Determination of creatinine with a sensor based on immobilized glutamate dehydrogenase and creatinine deiminase

Glutamate dehydrogenase and creatinine deiminase are immobilized by adsorption on wet poly(vinyl chloride) membranes. Creatinine is determined by a sensor consisting of the two membranes placed over an ammonia-sensing electrode. Endogenous ammonia is removed as it passes through the glutamate dehydrogenase layer. Creatinine (1–50 mg dl^?1) is converted to ammonia in the inner creatinine deiminase layer and is detected by the ammonia electrode. The assay requires 3 min, the minimum detectable concentration is 1 mg dl^?1 at pH 8.5, and the precision is ca. 5%. Endogenous ammonia can be tolerated up to 2 × 10^?4 M.     

Creatinine sensitive biosensor based on ISFETs and creatinine deiminase immobilised in BSA membrane

A creatinine sensitive biosensor based on ion sensitive field-effect transistors (ISFETs) with immobilised creatinine deiminase (CD) is developed. CD is immobilised on the transducer surface by classical cross-linking with bovine serum albumin (BSA) in a glutaraldehyde (GA) vapour. The linear dynamic ranges of biosensors are between 0 and 5 mM creatinine concentration, and the sensor sensitivity depends on the sample buffer concentration. Minimal detection limit for creatinine determination in model solution with 144 mM NaCl and 5% BSA, pH 7.4, is about 10 muM. Biosensor responses are reproducible and stable during continuous work at least for 8 h, and the relative standard deviation of sensor response is approximately 3% (n=48, for creatinine concentrations of 0.2 and 0.6 mM). An investigation about storage stability of creatinine sensitive ENFETs kept in dry at 4-6 degrees C shows that biosensors demonstrate an excellent storage stability for at least 6 months and more. Moreover, creatinine sensitive enzymatic field-effect transistors (ENFETs), demonstrating very good performances, are very selective and specific and well suitable for hemodialysis monitoring.     

Preparation of a novel aggregate like sugar-ball micelle composed of poly(methylglutamate) and poly(ethyleneglycol) modified by lactose and its molecular recognition by lectin

We report the preparation and characteristics of a novel micellar aggregate of an amphiphilic diblock copolymer, poly(methylglutamate) (PMG)-poly(ethyleneglycol) (PEG), whose terminus was modified by lactose lactone (LA). Due to the terminal LA moiety, this aggregate could be specifically recognized by RCA120 lectin. PMG-PEG-LA was synthesized by polymerizing the N-carboxy anhydride of L-glutamic acid gamma-methyl ester with H2N-PEG-LA as a polymerization initiator. By applying a fluorescence method using pyrene as a probe molecule, we found that PMG-PEG-LA could form the aggregate in aqueous solution. Fluorescence measurements showed that the critical aggregation concentration (C.A.C.) was 1.1 x 10(-5) M. The average diameter of the aggregate was 220 nm at 25 degrees C, as determined by the dynamic light scattering method. Circular dichroism measurements for the aggregate solution showed that the PMG residue took an alpha-helical structure, and that they associated to constitute the hydrophobic core of the aggregate. By adding RCA120 lectin to the aggregate solution, the turbidity of the solution increased rapidly, due to association of the aggregates. This implies that the aggregate could be recognized by lectin, and also suggests that sugar residues locate at the surface of the aggregates. From these findings, we concluded that the PMG-PEG-LA molecules form an aggregate like a "sugar ball" micelle, whose surface is covered by the sugar moieties. Application of the present aggregate system as a drug carrier is briefly discussed.     

Immobilization of porphyrins in poly(hydroxymethylsiloxane)

Three tetracationic porphyrins differing in the position of charged nitrogen atoms on the peripheral substituents — 5,10,15,20-tetrakis(N-methylpyridinium-4-yl)porphyrin (TMPyP4), 5,10,15,20-tetrakis(N-methylpyridinium-2-yl)porphyrin (TMPyP2), 5,10,15,20-tetrakis(4-trimethylammoniophenyl) porphyrin (TMAPP), and hydrophobic 5,10,15,20-tetraphenylporphyrin (TPP), were immobilized by adsorption and encapsulation in poly(hydroxymethylsiloxane) (PHOMS). The so prepared porphyrin-PHOMS composites were characterized by porosimetry, scanning electron microscopy, fluorescence and diffuse reflectance UV-VIS spectroscopy. It was found that porphyrins are immobilized in the PHOMS matrix in the free base monomer form Their irradiation produced singlet oxygen O₂(¹Δ_g) with the lifetime of 10–30 μs.     

