Direct electrochemistry and electrocatalysis of hemoglobin immobilized in a magnetic nanoparticles-chitosan film

Magnetic nanoparticles (Fe₃O₄) were synthesized by a chemical coprecipitation method. X-ray diffraction (XRD) and transmission electron microscope (TEM) were used to confirm the crystallite structure and the particle's radius. The Fe₃O₄ nanoparticles and chitosan (CS) were mixed to form a matrix in which haemoglobin (Hb) can be immobilized for the fabrication of H₂O₂ biosensor. The Fe₃O₄-CS-Hb film exhibited a pair of well-defined and quasi-reversible cyclic voltammetric peaks due to the redox of Hb-heme Fe (III)/Fe (II) in a pH 7.0 phosphate buffer. The formal potential of Hb-heme Fe(III)/Fe(II) couple varied linearly with the increase of pH in the range of 4.0-10.0 with a slope of 46.5 mV pH⁻¹, indicating that electron transfer was accompanied with single proton transportation in the electrochemical reaction. The surface coverage of Hb immobilized on Fe₃O₄-CS film glassy carbon electrode was about 1.13 × 10⁻¹⁰ mol cm⁻². The heterogeneous electron transfer rate constant (k_s) was 1.04 s⁻¹, indicating great facilitation of the electron transfer between Hb and magnetic nanoparticles-chitosan modified electrode. The modified electrode showed excellent electrocatalytic activity toward oxygen and hydrogen peroxide reduction. The apparent Michaelis-Menten constant for H₂O₂ was estimated to be 38.1 μmol L⁻¹.     

Amperometric biosensor for hydrogen peroxide based on hemoglobin entrapped in titania sol-gel film

Hemoglobin (Hb) was entrapped in a titania sol-gel matrix and used as a mimetic peroxidase to construct a novel amperometric biosensor for hydrogen peroxide. The Hb entrapped titania sol-gel film was obtained with a vapor deposition method, which simplified the traditional sol-gel process for protein immobilization. The morphologies of both titania sol-gel and the Hb films were characterized using scanning electron microscopy (SEM) and proved to be chemically clean, porous, homogeneous. This matrix provided a biocompatible microenvironment for retaining the native structure and activity of the entrapped Hb and a very low mass transport barrier to the substrates. H₂O₂ could be reduced by the catalysis of the entrapped hemoglobin at −300 mV without any mediator. The reagentless H₂O₂ sensor exhibited a fast response (less than 5 s) and sensitivity as high as 1.29 mA mM⁻¹ cm⁻². The linear range for H₂O₂ determination was from 5.0×10⁻⁷ to 5.4×10⁻⁵ M with a detection limit of 1.2×10⁻⁷ M. The apparent Michaelis-Menten constant of the encapsulated hemoglobin was calculated to be 0.18±0.02 mM. The stability of the biosensor was also evaluated.     

Hydrogen peroxide biosensor based on hemoglobin modified zirconia nanoparticles-grafted collagen matrix

A novel method for the immobilization of hemoglobin (Hb) and preparation of reagentless biosensor was proposed using a biocompatible non-toxic zirconia enhanced grafted collagen tri-helix scaffold. The formed membrane was characterized with UV-vis and FT-IR spectroscopy, scanning electron microscope and electrochemical methods. The Hb immobilized in the matrix showed excellent direct electrochemistry with an electron transfer rate constant of 6.46 s⁻¹ and electrocatalytic activity to the reduction of hydrogen peroxide. The apparent Michaelis-Menten constant for H₂O₂ was 0.026 mM, showing good affinity. Based on the direct electrochemistry, a new biosensor for H₂O₂ ranging from 0.8 to 132 μM was constructed. Owing to the porous structure and high enzyme loading of the matrix the biosensor exhibited low limit of detection of 0.12 μM at 3σ, fast response less than 5 s and high sensitivity of 45.6 mA M⁻¹ cm⁻². The biosensor exhibited acceptable stability and reproducibility. ZrO₂-grafted collagen provided a good matrix for protein immobilization and biosensing preparation. This method was useful for monitoring H₂O₂ in practical samples with the satisfactory results.     

