HPLC analysis of aspartame and saccharin in pharmaceutical and dietary formulations

Summary A reversed-phase HPLC method has been developed suitable for a reliable quality control of pharmaceutical and dietary formulations containing the synthetic sweeteners aspartame and saccharin. The proposed method is able to separate acesulfame, aspartame and saccharin, and their impurities such as 5-benzyl-3,6-dioxo-2-piperazineacetic acid (the major degradation product of aspartame) and 4-sulphamoylbenzoic acid,o- andp-toluenesulphonamides (the synthesis impurities of saccharin). A convenient solid-phase extraction (SPE) procedure using C-18 sorbent, was also developed for the determination of potential saccharin impurities.     

Separation and simultaneous determination of four artificial sweeteners in food and beverages by ion chromatography

In this paper, the separation and determination of four artificial sweeteners (aspartame, sodium cyclamate, acesulfame-K and sodium saccharin) by ion chromatography coupled with suppressed conductivity detector is reported. The four artificial sweeteners were separated using KOH eluent generator. Due to the use of eluent generator, very low conductance background conductivity can be obtained and sensitivity of sweeteners has been greatly improved. Under the experimental condition, several inorganic anions, such as F-, Cl-, NO3-, NO2-, Br-, SO4(2)-, PO4(3)- and some organic acid such as formate, acetate, benzoate, and citrate did not interfere with the determination. With this method, good linear relationship, sensitivity and reproducibility were obtained. Detection limits of aspartame, sodium cyclamate, acesulfame-K, sodium saccharin were 0.87, 0.032, 0.019, 0.045 mg/L, respectively. Rate of recovery were between 98.23 and 105.42%, 99.48 and 103.57%, 97.96 and 103.23%, 98.46 and 102.40%, respectively. The method has successfully applied to the determination of the four sweeteners in drinks and preserved fruits.     

Flow injection potentiometric determination of saccharin in dietary products with relocation of filtration unit

A flow injection potentiometric procedure for saccharin determination in dietary products is proposed. Saccharin is precipitated as mercurous saccharinate and the excess of the mercurous cation is potentiometrically measured using a silver wire coated with a mercury film as the working electrode. A filter unit is used to avoid contact between the precipitate and the electrode surface. With relocation in the flow manifold, the accumulated precipitate is removed on-line. Sucrose, glucose, aspartame, sodium cyclamate and sodium benzoate do not interfere when present in amounts similar to those observed in commercial products. Results are comparable with those obtained by UV-spectrophotometry and the correlation coefficient between methods is equal to 0.9930. A linear relationship between DeltaE (mV) and the logarithm of saccharin concentration was obtained in the saccharin concentration range 2 x 10(-3) - 1 x 10(-2)M. The sampling frequency is 60/hour and only 0.76 mg of Hg(2+)(2) is consumed in each determination.     

Ion Selective Electrode for Potentiometric Determination of Saccharin Using a Thin Film of Silsesquioxane 3-n-Propylpyridinium Chloride Polymer Coated-Graphite Rod

ABSTRACT

A polymer coated graphite rod ion selective electrode for saccharin was constructed and evaluated for the determination of saccharin in artificial table top sweeteners. The polymer consists of a thin film of silsesquioxane 3-n-propylpyridinium chloride. The electrode response was based on the ion pair formed between saccharinate and the 3-n-propylpyridinium cation from the silsequioxane polymer. The electrode exhibits a Nernstian response for saccharin concentrations between 6.9×10^-6 and 5.3×10^-3 mol L^-1 and a detection limit of 5.5×10^-6 mol L^-1. The electrode response for saccharin was fast (10-20 s) and the potential independent of pH in the range of 3 to 7. The selectivity coefficients K _{A, B} ^pot for several anions usually present in commercial table top sweeteners were determined following the IUPAC recommendations. The potentiometric method with the ion selective electrode was validated by the HPLC reference method, through t8543226 determination of saccharin in commercial samples of table top sweeteners. The ion selective electrode is proved suitable for the routine quality control of table top sweeteners by potentiometry.     

