The impact of sucralose and neotame on the safety of metal precipitation in electronic cigarettes.

This study investigated the impact of sweeteners on the release of heavy metals during the heating and atomization processes in electronic cigarettes. Based on a PG/VG base e-liquid with the addition of 2% and 5% neotame or sucralose, we quantitatively analyzed the impact of sweetener content on the levels of heavy metals such as Ni, Cr, and Fe in the e-liquid and aerosol after heating and atomization. Additionally, the heated e-liquid samples were used to culture SH-SY-5Y and Beas-2B cells, and their cytotoxic effects were assessed using the CCK-8 assay. The results indicated that the e-liquid with 5% sucralose had the highest average levels of heavy metals after heating and atomization, particularly nickel (13.36 ± 2.50 mg/kg in the e-liquid and 12,109 ± 3,229 ng/200 puffs in the aerosol), whereas the e-liquid with neotame had significantly lower average heavy metal content in comparison. Additionally, it was measured that the chloride ion concentration in the e-liquid with 5% sucralose reached 191 mg/kg after heating at 200°C for 1 h, indicating that heating sucralose generated chloride ions, Which might corrode metal parts components leading to heavy metal release. Cytotoxicity tests revealed that the base e-liquid without sweeteners exhibited the highest average cell viability after heating, at 64.80% ± 2.84% in SH-SY-5Y cells and 63.24% ± 0.86% in Beas-2B cells. Conversely, the e-liquid variant with 5% sucralose showed a significant reduction in average cell viability, reducing it to 50.74% ± 0.88% in SH-SY-5Y cells and 53.03% ± 0.76% in Beas-2B cells, highlighting its more pronounced cytotoxic effects compared to other tested e-liquids. In conclusion, sucralose in e-liquids should be limited preferably less than 2%, or replaced with neotame, a safer alternative, to minimize health risks.

A Pharmacological perspective on the temporal properties of sweeteners.

Several non-caloric sweeteners exhibit a delay in sweetness onset and a sweetness linger after sampling. These temporal properties are thought to be the result of non-specific interactions with cell membranes and proteins in the oral cavity. Data and analysis presented in this report also support the potential involvement of receptor affinity and binding kinetics to this phenomenon. In general, affected sweeteners exhibit distinctly higher binding affinity compared to carbohydrate sweeteners, which do not have temporal issues. In addition, binding kinetic simulations illustrate much slower receptor binding association and dissociation kinetics for a set of non-caloric sweeteners presenting temporal issues, in comparison to carbohydrate sweeteners. So, the higher affinity of some non-caloric sweeteners, dictating lower use levels, and affecting binding kinetics, could contribute to their delay and linger in sweetness perception. Simple pharmacology principles could explain, at least in part, some of the temporal issues of sweeteners.

The artificial sweetener neotame negatively regulates the intestinal epithelium directly through T1R3-signaling and indirectly through pathogenic changes to model gut bacteria.

Introduction: Recent studies have indicated considerable health risks associated with the consumption of artificial sweeteners. Neotame is a relatively new sweetener in the global market however there is still limited data on the impact of neotame on the intestinal epithelium or the commensal microbiota. Methods: In the present study, we use a model of the intestinal epithelium (Caco-2) and microbiota (Escherichia coli and Enterococcus faecalis) to investigate how physiologically-relevant exposure of neotame impacts intestinal epithelial cell function, gut bacterial metabolism and pathogenicity, and gut epithelium-microbiota interactions. Results: Our findings show that neotame causes intestinal epithelial cell apoptosis and death with siRNA knockdown of T1R3 expression significantly attenuating the neotame-induced loss to cell viability. Similarly, neotame exposure results in barrier disruption with enhanced monolayer leak and reduced claudin-3 cell surface expression through a T1R3-dependent pathway. Using the gut bacteria models, E. coli and E. faecalis, neotame significantly increased biofilm formation and metabolites of E. coli, but not E. faecalis, reduced Caco-2 cell viability. In co-culture studies, neotame exposure increased adhesion capacity of E. coli and E. faecalis onto Caco-2 cells and invasion capacity of E. coli. Neotame-induced biofilm formation, E.coli-specific Caco-2 cell death, adhesion and invasion was identified to be meditated through a taste-dependent pathway. Discussion: Our study identifies novel pathogenic effects of neotame on the intestinal epithelium or bacteria alone, and in co-cultures to mimic the gut microbiome. These findings demonstrate the need to better understand food additives common in the global market and the molecular mechanisms underlying potential negative health impacts.

