochemical enzyme glucose probe

Hg(II), methylmercury and ethylmercury have been determined with an electrochemical glucose probe. Mercury and its compounds inhibit the enzyme invertase which, in presence of its substrate, sucrose, produces glucose. When invertase is in presence of mercury its activity decreases; this causes a decrease of glucose production, which is monitored by the glucose sensor and correlated to the concentration of mercury in solution. Parameters such as pH, enzyme concentration, substrate concentration, and reaction and incubation time were optimized. Results showed that mercury, methylmercury and ethylmercury can be detected directly in aqueous solution in the range 2–10 ng/ml.     

Applications of enzyme-catalysed reactions in trace analysis--I. Determination of mercury and silver by their inhibition of invertase

Procedures are described for the accurate determination of silver (2-10 x 10(-7)M) and mercury(II) (2-10 x 10(-7)M) in the presence of each other and of most other metals. The methods are based on the inhibition by these metals of invertase catalysis of the hydrolysis of sucrose.     

Applications of enzyme-catalysed reactions in trace analysis-II Determination of cyanide, sulphide, and iodine with invertase

Methods are described for the determination of cyanide (10⁻⁸–10⁻⁵M and sulphide (10⁻⁷–10⁻⁵ M) based on the de-inhibitory effect of these ions on invertase inhibited by mercury(II) or by silver. Iodine (0.1–3 μg) may be determined by its inhibition of invertase.     

A dip-and-read test strip for the determination of mercury(II) ion in aqueous samples based on urease activity inhibition.

A sensitive dip-and-read test strip for the determination of mercury in aqueous samples based on the inhibition of urease reaction by the ion has been developed. The strip has a circular sensing zone that containing two layers: the top layer is a cellulose acetate membrane where urease is immobilized on it; the bottom layer is a pH indicator wafer that is impregnated with urea. The principle of the measurement is based on the disappearance of a yellow spot on the pH indicator wafer. The elapsing time until the disappearance of the spot which depends on the concentration of mercury(II) ion is measured with a stopwatch. Under the experimental conditions, as low as 0.2 ng/ml mercury can be observed with the detection range from 0.2 to 200 ng/ml in water. Organomercury compounds give essentially the same response as inorganic mercury. Heavy-metal ions such as Ag(I), Cu(II), Cd(II), Ni(II), Zn(II), and Pb(II) as well as other sample matrixes basically do not interfere with the mercury measurement.     

Screening of Fish Tissue for Methyl Mercury Using the Enzyme Invertase in a Solvent Interface

This paper describes a simple and rapid screening system for the extraction and determination of methyl mercury in tissue samples from fish. A novel clean-up procedure based on the use of two immiscible phases, an organic one containing methyl mercury and an aqueous one containing invertase, was developed. Methyl mercury was selectively extracted from the organic into the aqueous phase by its irreversible reaction with thiol groups of invertase, and the resulting inhibition of enzymatic activity served as a measure of methyl mercury concentration. Enzyme activity was measured with a spectrophotometric method using 3,5-dinitrosalicylic acid as a reagent. After parameter optimisation including pH, substrate concentration, and enzyme reaction time, it was possible to determine methyl mercury in the ppb range with this technique. Concentrations as low as 10ppb of methyl mercury in the extract, corresponding to 0.2ppm of methyl mercury in fish, were detected. The proposed procedure was successfully demonstrated as a simple screening method for methyl mercury in fish samples.     

Determination of trace chromium(VI) by an inhibition-based enzyme biosensor incorporating an electropolymerized aniline membrane and ferrocene as electron transfer mediator

A novel inhibition-based glucose oxidase (GOx) biosensor for environmental chromium(VI) detection is described. An electropolymerized aniline membrane has been prepared on a platinum electrode containing ferrocene as electron transfer mediator, on which GOx is cross-linked by glutaraldehyde. The mechanism of the redox reaction on the electrode and the performance of the sensor are studied. The sensor's response to glucose decreases when it is inhibited by chromium(VI), with a lower detection limit of 0.49?µg?L^?1, and the linear response range is divided into two parts, one of which is 0.49–95.73?µg?L^?1 and the other is 95.73?µg^?1 to8.05?mg?L^?1. The enzyme membrane is shown to be completely reactivated after inhibition, retaining 90% activity over more than forty days. Interference to chromium(VI) determination from lead(II), copper(II), cadmium(II), chromium(III), cobalt(II), tin(II) and nickel(II) is found to be minimal, while high concentrations of mercury(II) and silver(I) may interfere with the determination of trace chromium(VI). The sensor has been used for chromium(VI) determination in soil samples with good results.     

A Prospect for the Analysis of Species Acting as Enzyme Inhibitors as Illustrated by Heavy Metal Inhibition

Abstract

A flow system incorporating an amperometric glucose oxidase enzyme electrode has been used to study the inhibitory effects of 16 metal cations on glucose oxidase. Only copper(II), mercury(II) and silver(I) caused any significant inhibition. the enzyme electrode could be reactivated by EDTA, the reactivation being most effective for copper(II) and least so for silver(I). Other complexing agents were tried for reactivation but proved to be unsatisfactory.