Validation of a high-performance liquid chromatographic method for the simultaneous determination of tramadol and its impurities in oral drops as a pharmaceutical formulation

The novel, rapid high performance liquid chromatographic method for the determination of tramadol hydrochloride and its three impurities was developed and validated. The method can simultaneously assay potassium sorbate, used as preservative, and saccharin sodium, used as sweetener in tramadol pharmaceutical formulation. The separation was carried out on a C(18) XTerra (150 mm x 4.6 mm, 5 mm) column using acetonitrile-0.015 M Na(2)HPO(4) buffer (2:8, v/v) as mobile phase (pH value 3.0 was adjusted with orthophosphoric acid) at a flow rate 1.0 ml min(-1), temperature of the column 20 degrees C and UV detection at 218 nm. The method was found to be linear (r > 0.999) in the range of 0.05-0.8 mg ml(-1) for tramadol hydrochloride, 0.1-1.2 mg ml(-1) for impurities B and C and for impurity A (r > 0.995) in the range 0.15-2.4 mg ml(-1). The low RSD values indicate good precision and high recovery values indicate excellent accuracy of the HPLC method. Developed method was successfully applied to the determination of tramadol hydrochloride, its investigated impurities and potassium sorbate in commercial formulation. The recovery of tramadol hydrochloride was 98.25% and RSD was 1.80%. The method is rapid and sensitive enough to be used to analyse Trodon oral drops.     

Simultaneous determination of sweeteners and preservatives in preserved fruits by micellar electrokinetic capillary chromatography

A micellar electrokinetic capillary method for the simultaneous determination of the sweeteners dulcin, aspartame, saccharin, and acesulfame-K and the preservatives sorbic acid; benzoic acid; sodium dehydroacetate; and methyl-, ethyl-, propyl-, isopropyl-, butyl-, and isobutyl-p-hydroxybenzoate in preserved fruits is developed. These additives are ion-paired and extracted using sonication followed by solid-phase extraction from the sample. Separation is achieved using a 57-cm fused-silica capillary with a buffer comprised of 0.05 M sodium deoxycholate, 0.02 M borate-phosphate buffer (pH 8.6), and 5% acetonitrile, and the wavelength for detection is 214 nm. The average recovery rate for all sweeteners and preservatives is approximately 90% with good reproducibility, and the detection limits range from 10 to 25 microg/g. Fifty preserved fruit samples are analyzed for the content of sweeteners and preservatives. The sweeteners found in 28 samples was aspartame (0.17-11.59 g/kg) or saccharin (0.09-5.64 g/kg). Benzoic acid (0.02-1.72 g/kg) and sorbic acid (0.27-1.15 g/kg) were found as preservatives in 29 samples.     

A simple turbidimetric flow injection system for saccharin determination in sweetener products

A new method for saccharin determination in liquid sweetener products was developed. The method is based on the precipitation reaction of Ag(I) ions with saccharin in aqueous medium (pH 3.0), using a flow injection analysis system with merging zones, the suspension was stabilized with 5 g L⁻¹ Triton X-100. All experimental parameters influencing the flow injection system were optimized by means of chemometric approaches. The linear analytical curve was built from 2.4 g L⁻¹ up to 9.64 g L⁻¹ (r = 0.9968) with a quantification limit of 2.40 g L⁻¹. The precision assessed as relative standard deviation (n = 10) was found to be 1.75 % for the saccharin concentration of 7.20 g L⁻¹. Based on interference studies performed with the substances commonly found in liquid sweeteners, such as sodium cyclamate, methylparaben, sodium aspartame, and benzoic and citric acids, at the analyte to interferent mole ratio of up to 1: 10, no interference with the saccharin determination was observed. The presence of chloride ions interferes with the method, but a preceding liquid-liquid saccharin extraction with ethyl acetate was successfully employed to overcome this drawback. Accuracy of the method in sweetener products was evaluated by a comparison with the HPLC method.     