Determination of neotame in various foods by high-performance liquid chromatography coupled with ultraviolet and mass spectrometric detection.

A new analytical technique involving protein precipitation, heating, lipid degreasing, and SPE procedures combined with HPLC-UV and HPLC-MS/MS has been developed for the determination of neotame in a variety of food samples. This method is applicable for high-protein, high-lipid, or gum-based solid samples. The limit of detection of the HPLC-UV method was 0.5 µg/mL, while that of the HPLC-MS/MS method was 3.3 ng/mL. The spiked recoveries of neotame in 73 kinds of foods were in the range of 81.1-107.2 % with UV detection. The spiked recoveries obtained by HPLC-MS/MS in 14 kinds of foods ranged from 81.6 % to 105.8 %. This technique was successfully used to determine the contents of neotame in two positive samples, indicating its applicability in food analysis.

New strategies for identifying and masking the bitter taste in traditional herbal medicines: The example of Huanglian Jiedu Decoction.

Suppressing the bitter taste of traditional Chinese medicine (TCM) largely has been a major clinical challenge due to complex and diverse metabolites and high dispersion of bitter metabolites in liquid preparations. In this work, we developed a novel strategy for recognizing bitter substances, hiding their bitter taste, and elucidated the mechanism of flavor masking in TCM. Huanglian Jie-Du Decoction (HLJDD) with an intense bitter taste was studied as a typical case. UHPLC-MS/MS was used to analyze the chemical components in HLJDD, whereas the bitter substances were identified by pharmacophores. Additionally, the screening results of the pharmacophores were further validated by using experimental assays. The mask formula of HLJDD was effectively screened under the condition of clear bitter substances. Subsequently, computational chemistry, molecular docking, and infrared characterization (IR) techniques were then used to explicate the mechanism of flavor masking. Consequently, neotame, γ-CD, and mPEG2000-PLLA2000 significantly reduced the bitterness of HLJDD. Specifically, mPEG2000-PLLA2000 increased the colloid proportion in the decoction system and minimized the distribution of bitter components in the real solution. Sweetener neotame suppressed the perception of bitter taste and inhibited bitter taste receptor activation to eventually reduce the bitter taste. The γ-CD included in the decoction bound the hydrophobic groups of the bitter metabolites in real solution and "packed" all or part of the bitter metabolites into the "cavity". We established a novel approach for screening bitter substances in TCM by integrating virtual screening and experimental assays. Based on this strategy, the bitter taste masking of TCM was performed from three different aspects, namely, changing the drug phase state, component distribution, and interfering with bitter taste signal transduction. Collectively, the methods achieved a significant effect on bitter taste suppression and taste masking. Our findings will provide a novel strategy for masking the taste of TCM liquid preparation/decoction, which will in return help in improving the clinical efficacy of TCM.

Charge-transfer interaction of aspartame and neotame with several π-acceptors: Stoichiometric data.

This data article is related to a research paper entitled ``Correlations between spectroscopic data for charge-transfer complexes of two artificial sweeteners, aspartame and neotame, generated with several π-acceptors'' [J. Mol. Liq. 333 (2021) 115904] [1]. Herein we present stoichiometric data of charge-transfer (CT) complexes generated from the interaction between aspartame and neotame with three π-acceptors in methanol solvent at room temperature. The investigated π-acceptors were picric acid (PA), chloranilic acid (CA), and 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ), where the methods used to determine the stoichiometry of the CT interaction were the spectrophotometric titration method and the Job's continuous variation method.

