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Development of miniaturized tapered fiber optics diamine oxidase-based sensor for histamine detection


Usman, Hamza Boko (2015) Development of miniaturized tapered fiber optics diamine oxidase-based sensor for histamine detection. Masters thesis, Universiti Putra Malaysia.


Estimating the level of histamine in fish and fish products is very important because of their implication in fish poisoning in human; hence, ascertaining histamine levels in the afore-mentioned serves as a chemical index for spoilage. Factors such as expensive instrumentation, time consumption, size and weight are some of the problems with conventional analytical methods of detection. Tapered fiber optics-enzyme based sensors are prospective candidates towards resolving these quagmires due to their portability and simplicity vis-à-vis currently available detectors. Taking into cognizance immobilization as the backbone of any biosensor, this work, reports a technique to immobilize an ordered multilayer of diamine oxidase (DAO) by means of chemical cross-linking on the biconical tapered fiber surface step-wisely alternating between chitosan, glutaraldehyde (GA) and the enzyme. The optimum parameters for the fabricated biosensor included 160 mg/ml DAO, 0.5% chitosan, 2.5% GA, pH 7.0,and tapered fiber surface of waist diameter 12 μm and length 20 mm. A spectrophotometric signal resulted from horseradish peroxidase catalyzed reduction of H2O2, a secondary product of the oxidative deamination of histamine monitored at 450 nm in 0.1 M phosphate buffer (pH 7.0 and room temperature). Atomic force microscopy (AFM), scanning electron microscope (SEM) and spectrophotometric technique confirmed the functionality of the biosensor. The biosensor showed a response and recovery time of 14 sec, a linear response range up to 1.5 mM, a good sensitivity of 0.64 mM-1 with detection and quantification limits towards histamine of 0.086 mM (15.8 ppm) and 0.204 mM (37.7 ppm) and a linear response range of 0-1.5 mM. The sensor showed an excellent anti-interferents property towards the common interferents’ agents of <5%, with good recovery performance towards varying concentration of histamine ranging from 95.6 to 103.6% (RSD <5%). It showed operational stability to up to 40 repeated analyses without significant loss of sensitivity. The developed miniaturized biosensor has a good potential for use in quantitative measurement of histamine in seafood.

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Additional Metadata

Item Type: Thesis (Masters)
Subject: Chemical detectors
Subject: Optical fiber detectors
Subject: Design
Call Number: FBSB 2015 9
Chairman Supervisor: Abu Bakar Salleh, PhD
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Depositing User: Haridan Mohd Jais
Date Deposited: 26 Apr 2018 03:50
Last Modified: 26 Apr 2018 03:50
URI: http://psasir.upm.edu.my/id/eprint/60432
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