Effects of common artificial sweeteners at environmentally relevant concentrations on soil springtails and their gut microbiota.

Artificial sweeteners (AS) are extensively utilized as sugar substitutes and have been recognized as emerging environmental contaminants. While the effect of AS on aquatic organisms has garnered recent attention, their effects on soil invertebrates and gut microbial communities remain unclear. To address this knowledge gap, we exposed springtails (Folsomia candida) to both single and combined treatments of four typical AS (sucralose [SUC], saccharin [SAC], cyclamate [CYC], and acesulfame [ACE]) at environmentally relevant concentrations of 0.01, 0.1 and 1 mg kg-1 in soil. Following the first-generational exposure, the reproduction of juveniles showed a significant increase under all the AS treatments of 0.1 mg kg-1. The transcriptomic analysis revealed significant enrichment of several Kyoto Encyclopedia of Gene and Genome pathways (e.g., glycolysis/gluconeogenesis, pentose and glucuronate interconversions, amino sugar, and nucleotide sugar metabolism, ribosome, and lysosome) in springtails under all AS treatments. Analysis of gut bacterial microbiota indicated that three AS (SUC, CYC, and ACE) significantly decreased alpha diversity, and all AS treatments increased the abundance of the genus Achromobacter. After the sixth-generational exposure to CYC, weight increased, but reproduction was inhibited. The pathways that changed significantly (e.g., extracellular matrix-receptor interaction, amino sugar and nucleotide sugar metabolism, lysosome) were generally similar to those altered in first-generational exposure, but with opposite regulation directions. Furthermore, the effect on the alpha diversity of gut microbiota was contrary to that after first-generational exposure, and more noticeable disturbances in microbiota composition were observed. These findings underscore the ecological risk of AS in soils and improve our understanding of the toxicity effects of AS on living organisms.

Levels, consumption, and variations of eight artificial sweeteners in the wastewater treatment plants of Dalian city, China.

Artificial sweeteners (ASs) are emerging contaminants in the environment, primarily derived from wastewater treatment plant (WWTP) effluents. In this study, the influents and effluents of three WWTPs in the Dalian urban area, China, were analyzed for the distribution of 8 typical ASs to investigate their seasonal fluctuations in the WWTPs. The results showed that acesulfame (ACE), sucralose (SUC), cyclamate (CYC), and saccharin (SAC) were both detected in the influent and effluent water samples of WWTPs, with concentrations ranging from not detected (ND) to 14.02 µg·L-1. In addition, SUC was the most abundant ASs type, accounting for 40 %-49 % and 78 %-96 % of the total ASs in the influent and effluent water, respectively. The WWTPs revealed high removal efficiencies of CYC, SAC, and ACE, while the SUC removal efficiency was poor (26 % ± 36 %). The ACE and SUC concentrations were higher in spring and summer, and all ASs showed lower levels in winter, which may be caused by the high consumption of ice-cream in warmer months. The per capita ASs loads in the WWTPs were determined in this study based on the wastewater analysis results. The calculated per capita daily mas loads for individual ASs ranged from 0.45 g·d-1·1000p-1 (ACE) to 2.04 g·d-1·1000p-1 (SUC). In addition, the relationship between per capita ASs consumption and socioeconomic status showed no significant correlation.

Occurrence, distribution, and ecological risk assessment of artificial sweeteners in surface and ground waters of the middle and lower reaches of the Yellow River (Henan section, China).

As a new class of water contaminants, artificial sweeteners (ASs) have attracted much attention due to their environmental persistence and potential adverse effects to human and the environment. This study systematically investigated the occurrence and distribution of four commonly used ASs in the effluent of wastewater treatment plants (WWTPs), surface water and groundwater in the middle and lower reaches of the Yellow River (Henan section). Sucralose (SUC) was dominant in WWTP effluents and had the highest mass loading. Acesulfame (ACE), cyclamate (CYC), saccharin (SAC), and SUC were consistently detected in surface water at concentrations ranging from 1.364 ng/L (CYC) to 7786 ng/L (ACE). Spatial analysis showed that the pollution level of ASs in the trunk stream was lower than that in most tributaries. The total concentrations of ASs detected in surface water ranged between 308.7 and 10,498 ng/L, while in groundwater, the total concentration of ASs detected was between ND-4863 ng/L. ACE and SUC are the main pollutants in surface water and groundwater within this survey area. The risk assessment showed that the risks of the four target ASs to aquatic organisms were negligible (risk quotient (RQ) values < 0.1), and the maximum risk quotient of the mixtures (MRQ) values of all rivers were all much less than 0.1.

