Long-term Storage Stability Research Articles

Evaluation of a Flexible NOTA-RGD Kit Solution Using Gallium-68 from Different 68Ge/68Ga-Generators: Pharmacokinetics and Biodistribution in Nonhuman Primates and Demonstration of Solitary Pulmonary Nodule Imaging in Humans.

Radiopharmaceuticals containing the motive tripeptide arginyl-glycyl-asparatic acid (RGD) are known to target ανβ3 integrins during tumor angiogenesis. A more generic kit radiolabeling procedure accommodating Ga-68 from different generators was developed for NOTA-RGD and evaluated for its versatile use and safety in subsequent in vivo applications. The [68Ga]NOTA-RGD kit was further verified for its expected biodistribution and pharmacokinetics in nonhuman primates and its clinical sensitivity to detect solitary pulmonary nodules (SPN) in cancer patients. Single vial kits containing 28-56nmol of NOTA-cyclo-Arg-Gly-Asp-d-Tyr-Lys (NOTA-RGD) and sodium acetate trihydrate buffer were formulated. Versatility of the NOTA-RGD radiolabeling performance and adaption to a TiO2- and a SnO2-based generator type, characterization and long-term storage stability of the kits were carried out. The blood clearance and urine recovery kinetics as well as the image-guided biodistribution of [68Ga]NOTA-RGD was studied in a vervet monkey model. [68Ga]NOTA-RGD kits were further tested clinically to target solitary pulmonary nodules. The kits could be successfully formulated warranting integrity over 3-4months with a good [68Ga]NOTA-RGD radiolabeling performance (radiochemical purity >95%, decay corrected yield 76-94%, specific activity of 8.8-37.9GBq/μmol) The kits met all quality requirements to be further tested in vivo. [68Ga]NOTA-RGD cleared rapidly from blood and was majorly excreted via the renal route. The liver, spleen, heart and intestines showed initial uptake with steadily declining tissue activity concentration over time. In addition, the [68Ga]NOTA-RGD kit allowed for delineation of SPN from non-malignant lung tissue in humans. A more versatile radiolabeling procedure using kit-formulated NOTA-RGD and different generator types was achieved. The uncompromised in vivo behavior and efficient targeting of SPN warrants further investigations on the clinical relevance of [68Ga]NOTA-RGD derivatives to implement initial guidelines and management of patients, with regard to integrin targeted imaging.

Facile approach to the green synthesis of novel ternary composites with excellent superhydrophobic and thermal stability property: An expanding horizon

In present study, a novel and facile strategy of electrophoretic deposition (EPD) with following perfluorodecyltriethoxysilane (FAS-17) modification is successfully introduced to fabricate superhydrophobic ternary composites-AlxNiy(Bi2O3)z with excellent superhydrophobic and thermal stability property. The chemical composition, morphology and heat-release performance of the products were analyzed by XRD, FESEM, FT-IR, EDX, DSC and a high-speed camera, respectively. The stable suspension consisted of isopropyl alcohol and additive-polyethyleneimine. The deposition mechanism of ternary composite coating was firstly given, and EPD kinetics were investigated in detail. Moreover, the water contact angle and roll-off angle of superhydrophobic AlxNiy(Bi2O3)z with as well as significant water bouncing properties can reach to 169±2° and ca. 1°, respectively. In addition, superhydrophobicity and internal chemical energy (ca. 2MJkg−1) of products all keep almost stable after 300days, exhibiting a high value of practical application. Thus, this study provides a promising and low cost method for the synthesis of various superhydrophobic composites with long-term storage stability in different humidity environments and zones, even underwater.

Storage stability of exhaled breath on Tenax TA

Exhaled breath is coming to the forefront of non-invasive biomarker discovery efforts. Concentration of exhaled breath volatile organic compounds (VOCs) on thermal desorption (TD) tubes with subsequent analysis by gas chromatography-mass spectrometry (GC-MS) has dominated this field. As discovery experimentation increases in frequency, the need to evaluate the long-term storage stability of exhaled breath VOCs on thermal desorption adsorbent material is critical. To address this gap, exhaled breath was loaded on Tenax TA thermal desorption tubes and stored at various temperature conditions. 74 VOCs, 56 of which have been previously uncharacterized, were monitored using GC-MS over a period of 31 d. The results suggest that storage of exhaled breath at cold temperatures (4 °C) provides the most consistent retention of exhaled breath VOCs temporally. Samples were determined to be stable up to 14 d across storage conditions prior to gaining or losing 1–2 standard deviations in abundance. Through gene set enrichment analysis (GSEA), certain chemical classes were found to be positively (acids) or negatively (sulfur-containing) enriched temporally. By means of field sample collections, the effect of storage and shipping was found to be similar to those studies preformed in the laboratory at 4 °C. Collectively this study not only provides recommendations for proper storage conditions and storage length, but also illustrates the use of GSEA to exhaled breath based GC-MS data.