The ability to reactivate the enzyme on the electrode following copper(II) inhibition, and the linear response of the system to the level of this inhibitor according to I/A = -9.49 × 10^?7 log([Cu]/M) + 4.84 × 10^?8; r = 0.994 between 2.5 × 10^?4M and 5 × 10^?3M [Cu]²⁺ indicates a prospect for the use of a flow system for determining enzyme inhibitors in samples.     

An enzyme-catalysed method for the determination of mercury traces in carbonated soft drinks, by the Hg(2+) inhibition of beta-fructofuranosidase

This paper describes the utilization of the inhibition of beta-fructofuranosidase by Hg(II) during the hydrolysis of saccharose, for the determination of Hg(II) in waters and drinks. The mercury levels in the samples tested ranged from 100 to 270ng l .     

Synthesis and properties of a chelating resin containing triazolethiol groups

A macroreticular poly(acrylic acid)-based resin with triazolethiol as the functional group has been synthesized. The stability of the resin in acidic media and the behaviour in sorption and desorption of various metal ions have been investigated and compared with those of the acylthiosemicarbazide resin which is an intermediate in synthesis of the triazolethiol resin. Both resins show high affinity for copper(II) silver, cadmium and mercury(II), and high selectivity for silver and mercury(II) at low pH (1–2), and even at pH 7 if EDTA is present. The triazolethiol resin sorbs metal ions faster than the acylthiosemicarbazide resin does and sorbs mercury(II) from high concentrations of acids and neutral salt solutions. This resin has been applied to the concentration of silver and mercury(II) from sea-water samples by column operation.     

Improvements in the non-flame atomic fluorescence determination of mercury

A new non-flame atomic fluorescence system with a detection limit of 5 parts per trillion (p.p.t.) or 5 pg of mercury(II) has been developed. The relative precision is 5% or better above 50 p.p.t. Hg(II), and the dynamic range in terms of linearity is from 0–100 p.p.b. Hg(II). The system is useful for analysis of residual mercury levels in water at a rate of about thirty samples per h.     

The Color Reaction of Mercury(II) with the New Reagent 2-Methylthiophenyldiazoaminoazobenzene and Its Application

A new and very sensitive and selective chromogenic reagent, 2-methylthiophenyldiazoaminoa-zobenzene (MTDAA), was synthesized and studied in detail for the determination of trace mercury(II) in water samples. The method is based on the color reaction between MTDAA and mercury(II). It was found that mercury(II) reacts with MTDAA in Na₂B₄O₇-NaOH buffer solution (pH = 10.0) to form 1 : 2 red complexes; these show maximum absorption at 520 nm. Beer’s law is obeyed in 0–15 µg of mercury(II) in 25 mL of solution. The apparent molar absorptivity of the complex is 1.33 × 10⁵ L/(mol cm); its limit of quantification, limit of detection, and relative standard deviation are 0.75 ng/mL, 0.27 ng/mL, and 1.0%, respectively, giving better sensitivity. The influence of the reaction variables and the effect of interfering ions are reported; most of the metal ions in water samples can be tolerated in considerable amounts. Only a few ions can interfere with the determination of trace mercury(II), but these can be eliminated by prior extraction. The proposed method is sensitive, simple, and rapid. It has been applied to the determination of trace mercury(II) in water samples with satisfactory results.__________From Zhurnal Analiticheskoi Khimii, Vol. 60, No. 7, 2005, pp. 703–706.Original English Text Copyright © 2005 by Guo, Din, Tian, Liu, Chang, Meng.The text was submitted by the authors in English.     

Glucose Monitoring by a Biosensor Based on Mercury Thin Film Electrodes

Abstract

Mercury film electrodes consist of a thin film of mercury deposited on an electrode surface (typically glassy carbon) by reduction of a mercury (II) salt in solution. The surface area/volume ratio is larger for the mercury film electrode, and this electrode is more stable than mercury drop electrode, which allows a faster stirring rate to be used in the deposition step. An enzyme electrode is described, based on glucose oxidase immobilized by gelatin and glutaraldehyde and held over a glassy carbon electrode coated with a thin mercury film. This biosensor responds fast and linearly to glucose in a wide concentration range, which is significant because monitoring of glucose levels is a critical component of diabetes care. Certain optimization and characterization studies were carried out. Average value, standard deviation (SD), and variation coefficient (CV) were calculated with the help of the repeatability studies. Finally, glucose content of human blood samples was monitored with the help of the biosensor presented.     

Mercury speciation in environmental solid samples using thermal release technique with atomic absorption detection

A sensitive and very simple method for determination of mercury species in solid samples has been developed involving thermal release analysis in combination with atomic absorption (AAS) detection. The method allows determination of mercury(II) chloride, methylmercury and mercury sulfide at the level of 0.70, 0.35 and 0.20 ng with a reproducibility of the results of 14, 25 and 18%, respectively. The accuracy of the developed assay has been estimated using certified reference materials and by comparison of the results with those of an independent method. The method has been applied for Hg species determination in original samples of lake sediments and plankton.     

