Simultaneous determination of aspartame,acesulfame-K,saccharin, citric acid and sodium benzoate in various food products using HPLC–CAD–UV/DAD

A newly developed method for simultaneous determination of aspartame, acesulfame-K, saccharin, citric acid and sodium benzoate in various diet supplements and non-alcoholic beverages in a single run is presented. The analytes were analysed by high-performance liquid chromatography coupled to a charged aerosol (Corona CAD) and ultraviolet–diode array detectors simultaneously connected in series. Mass spectrometer MicrOTOF-QII from Bruker Daltonik (Bremen, Germany) was used to obtain the mass spectra for peak identifications. The method was validated using a Thermo Hypersil Gold-C18 column packed with 5 μm shell particles (150 × 4.6 mm) and methanol–water with 0.05 % TFA gradient mobile phase at a flow rate of 0.80 mL/min. The elaborated method was validated for linearity, precision and accuracy. The analytical results obtained in the validation study were highly satisfying; the recoveries for the analytes studied ranged between 98.1 and 101 % and the precision values from 0.11 to 1.73 %. By these procedures, the three sweeteners (aspartame, acesulfame-K and saccharin), citric acid and sodium benzoate could be well separated and quantitatively determined in varied food products. Hundred millilitres of soft drinks contained on average 5.50 mg of aspartame, 6.38 mg of saccharin, 8.94 mg of acesulfame-K, 9.05 mg of sodium benzoate and 111 mg of citric acid. Citric acid was the most abundant additive in all the samples analysed except for table sweeteners, and its highest concentration was determined in diet supplements, i.e. 347 mg/g. The percentage of adequate daily intake realisation in case of all additives is lower than 10 %, except for sodium benzoate in isotonic drinks (10.1 %).     

Intake of saccharin, aspartame, acesulfame K and cyclamate in Italian teenagers: present levels and projections

The intake of saccharin, aspartame, acesulfame K and cyclamate was assessed in 212 Italian teenagers aged 13-19 in 1996. Total daily intake of intense sweeteners was assessed on the basis of dietary records (14 consecutive days). The sweetener content of sugar-free products (soft drinks, candies, chewing gums, yoghurts, jam and table-top sweeteners) was provided by manufacturers. Sugar-free products were consumed by 77% of the subjects. Mean daily intake among consumers was 0.24mg/kg body weight (bw) for cyclamate (13 subjects), 0.21mg/kg bw for saccharin (9 subjects), 0.03mg/kg bw for aspartame (162 subjects), and 0.02mg/kg bw for acesulfame K (56 subjects). No subject exceeded the ADI (Acceptable Daily Intake) of an intense sweetener. Projections based on the present levels of use of intense sweeteners in sugar-free products and on the dietary pattern observed in the sample suggest that approaching the ADI could be possible only if subjects with high intakes of both soft drinks and table-top sugar substituted these items with respectively sugar-free beverages and table-top sweeteners containing either saccharin or cyclamate.     

超高效液相色谱-质谱联用法(UPLC-MS/MS)同时测定白酒中的微量甜味剂

以超纯水为提取溶剂,采用甲醇-0.02 mol/L乙酸铵溶液的流动相体系和负离子电离模式,用超高效液相色谱-三重四极杆质谱联用仪测定白酒中的安赛蜜、糖精钠、甜蜜素和三氯蔗糖。结果表明,安赛蜜、糖精钠、甜蜜素和三氯蔗糖线性范围在10～1000 ng/mL内R2大于0.9983,检出限分别为0.024 mg/kg、0.06 mg/kg、0.03 mg/kg、0.1 mg/kg,加标回收率大于82%,RSD小于2.01%。该方法简便灵敏,准确快速,可用于白酒中微量甜味剂的快速测定。     

Rapid separation of soft drinks ingredients using high performance liquid chromatography