Electrochemical aging of poly(aniline) and its ring substituted derivatives

Aniline and methyl, ethyl, propyl, and methoxy ring substituted derivatives have been polymerized by electrochemical methods. The reduction of the film produces aging of the polymers. This is followed by the changes in the current/potential profile of the voltammetric response. The effect of the substituents produces changes in both the extent and the rate of aging. The rate of aging can be represented by a Roginsky-Zeldovich or Elovich type of kinetics characterized by a pseudo zero order rate constant and a self-inhibiting parameter. Both parameters and the extent of aging are related to the free volume available for the different types of polymers.     

Novel flexible chemical gas sensor based on poly(3,4-ethylenedioxythiophene) nanotube membrane

Poly(3,4-ethylenedioxythiophene) nanotubes (PEDOT NTs) flexible membrane was successfully fabricated by vapor deposition polymerization (VDP) mediated electrospinning for ammonia gas detection. PVA nanofibers (NFs) were electrospun as a core part and polyvinyl alcohol (PVA)/PEDOT coaxial nanocables (NCs) were prepared by VDP method via EDOT monomer adsorption onto the electrospun PVA NFs as templates. To obtain the PEDOT NTs membrane, the PVA NFs were removed from PVA/PEDOT coaxial NCs with distilled water. PVA/PEDOT coaxial NCs and PEDOT NTs had the conductivities of 71 and 61 S cm⁻¹ and were applied as a transducer for ammonia gas detection in the range of 1-100 parts per million (ppm) of NH₃ gas. They exhibited the minimum detectable level of ca. 5 parts per million (ppm) and fast response time (less than 1 s) towards ammonia gas. In a recovery time, the PEDOT NTs membrane sensor was ca. 30 s and shorter compared to that of the membrane sensor based on the PVA/PEDOT NCs (ca. 50 s). In addition, sensor performance of PEDOT NTs membrane was also undertaken as a function of membrane thickness. Thick membrane sensor (30 μm) had the enhanced sensitivity and the sensitivity on the membrane thickness was in the order of 30 μm > 20 μm > 10 μm at 60 ppm of NH₃ gas.     

On the use of methyl substituted poly(vinylnaphthalene) as a reversible singlet oxygen carrier

Poly(1,4-dimethyl-6-vinylnaphthalene) and poly(1,2,4-trimethyl-6-vinylnaphthalene) react with singlet oxygen to give corresponding endoperoxide polymers which on warming generate singlet oxygen efficiently. Singlet oxygenations by the use of these polymers have been described.     

Fiber-optic sensor for carbon dioxide with a ph indicator dispersed in a poly(ethylene glycol) membrane

A fiber-optic sensor for carbon dioxide gas is constructed, without an inner buffer solution, by using a dispersion of fluorescein in poly(ethylene glycol) deposited on the distal end of an optical fiber. Evaporation of the solvent is thus negligible. The response range is 0–28% (v/v) for carbon dioxide, with a detection limit of 0.1%. The response time achieved is 10 s. The membrane (ca. 10 μm thick) is composed of poly(ethylene glycol)s with molecular weights of 200 and 1540 dalton in a 20:80% (w/w) ratio. The best concentration of fluorescein is 5 × 10^?7 mol g^?1 of poly(ethylene glycol). The response mechanism of the sensor is discussed.     

Immobilization of a hexaphyrin(1.0.1.0.0.0) derivative onto a tentagel-amino resin and its use in uranyl cation detection

The synthesis of an isoamethyrin derivative containing two CH(2)CH(2)CO(2)CH(3) moieties in the beta-pyrrolic positions and its use in the colorimetric detection of the uranyl cation after immobilization onto a solid support is reported.     