Cross-linked carbon nanotubes-based biocatalytic membranes for micro-pollutants degradation: Performance, stability, and regeneration

The presence of the micro-pollutants in the aqueous environment has become a major environmental challenge. The enzymatic membrane reactor (EMR) provides a promising approach to tackle this problem through benign enzymatic biodegradation, and the fabrication of highly efficient enzymatic membrane is the key to this process. By using a facile carbon nanotube (CNTs) coating technique, we prepared a stable CNTs coating on a polymer membrane support, which was then applied for laccase immobilization via both physical adsorption and covalent bonding. The biocatalytic membrane possessed stability for long-term storage and under hydraulic shear force. Micro-pollutant degradation was carried out with a mixture of five representative compounds (bisphenol-A (BPA), carbamazepine (CBZ), diclofenac (DCF), clofibric acid (CA) and ibuprofen (IBF)). The EMR exhibited substantial improvement in micro-pollutant removals compared with the CNTs-coated membrane having no enzyme. Furthermore, the effects of CNTs loading and operational flux on the EMR performance were examined, and the stability of the enzymatic membranes was studied. Active laccase coatings on CNTs-coated membrane could be renewed after simple cleaning and re-coating, offering new opportunities towards sustainable, long-term applications.

Au/Cu–Fe–La–Al2O3: A highly active, selective and stable catalysts for preferential oxidation of carbon monooxide

A series of gold-based catalysts supported by Fe–La–Al2O3 composite doped with various amounts of copper were prepared using a modified deposition-precipitation method. Investigations were carried out based on the reaction of the preferential carbon monoxide oxidation (CO-PROX) in H2-rich stream and CO oxidation at low temperatures in an O2-rich stream as a reference to examine the effects of copper on the catalytic performance. The high dispersion of gold nanoparticles in alumina-based support, the formation of Au–Cu alloy, and the close contacts of these neat or alloyed nanoparticles with CuO species, making Au partially positively charged, were examined with H2-TPR, XPS, and HRTEM-FFT techniques. Gold catalysts were obtained with very promising performance for polymer electrolyte membrane fuel cell with positive results in activity, CO2 selectivity, and thermal stability and long-term storage and on-line stability. Although H2 addition decreased the rate of CO oxidation over the studied gold catalysts, the activity and selectivity in CO2 formation were continuously improved with an increase in copper loading.

Chitosan as a potential stabilizing agent for titania nanoparticle dispersions for preparation of multifunctional cotton fabric

Titania (TiO2) nanoparticle dispersions in water were prepared using chitosan (CS) as the stabilizing agent. The dispersion stability was evaluated with respect to storage time, hydrodynamic particle size, and zeta potential. The effect of the molecular weight of CS and presence of non-ionic polymers (poly(vinyl alcohol) and poly(ethylene glycol)) as co-dispersants was investigated. Despite the increase in size of dispersed particles, the long-term storage stability of the dispersions improved with increasing concentration and molecular weight of CS. The TiO2/CS dispersions were applied on cotton fabric and characterized. The presence of CS did not seriously affect the photocatalytic self-cleaning activity (SCA) of TiO2; with CS, a SCA of 89% was achieved compared with a value of 96% without CS. In addition, the TiO2/CS-treated cotton fabrics provided UV protection and significant antimicrobial activity.

An Extrapolation of a Radical Equation More Accurately Predicts Shelf Life of Frozen Biological Matrices.