Light-shading reaction microfluidic PMMA/paper detection system for detection of cyclamate concentration in foods.

Cyclamate is an artificial sweetener with high sweetness and low calories, and is a common sugar substitute for weight control and diabetic patients. However, excessive cyclamate consumption is associated with various health disorders, and hence it is prohibited as a food additive in many countries around the world. The current research proposes a light-shading reaction microfluidic PMMA/paper detection (MPD) system for determining the cyclamate concentration in food. In the current system, inject 10 µL of the extracted sodium cyclamate sample into the sample chamber of the MPD device, perform the diazotization reaction under shading conditions, and then suck it into the detection area through a paper strip, which consists of a paper chip embedded with modified Bratton-Marshall reagent. Once the paper chip is thoroughly wetted, the MPD device is inserted into a microanalysis box, where a fuchsia azo reaction compound is produced through heating at 40 °C for 3 min. The reaction complex is observed by a camera and the reaction image is wirelessly transmitted to a smartphone, and the concentration of sodium cyclamate is measured through the self-developed grayscale software. The results obtained for the sodium cyclamate samples with a concentration in the range of 50-1000 ppm show that the measured gray value changes linearly with the sodium cyclamate concentration, and the correlation coefficient (R2) is 0.9898. By analyzing the concentration of sodium cyclamate in 10 real-world samples, the practical feasibility of the current MPD system is proved. The results showed that the concentration measurement value did not deviate by more than 4.8 % from the value obtained using the conventional liquid chromatography/tandem mass spectrometry (LC-MS/MS) method.

Seasonal occurrence, removal and mass loads of artificial sweeteners in the largest water reclamation plant in China.

Artificial sweeteners (ASs) are of growing concern as an emerging contaminant. In the study, the seasonal occurrence, removal and mass load of six ASs in sewage, suspended particulate matter (SPM) and sludge were investigated throughout the treatment process of the largest water reclamation plant in China. The highest ASs concentrations in the influent (13.0 µg/L), effluent (2.22 µg/L), SPM (4.48 µg/g) and sludge (0.15 µg/g) were observed in the dry season, which were 1.24- to 5.0-fold higher than in the normal season and 1.06- to 37.5-fold higher than the flood season. Following treatment, ASs concentrations decreased by 24.3 %, 51.7 % and 5.1 % (on average) in primary, secondary and reclaimed processes, respectively. Among the investigated ASs, acesulfame (93.1 %) and cyclamate (98.4 %) were removed most efficiently, with removal occurring mainly in secondary processes, while sucralose exhibited the lowest removal efficiency (38.7 %). Seasonal characteristics affect the consumption of ASs, which subsequently changes the input and discharge ASs loads of STPs. The maximum mass load of ASs occurred in the dry season, ranging from 0.002 (neotame) to 1.33 mg/d/person (cyclamate), while the maximum emission load occurred in the flood season, ranging from 0.003 (neotame) to 0.83 mg/d/person (sucralose). The mass and emission load of ASs in Beijing is significantly lower than in European or the United States, due to Beijing having low per capita consumption of ASs (5.50 mg/d/person). The highest ASs risk in the receiving water occurred in the flood season due to the input of other pollution sources by rainfall runoff. Meanwhile, attention should be paid to the risk of receiving water close to the STP outlet in the dry seasons for the highest ASs concentration in the STP effluent in the season. The present study provides important guidance on controlling the input and reducing the emission of ASs in different seasons.

Rapid and sensitive quantification of cyclamate in beverages by miniature microplasma optical emission spectrometry.