Separation and Removal of Mercury(II) from Water Samples Using (Acetylacetone)‐2‐Thiol‐Phenyleneimine Immobilized on Anion‐Exchange Resin Prior to Determination by Cold Vapor Inductively Coupled Plasma Atomic Emission Spectroscopy

Abstract

(Acetylacetone)‐2‐thiol‐phenyleneimine (H₂L) immobilized on an anion‐exchange resin (Dowex) was used for separation and removal of mercury from natural water samples and for preconcentration prior to its determination by cold vapor inductively coupled plasma atomic emission spectroscopy. The metal was eluted from the column using a solution of 10% thiourea in 0.1 M HCl. The modified resin is higly selective with an exchange capacity of 1.60 mmol g^?1. Various parameters like pH, column flow rate, and desorbing agents are optimized. The proposed method has a linear calibration range of 15–1000 ng/ml Hg(II), with a relative standard deviation at the 15 ng/ml level of 3.5%. The precision of the method (evaluated as the relative standard deviation obtained after analyzing six series of five replicates) was ±4.2% at the 50 ng/ml level of Hg(II). The method has been used for routine determination of trace levels of mercury species in natural waters. The potential application of modified resin for the removal of mercury(II) from two natural water samples (top water and lake water) spiked with 50 ng/ml of mercury (II) was studied by ICP‐AES, and the results proved that excellent percent extraction of mercury(II) from both natural water samples was obtained by column method using modified resin.     

On-line preconcentration and determination of mercury in biological samples by flow injection vapour generation inductively coupled plasma atomic emission spectrometry

An FI-ICP-AES method for the determination of trace levels of mercury in biological samples has been described, which is based on the extraction of the mercury complex with 1,5-bis (di-2-pyridyl)methylene thiocarbonohydrazide (DPTH) on-line into isobuthyl-methyl ketone (IBMK). The organic phase (containing the complex) has been mixed on-line with SnCl₂ in N,N-dimethylformamide. Thus, mercury vapour can be generated directly from the organic phase and separated in a gas-liquid separation device. The detection limit for mercury is 4 ng/ml and the calibration curve is linear at least from 10 to 2500 ng/ml. The relative standard deviation for 10 replicate measurements is ±1% for 100 ng/ml of Hg(II). Results from the analysis of some certified biological reference materials are given.     

Test Method for the Determination of Mercury(II) with Dithizone on the Solid Phase of a Fibrous Anion Exchanger

The sorption and determination of mercury(II) on the solid phase of a fibrous polyacrylonitrile material filled with the AV-17 anion exchanger is studied. Mercury is adsorbed in the form of the chloride complex from a solution with pH 5–7.5; it is determined on the solid phase by the reaction with dithizone either visually or by the measuring the color hue. A procedure for the test determination of mercury(II) has been developed and tested in the analysis of fresh natural waters by the added–found method. The detection limit is 0.02 mg/L. The time of analysis of 5 or 6 samples is no more than 15–20 min.     

Selective extraction of Hg(II) from aqueous mineral acid solution containing iodide ions using 2-benzylpyridine in benzene

A method for the selective extraction of mercury has been developed. The extraction of Hg(II) by 2-benzylpyridine (BPy) in benzene from dilute mineral acid solution containing iodide ions has been investigated, and variables such as concentration of acids, iodide and the extractant have been optimized. The optimum conditions for the extraction of Hg(II) by 0.1M BPy/benzene are: 0.01M (HCl, HNO₃, H₂SO₄)+0.01M KI. The distribution coefficients and separation factors of 19 elements relative to Hg(II), have been reported. Effect of anions such as ascorbate, acetate, citrate, oxalate and thiosulfate has also been studied. The method developed could find useful applications in selective extraction of small amounts of mercury from environmental samples.     

A new resin containing benzimidazolylazo group and its use in the separation of heavy metals

A new resin incorporating benzimidazolylazo group into a matrix of polystyrene divinylbenzene has been prepared. The exchange capacity of the resin for the ions mercury(II), silver(I) and palladium(II) as a function of pH has been determined. The resin exhibits no affinity for alkali or alkaline earth metals. It is highly selective for Hg(II), Ag(I) and Pd(II). In column operation, it has been observed that Hg(II), Ag(I) and Pd(II) in trace quantities can be selectively separated from geological, medicinal and environmental samples.     

Plasticized open-cell polyurethane foam as a universal matrix for organic reagents in trace element preconcentration : Part II. Collection of mercury traces on dithizone and diethyldithiocarbamate foams

The analytical suitability of zinc dithizonate foam for the collection and preconcentration of traces of mercury(II) has been examined. The effect of pH, plasticizer and chelating agent concentration on the collection rate of mercury has been critically investigated. The capacity for mercury(II) of a TBP-plasticized zinc dithizonate foam at pH 6 is 23.4 μeq g^?1. The effect of plasticizer on the rate of recovery of mercury with sodium thiosulphate solution is also discussed. The preparation of zinc diethyldithiocarbonate foam is described. Traces of mercury(II) can also be collected rapidly and quantitavely by this foam.