Acesulfame Research Articles

Association between non-nutritive sweeteners and obesity risk among university students in Latin America

The association between non-nutritive sweeteners and obesity is controversial. To determine whether the consumption of non-nutritive sweeteners is related to higher risk for overweight or obesity among university students in Chile, Panama, Guatemala and Peru. A total of 1,224 (472 from Chile, 300 from Panama, 248 from Guatemala and 204 from Peru) male and female university students aged between 18 and 26 years participated in the study. Each student reported their food intake (frequency of weekly consumption) in a survey that contained photos of foods containing non-nutritive sweeteners adapted for each country. Anthropometry was also measured. More than 80% of students consumed at least one product containing non-nutritive sweeteners. Females who ate acesulfame potassium and sucralose had a lower risk of overweight or obesity with an odds ratio (OR) of 0.5 (confidence intervals (CI) = 0.3-0.9; p = 0.003) and OR = 0.4 (IC = 0.2-0.8; p = 0.01), respectively. In this sample of Latinamerican university students, consumption of non-nutritive sweeteners was associated with lower risk of overweight only in females.

Design and Evaluation of Orally Disintegrating Tramadol Hydrochloride Tablets by Direct Compression Method

Aims: To design and evaluate an orally disintegrating tramadol hydrochloride tablets (ODT). Methods: Tramadol hydrochloride orally disintegrating tablets were designed and manufactured by direct compression method, using Cross povidone, Precirol, EPO, Sorbitol, PEG 6000, Aerosol, HCL, magnesium stearate, xylitol, acesulfame potassium, as key excipients, and peppermint flavor and sweetener, respectively. These formulations were then evaluated using pharmacopoeial and non-pharmacopoeial physical and chemical tests. Dissolution and assay tests were performed using USP apparatus II and ultraviolet (UV) spectrophotometry, respectively. Results: The tablet formulation prepared with crospovidone (F1) showed good flow properties, low disintegration time (13 s) and improved drug release (100% at 30 min) compared with those of the other formulations (84 % at 30 min). All the formulations exhibited satisfactory physicochemical characteristics. The results indicated that suitable ODT of tramadol could be prepared. Conclusion: A suitable preparation of tramadol Hcl ODT which contains Crospovidone (superdisintegrant) and sorbitol (bulking agent) was found to be the best among Tramadol hydrochloride ODT formulations prepared by direct compression method, because it has exhibited good disintegration time and good dissolution profile when compared to other formulations.

凝集沈殿剤を用いたサッカリンナトリウムおよびアセスルファムカリウムの新規分析法の開発

An HPLC method for determination of sodium saccharin and acesulfame potassium was newly developed, employing coagulant pretreatment to remove particles dispersed in the sample extract. The method showed recovery of 96-101% for both analytes with a repeatability of less than 1% and a reproducibility of less than 2%. The limit of quantification for sodium saccharin was 0.025 g/kg and that for acesulfame potassium was 0.025 g/kg. Only about 20 min was required for preparation of the test solution, whereas the dialysis method takes much longer.

Amygdalin in bitter and sweet seeds of apricots

Hydrogen cyanide (HCN) poisoning due to amygdalin (AMY) in apricot seeds is one of the public health issues in Turkey. The aim of this study was to investigate the AMY content of 13 different apricot seeds including bitter and sweet ones, and which are either sulfurized or roasted. The AMY content was determined by a high-performance liquid chromatography. Release of HCN was predicted and total amount of seeds which can cause poisoning was calculated. The mean AMY content of bitter seeds was 26 ± 14 mg g−1 and that of sweet seeds was 0.16 ± 0.09 mg g−1. The consumption of small amounts of bitter seeds may cause cyanide poisoning.