Direct electrochemistry with enhanced electrocatalytic activity of hemoglobin in hybrid modified electrodes composed of graphene and multi-walled carbon nanotubes

A graphene (GR) and multi-walled carbon nanotubes (MWCNT) hybrid was prepared and modified on a 1-hexylpyridinium hexafluorophosphate based carbon ionic liquid electrode (CILE). Hemoglobin (Hb) was immobilized on GR-MWCNT/CILE surface with Nafion as the film forming material and the modified electrode was denoted as Nafion/Hb-GR-MWCNT/CILE. Spectroscopic results revealed that Hb molecules retained its native structure in the GR-MWCNT hybird. Electrochemical behaviors of Hb were carefully investigated by cyclic voltammetry with a pair of well-defined redox peaks obtained, which indicated that direct electron transfer of Hb was realized in the hybrid modified electrode. The result could be attributed to the synergistic effects of GR-MWCNT hybrid with enlarged surface area and improved conductivity through the formation of a three-dimensional network. Electrochemical parameters of the immobilized Hb on the electrode surface were further calculated with the results of the electron transfer number (n) as 1.03, the charge transfer coefficient (a) as 0.58 and the electron-transfer rate constant (k_s) as 0.97 s⁻¹. The Hb modified electrode showed good electrocatalytic ability toward the reduction of different substrates such as trichloroacetic acid in the concentration range from 0.05 to 38.0 mmol L⁻¹ with a detection limit of 0.0153 mmol L⁻¹ (3σ), H₂O₂ in the concentration range from 0.1 to 516.0 mmol L⁻¹ with a detection limit of 34.9 nmol/L (3σ) and NaNO₂ in the concentration range from 0.5 to 650.0 mmol L⁻¹ with a detection limit of 0.282 μmol L⁻¹ (3σ). So the proposed electrode had the potential application in the third-generation electrochemical biosensors without mediator.     

Direct electrochemistry and electrocatalysis of cytochrome c immobilized on gold nanoparticles-chitosan-carbon nanotubes-modified electrode

A robust and effective nanohybrid film based on gold nanoparticles (GNPs)/chitosan (Chit)/multi-walled carbon nanotubes (MWNTs) was prepared by a layer-by-layer self-assembly technique. Cytochrome c (Cyt c) was successfully immobilized on the nanohybrid film modified glassy carbon (GC) electrode by cyclic voltammetry. The direct electron transfer between Cyt c and the modified electrode was investigated in detail. Cyt c shows a couple of quasi-reversible and well-defined cyclic voltammetry peaks with a formal potential (E^0′) of −0.16 V (versus Ag/AgCl) in pH 7.0 phosphate buffer solution (PBS). The Cyt c/GNPs/Chit/MWNTs modified GC electrode gives an improved electrocatalytic activity towards the reduction of hydrogen peroxide (H₂O₂). The sensitivity is 92.21 μA mM⁻¹ cm⁻² and the calculated apparent Michaelis-Menten constant () is 0.791 mM, indicating a high-catalytic activity of Cyt c. The catalysis currents increase linearly to the H₂O₂ concentration in a wide range of 1.5 × 10⁻⁶ to 5.1 × 10⁻⁴ M with a correlation coefficient 0.999. The detection limit is 9.0 × 10⁻⁷ M (at the ratio of signal to noise, S/N = 3). Moreover, the modified electrode displays rapid response (5 s) to H₂O₂, and possesses good stability and reproducibility.     