Determination of Saccharin in Plasma and Urine by High Performance Liquid Chromatography

Abstract

A sensitive and specific high performance liquid chromatographic (HPLC) assay for the determination of saccharin in plasma and urine was developed. Saccharin is extracted into diethyl ether at acid pH, evaporated, and reconstituted prior to instrumental analysis. Overall recovery of saccharin is 86.9 + 8.6% and the sensitivity limits of detection is 0.15 μg per ml of plasma or urine using a fluorescence detector. The sensitivity limit in plasma can be extended to 20 ng per ml by use of a 2 ml assay volume and detector attenuation. The assay was used for the determination of saccharin in plasma and urine of rats following oral doses of 5 mg/kg.     

Simultaneous determination of nine intense sweeteners in foodstuffs by high performance liquid chromatography and evaporative light scattering detection--development and single-laboratory validation

A high performance liquid chromatographic method with evaporative light scattering detection (HPLC-ELSD) has been developed for the simultaneous determination of multiple sweeteners, i.e., acesulfame-K, alitame, aspartame, cyclamic acid, dulcin, neotame, neohesperidine dihydrochalcone, saccharin and sucralose in carbonated and non-carbonated soft drinks, canned or bottled fruits and yoghurt. The procedure involves an extraction of the nine sweeteners with a buffer solution, sample clean-up using solid-phase extraction cartridges followed by an HPLC-ELSD analysis. The trueness of the method was satisfactory with recoveries ranging from 93 to 109% for concentration levels around the maximum usable dosages for authorised sweeteners and from 100 to 112% for unauthorised compounds at concentration levels close to the limit of quantification (LOQs). Precision measures showed mean repeatability values of <4% (expressed as relative standard deviation) for highly concentrated samples and <5% at concentration levels close to the LOQs. Intermediate precision was in most cases <8%. The limits of detection (LODs) were below 15 microg g(-1) and the LOQs below 30 microg g(-1) in all three matrices. Only dulcin showed slightly higher values, i.e., LODs around 30 microg g(-1) and LOQs around 50 microg g(-1)     

表面增强拉曼光谱法快速测定干果类食品中的糖精钠

建立了干果中糖精钠的表面增强拉曼光谱(SERS)快速分析方法。干果样品经水提取、C18固相萃取柱净化除杂后进行表面增强拉曼光谱分析。该方法的线性范围为50.0~250 mg/L,检出限为0.6 g/kg,回收率为80.0%~125%,相对标准偏差(RSD,n=5)不大于8.4%。结果表明该方法灵敏度高、杂质干扰小、准确度高,满足干果类食品中糖精钠的快速检测要求,在干果类食品安全质量监控方面具有良好的应用潜力。     

Potentiometric Membrane Sensor for Determination of Saccharin

A potentiometric poly(vinyl chloride) membrane sensor for determination of saccharin is described. It is based on the use of Aliquat 336S-saccharinateion-pair as an electroactive material in plasticized PVC membranes with o-nitrophenyloctylether or dioctylphthalate. The sensor is conditioned for at least two days in 0.1 mol L⁻¹ sodium saccharinate before use. It exhibits fast, stable and Nernstian response for saccharinate ions over the concentration range of 1.0 × 10⁻¹–5.0 × 10⁻⁵ mol L⁻¹ and pH range of 4.5–11. The sensor is used for determination of saccharin in some dosage forms. Results with an average recovery of 101% and a mean standard deviation of 0.2% are obtained which is compared favourably with data obtained using the British pharmacopoeia method. The sensor shows reasonable selectivity towards saccharin in presence of many anions and natural sweeteners.     