Molecularly imprinted sol-gel nanofibers based solid phase microextraction coupled on-line with high performance liquid chromatography for selective determination of acesulfame

Sol-gel based molecularly imprinted polymer (MIP) nanofiber was successfully fabricated by electrospinning technique on the surface of a stainless steel bar. The manufactured tool was applied for on-line selective solid phase microextraction (SPME) and determination of acesulfame (ACF) as an artificial sweetener with high performance liquid chromatography (HPLC). The selective ability of method for the extraction of ACF was investigated in the presence of some selected sweeteners such as saccharine (SCH), aspartame (ASP) and caffeine (CAF). Electrospinning of MIP sol-gel solution on the stainless steel bar provided an unbreakable sorbent with high thermal, mechanical, and chemical stability. Moreover, application of the MIP-SPME tool revealed a unique approach for the selective microextraction of the analyte in beverage samples. In this work, 3-(triethoxysilyl)-propylamine (TMSPA) was chosen as a precursor due to its ability to imprint the analyte by hydrogen bonding, Van der Walls, and dipole-dipole interactions. Nylon 6 was also added as a backbone and support for the precursor in which sol could greatly growth during the sol-gel process and makes the solution electrospinable. Various effective parameters in the extraction efficiency of the MIP-SPME tool such as loading time, flow rate, desorption time, selectivity, and the sample volume were evaluated. The linearity for the ACF in beverage sample was in the range of 0.78–100.5ng mL−1. Limit of detection (LOD) and quantification (LOQ) were 0.23 and 0.78ng mL−1 respectively. The RSD values (n=5) were all below 3.5%at the 20ng mL−1 level.

Consumption of carbonated beverages with nonnutritive sweeteners in Latin American university students

Consumption of carbonated beverages with nonnutritive sweeteners (NNS) is increasingly common in order to maintain a healthy weight, but the effect of NNS on body weight is controversial. University students (n=1,229) of both sexes aged 18 to 26, of which 472 were from Chile, 300 of Panama, 253 from Guatemala and 204 of Peru. Each student was applied a frequency survey of weekly food consumption supported by photographs of beverages with NNS from each country to determine the intake of them. Also they underwent anthropometric measurements. 80% of these students consumed carbonated beverages with NNS, none of them exceeded the acceptable daily intake for sucralose, potassium acesulfame and aspartame. Increased consumption in both men and women was observed in chilean students (p.

Functional relationship between oxytocin and appetite for carbohydrates versus saccharin.

Centrally acting oxytocin (OT) inhibits feeding. Recent evidence suggests a link between OT and control of carbohydrate and saccharin intake, but it is unclear whether OT affects appetite for only carbohydrates, especially sweet ones, or sweet tastants irrespective of their carbohydrate content. Therefore, a blood-brain barrier penetrant OT receptor antagonist, L-368,899, was administered in mice and intake of liquid diets containing carbohydrates sucrose, glucose, fructose, polycose, or cornstarch (CS) or the noncarbohydrate, noncaloric sweetener saccharin was studied in episodic intake paradigms: one in which only one tastant was available and the other in which a choice between a carbohydrate (sucrose, glucose, or fructose) and saccharin was provided. We also used real-time PCR to examine hypothalamic Ot mRNA levels in mice provided short-term access to sucrose, CS, or saccharin. In the no-choice paradigm, L-368,899 increased the intake of all carbohydrates, whereas its effect on saccharin consumption showed only a trend. A 10 times lower dose (0.3 mg/kg) stimulated intake of sucrose than other carbohydrates. In the choice test, a very low 0.1 mg/kg dose of L-368,899 doubled the proportion of sucrose consumption relative to saccharin, but did not affect fructose or glucose preference. Ot gene expression increased after sucrose and CS, but not saccharin exposure compared with the controls; however, a higher level of significance was detected in the sucrose group. We conclude that OT inhibits appetite for carbohydrates. Sucrose consumption considerably enhances Ot gene expression and is particularly sensitive to OT receptor blockade, suggesting a special functional relationship between OT and sugar intake.

Quantification of acesulfame potassium in processed foods by quantitative 1H NMR

Acesulfame potassium (AceK), a high-intensity and non-caloric artificial sweetener, is used in various processed foods as a food additive. In this study, we established and validated a method for determining the AceK content in various processed foods by solvent extraction and quantitative 1H NMR, using a certified reference material as the internal standard. In the recovery test, the proposed method gave satisfactory recoveries (88.4–99.6%) and repeatabilities (0.6–5.6%) for various processed foods. The limit of quantification was confirmed as 0.13gkg−1, which was sufficiently low for the purposes of monitoring AceK levels. In the analysis of commercially processed foods containing AceK, all AceK contents determined by the proposed method were in good agreement with those obtained by a conventional method based on dialysis and HPLC. Moreover, this method can achieve rapid quantification and yields analytical data with traceability to the International System of Units (SI) without the need for an authentic analyte standard. Therefore, the proposed method is a useful and practical tool for the determination of AceK in processed foods.