Poly(methylglutamate)-coated surfaces in HPLC and CE

Summary Thermal polymerization of the N-carboxyanhydride of glutamic acid-5-methylester was used to coat microparticulate silica and the surface of capillaries with poly(methylglutamate) (PMG). By increasing the thickness of the PMG layer the hydrophobicity of the stationary phases could be increased. However, total shielding of surface silanols was not achieved. The PMG-coated silicas for HPLC showed unexpected and strange behavior in the chromatography of proteins. Nevertheless, capillaries coated with PMG proved to be excellent for highly efficient CE separations of proteins at medium pH values.     

Facile fabrication of a novel high performance CO2 separation membrane: Immobilization of poly(amidoamine) dendrimers in poly(ethylene glycol) networks

Poly(amidoamine) (PAMAM) dendrimers showed high CO₂ separation properties and were successfully immobilized in a poly(ethylene glycol) (PEG) network upon photopolymerization of PEG dimethacrylate. The PAMAM dendrimer incorporation ratio was readily controlled, and a stable self-standing membrane containing up to 75 wt.% PAMAM dendrimer was obtained. The CO₂ separation properties over smaller H₂ were investigated by changing the PAMAM dendrimer content or generation and CO₂ partial pressure (ΔPCO₂$\mathrm{\Delta }{P}_{\text{C}{\text{O}}_2}$) under atmospheric conditions. Especially, a polymeric membrane containing 50 wt.% PAMAM dendrimer (0th generation) exhibited an excellent CO₂/H₂ selectivity of 500 with CO₂ permeability of 2.74 × 10⁻¹⁴ m³(STP)m/(m² s Pa) or 3.65 × 10³ barrer (1 barrer = 7.5 × 10⁻¹⁸ m³(STP)m/(m² s Pa)) when a mixture gas (CO₂/H₂: 5/95 by vol.) was fed at 25 °C and 100 kPa with 80% relative humidity. This polymeric materials are promising for a novel CO₂ separation membrane.     

Chemical modification of poly(lactic acid) and its use as matrix in poly(lactic acid) poly(butylene adipate-co-terephthalate) blends

Poly(lactic acid), PLA, was chemically modified with maleic anhydride (MA) by reactive extrusion. The effect of this modification on molar mass (MM) and acidity was assessed by means of size-exclusion chromatography (SEC) and titration, respectively. PLA MM decreased in the presence of MA solely and of MA and peroxide. Reduction in MM was monitored by the increase in acidity. PLA blends containing poly(butylene adipate-co-terephthalate) (PBAT) were prepared through different mixing protocols, PLA/PBAT, PLA-g-MA/PBAT and PLA/PBAT/MA/peroxide (PLA/PBAT in situ). SEC results and rheological properties revealed reduction in MM and viscosity of the modified blends. PLA/PBAT presented increase in MM and bimodal MM distribution. The calculated interfacial tension was significantly lower for the modified blends, despite the lower average particle area of PLA/PBAT. Surprisingly, the modified blends presented higher yield strength than that predicted by the rule of mixtures, which might indicate interfacial reactions.     

Immobilization of glucose oxidase with the blend of regenerated silk fibroin and poly(vinyl alcohol) and its application to a 1,1′- dimethylferrocene-mediating glucose sensor

The structure and properties of the blend of regenerated silk fibroin (RSF) and poly(vinyl alcohol) (PVA) were investigated. The two polymers in the blend are in the state of phase segregation. Infrared (IR) spectra indicate that the RSF in the blend maintains its intrinsic properties, thus, ethanol treatment can transfer silk I structure of RSF to silk II structure. The water absorption property and mechanical property of the blend are improved in comparison with those of RSF. The blend maintains the major merit of RSF, that is, it can immobilize glucose oxidase on the basis of the conformational transition from silk I structure to silk II structure. The properties of the immobilized enzyme are examined. Moreover, the second generation of glucose sensor based on the immobilized enzyme is fabricated and it has a variety of advantages including easy maintenance of enzyme, simplicity of construction, fast response time and high stability.     