Most of the available methods for the quantification of cyclamate depend on laboratory instruments and their application in the field was limited. Herein, a simple and sensitive method was developed for the determination of cyclamate in beverage samples based on chemical vapor generation and miniature point discharge optical emission spectrometry (µPD-OES). The combination of headspace sampling and µPD-OES not only simplifies the separation process of cyclamate, improves sensitivity, and alleviates matrix interference but also eliminates the use of a bulky and expensive instrument. Under the optimal conditions, this method provided a limit of detection of 0.1 mg L-1 comparable to or better than most reported methods. The method eventually was applied to 14 different beverages and cyclamate was found below the threshold set by Chinese Standards for Food Additives. The proposed method provides great potential for the field analysis of cyclamate in the supervision of food safety.

Suspect screening of pharmaceuticals, illicit drugs, pesticides, and other emerging contaminants in Argentinean Piaractus mesopotamicus, a fish species used for local consumption and export.

The widespread distribution of contaminants of emerging concern (CECs) is a major concern due to their potential effects on human health and the environment. The insufficient sewage treatment plant infrastructures is a global problem most accentuated in less developed countries and results in the discharge of CECs to water bodies. Pacu (Piaractus mesopotamicus) is a ray-finned freshwater fish species native to the Paraná basin. It is also the most produced aquaculture fish species in Argentina since 2012. Though uninvestigated to date, the occurrence of CECs in pacu may be of high relevance due to production volumes and relevance to human exposure through fish consumption. In this study, we applied a high-resolution mass spectrometry screening method to qualitatively analyze over 100 CECs in pacu. Four extraction/cleanup methods were tested on pooled pacu fillet, including solid-phase extraction and QuEChERS. The method that produced the highest number of detections was selected for further analysis of pacu purchased in supermarkets and fish markets in Argentina between 2017 and 2020. Residues of pesticides, antibiotics, pharmaceuticals, personal care products, plasticizers, sweeteners, drug metabolites, stimulants, and illegal drugs were detected in the samples. A total of 38 CECs were detected, ranging between 24 and 35 CECs per individual sample. 100% of the samples had positive detections of caffeine, 1,7-dimethylxanthine, xanthine, benzoylecgonine, methylparaben, ethylparaben, bis(2-ethylhexyl) phthalate (DEHP), metolachlor, carbendazim, salicylic acid, 2,4-D, saccharin, cyclamate, and dodecanedioic acid. Mappings generated with correspondence analysis were used to explore similarities/dissimilarities among the detected compounds. To our knowledge this is the first report of saccharin, cyclamate, 2,4 - D, carbendazim, metolachlor, ethylparaben, propylparben, bisphenol A, DEHP, and benzotriazole in fish from Argentina, and the first report on the presence of lisinopril, metropolol acid and dodecanedioic acid in fish worldwide.

Dietary exposure to low-calorie sweeteners in a sample of Brazilian pregnant women.

The dietary exposure to low-calorie sweeteners (LCS) was estimated in a sample of pregnant Brazilian women. Consumption data were obtained with a 24-h Dietary Recall interview. Because of the uncertainty in assessing foods with LCS, they were classified into three scenarios to ensure inclusion of the 15 LCS allowed for use in Brazil: ranging from a less to a more conservative scenario. The concentration of LCS was estimated using the amount declared on the label or the maximum permitted levels and analytical determination data for table-top sweeteners. The frequency of consumption was higher for acesulfame-K, aspartame, and cyclamate. The food groups contributing the most to the consumption of LCS were non-alcoholic beverages, table-top sweeteners, confectionary and desserts. The level of dietary exposure to LCS was within the safety limit. However, continued efforts to monitor their dietary exposure are necessary given the limitations highlighted in this study.

Artificial Sweeteners in Breast Milk: A Clinical Investigation with a Kinetic Perspective.

