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Abstract More than 200 different cultivars of durian exist worldwide but Durio zibethinus or Musang King (MK) is the most premium and prized durian fruit among the recommended varieties. Early identification of this premium variety is critical to protect from non-authentic MK durian cultivars. However, the MK variety's morphological traits are nearly identical to other varieties. Currently, the identification of durian varieties is mostly performed via evaluation of leaf shape, fruit shape, aroma, taste and seed shape and this requires trained personnel for the morphology observation. To enable the rapid identification of the MK variety, PCR amplification of ten durian varieties using six gene candidates from the chloroplast genome was first performed to obtain DNA probes that were specific to the MK durian variety. PCR amplification of ten durian varieties using primers designed confirmed that the nadhA gene sequence showed an obvious difference in the MK variety from other durian varieties. The unique sequence of MK was used as a DNA probe to develop an electrochemical biosensor for the direct identification of the MK durian variety. The electrochemical biosensor was based on the hybridization response of the immobilized DNA probe with the target DNA from the MK variety and was monitored via differential pulse voltammetry technique. Under optimal conditions, the DNA electrochemical biosensor showed a low detection limit at 10% of MK genomic DNA concentration with a wide linear calibration range of 0.05–1.5 µM (R2 = 0.9891) and RSD value of 3.77% (n = 3). The results of the developed DNA biosensor provide high promise for the development of portable sensors employed in the determination of MK variety in the field.

Histamine is commonly present in food containing proteins, like in mackerel. Consuming fish is imperative for the improvement of human muscles. Nevertheless, some studies reported ingesting fish containing histamine more than 50 mg•kg-1 can cause toxicity. This study analyzed and determined the composition of histamine in mackerel and its products commonly consumed in Malaysia, especially on the East Coast of Malaysia. These included processed mackerel such as canned products, satay (skewed fish) and keropok lekor (fish cake/ cracker). Histamine analysis was performed using High Performance Liquid Chromatography (HPLC) equipped with a fluorescence detector. A derivatizing reaction was applied to increase the sensitivity of HPLC to histamine using 9-flourenilmethylchloroformate (FMOC-Cl). The chromatographic separation was achieved in 15 min. Method validation was in accordance to Commission Decision 657/2002/CE. The linear range was at 0.16 – 5.00 µg•mL-1 (histamine) with the LOD at 0.10 µg•mL-1 and LOQ at 0.30 µg•mL-1. Method applicability was checked on seven real samples involving raw, cooked, and dry products, yielding acceptable recovery.

In view of the presence of pathogenic Vibrio cholerae (V. cholerae) bacteria in environmental waters, including drinking water, which may pose a potential health risk to humans, an ultrasensitive electrochemical DNA biosensor for rapid detection of V. cholerae DNA in the environmental sample was developed. Silica nanospheres were functionalized with 3-aminopropyltriethoxysilane (APTS) for effective immobilization of the capture probe, and gold nanoparticles were used for acceleration of electron transfer to the electrode surface. The aminated capture probe was immobilized onto the Si-Au nanocomposite-modified carbon screen printed electrode (Si-Au-SPE) via an imine covalent bond with glutaraldehyde (GA), which served as the bifunctional cross-linking agent. The targeted DNA sequence of V. cholerae was monitored via a sandwich DNA hybridization strategy with a pair of DNA probes, which included the capture probe and reporter probe that flanked the complementary DNA (cDNA), and evaluated by differential pulse voltammetry (DPV) in the presence of an anthraquninone redox label. Under optimum sandwich hybridization conditions, the voltammetric genosensor could detect the targeted V. cholerae gene from 1.0 × 10−17–1.0 × 10−7 M cDNA with a limit of detection (LOD) of 1.25 × 10−18 M (i.e., 1.1513 × 10−13 µg/µL) and long-term stability of the DNA biosensor up to 55 days. The electrochemical DNA biosensor was capable of giving a reproducible DPV signal with a relative standard deviation (RSD) of n = 5). Satisfactory recoveries of V. cholerae cDNA concentration from different bacterial strains, river water, and cabbage samples were obtained between 96.5% and 101.6% with the proposed DNA sandwich biosensing procedure. The V. cholerae DNA concentrations determined by the sandwich-type electrochemical genosensor in the environmental samples were correlated to the number of bacterial colonies obtained from standard microbiological procedures (bacterial colony count reference method).

Polyurethane (PU) is a unique polymer that has versatile processing methods and mechanical properties upon the inclusion of selected additives. In this study, a freestanding bio-based polyurethane film the screen-printed electrode (SPE) was prepared by the solution casting technique, using acetone as solvent. It was a one-pot synthesis between major reactants, namely, palm kernel oil-based polyol and 4,4-methylene diisocyanate. The PU has strong adhesion on the SPE surface. The synthesized bio-based polyurethane was characterized using thermogravimetry analysis, differential scanning calorimetry, Fourier-transform infrared spectroscopy (FTIR), surface area analysis by field emission scanning electron microscope, and cyclic voltammetry. Cyclic voltammetry was employed to study electrocatalytic properties of SPE-polyurethane towards oxidation of PU. Remarkably, SPE-PU exhibited improved anodic peak current as compared to SPE itself using the differential pulse voltammetry method. Furthermore, the formation of urethane linkages (-NHC(O) backbone) after polymerization was analyzed using FTIR and confirmed by the absence of peak at 2241 cm-1 attributed to the sp-hydridized carbons atoms of C≡C bonds. The glass transition temperature of the polyurethane was detected at 78.1°C.