Arrhenius modeling of analyte recovery at increased temperatures to predict long-term colder storage stability of biological raw materials, reagents, calibrators, and controls is standard practice in the diagnostics industry. Predicting subzero temperature stability using the same practice is frequently criticized but nevertheless heavily relied upon. We compared the ability to predict analyte recovery during frozen storage using 3 separate strategies: traditional accelerated studies with Arrhenius modeling, and extrapolation of recovery at 20% of shelf life using either ordinary least squares or a radical equation y = B1x(0.5) + B0. Computer simulations were performed to establish equivalence of statistical power to discern the expected changes during frozen storage or accelerated stress. This was followed by actual predictive and follow-up confirmatory testing of 12 chemistry and immunoassay analytes. Linear extrapolations tended to be the most conservative in the predicted percent recovery, reducing customer and patient risk. However, the majority of analytes followed a rate of change that slowed over time, which was fit best to a radical equation of the form y = B1x(0.5) + B0. Other evidence strongly suggested that the slowing of the rate was not due to higher-order kinetics, but to changes in the matrix during storage. Predicting shelf life of frozen products through extrapolation of early initial real-time storage analyte recovery should be considered the most accurate method. Although in this study the time required for a prediction was longer than a typical accelerated testing protocol, there are less potential sources of error, reduced costs, and a lower expenditure of resources.

Sensitive detection of maltose and glucose based on dual enzyme-displayed bacteria electrochemical biosensor

Glucoamylase-displayed bacteria (GA-bacteria) and glucose dehydrogenase-displayed bacteria (GDH-bacteria) were co-immobilized on multi-walled carbon nanotubes (MWNTs) modified glassy carbon electrode (GCE) to construct GA-bacteria/GDH-bacteria/MWNTs/GCE biosensor. The biosensor was developed by optimizing the loading amount and the ratio of GA-bacteria to GDH-bacteria. The as-prepared biosensor exhibited a wide dynamic range of 0.2–10mM and a low detection limit of 0.1mM maltose (S/N=3). The biosensor also had a linear response to glucose in the range of 0.1–2.0mM and a low detection limit of 0.04mM glucose (S/N=3). Interestingly, at the same concentration, glucose was 3.75-fold sensitive than that of maltose at the proposed biosensor. No interferences were observed for other possible mono- and disaccharides. The biosensor also demonstrated good long-term storage stability and repeatability. Further, using both GDH-bacteria/MWNTs/GCE biosensor and GA-bacteria/GDH-bacteria/MWNTs/GCE biosensor, glucose and maltose in real samples can be detected. Therefore, the proposed biosensor is capable of monitoring the food manufacturing and fermentation process.

Oxyresveratrol and ascorbic acid O/W microemulsion: Preparation, characterization, anti-isomerization and potential application as antibrowning agent on fresh-cut lotus root slices

The purpose of this study is to prepare an oxyresveratrol (Oxy) microemulsion (ME) with improved Oxy’s solubility and stability and to investigate its antibrowning effects on fresh-cut lotus root slices. The formula of OxyME consisted of ethyl butyrate, Tween 80, PEG400, and water with w/w of 4%, 10.67%, 5.33%, and 80%, respectively. Encapsulating Oxy into OxyME greatly increased its solubility and stability compared with that of in water. Strong antibrowning effects were observed on fresh-cut lotus root slices treated with OxyME, even better than 4-hexylresorcinol. The addition of ascorbic acid (VC) into OxyME greatly improved the Oxy stability in long-term storage and antibrowning effects on fresh-cut lotus root slices. However, the simultaneous addition of calcium chloride and VC did not obviously improve the antibrowning effects compared with the addition of VC alone. These results indicated that Oxy+VCME may be suitable as an antibrowning agent for fresh-cut vegetables.

Inkjet-Printed Multi-Bit Low-Voltage Fuse-Type Write-Once-Read-Many Memory Cell

Low-voltage fuse-type write-once-read-many memories were fabricated using a common material inkjet printer to form both the contact pads and the resistor tracks with the same silver ink and process settings. Based on the dependence of the fusing voltage on the length of the printed resistor track, a new cell structure was proposed to achieve multi-bit memories. The fabricated 2-b memory cell presents excellent long-term storage and operation stabilities, and 16-b memories were achieved using this cell structure with greatly saved contact pad numbers for easier external electrical connections than the conventional design. A system demonstration of using the memories was finally provided to prove the potential of this technology for practical use.

