Since ancient times, honey has been used for medical purposes and the treatment of various disorders. As a high-quality food product, the honey industry is prone to fraud and adulteration. Moreover, limited experimental studies have investigated the impact of adulterated honey consumption using zebrafish as the animal model. The aims of this study were: (1) to calculate the lethal concentration (LC50) of acid-adulterated Apis mellifera honey on embryos, (2) to investigate the effect of pure and acid-adulterated A. mellifera honey on hatching rate (%) and heart rate of zebrafish (embryos and larvae), (3) to elucidate toxicology of selected adulterated honey based on lethal dose (LD50) using adult zebrafish and (4) to screen the metabolites profile of adulterated honey from blood serum of adult zebrafish. The result indicated the LC50 of 31.10 ± 1.63 (mg/ml) for pure A. mellifera honey, while acetic acid demonstrates the lowest LC50 (4.98 ± 0.06 mg/ml) among acid adulterants with the highest mortality rate at 96 hpf. The treatment of zebrafish embryos with adulterated A. mellifera honey significantly (p ≤ 0.05) increased the hatching rate (%) and decreased the heartbeat rate. Acute, prolong-acute, and sub-acute toxicology tests on adult zebrafish were conducted at a concentration of 7% w/w of acid adulterants. Furthermore, the blood serum metabolite profile of adulterated-honey-treated zebrafish was screened by LC-MS/MS analysis and three endogenous metabolites have been revealed: (1) Xanthotoxol or 8-Hydroxypsoralen, (2) 16-Oxoandrostenediol, and (3) 3,5-Dicaffeoyl-4-succinoylquinic acid. These results prove that employed honey adulterants cause mortality that contributes to higher toxicity. Moreover, this study introduces the zebrafish toxicity test as a new promising standard technique for the potential toxicity assessment of acid-adulterated honey in this study and hazardous food adulterants for future studies.
Soy sauce taste has become a focus of umami taste research. Umami taste is a 5th basic taste, which is associated to a palatable and pleasurable taste of food. Soy sauce has been used as an umami seasoning since the ancient time in Asia. The complex fermentation process occurred to soy beans, as the raw material in the soy sauce production, gives a distinct delicious taste. The recent investigation on Japanese and Indonesian soy sauces revealed that this taste is primarily due to umami components which have molecular weights lower than 500 Da. Free amino acids are the low molecular compounds that have an important role to the taste, in the presence of sodium salt. The intense umami taste found in the soy sauces may also be a result from the interaction between umami components and other tastants. Small peptides are also present, but have very low, almost undetected umami taste intensities investigated in their fractions.
Safety and quality aspects of food products have always been critical issues for the food production and processing industries. Since conventional quality measurements are laborious, time-consuming, and expensive, it is vital to develop new, fast, non-invasive, cost-effective, and direct techniques to eliminate those challenges. Recently, non-destructive techniques have been applied in the food sector to improve the quality and safety of foodstuffs. The aim of this review is an effort to list non-destructive techniques (X-ray, computer tomography, ultraviolet-visible spectroscopy, hyperspectral imaging, infrared, Raman, terahertz, nuclear magnetic resonance, magnetic resonance imaging, and ultrasound imaging) based on the electromagnetic spectrum and discuss their principle and application in the food sector. This review provides an in-depth assessment of the different non-destructive techniques used for the quality and safety analysis of foodstuffs. We also discussed comprehensively about advantages, disadvantages, challenges, and opportunities for the application of each technique and recommended some solutions and developments for future trends.
The characterization and fat migration of palm kernel stearin (PKS) and desiccated coconut, used as base filling centre in dark chocolate were studied. C36 and C38 triglycerides of PKS decreased by 11% and 9.6% respectively, whereas C32 and C34 increased by 97% and 48% respectively. The change in the triglycerides composition of PKS shift the melting point of PKS from 33.2 to 31.4 degrees C. Solid fat content (SFC) of PK reduced by 40% at 30 degrees C. The rate of fat migration was very slow at 18 degrees C storage compared to 30 degrees C. The rate of change of C36 in the chocolate layer was 0.1% week-1 and 1.2% week-1 at 18 and 30 degrees C respectively. Chocolate stored at 18 degrees C showed post hardening during storage period and withstood bloom during the storage period, whereas that stored at 30 degrees C became soft and bloomed faster after 3 weeks of storage.
