Development of a zebrafish model for toxicity evaluation of adulterated Apis mellifera honey.

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.

Development of Methyl Ester Antibody-Based Competitive Indirect ELISA for Quantitative Detection of Mitragynine in Human Urine.

Mitragynine is the main psychoactive compound of Mitragyna speciosa Korth. (kratom). This alkaloid could render psychotropic effects and is often misused as a substitute for commercial drugs. Nowadays, the increasing popularity of kratom has led to the development of a rapid and effective detection method. The detection of mitragynine in a biological sample such as urine requires a highly sensitive and specific method due to the complex nature of mitragynine in urine. Enzyme-linked immunosorbent assay (ELISA) is well known as a rapid screening method for biological samples. In this study, a competitive indirect ELISA was successfully developed using MG-22-OCH3 IgG as a detection antibody for mitragynine in human urine. The mitragynine immunoassay showed a limit of detection and a limit of quantification of 0.412 and 1.25 µg/mL, respectively. The measurement range was between 0.01 and 100.0 µg/mL, with a minimal inhibition (IC50) value of 0.152 µg/mL. The developed ELISA was validated using a gold method such as high-performance liquid chromatography-mass spectrometry (HPLC-MS). The percentage of recovery and the coefficient of variation (CV) for the ELISA and LCMS/MS analyses were 84.0-95.70%, 99.20-112.0%, 7.69-9.78%, and 2.86-6.62%, respectively. This indicates that the developed ELISA is a reliable method that can be used as a rapid approach for quantifying mitragynine content in biological samples.

1H nuclear magnetic resonance-based metabolomics study of serum and pectoralis major for different commercial chicken breeds.

This study aimed to characterize the metabolic composition of four types of commercially available chicken breeds [village chicken, colored broiler (Hubbard), broiler (Cobb), and spent layers (Dekalb)] by 1H NMR coupling and discriminate them using multivariate analysis. Five chickens were collected for each chicken breed based on the marketing age from the respective commercial farms. The orthogonal partial least squares discriminant analysis (OPLS-DA) results showed an obvious separation of local village chickens from the other breeds based on the metabolites present in their serum and meat (pectoralis major). The cumulative values of Q 2, R 2 X, and R 2 Y of the OPLS-DA model for chicken serum were 0.722, 0.877, and 0.841. For the pectoralis major muscle, the cumulative values of Q 2, R 2 X, and R 2 Y of the OPLS-DA model were reported as 0.684, 0.781, and 0.786, respectively. The quality of both OPLS-DA models was accepted by the cumulative values of Q 2 ≥ 0.5 and R 2 ≥ 0.65. The 1H NMR result with multivariate analysis has successfully distinguished local village chicken from the other three commercial chicken breeds based on serum and pectoralis major muscle. Nonetheless, colored broiler (Hubbard) was not distinguished from broiler (Cobb) and spent layers (Dekalb) in serum and pectoralis major, respectively. The OPLS-DA assessment in this study identified 19 and 15 potential metabolites for discriminating different chicken breeds in serum and pectoralis major muscle, respectively. Some of the prominent metabolites identified include amino acids (betaine, glycine, glutamine, guanidoacetate, phenylalanine, and valine), nucleotides (IMP and NAD+), organic acids (lactate, malate, and succinate), peptide (anserine), and sugar alcohol (myo-inositol).

Cross-reactivity analysis of milk proteins from different goat breeds with cow's milk allergens using a proteomic approach.

INTRODUCTION: Goat's milk thought to be a good substitute for cow's milk protein allergic (CMPA) individuals. However, there is growing evidence that their proteins have cross-reactivities with cow's milk allergens. This study aimed to profile and compare milk proteins from different goat breeds that have cross-reactivity to cow's milk allergens. METHODOLOGY: Proteomics was used to compare protein extracts of skim milk from Saanen, Jamnapari, and Toggenburg. Cow's milk was used as a control. IgE-immunoblotting and mass spectrometry were used to compare and identify proteins that cross-reacted with serum IgE from CMPA patients (n = 10). RESULTS: The analysis of IgE-reactive proteins revealed that the protein spots identified with high confidence were proteins homologous to common cow's milk allergens such as α-S1-casein (αS1-CN), ß-casein (ß-CN), κ-casein (κ-CN), and beta-lactoglobulin (ß-LG). Jamnapari's milk proteins were found to cross-react with four major milk allergens: α-S1-CN, ß-CN, κ-CN, and ß-LG. Saanen goat's milk proteins, on the other hand, cross-reacted with two major milk allergens, α-S1-CN and ß-LG, whereas Toggenburg goat's milk proteins only react with one of the major milk allergens, κ-CN. CONCLUSION: These findings may help in the development of hypoallergenic goat milk through cross-breeding strategies of goat breeds with lower allergenic milk protein contents.