Morphological stability of microencapsulated vitamin formulations by AFM imaging

Abstract The morphological stability of microencapsulated lipophilic vitamin formulations directly influences the long-term storage stability and performance in final applications. The internal particle morphology of three commercial dry Vitamin A acetate formulations differing in their matrix composition (hydrocolloid/polysaccharide and plasticizer) was systematically characterized by atomic force microscopy (AFM). The evolution of the particle internal morphology after per-se storage at 25 and 40 °C for ten months was investigated. Furthermore, an accelerated stress test by in-situ heating and AFM imaging was developed. Surface roughness parameters from the AFM images were used to semi-quantitatively estimate the evolution of the particle internal microstructure, and thus the aging and destabilization of the lipophilic vitamin formulations. The influence of humidity level on the thermal morphological restructuring of one of the commercial samples was also investigated. This study has shown that such AFM investigations of microparticles are a novel analytical tool to rapidly estimate the physical stability of the lipophilic vitamin dried emulsions.

Hierarchically assembled ZnO nanosheets microspheres for enhanced glucose sensing performances

Nanostructures with higher surface specific area has great potential applications in sensing devices because higher surface specific area not only improve protein/enzyme immobilization efficiency, but also enhances charge transport and sensing performances. Herein, hierarchically assembled ZnO nanosheets microspheres (HAZNMs) were synthesized by facile one-pot solution process at low-temperature. Results showed that as-synthesized HAZNMs possessing higher specific surface area, significantly increased the enzyme loading efficiency which in turn improved the sensing performances. The as-fabricated biosensors showed a remarkably high sensitivity (210.8μA/mMcm2) in the wide-linear response range of 0.05–23mM, favorable stability for long-term storage, excellent anti-interference ability and high reliability for glucose detection in human blood serum samples. The improved sensing performances can be ascribed to the high glucose oxidase (GOx) enzyme immobilization on HAZNMs that provides a favorable microenvironment for the maintenance of GOx enzyme bioactivity.

Covalent biofunctionalization of chitosan nanofibers with trypsin for high enzyme stability

The electrospun chitosan nanofibers provide excellent material for immobilized proteolytic enzymes, and are biocompatible, nontoxic and hydrophilic matrices with large specific area. This paper deals with an application of electrospun chitosan nanofibers and optimizing conditions for their biofunctionalization by model proteolytic enzyme trypsin. Nanofibers from chitosan were prepared using NanospiderTM technology and covalent immobilization of trypsin followed. Three immobilization techniques preserving biocompatibility and utilizing amine and/or hydroxyl groups of chitosan were optimized and compared to simple adsorption to achieve maximum proteolytic activity per cm2 of the functionalized chitosan nanofibers (Tryp-NF). Significant differences were observed. Trypsin immobilized by the carbodiimide one-step protocol demonstrated the highest activity of the three procedures, ranging from 132 to 210IU/cm2 (i.e., 548–874IU/mg of nanofibers), depending on the initial amount of trypsin used. Long-term storage stability together with high reusability of Tryp-NF confirmed advantages of the immobilized enzyme. Tryp-NF showed no cytotoxicity toward growth of HeLa cells. The in vivo tests for irritation and skin sensitization demonstrated no undesirable skin reactions.

Green synthesized gold nanoparticles decorated graphene oxide for sensitive determination of chloramphenicol in milk, powdered milk, honey and eye drops

A simple and rapid green synthesis using Bischofia javanica Blume leaves as reducing agent was developed for the preparation of gold nanoparticles (AuNPs). AuNPs decorated graphene oxide (AuNPs/GO) was prepared and employed for the sensitive amperometric determination of chloramphenicol. The green biosynthesis requires less than 40s to reduce gold salts to AuNPs. The formations of AuNPs and AuNPs/GO were evaluated by scanning electron and atomic force microscopies, UV–Visible and energy dispersive X-ray spectroscopies, X-ray diffraction studies, and electrochemical methods. AuNPs/GO composite film modified electrode was fabricated and shown excellent electrocatalytic ability towards chloramphenicol. Under optimal conditions, the amperometric sensing platform has delivered wide linear range of 1.5–2.95μM, low detection limit of 0.25μM and high sensitivity of 3.81μAμM−1cm−2. The developed sensor exhibited good repeatability and reproducibility, anti-interference ability and long-term storage stability. Practical feasibility of the sensor has been demonstrated in food samples (milk, powdered milk and honey) and pharmaceutical sample (eye drops). The green synthesized AuNPs/GO composite has great potential for analysis of food samples in food safety measures.