Prussian Blue electrodeposited on MWNTs-PANI hybrid composites for H2O2 detection

A Prussian Blue (PB)/polyaniline (PANI)/multi-walled carbon nanotubes (MWNTs) composite film was fabricated by step-by-step electrodeposition on glassy carbon electrode (GCE). The electrode prepared exhibits enhanced electrocatalytic behavior and good stability for detection of H₂O₂ at an applied potential of 0.0 V. The effects of MWNTs thickness, electrodeposition time of PANI and rotating rate on the current response of the composite modified electrode toward H₂O₂ were optimized to obtain the maximal sensitivity. A linear range from 8 × 10⁻⁹ to 5 × 10⁻⁶ M for H₂O₂ detection has been observed at the PB/PANI/MWNTs modified GCE with a correlation coefficient of 0.997. The detection limit is 5 × 10⁻⁹ M on signal-to-noise ratio of 3. To the best of our knowledge, this is the lowest detection limit for H₂O₂ detection. The electrode also shows high sensitivity (526.43 μA μM⁻¹ cm⁻²) for H₂O₂ detection which is more than three orders of magnitude higher than the reported.     

The effects of carbon nanotubes on the electrocatalysis of hydrogen peroxide by metallo-phthalocyanines

The pre-grafted screen-printed gold electrode modified with phenyl-amino monolayer was investigated for covalent immobilization of phenyl-amine functionalized single-walled carbon nanotubes (PA-SWCNT) and metal tetra-amino phthalocyanine (MTAPc) using Schiff-base reactions with benzene-1,4-dicarbaldehyde (BDCA) as cross-linker. The PA-SWCNT and MTAPc modified electrodes were applied as hybrids for electrochemical sensing of H₂O₂. The step-by-step fabrication of the electrode was followed using electrochemistry, impedance spectroscopy, scanning electron microscopy and Raman spectroscopy and all these techniques confirmed the fabrication and the immobilization of PA-SWCNT, MnTAPc and CoTAPc onto gold surfaces. The apparent electron transfer constant (k_app) showed that the carbon nanotubes and metallo-phthalocyanines hybrids possess good electron transfer properties compared to the bare, pre-grafted and the MTAPc modified gold electrode surfaces without PA-SWCNT. The electrochemical sensing of hydrogen peroxide was successful with PA-SWCNT-MTAPc hybrid systems showing higher electrocatalytic currents compared to the other electrodes. The analytical parameters obtained using chronoamperometry gave good linearity at H₂O₂ concentrations ranging from 1.0 to 30.0 μmol L⁻¹. The values for the limit of detection (LoD) were found to be of the orders of 10⁻⁷ M using the 3δ for all the electrodes. The PA-SWCNT-MTAPc modified SPAuEs were much more sensitive compared to PA-MTAPc modified SPAuEs.     

Mesoporous silica hollow sphere (MSHS) for the bioelectrochemistry of horseradish peroxidase

In this work, novel mesoporous silica hollow spheres (MSHS) were chosen as an immobilization matrix, to construct a mediator-free third-generation HRP biosensor. UV-vis spectroscopy revealed that horseradish peroxidase (HRP) entrapped in MSHS could retain its native structure. FTIR spectroscopy and nitrogen adsorption-desorption isotherms indicated that HRP are intercalated into the mesopores. The direct electron transfer of HRP entrapped in MSHS was observed. A pair of stable and well-defined redox peaks of HRP with a formal potential of about −0.150 V (vs. Ag/AgCl) in 0.1 M pH 7.0 phosphate-buffered solution (PBS) were obtained. The biosensor exhibited a fast amperometric response to H₂O₂ with a linear range of 3.9 × 10⁻⁶ to 1.4 × 10⁻⁴ M (R = 0.997, N = 20). The detection limit was 1.2 × 10⁻⁶ M based S/N = 3.     