Aqueous degradation of artificial sweeteners saccharin and neotame by metal organic framework material.

The artificial sweeteners (ASs) saccharin (SAC) and neotame (NEO) are widely used across the globe and are considered as emerging contaminants in surface, ground, and drinking waters. To degrade SAC and NEO, the metal organic framework material Co-based bio-MOF-11 was prepared by hydrothermal reaction and used with peroxymonosulfate (PMS) activator. The effects of the initial concentration of SAC and NEO, bio-MOF-11-Co dosage, PMS concentration, initial pH, temperature, and competitive anions were determined. The results revealed that bio-MOF-11-Co effectively catalyzed the degradation of SAC and NEO and possessed good stability and recycling efficiency. The degradation reaction was effective from pH 3.6-9.8 and followed quasi-first-order kinetics with degradation rate constants of 0.001-0.013 min-1 for SAC and 0.03-0.52 min-1 for NEO. Increased temperature was conducive to the degradation of both artificial sweeteners. The presence of Cl- inhibited the degradation of SAC and NEO, while the presence of CO32- promoted their degradation. Electron paramagnetic resonance (EPR) and free radical quenching demonstrated that the primary free radicals were sulfate radicals ( [Formula: see text] ) and hydroxyl radicals (HO). The change of cobalt oxidation state and electron transfer in bio-MOF-11-Co mainly induces the production of [Formula: see text] . A plausible mechanism for degradation is [Formula: see text] and HO attack on CS bonds, NS bonds, and benzene rings.

Multi-spectroscopic and molecular docking studies on the interaction of neotame with calf thymus DNA.

In this paper, we have studied the in vitro binding of neotame (NTM), an artificial sweetener, with native calf thymus DNA using different methods including spectrophotometric, spectrofluorometric, competition experiment, circular dichroism (CD), and viscosimetric techniques. From the spectrophotometric studies, the binding constant (Kb) of NTM-DNA was calculated to be 2 × 103 M-1. The quenching of the intrinsic fluorescence of NTM in the presence of DNA at different temperatures was also used to calculate binding constants (Kb) as well as corresponding number of binding sites (n). Moreover, the obtained results indicated that the quenching mechanism involves static quenching. By comparing the competitive fluorimetric studies with Hoechst 33258, as a known groove probe, and methylene blue, as a known intercalation probe, and iodide quenching experiments it was revealed that NTM strongly binds in the grooves of the DNA helix, which was further confirmed by CD and viscosimetric studies. In addition, a molecular docking method was employed to further investigate the binding interactions between NTM and DNA, and confirm the obtained results.

Development of a highly sensitive liquid chromatography with tandem mass spectrometry method for the qualitative and quantitative analysis of high-intensity sweeteners in processed foods.

A new method for the simultaneous determination of two sweeteners (Advantame and Neotame) in processed foods using liquid chromatography (LC) with tandem mass spectrometry(MS/MS) was developed herein. Chromatographic separations were performed using an ACQUITY UPLC CSH C18 column at 40 °C via a mobile phase comprising 10-mmol/L ammonium formate and methanol. Samples were prepared via rapid dialysis using 30% methanol solution in a thermostatic shaker set at 160 rpm and 50 °C for 1 h. The matrix in the test solution had no effect on the identification and quantification of the compound without a clean-up step using solid-phase extraction (SPE). This method satisfied all validation criteria with a limit of quantification (LOQ) of 0.01 µg/g for all samples. Using this method, the amounts of Advantame and Neotame in 24 processed foods were subsequently investigated, with the results indicating their detection beyond the lower LOQ. Moreover, a multiple reaction monitoring information-dependent acquisition-enhanced product ion (MRM-IDA-EPI) method was developed and described to further enhance product-identification ability.

Effects of the Artificial Sweetener Neotame on the Gut Microbiome and Fecal Metabolites in Mice.