Palm kernel cake (PKC) is the solid residue following oil extraction of palm kernels and useful to fatten animals either as a single feed with only minerals and vitamins supplementation, or mixed with other feedstuffs such as corn kernels or soy beans. The occurrence of mycotoxins (aflatoxins, ochratoxins, zearalenone, and fumonisins) in feed samples affects the animal's health and also serves as a secondary contamination to humans via consumption of eggs, milk and meats. Of these, aflatoxin B₁ (AFB₁) is the most toxically potent and a confirmed carcinogen to both humans and animals. Methods such as High Performance Liquid Chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry (LC-MS/MS) are common in the determination of mycotoxins. However, these methods usually require sample pre-treatment, extensive cleanup and skilled operator. Therefore, in the present work, a rapid method of electrochemical immunosensor for the detection of AFB₁ was developed based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). Multi-walled carbon nanotubes (MWCNT) and chitosan (CS) were used as the electrode modifier for signal enhancement.N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC) andN-hydroxysuccinimide (NHS) activated the carboxyl groups at the surface of nanocomposite for the attachment of AFB₁-BSA antigen by covalent bonding. An indirect competitive reaction occurred between AFB₁-BSA and free AFB₁ for the binding site of a fixed amount of anti-AFB₁ antibody. A catalytic signal based on horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H₂O₂) and 3,3',5,5'-tetramethylbenzidine (TMB) mediator was observed as a result of attachment of the secondary antibody to the immunoassay system. As a result, the reduction peak of TMB(Ox)was measured by using differential pulse voltammetry (DPV) analysis. Based on the results, the electrochemical surface area was increased from 0.396 cm² to 1.298 cm² due to the electrode modification with MWCNT/CS. At the optimal conditions, the working range of the electrochemical immunosensor was from 0.0001 to 10 ng/mL with limit of detection of 0.1 pg/mL. Good recoveries were obtained for the detection of spiked feed samples (PKC, corn kernels, soy beans). The developed method could be used for the screening of AFB₁ in real samples.
A reversed-phase HPLC optimization strategy is presented for investigating the separation and retention behavior of aflatoxin B1, B2, G1, G2, ochratoxin A and zearalenone, simultaneously. A fractional factorial design (FFD) was used to screen the significance effect of seven independent variables on chromatographic responses. The independent variables used were: (X1) column oven temperature (20-40°C), (X2) flow rate (0.8-1.2 ml/min), (X3) acid concentration in aqueous phase (0-2%), (X4) organic solvent percentage at the beginning (40-50%), and (X5) at the end (50-60%) of the gradient mobile phase, as well as (X6) ratio of methanol/acetonitrile at the beginning (1-4) and (X7) at the end (0-1) of gradient mobile phase. Responses of chromatographic analysis were resolution of mycotoxin peaks and HPLC run time. A central composite design (CCD) using response surface methodology (RSM) was then carried out for optimization of the most significant factors by multiple regression models for response variables. The proposed optimal method using 40°C oven temperature, 1 ml/min flow rate, 0.1% acetic acid concentration in aqueous phase, 41% organic phase (beginning), 60% organic phase (end), 1.92 ratio of methanol to acetonitrile (beginning) and 0.2 ratio (end) for X1-X7, respectively, showed good prediction ability between the experimental data and predictive values throughout the studied parameter space. Finally, the optimized method was validated by measuring the linearity, sensitivity, accuracy and precision parameters, and has been applied successfully to the analysis of spiked cereal samples.
In the present work, supercritical fluid extraction (SFE) with CO2 as solvent and EtOH/water (v/v) as co-solvent was optimised by applying 23 factorial experimental design for the extraction of betacyanins from red pitaya fruit (Hylocereus polyrhizus) peel. Three independent variables of pressure (20-30 MPa), temperature (40-60°C) and co-solvent concentration (10-20%) were chosen for response variables. With the 2 mL/min flow rate of CO2, the dynamic time of extraction was found to be 90 min. The linear effects of main factors and interactions were evaluated. The calculated response surface model for the pressure/temperature was found to be significant for all the dependent variables. At optimal condition of SFE, the response variables were assessed as maximum extraction yield of 4.09 ± 0.69%, total betacyanins content of 25.49 ± 1.54 mg/100 mL, redness (a*) of 58.18 ± 0.82, and IC50 (antioxidant activity) of 1.34 ± 0.12 mg/mL for the experimental peel extracts. The optimal levels of independent variables were validated for the experimental responses as predicted by the mathematical model. The reliability of this method was confirmed as there was no significant difference between experimental and predicted values. The HPLC-MS profile of betacyanins extract comprised of both acylated and non-acylated betacyanins constituents.