Direct large volume injection ultra-high performance liquid chromatography-tandem mass spectrometry determination of artificial sweeteners sucralose and acesulfame in well water

Acesulfame (ACE) and sucralose (SUC) have become recognized as ideal domestic wastewater contamination indicators. Liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) analysis is commonly used; however, the sensitivity of SUC is more than two orders of magnitude lower than that of ACE, limiting the routine monitoring of SUC. To address this issue, we examined the ESI behavior of both ACE and SUC under various conditions. ACE is ionic in aqueous solution and efficiently produces simple [M−H]− ions, but SUC produces multiple adduct ions, limiting its sensitivity. The formic acid (FA) adducts of SUC [M+HCOO]− are sensitively and reproducibly generated under the LC-MS conditions. When [M+HCOO]− is used as the precursor ion for SUC detection, the sensitivity increases approximately 20-fold compared to when [M−H]− is the precursor ion. To further improve the limit of detection (LOD), we integrated the large volume injection approach (500μL injection) with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), which reduced the method detection limit (MDL) to 0.2ng/L for ACE and 5ng/L for SUC. To demonstrate the applicability of this method, we analyzed 100 well water samples collected in Alberta. ACE was detected in 24 wells at concentrations of 1–1534ng/L and SUC in 8 wells at concentrations of 65–541ng/L. These results suggest that wastewater is the most likely source of ACE and SUC impacts in these wells, suggesting the need for monitoring the quality of domestic well water.

Artificial sweeteners in beverages by ultra performance liquid chromatography with photodiode array and liquid chromatography tandem mass spectrometry

Two fast, accurate and sensitive liquid chromatography methods have been developed and optimized for a better control of the content of artificial sweeteners in industrial beverages. Ultra performance liquid chromatography coupled with photodiode array (UPLC-PDA) and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) methods were implemented for the monitoring of aspartame, neohesperidine dihydrochalcone, neotame, potassium acesulfame, saccharin, sodium cyclamate and sucralose in beverages marketed as sugar-free or diet, including soft and powdered drinks. Minimal sample preparation procedure consisting on a simple dilution and filtration is required before analysis. The methods showed excellent linearity (R2<0.9990) for target compounds. Limits of quantification (LOQs) were far below the legal requirements for all considered compounds (0.01-0.1μgmL-1 and 0.05-5ngmL-1 for UPLC-PDA and LC-MS/MS, respectively). Precision and recovery studies in real samples showed excellent results. The recoveries at two concentration levels ranged between 90.0 and 114.6%, with relative standard deviations lower than 9.4 RSD%. Finally, the proposed methodology was successfully applied to the analysis of artificial sweeteners in 66 beverage products commonly consumed in Spain. Different sample categories were evaluated, including energy drinks, soft drinks, juices, teas, soy beverages, dairy-based drinks, beers, and spirit alcoholic drink, and proved its suitability for quick and reliable application in quality control laboratories. © 2014.

Morphological and Molecular Characterization of Turkish Landraces of &lt;i&gt;Cucumis melo&lt;/i&gt; L.&lt;/span&gt;