高效液相色谱/二极管阵列法快速测定白酒、配制酒与葡萄酒中5种人工合成甜味剂

建立了一种快速检测白酒、配制酒和葡萄酒中安赛蜜、糖精钠、阿斯巴甜、阿力甜及纽甜5种人工合成甜味剂的高效液相色谱分析方法。采用C_(18)柱为分离柱,对流动相的组成和洗脱方式、检测波长等参数进行优化。结果表明,以乙腈和0.02 mol/L硫酸铵(p H 4.4)溶液为流动相,梯度洗脱,柱温40℃,可使5种人工合成甜味剂在15 min内实现基线分离。最佳检测波长下,5种甜味剂在4~200 mg/L浓度范围内呈良好的线性关系,相关系数均大于0.999。样品加标回收率为95.2%~103.2%,相对标准偏差(n=5)均不大于3.4%。该方法简便、快捷、准确、灵敏度较高,适用于白酒、配制酒和葡萄酒等各类酒中5种人工合成甜味剂的快速检测。     

Spectrophotometric Determination of Saccharin in Soft Drinks and Pharmaceuticals

Abstract

A simple, rapid and sensitive spectrophotometric method has been developed for the determination of saccharin using azure C. Saccharin reacts quantitatively with azure C in citric acid buffer forming a blue product. The blue product can be extracted into chloroform, Beer's law is valid over the concentration range 3–75 ppm of saccharin and the molar absorptivity is 3.1 × 10³ 1 mol^?1 cm^?1. Reasonable amounts of ingredients that are likely to be present in soft drinks and pharmaceuticals do not interfere. Recoveries of saccharin from soft drinks and pharmaceuticals were satisfactory.     

比值导数波谱-多波长最小一乘回归分光光度法同时测定饮料中糖精钠、苯甲酸和山梨酸

提出了比值导数波谱－多波长最小一乘回归分光光度法。该法能同时测定三组分混合物而毋需零交点波长和等吸收点，且回归稳健性好．可消除异常点的影响。方法用于合成样品和饮料中糖精钠、苯甲酸和山梨酸的同时测定，相对误差＜± ３． ５％，回收率为 ９６． ６％～１０４．４％，相对标准偏差为２．０％～３．１％．     

高效液相色谱-串联质谱法测定食品中6种人工合成甜味剂

建立了食品中6种人工合成甜味剂(甜蜜素、糖精钠、安赛蜜、阿斯巴甜、阿力甜、纽甜)的高效液相色谱-串联质谱检测方法。样品经甲醇-水溶液(1:1, v/v)提取,以C18柱为分离柱,0.1%(v/v)甲酸-5 mmol/L甲酸铵溶液/乙腈为流动相,经高效液相色谱分离,采用电喷雾串联四极杆质谱进行检测。结果表明,6种人工合成甜味剂在20～500 μg/L范围内定量离子对的响应峰面积和样品质量浓度之间有良好的线性关系(相关系数＞0.998)。在3个添加水平下,样品平均回收率为81.3%～106.0%,相对标准偏差小于11%。该方法简单、灵敏、准确,可用于食品中6种人工合成甜味剂的同时检测。     

HPLC-Bestimmung polarer Verunreinigungen in Saccharin und Saccharin-Natrium

Zusammenfassung Eine HPLC-Methode wird beschrieben zur direkten Bestimmung polarer Verunreinigungen in Saccharin und Saccharin-Natrium, o- und p-Sulfobenzoesäure sowie o- und p-Sulfamoylbenzoesäure lassen sich bis zu einer Nachweisgrenze von 1 mg/kg bestimmen. Für eine Konzentration von 10 mg/kg beträgt die Standardabweichung 0,4 mg/kg.Eine tabellarische Zusammenfassung der Ergebnisse von handelsüblichen Saccharin-Qualitäten wird unter Hinweis auf das Herstellungsverfahren gegeben.