Seafood is common item in the world diet; Asian countries have the highest rates of fish consumption in the world, which is higher than world average. Several studies have been conducted on the epidemiology and clinical characteristics of seafood allergy in different countries, and some of the fish and seafood allergens unique to those regions have been characterized. Review on published data showed that seafood allergy is very ubiquitous in some regions of the world. Fish and shellfish are the most common seafood that cause adverse allergic reactions among nations; the symptoms ranged from oral allergy syndromes to urticaria and anaphylaxis. The major identified allergens are parvalbumin in fish and tropomyosin in shellfish. Nevertheless, such studies are lacking from some regions with high fish and seafood consumption. Furthermore, the published data are mostly from small groups of populations, which large-scale epidemiological studies need to be performed.
As, Cd, Pb and Hg were analysed in commonly consumed spices and herbs in Malaysia. The range of As, Cd, Pb and Hg content was 0.24-2.54, 0.23-8.07, 1.54-8.94 and 0.06-0.52 µg g(-1), respectively. The highest concentration of Cd, Pb and Hg in spices and herbs exceeded the maximum permitted proportion, which are 1, 2 and 0.05 µg g(-1), respectively. This study suggests further monitoring of Cd, Pb and Hg on daily consumption of spices and herbs and its toxicological implication for consumers since only the amount of As was lower than the permitted concentration.
Roasting is an important process in cocoa production which may lead to formation of non-desirable compounds such as polycyclic aromatic hydrocarbons (PAHs). Therefore, PAH4 (sum of four different polycyclic aromatic hydrocarbons; benz[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene) in roasted cocoa beans was determined using a modified method (combination of QuEChERS and DLLME), and quantified by HPLC-FLD. The modified method was validated and met the performance criteria required by the EU Regulation (No. 836/2011). Results show a significant (p
Rice bran, a by-product of the rice milling process, has emerged as a functional food and being used in formulation of healthy food and drinks. However, rice bran is often contaminated with numerous mycotoxins. In this study, a method to simultaneous detection of aflatoxins (AFB1, AFB2, AFG1, and AFG2), ochratoxin A (OTA), deoxynivalenol (DON), fumonisins (FB1 and FB2), sterigmatocystin (STG), T-2 toxin, HT-2 toxin, diacetoxyscirpenol (DAS) and zearalenone (ZEA) in rice bran was developed, optimized and validated using dispersive liquid-liquid microextraction (DLLME) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In DLLME, using a solvent mixture of methanol/water (80:20, v/v) as the dispersive solvent and chloroform as the extraction solvent with the addition of 5% salt improved the extraction recoveries (63-120%). The developed method was further optimized using the response surface methodology (RSM) combined with Box-Behnken Design (BBD). Under the optimized experimental conditions, good linearity was obtained with a correlation coefficient (r2) ≥ 0.990 and a limit of detection (LOD) between 0.5 to 50 ng g-1. The recoveries ranged from 70.2% to 99.4% with an RSD below 1.28%. The proposed method was successfully applied to analyze multi-mycotoxin in 24 rice bran samples.
The aim of the present research was to evaluate the application, stability and suitability of ω3 polyunsaturated fatty acids (PUFAs) incorporated nanoliposomes in food enrichment. Nanoliposomal ω3 PUFAs was prepared by Mozafari method, and their application in bread and milk was compared with unencapsulated (fish oil) and microencapsulated ω3 PUFAs. Sensory evaluation was conducted to determine the perceptible sensory difference/similarity between control, unencapsulated, microencapsulated, and nanoliposomal ω3 PUFAs enriched foods. Results showed no significant (p=0.11) detectable difference between control and nanoliposomal ω3 PUFAs enriched samples while, samples enriched with unencapsulated or microencapsulated ω3 PUFAs showed significant (p=0.02) fishy flavor. Moreover, significantly (p<0.01) higher ω3 PUFAs % recovery and lower peroxide and anisidine values were observed in nanoliposomal ω3 PUFAs enriched samples in comparison with other samples. In conclusion, an effective and reproducible method for application of ω3 PUFAs in the food system was developed.
A novel magnetic graphene oxide modified with chitosan (MGO-CTS) was synthesised as an adsorbent aimed to examine the simultaneous removal of mycotoxins. The composite was characterised by various procedures, namely Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and a scanning electron microscope (SEM). The adsorption evaluation was considered via pH effects, initial mycotoxin concentration, adsorption time and temperature. Adsorption isotherm data and kinetics experiments were acquired at the optimum pH 5 fit Freundlich isotherm as well as pseudo-second-order kinetic models. The thermodynamic results indicated that the adsorption of the mycotoxins was spontaneous, endothermic and favourable.