Enzyme mimics of spinel-type CoxNi1−xFe2O4 magnetic nanomaterial for eletroctrocatalytic oxidation of hydrogen peroxide

A series of spinel-type Co_xNi_1−xFe₂O₄ (x = 0, 0.2, 0.4, 0.5, 0.6, 0.8, 1.0) magnetic nanomaterials were solvothermally synthesized as enzyme mimics for the eletroctrocatalytic oxidation of H₂O₂. X-ray diffraction and scanning electron microscope were employed to characterize the composition, structure and morphology of the material. The electrochemical properties of spinel-type Co_xNi_1−xFe₂O₄ with different (Co/Ni) molar ratio toward H₂O₂ oxidation were investigated, and the results demonstrated that Co_0.5Ni_0.5Fe₂O₄ modified carbon paste electrode (Co_0.5Ni_0.5Fe₂O₄/CPE) possessed the best electrocatalytic activity for H₂O₂ oxidation. Under optimum conditions, the calibration curve for H₂O₂ determination on Co_0.5Ni_0.5Fe₂O₄/CPE was linear in a wide range of 1.0 × 10⁻⁸–1.0 × 10⁻³ M with low detection limit of 3.0 × 10⁻⁹ M (S/N = 3). The proposed Co_0.5Ni_0.5Fe₂O₄/CPE was also applied to the determination of H₂O₂ in commercial toothpastes with satisfactory results, indicating that Co_xNi_1−xFe₂O₄ is a promising hydrogen peroxidase mimics for the detection of H₂O₂.     

Langmuir-Blodgett film of p-tert-butylthiacalix[4]arene modified glassy carbon electrode as voltammetric sensor for the determination of Hg(II)

The π-A isotherms and UV-vis spectra of the transferred films suggested that the monolayer of p-tert-butylthiacalix[4]arene can coordinate with Hg²⁺ at the air-water surface. From these observations, a glassy carbon electrode coated with Langmuir-Blodgett film of p-tert-butylthiacalix[4] arene as a new voltammetric sensor is designed for the determination of trace amounts of Hg²⁺. Compared with bare glassy carbon electrode and modified glassy carbon electrode using direct coating method, the Langmuir-Blodgett film-modified electrode can greatly improve the measuring sensitivity of Hg²⁺. Under the selected conditions, the Langmuir-Blodgett film-modified electrode in 0.1 mol L⁻¹ H₂SO₄ + 0.01 mol L⁻¹ KCl solution shows a linear voltammetric response for Hg²⁺ in the range of 5.0 × 10⁻¹⁰ to 1.5 × 10⁻⁷ mol L⁻¹, with a detection limit of 2.0 × 10⁻¹⁰ mol L⁻¹. The proposed method was also applied to determine Hg²⁺ in water samples (tap, lake and river water). In addition, the fabricated electrode exhibited a distinct advantage of simple preparation, non-toxicity, good reproducibility and good stability.     

A hydrogen peroxide biosensor based on Langmuir-Blodgett technique: Direct electron transfer of hemoglobin in octadecylamine layer

The present paper describes the modification of hemoglobin (Hb)-octadecylamine (ODA) Langmuir-Blodgett (LB) film on a gold electrode surface to develop a novel electrochemical biosensor for the detection of hydrogen peroxide. Atomic force microscopy (AFM) image of Hb-ODA LB film indicated Hb molecules existed in ODA layer in a well-ordered and compact form. The immobilized Hb displayed a couple of stable and well-defined redox peaks with an electron transfer rate constant of 4.58 ± 0.95 s⁻¹ and a formal potential of −185 mV (versus Ag/AgCl) in phosphate buffer (1.0 mM, pH 5.0) contain 0.1 M KCl at a scan rate of 200 mV s⁻¹, characteristic of Hb heme Fe(III)/Fe(II) redox couple. The formal potential of Hb heme Fe(III)/Fe(II) redox couple in ODA film shifted linearly between pH 5 and 8 with a slope of −23.8 mV pH⁻¹, suggesting that proton took part in electrochemical reaction. The ODA could accelerate the electron transfer between Hb and the electrode. This modified electrode showed an electrochemical activity to the reduction of hydrogen peroxide (H₂O₂) without the aid of any electron mediator.     