A method is described which uses high performance liquid chromatography in the reverse phase mode to separate amaranth, quinoline yellow, quinine sulphate, sunset yellow, caffeine, aspartame, saccharin, vanillin, sorbic acid, benzoic acid and green S in 4 min in samples of soft drinks. The final separation was achieved using 17·5% acetonitrile, 12·5% methanol, 70% buffer (0·85% v/v sulphuric acid and 17·5% mm potassium dihydrogen orthophosphate in water with the pH adjusted to 1·8), with a flow rate of 1·35 ml/min through a 100 mm × 4·6 mm column containing 3 μm Spherisorb octylsilane. The octylsilane material is fully capped with monolayer coverage and a carbon loading of 6% w/w, pore diameter of 8 nm and a surface area of 220 m²/g. This material was used in preference to the more widely used octadecylsilane as it provides shorter analysis time and allows for the use of more polar eluting solvents. Detection was at 220 nm, 0·1 AUFSD with an injection of 5 μl samples using a rotary injection valve. The standard solution concentration was 100 mg litre⁻¹ for aspartame; 20 mg litre⁻¹ for quinine sulphate, caffeine, vanillin, sorbic acid and benzoic acid and 5 mg litre⁻¹ for amaranth, quinoline yellow, sunset yellow, green S, tartrazine, indigo carmine and carmoisine.     

白酒中爱德万甜及其他甜味剂的检测方法研究

该研究建立了同时检测白酒中新型甜味剂爱德万甜和安赛蜜、糖精钠、阿斯巴甜、阿力甜、纽甜的高效液相色谱多通道法。结 果表明,试样经70 ℃水浴蒸干、2 mL纯水复溶,水相滤膜过滤进样,以20 mmol/L乙酸铵和乙腈为流动相,梯度洗脱,C18柱分离,二极 管阵列检测器（DAD）检测,采用外标法定量。 阿斯巴甜、阿力甜在1.0～50 μg/mL范围内线性关系良好（相关系数R2均＞0.999 5）;爱 德万甜、安赛蜜、糖精钠和纽甜在0.1～50 μg/mL范围内线性关系良好（相关系数R2均＞0.999 9）。 爱德万甜、纽甜、安赛蜜、糖精钠检 出限为0.02 mg/kg,阿斯巴甜、阿力甜检出限为0.2 mg/kg。 相对标准偏差（RSD）＜0.5%;加标回收率为98.35%～105.25%;表明该方法 准确度、精密度良好。     

Simultaneous Determination of Caffeine and Aspartame in Diet Supplements and Non-Alcoholic Beverages Using Liquid-Chromatography Coupled to Corona CAD and UV-DAD Detectors

A method for the simultaneous determination of caffeine and aspartame in various diet supplements and non-alcoholic beverages is presented. The analytes were analysed by high-performance liquid chromatography coupled to a charged aerosol detector (Corona CAD) and ultraviolet-diode array detector (UV-DAD) simultaneously connected in series. The method was validated using a Thermo Hypersil Gold-C18 column packed with 5 μm shell particles (150 × 4.6 mm) and acetonitrile–water (15/85% v/v) mobile phase at a flow rate of 1.00 mL/min. The elaborated method was validated for linearity, precision and accuracy. The rapid HPLC–CAD–UV-DAD technique is suitable for quantifying caffeine and aspartame in the range of 0.25–75 μg/mL in diet supplements and non-alcoholic beverages. The limit of detection for caffeine and aspartame was 62 and 43 ng/mL for Corona CAD and 31 and 30 ng/mL for UV-DAD detector, respectively. Each analyte calibration curve had a correlation coefficient of at least 0.999 and was linear in the defined range. The accuracies of CAD and UV-DAD detection were all acceptable, with the mean value of 100% for aspartame and 98.3% for caffeine. Precision values ranged from 0.09% to 1.12%. The work has demonstrated that charged aerosol detector can be successfully used in a rapid screening technique for biologically active substances in non-alcoholic beverages and diet supplements.     

HPLC-MS/MS analysis of 9 artificial sweeteners in imported foods

An analytical method for the simultaneous determination of 9 artificial sweeteners (acesulfame-K, cyclamate, sodium saccharin, aspartame, alitame, neotame, sucralose, dulcin, and neohesperidine dihydrochalcone) in imported foods by HPLC-MS/MS was developed. Samples were extracted with buffer solution (pH 4.5). The supernatant was analyzed by HPLC-MS/MS after centrifugation and filtration. The sample was separated on a thermo hypersil BPS C₁₈ (250×3 mm, 5-μm) column, and detected by MS/MS with selective reaction monitoring (SRM) mode. The correlation coefficients (r ²) of the calibration curve were >0.99. The recoveries were 90.0–107.5% with good coefficients of variation of 1.8–8.6%. The limits of detection and limits of quantification were between 0.001 and 0.375 mg/mL and between 0.003 and 1.125 mg/mL, respectively. The proposed method has been successfully applied to the determination of the 9 sweeteners in various foods.