Artificial sweeteners (ASs) are calorie-free chemical substances used instead of sugar to sweeten foods and drinks. Pregnant women with obesity or diabetes are often recommended to substitute sugary products with ASs to prevent an increase in body weight. However, some recent controversy surrounding ASs relates to concerns about the risk of obesity caused by a variety of metabolic changes, both in the mother and the offspring. This study addressed these concerns and investigated the biodistribution of ASs in plasma and breast milk of lactating women to clarify whether ASs can transfer from mother to offspring through breast milk. We recruited 49 lactating women who were provided with a beverage containing four different ASs (acesulfame-potassium, saccharin, cyclamate, and sucralose). Blood and breast milk samples were collected before and up to six hours after consumption. The women were categorized: BMI < 25 (n = 20), BMI > 27 (n = 21) and type 1 diabetes (n = 8). We found that all four ASs were present in maternal plasma and breast milk. The time-to-peak was 30−120 min in plasma and 240−300 min in breast milk. Area under the curve (AUC) ratios in breast milk were 88.9% for acesulfame-potassium, 38.9% for saccharin, and 1.9% for cyclamate. We observed no differences in ASs distributions between the groups.

Young population consume twice as much artificial sweetener than the general population - A wastewater-based assessment in China.

Understanding artificial sweetener consumption patterns and levels in different demographics is important for formulating public health policies on controlling sugar consumption. There is a considerable knowledge gap with respect to the pattern of artificial sweetener consumption in China. To narrow this gap, wastewater analysis was used to assess the temporal patterns of consumption of seven artificial sweeteners in an urban population and a university town in a megacity in South China over a one-year period. Daily influent wastewater samples were collected from an urban catchment and weekly samples collected from a university sub-catchment. Population normalized per capita consumption of the four detected artificial sweeteners (cyclamate, acesulfame, sucralose and saccharin) in the university catchment (1.0-5.9 mg d-1 p-1) was much higher than those in urban catchment (0.5-1.3 mg d-1 p-1), indicating younger population consume more artificial sweeteners than the general population. The daily consumption of artificial sweeteners was found to be stable throughout the week in the urban catchment. Time-series analysis showed that an average increase in temperature of 1 °C was associated with an increase consumption of 33 µg d-1 p-1 for acesulfame, 15 µg d-1 p-1 for sucralose and 14 µg d-1 p-1 for saccharin. This was the first study that objectively quantified the greater consumption of artificial sweeteners (proxy for consumption of artificially sweetened food and beverages) in a younger age group when compared to the general population, which could potentially pose a risk of health related diseases.

Tracking the fate of artificial sweeteners within the coastal waters of Shenzhen city, China: From wastewater treatment plants to sea.

Artificial sweeteners (ASs), as a new emerging pollutant, the fate from wastewater treatment plants (WWTPs) to sea is growing concerned. In this study, the distribution and polluted loading of six typical ASs were analyzed based on the measurement in influent, primary effluent, secondary effluent, tertiary effluent, suspended particulate matter (SPM), and sludge from four WWTPs and receiving waters near the coast of Shenzhen city, China. Elevated levels of ASs were detected in WWTPs located near a highly populated area (about 14,000 people km-2). Sucralose was predominant in all water samples (0.1-22.2 µg L-1), and had a 1.6-704.0 times higher concentration than the means of other ASs detected. Aspartame were mainly distributed in SPM and sludge, and the fractions in offshore water exceeded 45%. Acesulfame, sucralose, cyclamate and saccharin could be effectively removed by secondary biochemical treatment. The average daily loading (4.2 g d-1 1000people-1) and discharge loading (0.4 g d-1 1000people-1) of sucralose in WWTPs was higher than those of the other ASs. Dissolved organic carbon and UV254 can affect the fate of ASs, and SPM mainly affects the distributions of aspartame and neotame. As a potential sewage indicator, neotame deserves further attention.

Impact assessment of anthropogenic activities on the ecological systems in the Xiongan New Area in the North China Plain.