A highly sensitive hydrogen peroxide amperometric sensor based on MnO2-modified vertically aligned multiwalled carbon nanotubes

In this report, a highly sensitive amperometric sensor based on MnO₂-modified vertically aligned multiwalled carbon nanotubes (MnO₂/VACNTs) for determination of hydrogen peroxide (H₂O₂) was fabricated by electrodeposition. The morphology of the nanocomposite was characterized by scanning electron microscopy, energy-dispersive X-ray spectrometer and X-ray diffraction. Cyclic voltammetry, chronoamperometry and electrochemical impedance spectroscopy were applied to investigate the electrochemical properties of the MnO₂/VACNTs nanocomposite electrode. The mechanism for the electrochemical reaction of H₂O₂ at the MnO₂/VACNTs nanocomposite electrode was also discussed. In borate buffer (pH 7.8, 0.20 M), the MnO₂/VACNTs nanocomposite electrode exhibits a linear dependence (R = 0.998) on the concentration of H₂O₂ from 1.2 × 10⁻⁶ M to 1.8 × 10⁻³ M, a high sensitivity of 1.08 × 10⁶ μA M⁻¹ cm⁻² and a detection limit of 8.0 × 10⁻⁷ M (signal/noise = 3). Meanwhile, the MnO₂/VACNTs nanocomposite electrode is also highly resistant towards typical inorganic salts and some biomolecules such as acetic acid, citric acid, uric acid and d-(+)-glucose, etc. In addition, the sensor based on the MnO₂/VACNTs nanocomposite electrode was applied for the determination of trace of H₂O₂ in milk with high accuracy, demonstrating its potential for practical application.     

Synthesis of a novel bistriazene reagent 4,4′-bis[3-(4-phenylthiazol-2-yl)triazenyl]biphenyl and its highly sensitive color reaction with mercury(II)

We report the synthesis of a novel bistriazene, 4,4′-bis(3-(4-phenylthiazol-2-yl)triazenyl)biphenyl (BPTTBP), and its highly sensitive color reaction with Hg²⁺. The new reagent was synthesized in good yield by coupling 2-amino-4-phenylthiazole with 4,4′-biphenyldiamine bisdiazonium salt. Using a blend of surfactants N-cetylpyridinium chloride (CPC) and polyethylene glycol n-octanoic phenyl ether (OP) as a micelle sensitizer, the red colored reagent assembles with Hg²⁺ in pH 9.8 borate buffer according to a 1:1 stoichiometry, forming a blue oligomeric/polymeric chelating complex with a high apparent stability constant (1.1 × 10⁸ M⁻¹). Whereas the maximum absorption of reagent occurs at 510 nm with an extinct coefficient of 1.35 × 10⁴ M⁻¹ cm⁻¹, the complex absorbs at 611 nm, with an apparent extinct coefficient of 1.04 × 10⁵ M⁻¹ cm⁻¹. Beer's law is obeyed in the range of 0-15 μg/25 mL Hg²⁺, and Sandell's sensitivity is 1.92 × 10⁻³ μg/cm². In the presence of thiourea and Na₄P₂O₇ as masking agents, the method was found free from interferences of foreign ions commonly occurring with mercury. The optimized protocol has been successfully applied to spectrophotometric determination of mercury in waste water samples. The features of the new reagent associated with its special structure were discussed, and an unprecedented “domino effect” was proposed to account for its unique chelating stoichiometry with Hg²⁺.     