Methyl ester and aromatic ether modification of mitragynine for generation of mitragynine-specific polyclonal antibodies.

Mitragynine is an alkaloid from Mitragyna speciosa Korth. (kratom), a native tropical plant in Southeast Asia. It could render psychotropic effects and is often misused in substitution for commercial drugs. In recent years, the consumption of kratom has grown rapidly and has led some countries to ban its use. The misuse of kratom can be detected and monitored through the determination of mitragynine from biological samples of the users. Therefore, the development of a rapid and effective detection method is needed. In this study, polyclonal antibodies were produced using mitragynine coupled to a carrier protein (cationic bovine serum albumin, cBSA) as an immunogen, which was prepared with coupling agents (i.e., N, N- dicyclohexylcarbodiimide, DCC and N-hydroxysuccinimide, NHS). It was conjugated to different mitragynine structure, 16-COOCH3 (methyl ester) and 9-OCH3 (aromatic ether). 2,4,6-Trinitrobenzenesulfonic acid (TNBS) method showed that 45 and 46 amino groups were bound to C22-MG-cBSA and C9-MG-cBSA, respectively. Fourier-transform infrared spectroscopy (FTIR) spectral changes at C22- and C9-hydroxymitragynine indicated reduction and demethylation process. In UV-Vis spectra, conjugated mitragynine to cBSA and OVA were displayed at a spectral region at 240-300 nm. For the antibody titre, the C22-MG-cBSA anti-serum showed a significantly higher titre than the C9-MG-cBSA at 1/128000 and 1/32000 dilutions, respectively. The detection range of the developed competitive indirect ELISA (CI-ELISA) was 0.01 to 10.00 µg/mL (R2 = 0.9964). The assay exhibited a limit of detection (LOD) and limit of quantification (LOQ) at 0.041 and 0.124 µg/mL, respectively. The antibody produced is a high-value biorecognition molecule that can be further used in developing immuno-based detection methods such as immunosensors and immunochromatographic lateral flow assays. This will benefit the task force or forensic agencies for toxicological screening with high speed and efficiency.

Simultaneous kinetics formation of heterocyclic amines and polycyclic aromatic hydrocarbons in phenylalanine model system.

A combination of chemical model system with kinetics study was used to investigate the simultaneous formation of heterocyclic amines (HCAs) and polycyclic aromatic hydrocarbons (PAHs). Heating a mixture of phenylalanine, creatinine, and glucose at a commonly practiced household cooking time and temperature successfully differentiated the rate formation (k) of HCAs and PAHs. The good fit suggested that the simultaneous formation was an endothermic bimolecular reaction, and followed the first-order model. The rate formation (k) of HCAs and PAHs significantly increased with increasing heating time and temperature. Only 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) showed degradation rate (k) at higher heating temperatures of 210 °C and 240 °C respectively. Increasing phenylalanine concentration increased the possibility of higher HCAs and PAHs formation. The activation energy (Ea) showed that heating phenylalanine mixture resulted in higher rate of formation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and benzo[b]fluoranthen (BbF).

In Vivo Toxicity Evaluation of Sugar Adulterated Heterotrigona itama Honey Using Zebrafish Model.

