Immobilization of laccase on the functionalized methacrylate-acrylate copolymer microspheres was studied. Poly(glycidyl methacrylate-co-n-butyl acrylate) microspheres consisting of epoxy groups were synthesized using facile emulsion photocuring technique. The epoxy groups in poly(GMA-co-nBA) microspheres were then converted to amino groups. Laccase immobilization is based on covalent binding via amino groups on the enzyme surface and aldehyde group on the microspheres. The FTIR spectra showed peak at 1646 cm⁻¹ assigned to the conformation of the polymerization that referred to GMA and nBA monomers, respectively. After modification of the polymer, intensity of FTIR peaks assigned to the epoxy ring at 844 cm⁻¹ and 904 cm⁻¹ was decreased. The results obtained from FTIR exhibit a good agreement with the epoxy content method. The activity of laccase-immobilized microspheres increased upon increasing the epoxy content. Furthermore, poly(GMA-co-nBA) microspheres revealed uniform size below 2 µm that contributes to large surface area of the microspheres to be used as a matrix, thus increasing the enzyme capacity and enzymatic reaction. Immobilized enzyme also shifted to higher pH and temperature compared to free enzyme.

A developed colorimetric pH sensor film based on edible materials for real-time monitoring of food freshness is described. The mixed natural dyes from edible plants Clitoria sp and Brassica sp were extracted and incorporated into ι-carrageenan film as a colorimetric pH sensor film for monitoring food spoilage and its freshness. The color changes of the developed colorimetric sensor film were measured with chromametry and UV-vis spectroscopy, respectively. Experimental results show that colorimetric pH sensor film demonstrated statistically significant differences (p < 0.05) between CIE-L*a*b* coordinates color system indicated that the developed colorimetric sensor film was able to give a gradual change in color over a wide pH range. The color of the colorimetric sensor film also changes discretely and linearly with factors that contribute to food spoilage using shrimp and durian samples. Moreover, the developed colorimetric pH sensor film has the potential to be used as a safe, non-destructive testing and also a flexibly visual method for direct assessment of food freshness indicator during storage.

For the past decade, much attention was focused on polysaccharide natural resources for various purposes. Throughout the works, several efforts were reported to prepare new function of chitosan by chemical modifications for renewable energy, such as fuel cell application. This paper focuses on synthesis of the chitosan derivative, namely, O-nitrochitosan which was synthesized at various compositions of sodium hydroxide and reacted with nitric acid fume. Its potential as biopolymer electrolytes was studied. The substitution of nitro group was analyzed by using Attenuated Total Reflectance Fourier Transform Infra-Red (ATR-FTIR) analysis, Nuclear Magnetic Resonance (NMR) and Elemental Analysis (CHNS). The structure was characterized by X-ray Diffraction (XRD) and its thermal properties were examined by using differential scanning calorimetry (DSC) and thermal gravimetric analysis (TGA). Whereas, the ionic conductivity of the samples was analyzed by electrochemical impedance spectroscopy (EIS). From the IR spectrum results, the nitro group peaks of O-nitrochitosan, positioned at 1646 and 1355 cm^-1, were clearly seen for all pH media. At pH 6, O-nitrochitosan exhibited the highest degree of substitution at 0.74 when analyzed by CHNS analysis and NMR further proved that C-6 of glucosamine ring was shifted to the higher field. However, the thermal stability and glass transition temperatures were decreased with acidic condition. The highest ionic conductivity of O-nitrochitosan was obtained at ~10⁻⁶ cm^-1. Overall, the electrochemical property of new O-nitrochitosan showed a good improvement as compared to chitosan and other chitosan derivatives. Hence, O-nitrochitosan is a promising biopolymer electrolyte and has the potential to be applied in electrochemical devices.

A simple and sensitive aptasensor based on conductive carbon nanodots (CDs) was fabricated for the detection of 17ß-Estradiol (E2). In the present study, the hydrothermal synthesis of carbon nanodots was successfully electrodeposited on a screen-printed electrode (SPE) as a platform for immobilization of 76-mer aptamer probe. The morphology and structure of the nanomaterial were characterized by UV-visible absorption spectra, Fluorescence spectra, Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR). Moreover, cyclic voltammetry and electrochemical impedance spectroscopy were used to investigate the electrochemical performance of the prepared electrodes. Subsequently, impedimetric (EIS) measurements were employed to investigate the relative impedances changes before and after E2 binding, which results in a linear relationship of E2 concentration in the range of 1.0 × 10⁻⁷ to 1.0 × 10 ⁻¹² M, with a detection limit of 0.5 × 10⁻¹² M. Moreover, the developed biosensor showed high selectivity toward E2 and exhibited excellent discrimination against progesterone (PRG), estriol (E3) and bisphenol A (BPA), respectively. Moreover, the average recovery rate of spiked river water samples with E2 ranged from 98.2% to 103.8%, with relative standard deviations between 1.1% and 3.8%, revealing the potential application of the present biosensor for E2 detection in water samples.

Laccase enzyme, a commonly used enzyme for the construction of biosensors for phenolic compounds was used for the first time to develop a new biosensor for the determination of the azo-dye tartrazine. The electrochemical biosensor was based on the immobilization of laccase on functionalized methacrylate-acrylate microspheres. The biosensor membrane is a composite of the laccase conjugated microspheres and gold nanoparticles (AuNPs) coated on a carbon-paste screen-printed electrode. The reaction involving tartrazine can be catalyzed by laccase enzyme, where the current change was measured by differential pulse voltammetry (DPV) at 1.1 V. The anodic peak current was linear within the tartrazine concentration range of 0.2 to 14 μM (R² = 0.979) and the detection limit was 0.04 μM. Common food ingredients or additives such as glucose, sucrose, ascorbic acid, phenol and sunset yellow did not interfere with the biosensor response. Furthermore, the biosensor response was stable up to 30 days of storage period at 4 °C. Foods and beverage were used as real samples for the biosensor validation. The biosensor response to tartrazine showed no significant difference with a standard HPLC method for tartrazine analysis.

An enzyme-based electrochemical biosensor was investigated for the analysis of Sunset Yellow synthetic food dye. A glassy carbon electrode was coated with a poly(acrylamide-co-ethyl methacrylate) membrane to immobilize laccase using a single-step photopolymerization procedure. Poly(acrylamide-co-ethyl methacrylate) membrane was demonstrated to have acceptable water absorption and suitable for biosensor application. Sunset Yellow biosensor exhibited a linear response range from 0.08 to 10.00 µM with a detection limit of 0.02 µM. This biosensor was successfully used to determine Sunset Yellow in soft drinks with recoveries of 99.0–101.6%. The method was validated using high-performance liquid chromatography, indicating the biosensor can be as a promising alternative method for Sunset Yellow detection.