Chronic exposure of 17β-estradiol (E2) even at low concentration can disorganize the endocrine system and lead to undesirable health problems in the long run. An electrochemical biosensor for rapid detection of E2 in water samples was successfully developed. The biosensor was based on split DNA aptamers attached onto poly (methacrylic acid-co-n butyl acrylate-succinimide) microspheres deposited on polypyrrole nanowires coated electrode (PPY/PMAA-NBA). The sandwich paired of split DNA aptamers used were truncated from 75 mer parent aptamers. These two strands of 12-mer and 14-mer split DNA aptamers were then immobilized on the PMAA-NBA microspheres. In the presence of E2, the split DNA aptamers formed an apt12-E2-apt14 complex, where the binding reaction on the electrode surface led to the detection of E2 by differential pulse voltammetry using ferrocyanide as a redox indicator. Under optimum conditions, the aptasensor detected E2 concentrations in the range of 1 × 10−4 M to 1 × 10−12 M (R2 = 0.9772) with a detection limit of 4.8 × 10−13 M. E2, which were successfully measured in a real sample with 97–104% recovery and showed a good correlation (R2 = 0.9999) with the established method, such as high-performance liquid chromatography. Interactions between short and sandwich-type aptamers (split aptamers) demonstrated improvement in aptasensor performance, especially the selectivity towards several potential interferents.

A new electrochemical DNA biosensor based on mercaptopropionic acid (MPA)-capped ZnS quantum dots (MPA-ZnS QDs) immobilization matrix for covalent binding with 20-base aminated oligonucleotide has been successfully developed. Prior to the modification, screen-printed carbon paste electrode (SPE) was self-assembled with multilayer gold nanoparticles (AuNPs) and cysteamine (Cys). The inclusion of MPA-ZnS QDs semiconducting material in modified electrodes has enhanced the electron transfer between the SPE transducer and DNA leading to improved bioanalytical assay of target biomolecules. Electrochemical studies performed by cyclic voltammetry (CV) and differential pulsed voltammetry (DPV) demonstrated that the MPA-ZnS QDs modified AuNPs electrode was able to produce a lower charge transfer resistance response and hence higher electrical current response. Under optimal conditions, the immobilized synthetic DNA probe exhibited high selectivity towards synthetic target DNA. Based on the DPV response of the reduction of anthraquinone monosulphonic acid (AQMS) redox probe, the MPA-ZnS QDs-based electrochemical DNA biosensor responded to target DNA concentration from 1 × 10−9 μM to 1 × 10−3 μM with a sensitivity 1.2884 ± 0.12 µA, linear correlation coefficient (R2) of 0.9848 and limit of detection (LOD) of 1 × 10−11 μM target DNA. The DNA biosensor exhibited satisfactory reproducibility with an average relative standard deviation (RSD) of 7.4%. The proposed electrochemical transducer substrate has been employed to immobilize the aminated Arowana fish (Scleropages formosus) DNA probe. The DNA biosensor showed linearity to target DNA from 1 × 10−11 to 1 × 10−6 µM (R2 = 0.9785) with sensitivity 1.1251 ± 0.243 µA and LOD of 1 × 10−11 µM. The biosensor has been successfully used to determine the gender of Arowana fish without incorporating toxic raw materials previously employed in the hazardous processing conditions of polypyrrole chemical conducting polymer, whereby the cleaning step becomes difficult with thicker films due to high levels of toxic residues from the decrease in polymerization efficacy as films grew.

In the last decade, there has been a steady stream of information on the methods and techniques available for detecting harmful algae species. The conventional approaches to identify harmful algal bloom (HAB), such as microscopy and molecular biological methods are mainly laboratory-based and require long assay times, skilled manpower, and pre-enrichment of samples involving various pre-experimental preparations. As an alternative, biosensors with a simple and rapid detection strategy could be an improvement over conventional methods for the detection of toxic algae species. Moreover, recent biosensors that involve the use of nanomaterials to detect HAB are showing further enhanced detection limits with a broader linear range. The improvement is attributed to nanomaterials’ high surface area to volume ratio, excellent biological compatibility with biomolecules, and being capable of amplifying the electrochemical signal. Hence, this review presents the potential usage of biosensors over conventional methods to detect HABs. The methods reported for the detection of harmful algae species, ranging from conventional detection methods to current biosensor approaches will be discussed, along with their respective advantages and drawbacks to indicate the future prospects of biosensor technology for HAB event management.

Histamine is commonly present in food containing proteins, like in mackerel. Consuming fish is imperative for the improvement of human muscles. Nevertheless, some studies reported ingesting fish containing histamine more than 50 mg·kg-1 can cause toxicity. This study analyzed and determined the composition of histamine in mackerel and its products commonly consumed in Malaysia, especially on the East Coast of Malaysia. These included processed mackerel such as canned products, satay (skewed fish) and keropok lekor (fish cake/ cracker). Histamine analysis was performed using High Performance Liquid Chromatography (HPLC) equipped with a fluorescence detector. A derivatizing reaction was applied to increase the sensitivity of HPLC to histamine using 9-flourenilmethylchloroformate (FMOC-Cl). The chromatographic separation was achieved in 15 min. Method validation was in accordance to Commission Decision 657/2002/CE. The linear range was at 0.16 – 5.00 μg·mL-1 (histamine) with the LOD at 0.10 μg·mL-1 and LOQ at 0.30 μg·mL-1. Method applicability was checked on seven real samples involving raw, cooked, and dry products, yielding acceptable recovery. [ABSTRACT FROM AUTHOR]