     

Metabolic effects of aspartame in adulthood: A systematic review and meta-analysis of randomized clinical trials

Data about harms or benefits associated with the consumption of aspartame, a nonnutritive sweetener worldwide consumed, are still controversial. This systematic review and meta-analysis of randomized controlled clinical trials aimed to assess the effect of aspartame consumption on metabolic parameters related to diabetes and obesity. The search was performed on Cochrane, LILACS, PubMed, SCOPUS, Web of Science databases, and on a gray literature using Open Grey, Google Scholar, and ProQuest Dissertations &; Theses Global. Searches across all databases were conducted from the earliest available date up to April 13, 2016, without date and language restrictions. Pooled mean differences were calculated using a random or fixed-effects model for heterogeneous and homogenous studies, respectively. Twenty-nine articles were included in qualitative synthesis and twelve, presenting numeric results, were used in meta-analysis. Fasting blood glucose (mmol/L), insulin levels (μU/mL), total cholesterol (mmol/L), triglycerides concentrations (mmol/L), high-density lipoprotein cholesterol (mmol/L), body weight (kg), and energy intake (MJ) were considered as the main outcomes in subjects that consumed aspartame, and results were presented as mean difference; % confidence interval, range. Aspartame consumption was not associated with alterations on blood glucose levels compared to control (?0.03 mmol/L; 95% CI, ?0.21 to 0.14) or to sucrose (0.31 mmol/L; 95% CI, ?0.05 to 0.67) and on insulin levels compared to control (0.13 μU/mL; 95% CI, ?0.69 to 0.95) or to sucrose (2.54 μU/mL; 95% CI, ?6.29 to 11.37). Total cholesterol was not affected by aspartame consumption compared to control (?0.02 mmol/L; 95% CI, ?0.31 to 0.27) or to sucrose (?0.24 mmol/L; 95% CI, ?0.89 to 0.42). Triglycerides concentrations were not affected by aspartame consumption compared to control (0.00 mmol/L; 95% CI, ?0.04 to 0.05) or to sucrose (0.00 mmol/L; 95% CI, ?0.09 to 0.09). High-density lipoprotein cholesterol serum levels were higher on aspartame compared to control (?0.03 mmol/L; 95% CI, ?0.06 to ?0.01) and lower on aspartame compared to sucrose (0.05 mmol/L; 95% CI, 0.02 to 0.09). Body weight did not change after aspartame consumption compared to control (5.00 kg; 95% CI, ?1.56 to 11.56) or to sucrose (3.78 kg; 95% CI, ?2.18 to 9.74). Energy intake was not altered by aspartame consumption compared to control (?0.49 MJ; 95% CI, ?1.21 to 0.22) or to sucrose (?0.17 MJ; 95% CI, ?2.03 to 1.69). Data concerning effects of aspartame on main metabolic variables associated to diabetes and obesity do not support a beneficial related to its consumption.     

超高效液相色谱串联质谱同时测定酒类产品中多种人工合成甜味剂

建立超高效液相色谱-电喷雾电离串联质谱（ultra performance liquid chromatography-electro spray ionizationtandem mass spectrometry,UPLC-ESI-MS-MS）同时测定酒类产品中安赛蜜、糖精钠、甜蜜素、三氯蔗糖、阿斯巴甜、阿力甜和纽甜人工合成甜味剂的分析方法。方法采用ZORBAX Eclipse XDB-C18柱（100 mm×4.6 mm,1.8 μm）色谱柱,柱温50 ℃。流动相由A（甲醇）和B（体积分数0.1%甲酸和20 mmol/L甲酸铵溶液,pH 4.0）组成,流速为0.5 mL/min,梯度洗脱。在ESI负离子模式下,采用多反应监测模式进行测定,可以在12.5 min内完成7 种人工合成甜味剂的检测。安赛蜜、糖精钠、甜蜜素、三氯蔗糖、阿斯巴甜、阿力甜和纽甜在10～5 000 μg/L范围内有良好的线性关系,相关系数大于0.999。安赛蜜、糖精钠、甜蜜素、三氯蔗糖、阿斯巴甜、阿力甜和纽甜检出限分别为4、6、4、8、4、4、2 μg/L,回收率在88.3%～98.3%之间,相对标准偏差为2.1%～6.7%。该方法快速、准确,灵敏度高,适用于酒类产品中低含量人工合成甜味剂的测定。     