Honey is prone to be adulterated through mixing with sugars, cheap and low-quality honey, and other adulterants. Consumption of adulterated honey may cause several health issues such as weight gain, diabetes, and liver and kidney dysfunction. Therefore, studying the impact of consumption of adulterated honey on consumers is critical since there is a lack of study in this field. Hence, the aims of this paper were: (1) to determine the lethal concentration (LC50) of adulterated honey using zebrafish embryo, (2) to elucidate toxicology of selected adulterated honey based on lethal dose (LD50) using adult zebrafish, (3) to determine the effects of adulterated honey on histological changes of zebrafish, and (4) to screen the metabolites profile of adulterated honey by using zebrafish blood serum. The LC50 of Heterotrigona itama honey (acacia honey) and its sugar adulterants (light corn sugar, cane sugar, inverted sugar, and palm sugar in the proportion of 1-3% (w/w) from the total volume) was determined by the toxicological assessment of honey samples on zebrafish embryos (different exposure concentrations in 24, 48, 72, and 96 h postfertilization (hpf)). Pure H. itama honey represents the LC50 of 34.40 ± 1.84 (mg/mL) at 96 hpf, while the inverted sugar represents the lowest LC50 (5.03 ± 0.92 mg/mL) among sugar adulterants. The highest concentration (3%) of sugar adulterants were used to study the toxicology of adulterated honey using adult zebrafish in terms of acute, prolong-acute, and sub-acute tests. The results of the LD50 from the sub-acute toxicity test of pure H. itama honey was 2.33 ± 0.24 (mg/mL). The histological studies of internal organs showed a lesion in the liver, kidney, and spleen of adulterated treated-honey groups compared to the control group. Furthermore, the LC-MS/MS results revealed three endogenous metabolites in both the pure and adulterated honey treated groups, as follows: (1) S-Cysteinosuccinic acid, (2) 2,3-Diphosphoglyceric acid, and (3) Cysteinyl-Tyrosine. The results of this study demonstrated that adulterated honey caused mortality, which contributes to higher toxicity, and also suggested that the zebrafish toxicity test could be a standard method for assessing the potential toxicity of other hazardous food additives. The information gained from this research will permit an evaluation of the potential risk associated with the consumption of adulterated compared to pure honey.

Muscle and Serum Metabolomics for Different Chicken Breeds under Commercial Conditions by GC-MS.

Globally, village chicken is popular and is known as a premium meat with a higher price. Food fraud can occur by selling other chicken breeds at a premium price in local markets. This study aimed to distinguish local village chicken from other chicken breeds available in the market, namely, colored broiler (Hubbard), broiler (Cobb), and spent laying hen (Dekalb) in pectoralis major and serum under commercial conditions using an untargeted metabolomics approach. Both pectoralis major and serum were analyzed using gas chromatography-mass spectrometry (GC-MS). The principal component analysis (PCA) results distinguished four different chicken breeds into three main groups for pectoralis major and serum. A total of 30 and 40 characteristic metabolites were identified for pectoralis major and serum, respectively. The four chicken breeds were characterized by the abundance of metabolites such as amino acids (L-glutamic acid, L-threonine, L-serine, L-leucine), organic acids (L-lactic acid, succinic acid, 3-hydroxybutyric acid), sugars (D-allose, D-glucose), sugar alcohols (myo-inositol), and fatty acids (linoleic acid). Our results suggest that an untargeted metabolomics approach using GC-MS and PCA could discriminate chicken breeds for pectoralis major and serum under commercial conditions. In this study, village chicken could only be distinguished from colored broiler (Hubbard) by serum samples.

Determination of minimal sequence for zearalenone aptamer by computational docking and application on an indirect competitive electrochemical aptasensor.

Aptamers are short single-stranded oligonucleotides (either DNA or RNA) that can fold into well-defined three-dimensional (3D) spatial structures which enable them to capture their specific target by complementary shape interactions. Aptamers are selected from large random libraries through the SELEX process and only a small fraction of the sequence is involved in direct docking with the target. In this paper, we describe the possible truncation variants of zearalenone (ZEA) aptamer which might be an effective binding region for the target. The originally selected zearalenone (ZEA) aptamer was 80-mer in length and shown to bind the target with a high affinity (Kd = 41 ± 5 nM). Herein, computational docking simulation was performed with 15 truncated variants to determine the predicted binding energy and responsible binding site of the aptamer-analyte complex. The results revealed that 5 truncated variants had binding energy lower than - 7.0 kcal/mol. Circular dichroism analysis was performed on the shortlisted aptamer and the conformational change of aptamers was observed with the presence of an analyte. Aptamer Z3IN (29-mer) was chosen as the most enhanced affinity for its target with a dissociation constant of 11.77 ± 1.44 nM. The aptamer was further applied in the electrochemical aptasensor of ZEA based on an indirect competitive format. The results demonstrated that the truncated aptamer leads to an enhancement of the sensitivity of the biosensor.

Dispersive Liquid-Liquid Microextraction (DLLME) and LC-MS/MS Analysis for Multi-Mycotoxin in Rice Bran: Method Development, Optimization and Validation.