A comparative assessment of long-term storage stability and quality attributes of orange juice in response to pulsed electric fields and heat treatments

Processing of orange juice by pulsed electric fields (PEF) and heat treatments was carried out to determine the quality variables during storage of 180 days at 4°C. Depending on the magnitude of applied electric field strength (13.82–25.26kVcm−1), energy (10.89–51.32J), in comparison with heat treatment (90°C for 10 and 20s); significant changes in pH, titration acidity, total dry matter, and browning index were found during the storage period. With no significant change in shelf-life period, the PEF processing better preserved orange juice quality than did the heat processing. Ascorbic acid content peaked with PEF-treated orange juice under the electric field strength of 17kVcm−1, the treatment time of 1034μs, and the energy of 17.37J through which no significant degradation occurred during the storage with the longest half-life (1112 days). Although ascorbic acid retention of the most of the PEF processed samples was higher than that of the heat processed ones; samples processed by 10.89, 12.70 and 29.57J energies resulted in lower ascorbic acid content than did the heat processing (90°C for 20s). Except for the samples treated by 43.99 and 51.32J energies on 180th day, the PEF processing led to no hydroxyl methyl furfural formation during and at the end of the storage, unlike the heat treatment.

Polysorbates 20 and 80 Degradation by Group XV Lysosomal Phospholipase A2 Isomer X1 in Monoclonal Antibody Formulations

Decreases in polysorbate (PS80) content were observed while evaluating the long-term storage stability of Chinese hamster ovary–derived, purified monoclonal antibodies. It was determined that polysorbate had been enzymatically degraded; therefore, studies were performed to identify and characterize the protein(s) responsible. Polysorbate degrading activity was enriched from Chinese hamster ovary media leading to the identification of group XV lysosomal phospholipase A2 isomer X1 (LPLA2) by shotgun proteomics. Recombinant LPLA2 was over expressed, purified, and functional integrity confirmed against a diheptanoyl phosphatidylcholine substrate. Incubation of recombinantly produced LPLA2 with PS20 and PS80 resulted in hydrolysis of PS20 and PS80 monoester but a much slower rate was observed for higher-order PS80. Endogenous LPLA2 was detected and quantitated at less than 1 ppm in 3 formulated antibodies while LPLA2 was not detected (or less than 0.1 ppm) in a fourth formulated antibody. Furthermore, antibodies with detectable quantities of endogenous LPLA2 demonstrated polysorbate hydrolysis while in contrast the antibody without detectable LPLA2 did not show polysorbate hydrolysis. Comparison of polysorbate degradation products generated from the formulated antibody and samples of polysorbate incubated with recombinant LPLA2 resulted in similar elution profiles by liquid chromatography–mass spectrometry. These results suggest that LPLA2 may play a key role in polysorbate degradation in some antibody preparations.

Spray Application and Release of Microalgae from Water-in-Oil Emulsions

Background: This study was completed as part of a larger effort to develop emulsions that have high stability for long-term algal cell storage and that release cells rapidly when applied to a pond surface as inoculum. Past research has shown that including silica nanoparticles in the oil phase yields stable emulsions but that cell release upon application takes place on the order of days. Methods: Water-in-oil (W/O) emulsions were prepared with Chlorella sorokiniana and brilliant sulfoflavine (BSF) in the internal phase to investigate the effects of oil phase thickeners on BSF and cell release upon spray application of the emulsion to a water surface. Three oil phase thickeners: Aerosil R974, Bentone 38, and Bentone 150, were tested at concentrations ranging from 0.25-1% (w/w) in the oil phase. Results: The oil phase thickeners had a modest impact on the release of BSF but no significant effect on cell release. In particular, emulsions with Bentone 150 had significantly faster release of BSF than other thickeners. Spray application of the emulsion onto a water surface, however, had a dramatic effect on cell release rate without negatively affecting cell viability. Average spray droplet size was found to be ~45 μm and nearly 100% release of both cells and BSF could be achieved within ~30 minutes. Conclusion: Spray application resulted in rapid release of algae and dye even for emulsions with a high concentration of silica nanoparticles. The fact that thickeners affected the release rate of soluble BSF but not cells suggests the potential to design emulsions with differential release rates of cells and compounds upon spray application. Keywords: Chlorella, controlled release, emulsion, oil phase thickener, silica nanoparticles, spray application.