The aim of this study is to assess the impact of anthropogenic activities on the ecological systems in the Xiongan New Area (XNA), China. Artificial sweeteners (ASs) were used as anthropogenic tracers to investigate how anthropogenic activities affect water, soil, and plant layers. For this purpose, the concentrations of 4 typical ASs, acesulfame (ACE), sucralose (SUC), saccharin (SAC), and cyclamate (CYC) were measured in water, soil, and plant layers. Moreover, a tracer test was conducted in the critical zone observatory (CZO) in the southern part of the study area to investigate the migration path and time of anthropogenic activities on the ecological systems. Acesulfame, SUC, SAC, and CYC were widely detected in all of the sample types, and their average values were 0.22, 0.53, 0.06, and 0.08 µg/L in the water layer; 0.04, 0.18, 0.06, and 0.06 µg/kg in the soil layer; and 1.79, 0.11, 4.42, and 0.95 µg/kg in the plant layer, respectively. The tracer test showed that 1.1% to 1.8% of the impacts of the anthropogenic activities migrated from the surface to the shallow aquifer within 13 d, 6% to 19% migrated to the maize layer within 28 d, and 79% to 93% were adsorbed and biodegraded in the soil layer. Therefore, the scope and strength of the influence of the anthropogenic activities on the environmental media exhibit a decreasing sequence of maize > water > soil. This study not only quantitatively characterizes the impact of anthropogenic activities on the ecological environment but also presents the first comprehensive overview of the environmental fate of ASs in the critical zone of the XNA, China. Integr Environ Assess Manag 2021;17:866-876. © 2021 SETAC.

FTIR spectroscopy coupled with machine learning approaches as a rapid tool for identification and quantification of artificial sweeteners.

Fourier transform infrared (FTIR) spectroscopy calibrations were developed to simultaneously determine the multianalytes of five artificial sweeteners, including sodium cyclamate, sucralose, sodium saccharin, acesulfame-K and aspartame. By combining the pretreatment of the spectrum and principal component analysis, 131 feature wavenumbers were extracted from the full spectral range for modelling to qualitative and quantitative analysis. Compared to random forest, k nearest neighbour and linear discriminant analysis, support vector machine model had better predictivity, indicating the most effective identification performance. Furthermore, multivariate calibration models based on partial least squares regression were constructed for quantifying any combinations of the five artificial sweeteners, and validated by prediction data sets. As shown by the good agreement between the proposed method and the reference HPLC for the determination of the sweeteners in beverage samples, a promising and rapid tool based on FTIR spectroscopy, coupled with chemometrics, has been performed to identify and objectively quantify artificial sweeteners.

[Validation Study of Rapid Analytical Method for Determination of Cyclamate in Various Kinds of Foods by LC-MS/MS].

A rapid, sensitive and selective analytical method by LC-MS/MS was developed and validated for the determination of cyclamate in various kinds of foods. The Preparation of test solutions was performed by heat extraction technique in accordance with an official notification method in Japan. We aimed to reduce the matrix effects in LC-MS/MS only by diluting extracts without clean-up using solid phase column. This method was assessed for 30 kinds of foods fortifying cyclamate at the concentration level of 0.5 µg/g. As a result, trueness was 85.0 to 106.6%, repeatability (RSDr) ranged from 1.7 to 9.4%, and within-laboratory reproducibility (RSDwr) ranged from was 4.1 to 9.7%. These date supported the reliability our method. Thus, this method could be useful for a rapid determination of cyclamate in various kinds of foods.

Simultaneous determination of aspartame, cyclamate, saccharin and acesulfame-K in powder tabletop sweeteners by FT-Raman spectroscopy associated with the multivariate calibration: PLS, iPLS and siPLS models were compared.

For the first time, a procedure for simultaneous determination of the main artificial sweeteners, aspartame (ASP), cyclamate (CYC), saccharin (SAC), and acesulfame-K (ACSK) by a spectroscopic method associated with the multivariate calibration is proposed. These analytes were quantified in tabletop sweeteners samples using FT-Raman spectroscopy. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used as reference method. Partial least squares (PLS), interval PLS (iPLS), and synergism PLS (siPLS) methods were evaluated in a comparative study where the selected interval models presented better results. Multivariate regression models, such as PLS, iPLS and siPLS were built and the lower root mean square errors for prediction (RMSEP) found were 0.027-0.031% w/w, 0.316-0.363% w/w, 0.082-0.184% w/w, and 0.040-0.049% w/w to ASP, CYC, SAC, and ACSK, respectively. The coefficient of determination for prediction (R2p) varied between 0.978 and 0.979, 0.969-0.977, 0.952-0.994, and 0.959-0.965 for ASP, CYC, SAC and ACSK, respectively. The analysis of model's residues was made by bias and permutation tests to evaluate systematic and trend errors. The selected intervals by iPLS and siPLS were evaluated and the bands related to the vibrational modes of the analytes were assigned with the aid of density functional theory calculations (DFT).