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 Toxic Impact of Honey Adulteration: A Review.

Honey is characterized as a natural and raw foodstuff that can be consumed not only as a sweetener but also as medicine due to its therapeutic impact on human health. It is prone to adulterants caused by humans that manipulate the quality of honey. Although honey consumption has remarkably increased in the last few years all around the world, the safety of honey is not assessed and monitored regularly. Since the number of consumers of honey adulteration have increased in recent years, their trust and interest in this valuable product has decreased. Honey adulterants are any substances that are added to the pure honey. In this regard, this paper provides a comprehensive and critical review of the different types of adulteration, common sugar adulterants and detection methods, and draws a clear perspective toward the impact of honey adulteration on human health. Adulteration increases the consumer's blood sugar, which can cause diabetes, abdominal weight gain, and obesity, raise the level of blood lipids and can cause high blood pressure. The most common organ affected by honey adulterants is the liver followed by the kidney, heart, and brain, as shown in several in vivo research designs.

A highly selective two-way purification method using liquid chromatography for isolating αS2-casein from goat milk of five different breeds.

The main challenges in the purification of αS2-casein are due to the low quantity in milk and high homology with other casein subunits, i.e., αS1-casein, ß-casein, and κ-casein. To overcome these challenges, the aim of this study was to develop a two-step purification to isolate native αS2-casein in goat milk from five different breeds; British Alpine, Jamnapari, Saanen, Shami, and Toggenburg. The first step of the purification was executed by anion-exchange chromatography under optimal elution conditions followed by size exclusion chromatography. Tryptic peptides from in-gel digestion of purified αS2-casein were sequenced and analyzed by LC-ESI-MS/MS. From 1.05 g of whole casein, the highest yield of αS2-casein (6.7 mg/mL) was obtained from Jamnapari and the lowest yield (2.2 mg/mL) was from Saanen. A single band of pure αS2-casein was observed on SDS-PAGE for all breeds. The αS2-casein showed coverage percentage of amino acid sequence from 76.68 to 92.83%. The two-step purification process developed herein was successfully applied for isolating native αS2-casein from goat milk with high purity, which will allow for future in vitro studies to be conducted on this protein.

The Inhibitory Effects of Heterotrigona Itama Honey Marinades on the Formation of Carcinogenic Heterocyclic Amines in Grilled Beef Satay.

Heterocyclic amines (HCAs) are carcinogenic food toxicants formed in cooked meats, which may increase the risk of cancer development in humans. Therefore, in this study, the effect of stingless bee honey from different botanical origins on the formation of HCAs in grilled beef satay was investigated. HCAs concentration in grilled beef satay was determined by using high performance liquid chromatography (HPLC). In total, six of the most toxigenic HCAs representing aminoimidazo-azaarenes (AIAs) (MeIQx, 4,8-DiMeIQx, and PhIP) and amino carbolines (norharman, harman, and AαC) groups were identified in all the beef samples investigated. A significant reduction in HCAs was observed in grilled beef marinated in honey as compared to beef samples marinated in table sugar (control), in which the reduction of 95.14%, 88.45%, 85.65%, and 57.22% was observed in gelam, starfruit, acacia, and Apis honey marinades, respectively. According to the partial least squares regression (PLS) model, the inhibition of HCAs in grilled beef was shown to be significantly correlated to the antioxidant activity (IC50) of the honey samples. Therefore, the results of this study revealed that the addition of stingless bee honey could play an important role in reducing HCAs in grilled beef.

Assessment of Fumonisin B1 Concentrations in Wheat and Barley Products in the Punjab Region of Pakistan.

ABSTRACT: A total of 133 samples of whole wheat and barley grains and wheat and barley flour collected from retail markets in the main cities of Punjab, Pakistan, were analyzed for the mycotoxin fumonisin B1 (FB1) using reverse phase high-performance liquid chromatography with fluorescence detection. Of these samples, 120 (90%) were positive for FB1, and 75 (63%) of the 120 positive samples had FB1 concentrations higher than the European Union maximum (200 µg/kg). The limit of detection was 4 µg/kg. The highest mean (±SD) concentration of FB1 was found in whole wheat samples, 980.5 ± 211.4 µg/kg. The calculated dietary intakes of FB1 from wheat and barley flours were 4,456 and 503.7 ng/g of body weight per day, respectively.