A highly soluble poly(3,4-ethylenedioxythiophene)-poly(styrene sulfonic acid)/Au nanocomposite for horseradish peroxidase immobilization and biosensing

A highly soluble poly(3,4-ethylenedioxythiophene)-poly(styrene sulfonic acid)/Au (PEDOT-PSS/Au) nanocomposite was prepared via one-step chemical synthesis and the matrix was characterized by UV-vis spectroscopy (UV-vis), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM) and transmission electron microscope (TEM). Due to the excellent aqueous compatibility and biocompatibility, the PEDOT/PSS-Au nanocomposite can be used as biomaterial for enzymes immobilization. In this system, redox enzyme, horseradish peroxidase (HRP) was integrated with PEDOT/PSS-Au nanocomposite and the direct electron transfer of HRP was observed. Moreover, we find that the HRP/PEDOT-PSS/Au modified electrode shows excellent electrocatalytic ability for H₂O₂ and the formal Michaelis-Menten constant was 0.78 mmol/L. The response currents have good linear relation with the concentrations of H₂O₂ with a linear range from 2.0 × 10⁻⁷ to 3.8 × 10⁻⁴ mol/L.     

Direct electrochemistry and electrocatalysis of hemoglobin in gelatine film modified glassy carbon electrode

Direct electrochemical and electrocatalytic behavior of hemoglobin (Hb) immobilized on glass carbon electrode (GCE) containing gelatine (Gel) films was investigated. The characteristics of Hb/Gel film modified GC electrode were performed by using SEM microscopy, UV-vis spectroscopy and electrochemical methods. The immobilized Hb showed a couple of quasi-reversible redox peak with a formal potential of −0.38 V (versus SCE) in 0.1 M pH 7.0 PBS. The formal potential changed linearly from pH 4.03 to 8.41 with a slope value of −52.0 mV pH⁻¹, which suggested that a proton transfer was accompanied with each electron transfer (ET) in the electrochemical reaction. The Hb/gelatine/GCE displayed a rapid amperometric response to the reduction of H₂O₂ and nitrite.     

Gamma radiation induced synthesis of gold nanoparticles in aqueous polyvinyl pyrrolidone solution and its application for hydrogen peroxide estimation

Gold nanoparticles (Au nps) have been synthesized in aqueous solution of polyvinyl pyrrolidone (PVP) by gamma radiolysis from HAuCl₄·3H₂O precursor and in presence of small concentrations of Ag⁺, 2-propanol and acetone. The effect of different experimental parameters, such as concentration of reactant, molecular weight of PVP on nanoparticle formation was studied. TEM image confirmed that spherical Au nps were formed when PVP of molecular weight 360,000 Da was used as capping agent. H₂O₂ is a reactant in the enzyme catalyzed reaction of o-phenylene diamine (o-PDA). The reaction product has a weak absorption in the yellow region of the spectrum. When this product interacts with Au nps, it leads to enhancement of the absorption peak. The nanoparticles synthesized by radiation method were used for estimation of H₂O₂. The absorbance value of this peak at λ_max was observed to change with H₂O₂ concentration, which was monitored for estimation of H₂O₂. The response is linear in the range of 2.5×10⁻⁶ mol dm⁻³ to 2×10⁻⁴ mol dm⁻³ and 1×10⁻⁷ mol dm⁻³ to 3×10⁻⁶ mol dm⁻³ H₂O₂ in two separate sets of experimental parameters with detection limit 1×10⁻⁷ mol dm⁻³.     

Myoglobin/arylhydroxylamine film modified electrode: Direct electrochemistry and electrochemical catalysis