Mass loading of typical artificial sweeteners in a pig farm and their dissipation and uptake by plants in neighboring farmland.

Pig farm is an important potential source for artificial sweeteners (ASs) in the environment due to their wide use as additives in pig feed. The objective of this study was to evaluate the fate of typical ASs in pig farm and neighboring farmland. For this purpose, the levels of four typical artificial ASs, i.e. saccharin (SAC), cyclamate (CYC), acesulfame (ACE) and sucralose (SUC), in pig feed and manure from a pig farm and water samples from an on-farm wastewater treatment plant (WWTP) in Tianjin, China were measured and the mass loadings and removal efficiencies were assessed. Moreover, the levels of ASs in different layers of soil and vegetables in neighboring farmland that received manure fertilizers and wastewater from the farm were consecutively monitored for 60-80days. The SAC, CYC and ACE were widely determined in all kinds of the samples, while SUC was only found in few soil samples. The mass loadings of the ASs in pig feed were estimated up to 311kg/year for SAC, 59.1kg/year for CYC, and 17.1kg/year for ACE, respectively. The fractions of the total mass of ASs excreted via manure were estimated to be 36.0% for SAC, 59.4% for CYC, and 36.7% for ACE as compared to those in pig feed. High removal efficiencies (>90%) of ASs in the on-farm WWTP was achieved. In greenhouse soils, CYC, SAC, ACE, and SUC were degraded quickly, with half-lives of 4.3-5.9 d, 2.7-4.2 d, 8.4-12.3 d, and 7.3-10.8 d, respectively. Lower levels of ASs were found in deeper soil layer (20-30cm). The ASs were considerably absorbed by plants when the ASs' concentrations were high in soil. This study presents the first comprehensive overview of ASs fate from a pig farm to the neighboring agricultural ecosystem.

Application of Liquid Chromatography-Tandem Mass Spectrometry To Determine Urinary Concentrations of Five Commonly Used Low-Calorie Sweeteners: A Novel Biomarker Approach for Assessing Recent Intakes?

Although the use of low-calorie sweeteners (LCSs) is widespread, methods of assessing consumption within free-living populations have inherent limitations. Five commonly consumed LCSs, namely, acesulfame-K, saccharin, sucralose, cyclamate, and steviol glycosides, are excreted via the urine, and therefore a urinary biomarker approach may provide more objective LCS intake data. A LC-ESI-MS/MS method of simultaneously determining acesulfame-K, saccharin, sucralose, cyclamate, and the excretory metabolite of steviol glycosides, steviol glucuronide, in human urine was developed and validated. Linearity was observed over a concentration range of 10-1000 ng/mL with coefficients of determination ranging from 0.9969 to 0.9997. Accuracy ranged from 92 to 104%, and intrabatch and interday precisions were within acceptable limits with %CV below 8% for all compounds. A double-blind, randomized crossover dose-response study was conducted to assess the usefulness of urinary LCS excretions (from both fasting spot and a full 24-h urine collection) for investigating recent intakes. Both modes of sampling were useful for distinguishing between the three short-term intakes of acesulfame-K, saccharin, cyclamates, and steviol glycosides (p < 0.001), whereas for sucralose, urinary concentrations were useful for distinguishing between low (0.1% ADI) and high doses (10% ADI) only (p < 0.001). In summary, this biomarker approach may be useful for assessing intakes of five commonly consumed LCSs.

On-cartridge derivatisation using matrix solid phase dispersion for the determination of cyclamate in foods.