Generation of High Affinity Anti-Peptide Polyclonal Antibodies Recognizing Goat αs1-Casein.

The chemical, technological and allergy properties of goat's milk are significantly affected by the level of αs1-casein. Detection and quantification of αs1-casein requires high-specificity methods to overcome high-sequence similarity between this protein and others in the casein family. Unavailability of antibodies with high affinity and specificity towards goat αs1-casein hinders the development of immuno-based analytical methods such as enzyme-linked immunosorbent assay (ELISA) and biosensors. Here, we report the generation of polyclonal antibodies (or immunoglobulins, IgGs) raised towards goat αs1-casein N- (Nter) and C-terminal (Cter) peptide sequences. The Nter and Cter peptides of goat αs1-casein were immunized in rabbits for the generation of antisera, which were purified using protein G affinity chromatography. The binding affinity of the antisera and purified IgGs were tested and compared using indirect ELISA, where peptide-BSA conjugates and goat αs1-casein were used as the coating antigens. The Nter antiserum displayed higher titer than Cter antiserum, at 1/64,000 and 1/32,000 dilutions, respectively. The purification step further yielded 0.5 mg/mL of purified IgGs from 3 mL of antisera. The purified Nter IgG showed a significantly (p < 0.05) higher binding affinity towards peptide-BSA and goat αs1-casein, with lower Kd value at 5.063 × 10-3 µM compared to 9.046 × 10-3 µM for the Cter IgG. A cross-reactivity test showed that there was no binding in neither Nter nor Cter IgGs towards protein extracts from the milk of cow, buffalo, horse and camel. High-quality antibodies generated will allow further development of immuno-based analytical methods and future in vitro studies to be conducted on goat αs1-casein.

Effect of Synthesis Temperature on the Size of ZnO Nanoparticles Derived from Pineapple Peel Extract and Antibacterial Activity of ZnO-Starch Nanocomposite Films.

This research investigated the effect of synthesis temperature on the size and shape of zinc oxide (ZnO) nanoparticles (NPs) synthesized using pineapple peel waste and antibacterial activity of ZnO NPs in starch films. Zinc oxide NPs synthesized at different temperatures were characterized by Fourier transform infrared spectroscopy, X-ray diffraction analysis, field-emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, and transmission electron microscopy. Micrographs of ZnO NPs synthesized at 28 and 60 °C showed that synthesis temperature affected the sizes and shapes of ZnO NPs. The non-heated (28 °C) condition resulted in NPs with diameters in the range of 8-45 nm with a mixture of spherical and rod shapes, whereas the heated (60 °C) condition led to NPs with diameters in the range of 73-123 nm with flower rod shapes. The ZnO-starch nanocomposite films incorporated with 1, 3, and 5 wt.% ZnO NPs were prepared via a film casting method. The antibacterial activity of the films against Gram-positive and Gram-negative bacteria was investigated using the disc diffusion method. The results showed an increase in the inhibition zone for Gram-positive bacteria, particularly Bacillus subtilis, when the concentration of ZnO NPs incorporated in the film was increased from 1 to 5 wt.%.

Electrochemical determination of zearalenone using a label-free competitive aptasensor.

An electrochemical aptasensor is described for determination of the phytohormone of zearalenone (ZEA). The gold electrode was modified with ZEA via covalent attachment using cysteamine-hydrochloride and 1,4-phenylene diisocyanate linker. A truncated ZEA aptamer with a dissociation constant of 13.4 ± 2.1 nM was used in an aptasensor. The electrochemical property was investigated using square wave voltammetry for monitoring the change in the electron transfer using the ferro/ferricyanide system as redox probe. Under optimal experimental conditions, the response was best measured at a potential of 0.20 V (vs. Ag/AgCl). The signals depended on the competitive mechanism between the immobilised ZEA and free ZEA for the aptamer binding site. The aptasensor works in the range 0.01 to 1000 ng·mL-1 ZEA concentration, with a detection limit of 0.017 ng·mL-1. High degree of cross-reactivity with the other analogues of ZEA was observed, whereas none towards other mycotoxins. The aptasensor was further applied for the determination of ZEA in the extract of maize grain and showed good recovery percentages between 87 and 110%. Graphical abstract Schematic representation of the electrochemical determination of zearalenone based on indirect competitive assay. Step a Immobilisation of ZEA on the surface of gold electrode via covalent attachment, b competition for the ZEA aptamer binding site between immobilised and free ZEA, and c current signal of the binding event based on SWV technique.