Recently, cultivation of high-yielding hybrid maize varieties has revolutionized maize production in Pakistan. Analyses of nutritional traits and aflatoxin (AF) contamination in these varieties can aid in the identification of susceptible and resistant varieties, particularly for cultivation in the Pakistani agro-climatic environment. Five spring maize varieties-Pioneer, Neelam, DK-919, Desi, and Hi-maize-were selected for analyses of their nutritional, tocopherol, and AF contents. Protein, carbohydrate, oil, ash, fiber, and moisture contents ranged between 8.7 and 10.8%, 68 and 71%, 3.72 and 5.56%, 1.09 and 1.81%, 1.1 and 3.1%, and 11.7 and 14.2%, respectively. Tocopherol levels in selected varieties were in the range of 461 to 1,430 μg/g. Hi-maize exhibited significantly higher protein and tocopherol contents than the other varieties, indicating its better suitability for feed and silage applications. The highest mean level of total AFs, 14.5 ± 0.12 μg/kg, was found in Desi, and results showed that the most dominant AF found in the maize varieties was AFB1. Furthermore, the results showed that the higher the level of tocopherol, the lower the concentration of total AFs and vice versa in maize varieties. The results can be used to investigate additional susceptible maize varieties that are resistant to fungal attack.
Nanoparticles (NPs) are, frequently, being utilized in multi-dimensional enterprises. Silver nanoparticles (AgNPs) have attracted researchers in the last decade due to their exceptional efficacy at very low volume and stability at higher temperatures. Due to certain limitations of the chemical method of synthesis, AgNPs can be obtained by physical methods including sun rays, microwaves and ultraviolet (UV) radiation. In the current study, the synthesis of pullulan mediated silver nanoparticles (P-AgNPs) was achieved through ultraviolet (UV) irradiation, with a wavelength of 365 nm, for 96 h. P-AgNPs were formed after 24 h of UV-irradiation time and expressed spectra maxima as 415 nm, after 96 h, in UV-vis spectroscopy. The crystallographic structure was "face centered cubic (fcc)" as confirmed by powder X-ray diffraction (PXRD). Furthermore, high resolution transmission electron microscopy (HRTEM) proved that P-AgNPs were covered with a thin layer of pullulan, with a mean crystalline size of 6.02 ± 2.37. The average lattice fringe spacing of nanoparticles was confirmed as 0.235 nm with quasi-spherical characteristics, by selected area electron diffraction (SAED) analysis. These green synthesized P-AgNPs can be utilized efficiently, as an active food and meat preservative, when incorporated into the edible films.
High-quality fish oil for human consumption requires low levels of toxic elements. The aim of this study was to compare different oil extraction methods to identify the most efficient method for extracting fish oil of high quality with the least contamination. The methods used in this study were Soxhlet extraction, enzymatic extraction, wet reduction, and supercritical fluid extraction. The results showed that toxic elements in fish oil could be reduced using supercritical CO2 at a modest temperature (60°C) and pressure (35 MPa) with little reduction in the oil yield. There were significant reductions in mercury (85 to 100%), cadmium (97 to 100%), and lead (100%) content of the fish oil extracted using the supercritical fluid extraction method. The fish oil extracted using conventional methods contained toxic elements at levels much higher than the accepted limits of 0.1 μg/g.
This study investigated the simultaneous formation of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatic amines (HCAs) in gas-grilled beef satay at different temperatures (150, 200, 250, 300, and 350°C). Solid-phase extraction (SPE) was used for sample clean-up. Fifteen PAHs were determined using high performance liquid chromatography with fluorescence detection (HPLC-FLD) and nine HCAs were quantified using liquid chromatography tandem-mass spectrometry (LC-MS/MS) with a gradient programme. The lowest significantly concentrations of PAHs and HCAs were generated at 150°C; the formation of PAHs and HCAs simultaneously increased with temperatures. Benzo[a]pyrene was detected in all samples and increased markedly at 300 and 350°C. The sums of 4 PAHs (PAH4) in marinated beef satay at 300 and 350°C exceeded the maximum level in Commission Regulation (EU) 2015/1125. Significant reductions of polar and non-polar HCAs (except PhIP) were found in marinated beef satay across all temperatures. Overall, PAHs and HCAs showed opposite trends of formation in beef satay with marination.