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Determination of polar impurities in saccharin and saccharin sodium by HPLC

Summary A HPLC method for the direct determination of polar impurities in saccharin and saccharin sodium is described, o- and p-Sulphobenzoic acid as well as o- and p-sulphamoylbenzoic acid can be detected down to a limit of 1 mg/kg. For a concentration of 10 mg/kg the standard deviation is 0.4 mg/kg.A tabulated summary of the results obtained from commercial saccharin qualities is given with reference to the manufacturing process.

     

高效离子排斥色谱法测定饮料中的糖精钠

提出了一种用高效离子排斥色谱法测定饮料中糖精钠的新方法。以ＩｏｎＰａｃ ＩＣＥ－ＡＳ ６柱为分离柱，０．１０ ｍｍｏｌ／ＬＨ２ＳＯ４＋甲醇（９＋１）为淋洗液，２０２ｎｍ波长下紫外检测。在１－１００ｍｇ／Ｌ范围内，糖精钠的含量与峰面积呈 良好线性关系。检出限为０．６０ｎｇ，灵敏度优于以往采用离子交换色谱－电导检测法的结果。在实验条件下，饮料 中常见有机酸以及其它人工合成甜味剂等均不产生干扰。方法用于饮料中糖精钠的测定，加标回收率为９８％～ １０５％。此外还比较了电导检测、紫外检测和经化学抑制系统后紫外检测等三种检测方式对测定灵敏度的影响。     

高效液相色谱法测定镍镀液中的糖精钠和苯亚磺酸钠

建立了高效液相色谱法同时测定镍镀液中常用光亮剂苯亚磺酸钠和糖精钠的定量检测方法.为避免高盐背景对色谱分析的干扰,研究了样品的除盐处理方法.对方法的线性关系、回收率、精密度进行验证,并利用方法测定电镀液中苯亚磺酸钠和糖精钠,得到其实际消耗曲线:在400 A/m2的电流密度条件下苯亚磺酸钠4 h内平均消耗速率为18.25 ...     

Alkoxide-induced reactions of N-substituted saccharins. Synthesis of 1,2-benzothiazocine 1,1-dioxide and 2,3-dihydropyrrolo[1,2-b]-[1,2]benzisothiazole 5,5-dioxide derivatives

The reactions of saccharin derivatives 1 with sodium alkoxides were studied. Under mild conditions, compounds 1a-f gave the corresponding open sulfonamides 5a-f . Under drastic conditions, β-(saccharin-2)propionic acid derivatives 1a,b reacted with sodium ethoxide affording saccharin and β-ethoxypropionic acid derivatives 4a,b . γ-(Saccharin-2)butyric acid derivatives 1c,d and γ-(saccharin-2)-butyrophenone 1f reacted with sodium t-butoxide in dimethyl sulfoxide affording 5-substituted 6-hydroxy-3,4-dihydro-2H-1,2-benzothiazocine 1,1-dioxides 9 . From mother liquors, 1-substituted 2,3-dihydro-pyrrolo[1,2-b][1,2]benzisothiazole 5,5-dioxides 10 were isolated several hours later, though not detected immediately after completing the reaction. When the reactions were carried out in t-butyl alcohol, the yields of 9 diminished and those of 10 increased with product ratio inversion. Different experimental observations on the possible pathway generating 9 and 10 are discussed.     

Facile One‐Pot Synthesis of 4‐Hydroxy‐2‐methyl‐(2H)‐1,2‐benzothiazine‐3‐sulfonic Acid 1,1‐Dioxide

We report a convenient synthesis of 4‐hydroxy‐2‐methyl‐(2H)‐1,2‐benzothiazine‐3‐sulfonic acid‐1,1‐dioxide (6a) prepared in a novel one‐pot reaction. The synthesis involves two transformations starting from 2‐methyl‐2H‐1,2‐benzothiazin‐4‐(3H)‐one 1,1‐dioxide (7) with an overall yield better than that from the stepwise process, as well as the alternate procedure starting from saccharin (1). One‐pot synthesis of an important intermediate, saccharin‐N‐methane sulfonic acid (4), is also described.