Caffeine intake and its sources: A review of national representative studies

Aim of this review is to summarize current daily caffeine intake of children, adolescents, and adults, and trends in caffeine intake over the past decade. A literature search was conducted (1997–2015) which yielded 18 reports on nationally representative studies, describing caffeine consumption of over 275,000 children, adolescents and adults. The data revealed that mean total daily caffeine intake in children, adolescents, and adults is below caffeine intake recommendations such as those stated by Health Canada (2.5 mg/kg bw/day for children and adolescents, and 400 mg/day for adults) and the European Food Safety Authority, EFSA (3 mg/kg bw/day for children and adolescents, and 400 mg/day for adults). Total daily caffeine intake has remained stable in the last 10–15 years, and coffee, tea and soft drinks are the most important caffeine sources. Across all age groups, energy drinks contribute little to total caffeine intake. The highest potential for reducing daily caffeine intake is by limiting coffee consumption, and in some countries and age groups, by reducing tea and soft drink consumption.     

Sensory Studies of High Potency Multiple Sweetener Systems for Shortbread Cookies with and without Polydextrose

Time-intensity (TI) sweetness and bitterness curves determined for six potent sweetener combinations with and without polydextrose were compared to sucrose in shortbread cookies. Hardness, fracturability and cohesiveness of shortbreads were determined. Sweetener combinations of aspartame/cyclamate, aspartame/cyclamate/saccharin, acesulfame K/saccharin, aspartame/saccharin/acesulfame K, acesulfame K/aspartame and aspartame/saccharin gave sweetener TI profiles similar to that of sucrose. Bitterness TI profiles were similar for all potent sweetener combinations but higher than for sucrose. Polydextrose increased hardness and fracturability and decreased cohesiveness of cookies compared to those made with high potency sweeteners without polydextrose. Textural characteristics of polydextrose cookies approached those of sucrose shortbreads.     

A deep eutectic solvent based liquid phase microextraction for the determination of caffeine in Turkish coffee samples by HPLC-UV

ABSTRACT

A novel liquid-phase microextraction procedure for the determination and extraction of caffeine using a deep eutectic solvent based liquid-phase microextraction method (DES-LPME) was developed. A deep eutectic solvent consisting of choline chloride-phenol (1:3) and tetrahydrofuran as a dilution solvent were used for the extraction of caffeine from Turkish coffee samples. The quantitative recoveries were obtained when the DES volume was 400 µL, and the volume of tetrahydrofuran that was used as an emulsifier solvent was 800 µL. The limit of detection and the limit of quantification were found to be 0.12 and 0.40 µg mL^?1, respectively. Linear dynamic range was 0.5–100 µg mL^?1, and coefficient of determination (R²) was 0.9998. The relative standard deviation (RSD) was 2.20% when the standard solution concentration was 1.0 µg mL^?1. The fact that the determination coefficient obtained from the calibration curve was greater than 0.9998 gives information about the accuracy of the method. Also, in order to determine the accuracy of the developed method, selected coffee samples were spiked with 25 and 50 µg mL^?1 caffeine.     

Racemization of amino acids during classical and microwave oven hydrolysis — application to aspartame and a Maillard reaction system

The conventional acid hydrolysis routinely used in protein analysis induces partial racemization of the amino acids. An alternative method, using microwave heating, was evaluated with the aim of assessing its efficiency in preserving the amino acids in their original configuration. The method was employed on the dipeptide aspartame. The results showed that some advantages, implying less racemization, could be gained by using the microwave technique, but the method was applicable only at temperatures under 150°C. Mild, conventional hot oven-hydrolysis, involving a shorter hydrolysis time, could have the same mild effect. The different techniques were applied to aspartame treated at different pH, a soft drink and a Maillard reaction model system. The results showed that aspartame was insusceptible to racemization in acid and neutral environments, and not until very high pH did racemization start. The soft drink contained a small amount of d-amino acids, 0.27 mg d-asp and 0.14 mg d-phe/100 ml. The model system exhibited no racemization.     