The adsorption processes and electrochemical behavior of 4-nitroaniline (4-NA) adsorbed onto glassy carbon electrodes (GCE) have been investigated in aqueous 0.1 M nitric acid (HNO₃) electrolyte solutions using cyclic voltammetry (CV). 4-NA adsorbs onto GCE surfaces, and upon potential cycling past −0.2 V, is transformed into the arylhydroxylamine (ArHA) derivative which exhibits a well-behaved pH dependent redox couple centered at 0.32 V at pH 1.5. It is noted as arylhydroxylamine modified glassy carbon electrodes (HAGCE). This modified electrode can be readily used as an immobilization matrix to entrap proteins and enzymes. In our studies, myoglobin (Mb) was used as a model protein for investigation. A pair of well-defined reversible redox peaks of Mb (Fe(III)-Fe(II)) was obtained at the Mb/arylhydroxylamine modified glassy carbon electrode (Mb/HAGC) by direct electron transfer between the protein and the GCE. The formal potential (E^0′), the apparent coverage (Γ^*) and the electron-transfer rate constant (k_s) were calculated as −0.317 V, 8.26 × 10⁻¹² mol/cm² and 51 ± 5 s⁻¹, respectively. Dramatically enhanced biocatalytic activity was exemplified at the Mb/HAGC electrode by the reduction of hydrogen peroxide (H₂O₂), trichloroacetic acid (TCA) and oxygen (O₂). The Mb/arylhydroxylamine film was also characterized by UV-visible spectroscopy (UV-vis), scanning electron microscope (SEM) indicating excellent stability and good biocompatibility of the protein in the arylhydroxylamine modified electrode. This new Mb/HAGC electrode exhibited rapid electrochemical response (2 s) for H₂O₂ and had good stability in physiological condition, showing the potential applicability of the films in the preparation of third generation biosensors or bioreactors based on direct electrochemistry of the proteins.     

Ultrasensitive fluorometric determination of hydrogen peroxide and glucose by using multiferroic BiFeO3 nanoparticles as a catalyst

BiFeO₃ magnetic nanoparticles (BFO MNPs) are used as a catalyst to develop an ultrasensitive method for the determination of H₂O₂. It is found that BFO MNPs can catalyze the decomposition of H₂O₂ to produce OH radicals, which in turn oxidize the weakly fluorescent benzoic acid to a strongly fluorescent hydroxylated product with a maximum emission at 405 nm. This makes it possible to sensitively quantify traces of H₂O₂. Under optimized conditions, the fluorescence intensity is observed to be well linearly correlated with H₂O₂ concentration from 2.0 × 10⁻⁸ to 2.0 × 10⁻⁵ mol L⁻¹ with a detection limit of 4.5 × 10⁻⁹ mol L⁻¹ (S/N = 3). In addition, a selective method for glucose determination is developed by using both glucose oxidase and BFO MNPs, which has a linear range for glucose concentration from 1.0 × 10⁻⁶ to 1.0 × 10⁻⁴ mol L⁻¹ with a detection limit of 5.0 × 10⁻⁷ mol L⁻¹. These new methods have been successfully applied for the determination of H₂O₂ in rainwater and glucose in human serum samples.     

Layer by layer assembly of catalase and amine-terminated ionic liquid onto titanium nitride nanoparticles modified glassy carbon electrode: Study of direct voltammetry and bioelectrocatalytic activity

A novel, simple and facile layer by layer (LBL) approach is used for modification of glassy carbon (GC) electrode with multilayer of catalase and nanocomposite containing 1-(3-Aminopropyl)-3-methylimidazolium bromide (amine terminated ionic liquid (NH₂-IL)) and titanium nitride nanoparticles (TiNnp). First a thin layer of NH₂-IL is covalently attached to GC/TiNnp electrode using electro-oxidation method. Then, with alternative self assemble positively charged NH₂-IL and negatively charged catalase a sensitive H₂O₂ biosensor is constructed, whose response is directly correlated to the number of bilayers. The surface coverage of active catalase per bilayer, heterogeneous electron transfer rate constant (k_s) and Michaelis–Menten constant (K_M) of immobilized catalase were 3.32 × 10⁻¹² mol cm⁻², 5.28 s⁻¹ and 1.1 mM, respectively. The biosensor shows good stability, high reproducibility, long life-time, and fast amperometric response with the high sensitivity of 380 μA mM⁻¹ cm⁻² and low detection limit of 100 nM at concentration range up to 2.1 mM.