Although artificial sweeteners are widely used in food industry, their effects on human health remain a controversy. It is known that the gut microbiota plays a key role in human metabolism and recent studies indicated that some artificial sweeteners such as saccharin could perturb gut microbiome and further affect host health, such as inducing glucose intolerance. Neotame is a relatively new low-caloric and high-intensity artificial sweetener, approved by FDA in 2002. However, the specific effects of neotame on gut bacteria are still unknown. In this study, we combined high-throughput sequencing and gas chromatography-mass spectrometry (GC-MS) metabolomics to investigate the effects of neotame on the gut microbiome and fecal metabolite profiles of CD-1 mice. We found that a four-week neotame consumption reduced the alpha-diversity and altered the beta-diversity of the gut microbiome. Firmicutes was largely decreased while Bacteroidetes was significantly increased. The Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis also indicated that the control mice and neotame-treated mice have different metabolic patterns and some key genes such as butyrate synthetic genes were decreased. Moreover, neotame consumption also changed the fecal metabolite profiles. Dramatically, the concentrations of multiple fatty acids, lipids as well as cholesterol in the feces of neotame-treated mice were consistently higher than controls. Other metabolites, such as malic acid and glyceric acid, however, were largely decreased. In conclusion, our study first explored the specific effects of neotame on mouse gut microbiota and the results may improve our understanding of the interaction between gut microbiome and neotame and how this interaction could influence the normal metabolism of host bodies.

Stability of aspartame and neotame in pasteurized and in-bottle sterilized flavoured milk.

Analytical high performance liquid chromatography (HPLC) conditions were standardized along with the isolation procedure for separation of aspartame and neotame in flavoured milk (pasteurized and in-bottle sterilized flavoured milk). The recovery of the method was approximately 98% for both aspartame and neotame. The proposed HPLC method can be successfully used for the routine determination of aspartame and neotame in flavoured milk. Pasteurization (90 °C/20 min) resulted in approximately 40% loss of aspartame and only 8% of neotame was degraded. On storage (4-7°C/7 days) aspartame and neotame content decreased significantly (P<0.05) from 59.70% to 44.61% and 91.78% to 87.18%, respectively. Sterilization (121 °C/15 min) resulted in complete degradation of aspartame; however, 50.50% of neotame remained intact. During storage (30 °C/60 days) neotame content decreased significantly (P<0.05) from 50.36% to 8.67%. Results indicated that neotame exhibited better stability than aspartame in both pasteurized and in-bottle sterilized flavoured milk.

Low-calorie sweeteners in food and food supplements on the Italian market.

This study determines the occurrence and concentration levels of artificial low-calorie sweeteners (LCSs) in food and food supplements on the Italian market. The analysed sample set (290 samples) was representative of the Italian market and comprised of beverages, jams, ketchups, confectionery, dairy products, table-top sweeteners and food supplements. All samples were analysed via UPLC-MS/MS. The method was in-house validated for the analysis of seven LCSs (aspartame, acesulfame-K, saccharin, sucralose, cyclamate, neotame and neohesperidin dihydrochalcone) in food and for five LCSs (aspartame, acesulfame-K, saccharin, cyclamate and sucralose) in food supplements. Except for cyclamate in one beverage which exceeded the maximum level (ML) with 13%, all concentrations measured in food were around or below the ML. In food supplements, 40 of the 52 samples (77%) were found to be above the ML, with exceedances of up to 200% of the ML.

Fabrication of copper nanoparticles decorated multiwalled carbon nanotubes as a high performance electrochemical sensor for the detection of neotame.

A highly sensitive and novel electrochemical sensor for the detection of neotame using differential pulse voltammetry with a modified glassy carbon electrode is presented. The method was further customized by the fabrication of the electrode surface with copper nanoparticles-ammonium piperidine dithiocarbamate-mutiwalled carbon nanotubes assimilated with ß-cyclodextrin. The multiwalled carbon nanotubes assimilated with ß-cyclodextrin/glassy carbon electrode exhibited catalytic activity towards the oxidation of neotame at a potential of 1.3 V at pH 3.0. The transmission electron microscopy, thermogravimetric analysis, frontier transform infrared spectroscopy and cyclic voltammetry were employed to characterize the electrochemical sensor. The sensitivity and detection limits of the electrode increased two-fold in contrast to the ß-CD-MWCNTs/GCE sensor. The developed method was successfully applied for the determination of neotame in food samples, with results similar to those achieved by our modified capillary electrophoresis method with a 96% confidence level.