Development of sulfonated poly (ether ether ketone) electrolyte membrane for applications in hydrogen sensor

The future economy is projected as hydrogen economy and fuel cells are set to become the energy source either replacing or augmenting the present oil based technology. A sulfonated poly ether ether ketone (SPEEK) membrane as the electrolyte for hydrogen sensor that operates at room temperature was developed in our lab. The electrolyte used was SPEEK, which is a proton conducting solid polymer membrane. The membranes were characterized using various available techniques like TGA, XRD, SEM, etc. The durability was studied using the Fenton’s reagent. The proton conducting ability was analyzed using impedance spectroscopy. The catalysts considered were platinum for the cathode and three different catalysts (Pt, Pt/Pd and Pd) for the anode. The MEAs were evaluated for their performance in hydrogen sensor and the one with platinum catalyst at the anode gave the best response among the three indicating its suitability for the SPEEK membrane for hydrogen sensor.     

Immobilization of urease in poly(1-vinyl imidazole)/poly(acrylic acid) network

In this study, urease was immobilized in a polymer network obtained by complexation of poly(1-vinyl imidazole) (PVI) with poly(acrylic acid) (PAA). Preparation of the polymer network was monitored by FT-IR spectroscopy. Scanning electron microscopy (SEM) revealed that enzyme immobilization had a strong effect on film morphology. Proton conductivity of the PVI/PAA network was measured via impedance spectroscopy under humidified conditions. Values of the Michaelis-Menten constant (K _M) for immobilized urease were higher than for the free enzyme, indicating a decreased affinity of the enzyme to its substrate. The basic characteristics (pH_opt, pH_stability, T _opt, T _stability, reusability, and storage stability) of immobilized urease were determined. The results show that the PAA/PVI polymer network is suitable for enzyme immobilization.     

Immobilization of poly(ethylene oxide) on polyethylene using a plasma-induced graft copolymerization process

A radio-frequency glow discharge induced graft copolymerization process has prepared a PAAc-grafted PE film in this present study. Poly(ethylene oxide) (PEO) was then covalently immobilized onto PAAc-grafted PE film by a chemical coupling reaction. Carboxylic groups were introduced onto the surface of PE film by exposing the film to an Ar plasma, and the plasma-treated film was then next reacted with a 50% acrylic acid aqueous solution. These functional groups were reacted with bisamino PEO by a coupling agent CMC. ESCA, IR, and water contact angle studies were conducted for characterizing the surfaces of these films during the varied stages of the reaction. Increasing the molecular weight of the PEO attached to the PAAc-grafted PE resulted in an increased wettability of the films. © 1993 John Wiley & Sons, Inc.     

Homogeneous blend membrane made from poly(ether sulphone) and poly(vinylpyrrolidone) and its application to water electrolysis

A homogeneous blend membrane has been successfully prepared from poly(ether sulphone) (PES) and poly(vinylpyrrolidone) (PVP), which have been usually thought to be immiscible to each other. The PES–PVP membrane possesses combined advantages of the hydrophobic and hydrophilic components; specifically, both good mechanical strength and excellent hydrophilicity have been achieved simultaneously. The success of achieving such special properties can be ascribed to the appropriate choice of solvent for membrane casting, N,N-dimethylformamide (DMF) in this case, and the post-treatment of the membrane in alkaline ethanol solution for obtaining hydrophilicity. Membranes with different blending ratios were characterized with tensile test, scanning electron microscopy (SEM), transmission electron microscopy (TEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and electrochemical measurements. The membrane was found to function well as a diaphragm in water electrolysis. In comparison with the traditional asbestos diaphragm, an energy saving of ca. 10% can be reached upon employing PES–PVP membranes in water electrolysis.     

Determination of glutamate-pyruvate transaminase activity in blood serum with a pyruvate oxidase/poly(vinyl chloride) membrane sensor

Pyruvate oxidase (E.C. 1.2.3.3.) is immobilized by adsorption on a wet PVC membrane. Glutamate-pyruvate transaminase activity (5–1600 IU l^?1) in serum is determined by a pyruvate oxidase sensor consisting of the immobilized pyruvate oxidase coupled to a platinum electrode for measuring hydrogen peroxide, after an l-alanine—α-ketoglutarate reaction. The assay requires ?60 s, and has a precision of 2–3%. Endogenous pyruvate should not interfere if measurements are made > 30 s after starting the reaction.