钛胶柱反相高效液相色谱法测定饮料中的阿斯巴甜

建立基于钛胶柱对无糖可乐中甜味剂阿斯巴甜的高效液相色谱快速测定方法。色谱条件：分离柱为Titania Sachtopore-RP柱(250mm×4.6m,5μm),柱温70℃,流动相为V[15mmol/L磷酸铵缓冲溶液,(NH4)2HPO4:NH4H2PO4＝6:4;10mmol/L氟化铵]-V(甲醇)＝70:30,流速1mL/min,检测波长217nm。线性范围为10～2500μg/mL,最低检出限为0.08μg/mL,回收率为92.5%～97.4%,相对标准偏差(n＝8)小于0.72%。本法准确度和精密度均可满足饮料中阿斯巴甜的测定要求。     

Estimated intake of intense sweeteners from non-alcoholic beverages in Denmark, 2005

In 2005 International Standards Organization. ISO 17025. 2005. General requirements for the competence of testing and calibration laboratories, Copenhagen: Danish Standard [Google Scholar], 76 out of 177 analysed samples of non-alcoholic beverages were found to contain the intense sweeteners cyclamate, acesulfame-K, aspartame, and saccharin. The content of cyclamate did not exceed the now permitted maximum level in the European Union of 250 mg l^?1 in soft drinks. The estimated intake of the sweeteners was calculated using the Danish Dietary Survey based on 3098 persons aged 1–80 years. The estimated intake with 90th percentiles of 0.7, 0.8 and 0.2 mg kg^?1 body weight day^?1 for acesulfame-K, aspartame, and saccharin, respectively, was much lower than the acceptable daily intake values of 15, 40, 7, and 2.5 mg kg^?1 body weight day^?1 for acesulfame-K, aspartame, and saccharin, respectively, and on the same level as in the similar investigation from 1999. In contrast to the 1999 investigation, the 90th percentile of the estimated cyclamate intake in 1–3 year olds with 3.7 mg kg^?1 body weight day^?1 was in 2005 lower than the acceptable daily intake of 7 mg kg^?1 body weight day^?1. However, the 99th percentile for 1–3 year olds with 7.4 mg kg^?1 body weight day^?1 still exceeded the acceptable daily intake slightly. The 90th percentile for the whole population with 0.9 mg kg^?1 body weight day^?1 was halved compared with 1999. The reduction in the European Union of the maximum permitted level for cyclamate from 400 to 250 mg l^?1 has brought the intake of cyclamate in small children down to well below the acceptable daily intake value.     

液相色谱-串联质谱法同时测定食品中9种甜味剂

建立了高效液相色谱-串联质谱法同时测定食品中9种甜味剂(安赛蜜、糖精钠、甜蜜素、阿斯巴甜、三氯蔗糖、阿力甜、甜菊糖苷、爱德万甜和纽甜)含量的分析方法。样品经超声、离心等前处理后过0.22μm滤膜后上机分析。采用AQ-C18(100 mm×2.1 mm×3μm)为分离柱,c(乙酸铵)为20 mmol/L的水溶液-φ(甲酸)为0.1%的乙腈溶液为流动相,梯度洗脱,在ESI负离子模式下,采用多反应监测模式进行测定,以质量数和保留时间定性、峰面积定量。结果表明,9种甜味剂在工作曲线范围内相关系数均大于0.9990,方法检出限在0.020~0.055 mg/kg,食品加标回收率为90.6%~111.5%,其相对标准偏差(n=6)均小于6.0%。应用此方法对食品(白酒、饮料、果冻、酸奶、糖果)中9种甜味剂进行检测,前处理步骤简单,测定快速、准确度和灵敏度高,能满足国家标准对食品中9种甜味剂的分析要求,与国家标准方法相比,提高了分析效率。     