Cucumis melo L. from Eastern and South-eastern Anatolian regions of Turkey were characterized by using 43 morphological traits and 207 markers obtained from 31 ISSR and 16 SSR primers. The genetic relatedness was studied by examining the Euclidian/UPGMA dendrogram obtained from the combined phenotypic-molecular data. In the combined morphological-molecular dendrogram, there were two main clusters. Sweet and non-sweet melon groups were separated and the flexuosus group accessions were discriminated from the sweet ones, but the momordica group accession was clustered with the sweet ones. Unclear South-eastern Anatolian accessions were sub-clustered separately among the sweet ones. Principle component analysis (PCA) of morphological characters was used in detail to discriminate melon accessions. The cumulative proportion of variation reached 44% by first three PCA axes. The first component was mainly based on sex expression, ovary index, ovary shape, flesh thickness, seed cavity length, seed cavity width, soluble solids content, fruit shape, aroma, netting, and taste. The PCA plot based on all measured traits allowed distinction between flexuosus group, subsp.agrestis and reticulatus group. A high variation among groups was observed for the fruits characters. Netting, aroma and abscission of peduncle represent reticulatus group; a small fruit size, strong typical aroma and secondary colour distribution characterize dudaim group. Monoecy, very long fruit shape and mature fruit rind colour discriminate flexuous group; Ovary index, fruit size and flesh width distinguish subsp. agrestis group. These findings indicated wide range of variations for investigated characteristics in Turkish gene pool that provides a good source of diversity to use in melon improvement program for better yield and other traits of interest.

Fast Detection of Illegal Sweeteners in Liquor and Wine by Laser Raman Spectroscopy

Efficient method was proposed for the qualitative detection of 6 illegal sweeteners in commercial liquor and wine by laser Raman spectroscopy. The method was based on the comparative analysis of Raman spectrogram of pure sweeteners and alcohol samples. For the 6 kinds of sweeteners tested, saccharin sodium, acesulfame potassium impurity and cyclamate, were present as additives in the samples. The results showed that the proposed method was fast and simple with high repeatability and sensitivity, and needed little samples and cost less. Keywords: Sweeteners, Qualitative Analysis, Micro-Laser Raman, Liquor and Wine

Development of sweetness sensor with selectivity to negatively charged high-potency sweeteners

Objectivetaste evaluation has been much in demand in the food, beverage and pharmaceutical industries. A taste-sensing system, which is an electronic tongue with “global selectivity,” is one of the methods used for objective taste evaluation. A taste sensor electrode responds to only one of the basic tastes (saltiness, sourness, sweetness, bitterness and umami) as a change in membrane potential caused by interactions with tastants. Sweet substances are compounds with diverse chemical structures and sizes. Since the taste-sensing system is a potentiometric measurement system using a change in membrane potential, three types of sweetness sensors are required, one for sweeteners with each type of electric charge (uncharged, positively charged and negatively charged). A sweetness sensor for uncharged sweeteners has been developed. Therefore, negatively charged sweeteners, such as saccharine sodium and acesulfame potassium, were chosen as the target substances in this study. We investigated the responses of various sensor membranes using a lipid and nine kinds of plasticizers to each basic taste sample. Furthermore, not only the selectivity of the membranes but also the concentration dependence of their response to sweeteners was investigated. As a result, one of the developed sensors showed responses of over 20mV to 5mM saccharine sodium and 10mM acesulfame potassium in CPA value measurement (CPA: change in membrane potential caused by adsorption). On the other hand, the sensor also showed nearly zero responses to other basic taste samples. In addition, saltiness was the only interfering taste, and the responses to target substances in relative value measurement were over 140mV. The developed sweetness sensor had high selectivity and concentration-dependent responses. Hence, we concluded that the sensor is suitable for use as a sweetness sensor for high-potency sweeteners with a negative electric charge.

Rapid and Sensitive Method for the Determination of Eight Food Additives in Red Wine by Ultra-performance Liquid Chromatography Tandem Mass Spectrometry

A simple and highly sensitive method for the simultaneous determination of eight water-soluble food additive residues, including acesulfame potassium (ACS-K), sodium cyclamate (CYC), sodium saccharin (SAC), tartrazine (E102), amaranthus red (E123), ponceau 4R (E124), sunset yellow (E110), and brilliant blue (E133), in red wine has been developed using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The compounds were separated using a BEH C18 column with a linear gradient elution program within 6 min. The mobile phase A was 10 mmol/L ammonium acetate solution and the mobile phase B was methanol-acetonitrile (v/v, 9:1) mixed organic solvent, which contained 0.002 % trifluoroacetic acid (TFA). The optimized method, based on MS/MS in multiple reaction monitoring (MRM) mode, gave good linearity, precision, and accuracy and limits of detection (LOD, S/N = 3) for the target analytes which ranged from 0.01 to 0.5 μg/L. The method was successfully applied to the analysis of red wines from different countries of origin.