Development and validation of a reliable and rapid LC-MS/MS method for simultaneous quantification of sacubitril and valsartan in rat plasma and its application to a pharmacokinetic study.

A selective, sensitive and rapid liquid chromatographic method with electrospray ionization tandem mass spectrometric detection has been developed and validated for simultaneous quantification of sacubitril and valsartan in rat plasma using telmisartan as internal standard (IS). The analytes were extracted by deprotenization of 50 μL of plasma sample using 200 μL of acetonitrile. In a short chromatographic run of 1.50 min run time, separation was achieved on a Hypersil Gold C18 column using a mobile phase composed of 0.1% formic acid in Milli-Q water-0.1% formic acid in acetonitrile in gradient elution mode. The quantification of target compounds was performed in a positive electrospray ionization mode and multiple reaction monitoring. Response was a linear function of concentration in the ranges of 0.5-20,000 ng/mL for both analytes, with r(2) > 0.9997. The intra- and inter-day precision and accuracy results were <15% and acceptable as per US Food and Drug Administration guidelines. Stability of compounds were established in a battery of stability studies, i.e. bench-top, autosampler and long-term storage stability as well as freeze-thaw cycles. The validated method can be used as a routine method to support pharmacokinetic studies. Copyright © 2016 John Wiley & Sons, Ltd.

Development and validation of a high throughput UPLC–MS/MS method for simultaneous quantification of esomeprazole, rabeprazole and levosulpiride in human plasma

A high throughput ultra pressure liquid chromatography–mass spectrometry (UPLC–MS/MS) method with good sensitivity and selectivity has been developed and validated for simultaneous quantification of esomeprazole, rabeprazole and levosulpiride in human plasma using lansoprazole as internal standard (IS). The extraction method based on liquid–liquid extraction technique was used to extract the analytes and IS from of 50µL of human plasma using methyl tert-butyl ether:ethyl acetate (80:20, v/v), which offers a high recovery. Chromatographic separation of analytes and IS was achieved on a Hypersil gold C18 column using gradient mobile phase consisting of 2mM ammonium formate/acetonitrile. The flow rate was set at 0.5mL/min to elute all the analytes and IS within 1.00min runtime. Detection of target compounds was performed on a triple quadruple mass spectrometer by multiple reaction monitoring (MRM) mode via positive electrospray ionization (ESI). Method validation results demonstrated that the developed method has good precision and accuracy over the concentration ranges of 0.1–2000ng/mL for each analyte. Stability of compounds was established in a battery of stability studies, i.e., bench top, autosampler, dry extract and long-term storage stability as well as freeze-thaw cycles. The validated method has been successfully applied to analyze human plasma samples for application in pharmacokinetic studies.

Size-controllable ultrathin carboxylated polypyrrole nanotube transducer for extremely sensitive 17β-estradiol FET-type biosensors.

17β-Estradiol is known as a steroid hormone in the human body but it is also known as a disruptor that can cause disequilibrium and dysfunction of the human immune system. Recently, there has been much interest in developing biosensors to detect low concentrations of 17β-estradiol. In this work, size-controllable aptamer conjugated ultrathin carboxylated polypyrrole nanotubes (A-UCPPyNTs) were fabricated as transducers in 17β-estradiol field-effect transistor (FET)-type biosensors. They were manufactured via a self-degradation method under several different conditions to control the diameter of the nanotubes. For targeting 17β-estradiol, the binding aptamers were immobilized through covalent bonding on its surface. The resulting A-UCPNT FET-type biosensor demonstrated p-type behavior with outstanding electrical conductivity, and exhibited Ohmic contacts between the samples and electrodes. The smaller diameter (40 nm) of ultrathin carboxylated polypyrrole nanotubes (UCPPyNTs) contributed to the biosensor's enhanced performance by generating a larger surface area, thereby increasing the number of conjugated binding aptamers. In conclusion, the A-UCPPyNT FET-type biosensor showed extremely high sensitivity (∼1 fM) toward 17β-estradiol, approximately 103 times more sensitive than the results found in other reports. Moreover, the A-UCPPyNT FET-type biosensor showed unique selectivity to the 17β-estradiol molecule, in addition to outstanding reusability and long-term storage stability (4 weeks of duration achieved in this work). These performances concerned with reusability and stability were achieved by the formation of covalent bonding in the anchorage to the substrate electrode. Thus this study can be effectively applied in biological and environmental fields.