Bitterness-masking Effects of Neotame on Five Bitter Chinese Herbal Ingredients / 世界科学技术-中医药现代化

This study was ai med to observe the taste-masking effects of Neotame on bitter Chinese herbal ingredients. Five kinds of herbal ingredients, which include Scutellaria baicalensis Georgi, Cortex Phellodendri chinensis, Coptis chinensis Franch, Gentiana scabra Bunge, Andrographis paniculata, were selected to measure the bitterness degree of decoctions with berberine solution as the benchmark. The decreasing of bitterness degree was used as index. Healthy volunteers were recruited to taste and compare the changes of bitterness of decoctions with the taste-masking effects of Neotame. Different concentrations of Neotame were selected in the determination of the influence on changes of bitterness. The results showed that when the concentration of Neotame was at 0.012 5‰-0.4‰, taste-masking effects of Neotame on selected herbal decoctions were in a concentration-dependent fashion. When the concentration of Neotame was 0.4‰, the reduced bitterness of S. baicalensis Georgi and Cortex P. chinensis decoctions were 1.22 and 1.77, by 70.11% and 71.88%, respectively. Three highly-bitter herbal ingredients C. chinensis Franch, G. scabra Bunge and A . paniculata were also reduced in bitter taste by 49.12%, 50.87% and 38.39%, with the bitter reduced value (△I) of 1.78, 2.02 and 1.43, respectively. It was concluded that Neotame exerted taste masking potential on bitter herbal ingredients with different bitter degrees.

Determination of neotame in non-alcoholic beverage by capillary zone electrophoresis.

BACKGROUND: Neotame (NEO) is a new artificial sweetener approved as a food additive in many countries. The present method for the determination of NEO in various foodstuffs is high-performance liquid chromatography (HPLC). There are no reports on the determination of NEO in foods by capillary zone electrophoresis (CZE). Therefore a simple and rapid method to determine NEO is desired for quality control. RESULTS: A CZE method combined with solid phase extraction was developed for the determination of NEO in non-alcoholic beverage. The optimum separation conditions obtained were 20 mmol L(-1) sodium borate buffer, pH 8, 25 kV applied voltage, 5 s hydrodynamic injection at 30 mbar and ultraviolet detection at 191 nm. The calibration curve showed good linearity (R(2) = 1.000) in the range 0.5-100 µg mL(-1) , and the limit of detection was 0.118 µg mL(-1) . The method was successfully applied to the determination of NEO in two kinds of beverage with migration time less than 5 min, relative standard deviation (n = 3) less than 2% and recoveries ranging from 90 to 95%. CONCLUSION: The proposed CZE method has the advantages of shorter analysis time and lower cost compared with HPLC, indicating that it may be a good alternative to the HPLC method.

Gain weight by "going diet?" Artificial sweeteners and the neurobiology of sugar cravings: Neuroscience 2010.

America's obesity epidemic has gathered much media attention recently. A rise in the percent of the population who are obese coincides with an increase in the widespread use of non-caloric artificial sweeteners, such as aspartame (e.g., Diet Coke) and sucralose (e.g., Pepsi One), in food products (Figure 1). Both forward and reverse causalities have been proposed. While people often choose "diet" or "light" products to lose weight, research studies suggest that artificial sweeteners may contribute to weight gain. In this mini-review, inspired by a discussion with Dr. Dana Small at Yale's Neuroscience 2010 conference in April, I first examine the development of artificial sweeteners in a historic context. I then summarize the epidemiological and experimental evidence concerning their effects on weight. Finally, I attempt to explain those effects in light of the neurobiology of food reward.