Role of nitrification in the biodegradation of selected artificial sweetening agents in biological wastewater treatment process

The biodegradation of the six artificial sweetening agents including acesulfame (ACE), aspartame (ASP), cyclamate (CYC), neohesperidindihydrochalcone (NHDC), saccharin (SAC), and sucralose (SUC) by nitrifying activated sludge was first examined. Experimental results showed that ASP and NHDC were the most easily degradable compounds even in the control tests. CYC and SAC were efficiently biodegraded by the nitrifying activated sludge, whereas ACE and SUC were poorly removed. However, the biodegradation efficiencies of the ASs were increased with the increase in initial ammonium concentrations in the bioreactors. The association between nitrification and co-metabolic degradation was investigated and a linear relationship between nitrification rate and co-metabolic biodegradation rate was observed for the target artificial sweeteners (ASs). The contribution of heterotrophic microorganisms and autotrophic ammonia oxidizers in biodegradation of the ASs was elucidated, of which autotrophic ammonia oxidizers played an important role in the biodegradation of the ASs, particularly with regards to ACE and SUC.

Precise measurement for the purity of amino acid and peptide using quantitative nuclear magnetic resonance

Precise measurement for the purity of organic compounds will fundamentally improve the capabilities and measurement services of the organic chemical analysis. Quantitative nuclear magnetic resonance (qNMR) is an important method to assess the purity of organic compounds. We presented a precise measurement method for the purity of small molecule with identification of impurities. In addition, the qNMR was rarely applied to purity of large compounds such as peptide, for which qNMR peaks are too crowded. Other than general idea of qNMR, we removed unwanted exchangeable peaks by proton exchange, as a new approach for qNMR, to make the quantitative protons of peptide isolated, which can ensure precise measurement. Moreover, a suitable internal standard, acesulfame potassium, was applied. The analytes were valine and peptide T5, due to their importance for protein analysis. For valine, the intraday CV was 0.052%, and the interday CV during 8 months was 0.071%. For peptide T5, simpler operation, shorter analytical time (1h vs. 3 days) and smaller CV (0.36% vs. 0.93%) were achieved by qNMR, compared with a traditional method (amino acid based isotope labeled mass spectrometry) via a hydrolysis reaction. This method has greatly increased the quantitative precision of qNMR for small compounds, and extended application scope of qNMR from small compounds to peptides.

Comprehensive Evaluation of Palatability for Commercial Medicine by Taste Sensing System

The bitterness of 10 basic medicines was evaluated using a multichannel taste sensor (Intelligent Sensor Technology). Three variables were obtained from the taste sensor data: sensor output (S), the change of membrane potential caused by adsorption, corresponding to aftertaste (C), and the ratio C/S. These variables were used to predict an estimated bitterness score in multiple regression analysis. There was correlation between the bitterness score predicted by the taste sensor and the score obtained by human gustatory sensation. The method showed quantitative predictability for the evaluation of bitterness. Secondly, bitterness intensities of eight H1-antihistamines were assessed by comparing the Euclidean distances between the drug and water using taste sensing system Astree II (α-MOS). Two sensors were ultimately selected as best suited to bitterness evaluation, and the data obtained from the two sensors depicted the actual taste map of the eight drugs. Also the bitterness masking efficiency of epinastine hydrochloride with acesulfame potassium was successfully predicted. Finally, Vesicare(®) tablets, whose main component is solifenacin succinate, are known to be extremely bitter. Recently, Vesicare(®) orally disintegrating tablets (ODTs), which contain a salting-out taste-masking system, have appeared on the market. To evaluate the effect of crushing on the bitterness of the tablets, Vesicare(®) ODTs and conventional Vesicare(®) tablets (CTs) were crushed either heavily or lightly. The bitterness scores of sample solutions were predicted using taste sensor. The lightly crushed ODT was predicted to be less bitter than CT. When tablets must be crushed, it is strongly recommended that ODT be crushed gently.