The present work aimed to determine the prevalence of aflatoxigenic Aspergillus section Flavi on different types of dairy goat's feed samples obtained from four dairy goat's farms around the central region of Peninsular Malaysia, and to examine the effects of climatic conditions (temperature, relative humidity) of the dairy goat's farms, and their feeding and storage practices on the fungal prevalence of different types of dairy goat's feed. A total of 60 goat's feed samples were obtained, and their proximate composition and water activity were determined, following which they were cultivated on DRBC and AFPA for total fungal load and Aspergillus section Flavi load determination, respectively. Fungal isolates were identified morphologically, and toxigenicity potentials of Aspergillus section Flavi isolates were determined using CCA. The temperature and relative humidity data of all farms were obtained from the Malaysian Meteorological Department. The total fungal loads (on DRBC) of the goat's feed samples were log 0.767 to 7.071 CFU/g which included the common feed contaminants such as Aspergillus, Fusarium, and Penicillium. The Aspergillus section Flavi loads (on AFPA) were log 0.667 to 3.206 CFU/g. Farm A yielded the highest number of Aspergillus section Flavi isolates as well as the highest number of aflatoxigenic isolates. It was found that climatic conditions and different practices between farms positively influenced the fungal prevalence on goat's feed samples based on the Pearson correlation analysis. The prevalence of mycotoxigenic isolates on goat's feed warrants for urgent intervention to ensure that goats are being fed with nutritionally adequate and safe feed. The presence of aflatoxigenic Aspergillus section Flavi isolates indicates the risk of aflatoxin B1 contamination on the goat's feed, aflatoxicosis development in the goats, and aflatoxin M1 bio-transformation in the goat's milk. This is a potential threat to the flourishing goat's milk industry in Malaysia.
In studying the ecophysiology of fungal phytopathogens, several stages are involved (in vitro, greenhouse, in planta). Most in vitro studies extensively utilise the general growth media such as Potato Dextrose Agar and Malt Extract Agar. Although the crop components in these media serve as excellent carbon sources and yield luxuriant growth, they are not naturally contaminated with Aspergillus flavus and thus might result in under- or overestimation of its actual toxigenic potentials. Empirical data on the formulation of semi-synthetic growth medium mimicking the natural crop commonly contaminated by A. flavus for the ecophysiological studies in vitro are scarce. The present work was aimed at investigating the ecophysiology of A. flavus on commercial growth media (PDA, MEA); formulating maize- and peanut-based semi-synthetic growth media using two methods of raw material preparation (milling, hot water extraction) at different concentrations (1, 3, 5, 7, 9% w/v), and comparing the ecophysiological parameters between commercial and formulated growth media. Growth rates were obtained by computing the hyphal expansion data into y = mx + c equation. AFB1 was quantified using high performance liquid chromatography with fluorescence detector. Formulated media were found to yield significantly higher growth rates when compared to commercial media. However, commercial media yielded significantly higher AFB1 when compared to all formulated media. Between the two formulations, milling yielded significantly higher growth rates and AFB1 when compared to hot water extraction. Although in vitro data cannot directly extrapolate in planta performance, results obtained in the present work can be used to gauge the actual toxigenic potential of A. flavus in maize and peanut agro-ecosystems.
The present work investigated the potential of fungal species from grain maize farms in Malaysia as antagonists against the indigenous mycotoxigenic fungal species and their subsequent mycotoxin production. Dual-culture assay was conducted on grain maize agar (GMA) with 12 strains of potential fungal antagonists namely Bjerkandra adusta, Penicillium janthinellum, Schizophyllum commune, Trametes cubensis, Trichoderma asperelloides, Trichoderma asperellum, Trichoderma harzianum, and Trichoderma yunnanense against seven mycotoxigenic strains namely Aspergillus flavus, Aspergillus niger, Fusarium verticillioides, and Fusarium proliferatum producing aflatoxins, ochratoxin A, and fumonisins, respectively. Based on fungal growth inhibition, Trichoderma spp. showed the highest inhibitory activity (73-100% PIRG, Percentage Inhibition of Radial Growth; 28/0 ID, Index of Dominance) against the tested mycotoxigenic strains. Besides, B. adusta and Tra. cubensis showed inhibitory activity against some of the tested mycotoxigenic strains. All fungal antagonists showed varying degrees of mycotoxin reduction. Aflatoxin B1 produced by A. flavus was mainly reduced by P. janthinellum, Tra. cubensis, and B. adusta to 0 ng/g. Ochratoxin A produced by A. niger was mainly reduced by Tri. harzianum and Tri. asperellum to 0 ng/g. Fumonisin B1 and FB2 produced by F. verticillioides was mainly reduced by Tri. harzianum, Tri. asperelloides, and Tri. asperellum to 59.4 and 0 µg/g, respectively. Fumonisin B1 and FB2 produced by F. proliferatum were mainly reduced by Tri. asperelloides and Tri. harzianum to 244.2 and 0 µg/g, respectively. This is the first study that reports on the efficacy of Tri. asperelloides against FB1, FB2, and OTA, P. janthinellum against AFB1, and Tra. cubensis against AFB1.