白酒和调香剂中4 种人工合成甜味剂的UPLC-MS-MS 测定

目的：建立白酒及调香剂中人工合成的非营养性甜味剂--甜蜜素、糖精钠、安赛蜜、阿斯巴甜的超高效液相色谱- 串联质谱检测法(UPLC-MS-MS),并用于市售白酒和调香剂中4 种甜味剂的测定。方法：采用WatersACQUITY UPLC BEH C18 色谱柱(50mm × 2.1mm,1.7μm),以乙腈-0.01g/100mL 氨水为流动相,梯度洗脱,流速0.2mL/min,待测物经电喷雾离子源负离子化后,用三重四级杆串联质谱仪的多反应离子监测(MRM)方式进行定量分析。结果：4 种甜味剂质量浓度在10～1000μg/L 范围内均与峰面积呈良好线性关系(r ≥ 0.997),检出限为0.03～0.35μg/L,平均加标回收率在75.2%～115.3% 之间,相对标准偏差在8% 以内。市售白酒和调香剂中甜蜜素、糖精钠和阿斯巴甜的含量分别为0～287、0～67.2、0～104.8μg/L,样品中没有发现安赛蜜。结论：本方法快速简便、检出限低,适合于白酒和调香剂中微量甜味剂的同时检测。     

SWEETNESS EQUIVALENCE OF DIFFERENT SWEETENERS IN STRAWBERRY‐FLAVORED YOGURT

ABSTRACT

To successfully substitute sucrose for sweeteners, further studies must be carried out based on previous knowledge of sweetener concentration to determine the equivalent sweetness of such compounds. In this work, sweetness equivalence of strawberry‐flavored yogurt with different sweeteners and/or their combinations (aspartame, acesulfame‐K, cyclamate, saccharin, stevia and sucralose) and yogurt sweetened with 11.5% w/w sucrose was measured using the sensory magnitude estimation method. The sweetness concentrations equivalent to strawberry yogurt sweetened with 11.5% w/w sucrose in the tested sweeteners were 0.072% w/w for aspartame, 0.042% w/w for aspartame/acesulfame‐K (2:1), 0.064% w/w for cyclamate/saccharin, 0.043% w/w for cyclamate/saccharin (2:1)/stevia (1.8:1) and 0.30% w/w for sucralose. These results can promote the use of different sweetener combinations in strawberry‐flavored yogurt, specially acesulfame‐K and stevia, once they produce more pleasing in this product.

PRACTICAL APPLICATIONS

This study provides some useful information, since there is no data in the literature about sweetness equivalence of sweeteners in yogurt, but only in simpler matrices such as pure water, juices, coffee and teas. The use of stevia blend presented several advantages such as increased sweetening power, demonstrating the potential of this natural sweetener. The magnitude estimation method has been successful in this study, being an important tool for development of new low‐calorie products. It may be noted that when evaluating different types of food using the same kinds of sweeteners, these promote distinct characteristics and that reflect directly on the sensory quality of the final product. Thus, such studies generate important information for the food industries working with dietetic food.     

Assessment of Korean consumer exposure to sodium saccharin,aspartame and stevioside

The dietary intakes of sodium saccharin, aspartame and stevioside were estimated on the basis of food consumption data of the Korean consumer and the concentration of sweeteners in processed foods. Results were compared with the acceptable daily intake (ADI) of sweeteners. Among the 28 food categories for which the application of sodium saccharin, aspartame and stevioside is permitted in Korea, they were detected in 5, 12 and 13 categories, respectively. The estimated daily intake (EDI) of sodium saccharin and aspartame were high in infants and children, whereas the EDI of stevioside was high in adolescents and adults. The most highly consumed sweetener was aspartame, and the highest EDI/ADI ratio was found for sodium saccharin. The main food categories contributing to sweetener consumption were beverages, including alcoholic beverages. For most Korean consumers, the EDIs were no greater than 20% of their corresponding ADI; however, the EDI of sodium saccharin for conservative consumers aged 1–2 years reached 60% of their ADI.     

反相液相色谱同时测定糖品中的防腐剂和甜味剂

本文建立了用反相液相色谱同时测定糖品中的苯甲酸、山梨酸、脱氢乙酸、安赛蜜、糖精钠等防腐剂和甜味剂的方法。方法采用VenusilXBP—C18（5gm,4．6x250mm）反相色谱柱,流动相为20mmol／L乙酸铵一甲醇,流速为1．0mL／min,柱温为30℃,进样量为10gL。该方法的测定低限为1．0mg／L,线性范围为10～100mg／L,加标回收率为89．85％～103．60％,相对标准偏差为1．50％～3．26％（n=4）。结果表明,该方法简便、可靠,适合对糖品中防腐剂和甜味剂的同时检测。