Detection of 10 sweeteners in various foods by liquid chromatography/tandem mass spectrometry

The analytical method for sweeteners in various food matrixes is very important for food quality control and regulation enforcement. A simple and rapid method for the simultaneous determination of 10 sweeteners [acesulfame potassium (ACS-K), aspartame (ASP), cyclamate (CYC), dulcin (DUL), glycyrrhizic acid (GA), neotame (NEO), neohesperidin dihydrochalcone (NHDC), saccharin (SAC), sucralose (SCL), and stevioside (STV)] in various foods by liquid chromatography/tandem mass chromatography (LC–MS/MS) was developed. The chromatographic separation was performed on a Phenomenex Luna Phenyl-Hexyl (5 μm, 4.6 mm × 150 mm) column with gradient elution of 10 mM ammonium acetate in water and 10 mM ammonium acetate in methanol. The recoveries of the 10 sweeteners were between 75% and 120%, and the coefficients of variation were less than 20%. The limits of quantification were 0.5 μg/kg for NHDC and SCL. For the other sweeteners, the limits of quantification were 0.1 μg/kg. Compared to the traditional high-performance liquid chromatography method, the LC–MS/MS method could provide better sensitivity, higher throughput, enhanced specificity, and more sweeteners analyzed in a single run. The samples included 27 beverages (16 alcoholic and 11 nonalcoholic beverages) and 15 pickled foods (1 pickled pepper, 3 candies, and 11 candied fruits). Two remanufactured wines were found to contain 7.2, 8.5 μg/g SAC and 126.5, 123 μg/g CYC, respectively. ACS-K, ASP, SCL, and NEO were detected in five beverages and drinks. The pickled peppers and candied fruits were found to contain SAC, GA, CYC, ASP, STV, NEO, and ACS-K. The wine with sweeteners detected was remanufactured wine, not naturally fermented wine. Therefore, the ingredient label for the sweeteners of remanufactured wine should be regulated by the proper authority for inspection of sweeteners.

Sweet Taste-Sensing Receptors Expressed in Pancreatic β-Cells: Sweet Molecules Act as Biased Agonists.

The sweet taste receptors present in the taste buds are heterodimers comprised of T1R2 and T1R3. This receptor is also expressed in pancreatic β-cells. When the expression of receptor subunits is determined in β-cells by quantitative reverse transcription polymerase chain reaction, the mRNA expression level of T1R2 is extremely low compared to that of T1R3. In fact, the expression of T1R2 is undetectable at the protein level. Furthermore, knockdown of T1R2 does not affect the effect of sweet molecules, whereas knockdown of T1R3 markedly attenuates the effect of sweet molecules. Consequently, a homodimer of T1R3 functions as a receptor sensing sweet molecules in β-cells, which we designate as sweet taste-sensing receptors (STSRs). Various sweet molecules activate STSR in β-cells and augment insulin secretion. With regard to intracellular signals, sweet molecules act on STSRs and increase cytoplasmic Ca2+ and/or cyclic AMP (cAMP). Specifically, when an STSR is stimulated by one of four different sweet molecules (sucralose, acesulfame potassium, sodium saccharin, or glycyrrhizin), distinct signaling pathways are activated. Patterns of changes in cytoplasmic Ca2+ and/or cAMP induced by these sweet molecules are all different from each other. Hence, sweet molecules activate STSRs by acting as biased agonists.

食品中の甘味料分析における迅速な改良透析法の開発

A simple and rapid dialysis method was developed for the extraction and purification of four artificial sweeteners, namely, sodium saccharin (Sa), acesulfame potassium (AK), aspartame (APM), and dulcin (Du), which are present in various foods. Conventional dialysis uses a membrane dialysis tube approximately 15 cm in length and is carried out over many hours owing to the small membrane area and owing to inefficient mixing. In particular, processed cereal products such as cookies required treatment for 48 hours to obtain satisfactory recovery of the compounds. By increasing the tube length to 55 cm and introducing efficient mixing by inversion at half-hour intervals, the dialysis times of the four artificial sweeteners, spiked at 0.1 g/kg in the cookie, were shortened to 4 hours. Recovery yields of 88.9-103.2% were obtained by using the improved method, whereas recovery yields were low (65.5-82.0%) by the conventional method. Recovery yields (%) of Sa, AK, APM, and Du, spiked at 0.1 g/kg in various foods, were 91.6-100.1, 93.9-100.1, 86.7-100.0 and 88.7-104.7 using the improved method.