Sains Malaysiana 48(7)(2019): 1409–1416

http://dx.doi.org/10.17576/jsm-2019-4807-09

A Hydrogen Peroxide Biosensor from Horseradish peroxidase Immobilization onto Acrylic Microspheres

(Biosensor Hidrogen Peroksida Berasaskan Pemegunan Peroksidase Lobak Putih pada Mikrosfera Akrilik)

EDA YUHANA ARIFFIN¹, NIK NURHANAN NIK MANSOR¹, EKA SAFITRI², LEE YOOK HENG¹ & NURUL IZZATY HASSAN¹*

¹Centre for Advanced Materials and Renewable Resources, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

²Department of Chemistry, Faculty of Mathematics and Natural Sciences, Syiah Kuala University (USK), Darussalam-Banda Acheh 23111, Indonesia

Received: 19 November 2018/Accepted: 8 April 2019

ABSTRACT

The sensitive and rapid detection of hydrogen peroxide is very important in the areas of clinical and environmental analyses. A sensitive and selective Horseradish peroxidase (HRP)-hydrogen peroxide (H2O2) biosensor was developed based on acrylic microspheres. Hydrophobic poly(n-butyl acrylate-N-acryloxysuccinimide) [poly(nBA-NAS)] microspheres were synthesized using photopolymerization in an emulsion to form an enzyme immobilization matrix. The HRP enzyme was covalently immobilized onto the acrylic microspheres via the succinimide functionality. Field emission scanning electron microscope (FESEM) has been utilized to characterize the screen-printed carbon paste electrode (SPE) constructed from enzyme conjugated acrylic microspheres and gold nanoparticles (AuNPs) composite (HRP/nBA-NAS/AuNPs/SPE). Differential pulse voltammetry was used to assess the biosensor performance. The linear response range of the hydrogen peroxide biosensor obtained was from 1.0 × 10-2 to 1.0 × 10-10 M (R2 = 0.99) with the limit of detection (LOD) approximately at 1.0 × 10-10 M. This is an improvement over many hydrogen peroxide biosensors reported so far. Such improvement may be attributed to the large surface area provided by the acrylic microspheres as a matrix for immobilization of the HRP enzyme.

Keywords: Acrylic microsphere; biosensor; Horseradish peroxidase; hydrogen peroxide

ABSTRAK

Pengesanan hidrogen peroksida yang sensitif dan cepat adalah sangat penting dalam bidang analisis persekitaran dan klinikal. Biosensor hidrogen peroksida (H2O2)-peroksidase lobak putih (HRP) yang sensitif dan memilih telah dibangunkan berasaskan mikrosfera akrilik. Mikrosfera hidrofobik poli(n-butil akrilat-N-akriloksuksinimida)[poli(nBA-NAS)] disintesis dengan menggunakan proses fotopempolimeran dalam bentuk emulsi dan ia bertindak sebagai matriks pemegun enzim. Enzim HRP dipegunkan secara kovalen pada mikrosfera akrilik melalui pengfungsian suksinimida. Mikroskop elektron imbasan pancaran medan (FESEM) telah digunakan bagi mencirikan elektrod karbon permukaan bercetak (SPE) yang dibina daripada komposit mikrosfera akrilik berkonjugat enzim dan nanosfera emas (AuNPs) (HRP/nBA-NAS/AuNPs/SPE). Voltametri denyutan pembezaan digunakan untuk penilaian prestasi biosensor. Julat keupayaan linear bagi biosensor hidrogen peroksida diperoleh daripada 1.0 × 10-2 hingga 1.0 × 10-10 M (R2 = 0.99) dengan had pengesanan (LOD) ditemui pada 1 × 10-10 M. Ini merupakan penambahbaikan berbanding biosensor hidrogen peroksida yang telah terlebih dahulu dilaporkan. Penambahbaikan ini mungkin ditentukan oleh luas permukaan yang besar yang disediakan oleh mikrosfera akrilik sebagai tapak pemegunan enzim HRP.

Kata kunci: Biosensor; hidrogen peroksida; mikrosfera akrilik; peroksidase lobak putih

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*Corresponding author; email: drizz@ukm.edu.my