A novel method for determination of sodium cyclamate in foods was developed. In this method, a syringe was loaded with the homogeneous mixture of the sample, KMnO4 powder and silica dispersant and used as a matrix solid phase dispersion (MSPD) reactor. As the reactor was infiltrated with small amounts of concentrated HCl, cyclamate was converted to 2-chlorocyclohexanone quickly and effectively within 5 min and determined by HPLC on a reversed-phase column using UV detection at a wavelength of 310 nm. Comparing with the traditional derivatisation in solution, the better clean-up was provided using on-cartridge derivatisation of MSPD, and much time, labor, and expense were saved. The results showed good linearity (r2 = 0.9998) over the concentration range of 1-500 mg/L. The limit of detection (LOD) and limits of quantification (LOQ) of the cyclamate were 0.3 mg/L and 1 mg/L respectively. The recoveries ranged from 91.6% to 101.3% with the relative standard deviations (RSDs) in the range of 2.5%-4.3%.

Validation of an HPLC-DAD/UV method for the quantification of cyclamate in tabletop sweeteners: risk of exceeding the acceptable daily intake.

Cyclamate is widely used as intense sweetener in the European Union. The absence of a maximum limit for the use of cyclamate in tabletop sweeteners and the growing demand for this type of product highlights the importance of developing robust analytical methods for the determination of its content to understand if the consumption of tabletop sweeteners can have a negative impact on human health. The present work aimed at the optimisation and validation of an high-performance liquid chromatography (HPLC) analytical method for cyclamate determination in tabletop sweeteners based on the procedure of European Standard EN 12857. The validated method was then applied to the determination of this sweetener in different types of tabletop sweeteners (liquid, powder and tablets). Both standards and samples solutions were submitted to a derivatisation procedure which converted cyclamate to N,N-dichlorocyclohexylamine. The derivatised product was separated and quantified using a reversed-phase column, a mobile phase composed of water (20%) and methanol (80%), isocratic flow of 1 ml min-1, and detection by ultraviolet spectrophotometry at a wavelength of 314 nm. The analytical method was internally validated according to the following validation parameters: working range, linearity, limits of detection and quantification, sensitivity, precision (repeatability and intermediate precision), and uncertainty. This method proved to be specific and selective for the determination of this sweetener, showing repeatability, RSDr ≤ 3%, intermediate precision, RSDR ≤ 3.3%, and recovery rates from 92% to 108% for the different tabletop sweeteners. The method uncertainty was 9.4%. The concentration of cyclamate in the samples varied significantly, from 2.9% to 73.9%, which demonstrated that a possible excessive consumption of one of the analysed sweeteners can lead to exceeding the acceptable daily intake for cyclamate.

[Simultaneous determination of 6 kinds of food additives in drink by HPLC-MS/MS].

OBJECTIVE: To establish a high performance liquid chromatographytandem mass spectrometry( HPLC-MS/MS) method for simultaneous determination of caffeine, benzoic acid, sorbic acid, acesulfame potassium, saccharin sodium and sodium cyclamate in drink. METHODS: The sample was extracted by ASPEC using extraction column( Pro Elut C_(18)500 mg/6 m L), dried by Turbo VapⅡ and dissolved in mobile phase. The supernatant was separated on Agilent SB-C_(18) column( 2. 1 mm × 50 mm, 1. 8 µm)using 0. 02 mmol/L ammonium acetate/methyle alcohol as mobile phase and then detected by HPLC-MS/MS using multiple reaction monitoring( MRM) in positive ionization and negative ionization mode. RESULTS: The average recoveries were from 72. 6% to 100. 5%and the relative standard deviations( RSD) were from 0. 3% to 3. 1%( n = 6). The calibration curves showed a good linearity with correlation coefficients r > 0. 9990. The linear range for saccharin sodium was 8-800 µg/L and others were 2-200 µg/L. The detection limits for caffeine, benzoic acid, sorbic acid, acesulfame potassium and sodium cyclamate were 0. 001 mg/kg and 0. 004 mg/kg for saccharin sodium. CONCLUSION: The method is specific, sensitive, easy, fast and suitable for the confirmation and quantification of caffeine, benzoic acid, sorbic acid, acesulfame potassium, saccharinsodium and sodium cyclamate in drink.