Effect of Different Amino Acids and Heating Conditions on the Formation of 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and Its Kinetics Formation Using Chemical Model System.

The formation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) was investigated using a kinetic study approach as described by first-order, Arrhenius, and Eyring equations. Chemical model systems with different amino acid precursors (proline, phenylalanine, and glycine) were examined at different times (4, 8, 12, and 16 min) and temperatures (150, 180, 210, 240, and 270 °C). PhIP was detected using high-performance liquid chromatography equipped with fluorescence detector (HPLC-FLD). The good fit in first-order suggested that PhIP formation was influenced by the types of amino acids and PhIP concentration significantly increased with time and temperature (up to 240 °C). PhIP was detected in proline and phenylalanine model systems but not in the glycine model system. The phenylalanine model system demonstrated low activation energy (Ea) of 95.36 kJ/mol that resulted in a high rate of PhIP formation (great amount of PhIP formed). Based on the ∆S‡ values both proline and phenylalanine demonstrated bimolecular rate-limiting steps for PhIP formation. Altogether these kinetic results could provide valuable information in predicting the PhIP formation pathway.

Etlingera elatior-Mediated Synthesis of Gold Nanoparticles and Their Application as Electrochemical Current Enhancer.

This work presents a simple green synthesis of gold nanoparticles (AuNPs) by using an aqueous extract of Etlingera elatior (torch ginger). The metabolites present in E. elatior, including sugars, proteins, polyphenols, and flavonoids, were known to play important roles in reducing metal ions and supporting the subsequent stability of nanoparticles. The present work aimed to investigate the ability of the E. elatior extract to synthesise AuNPs via the reduction of gold (III) chloride hydrate and characterise the properties of the nanoparticles produced. The antioxidant properties of the E. elatior extract were evaluated by analysing the total phenolic and total flavonoid contents. To ascertain the formation of AuNPs, the synthesised particles were characterised using the ultraviolet-visible (UV-Vis) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, high-resolution transmission electron microscopy (HRTEM), energy-dispersive X-ray (EDX) microscopy, and dynamic light scattering (DLS) measurement. The properties of the green synthesised AuNPs were shown to be comparable to the AuNPs produced using a conventional reducing agent, sodium citrate. The UV-Vis measured the surface plasmon resonance of the AuNPs, and a band centered at 529 nm was obtained. The FTIR results proved that the extract contained the O-H functional group that is responsible for capping the nanoparticles. The HRTEM images showed that the green synthesized AuNPs were of various shapes and the average of the nanoparticles' hydrodynamic diameter was 31.5 ± 0.5 nm. Meanwhile, the zeta potential of -32.0 ± 0.4 mV indicates the high stability and negative charge of the AuNPs. We further successfully demonstrated that using the green synthesised AuNPs as the nanocomposite to modify the working surface of screen-printed carbon electrode (SPCE/Cs/AuNPs) enhanced the rate of electron transfer and provided a sensitive platform for the detection of Cu(II) ions.

Effects of simultaneous UV-C radiation and ultrasonic energy postharvest treatment on bioactive compounds and antioxidant activity of tomatoes during storage.

The effects of a novel technology utilizing a simultaneous combination of Ultraviolet-C radiation and ultrasound energy postharvest treatment on tomato bioactive compounds during 28 days' storage period was investigated by varying Ultraviolet-C radiation intensities of 639.37 or 897.16 µW/cm2 at a constant ultrasound intensity of 13.87 W/L from a 40 kHz-1 kW transducer. A minimal treatment time of 240 s at Ultraviolet-C dosage of 2.15 kJ/m2 was observed to provoke a considerable increase in bioactive compounds content, proportionated to treatment time. Although treatment led to temperature increase in the system reaching 39.33 °C due to heat generation by ultrasonic cavitation, the extractability and biosynthesis of phytochemicals were enhanced resulting in 90%, 30%, 60%, 20%, and 36% increases in lycopene, total phenols, vitamin C, hydrophilic and lipophilic antioxidant activities respectively. Results present the potential use of the combined non-thermal technologies as post-harvest treatment to improve bioactive compounds and antioxidant activity during storage.