Sweeteners detection systems

Since cyclamate has poor UV absorbing characteristics, HPLC methods for the analysis of this sweetener require specific detection systems, such as indirect photometry or conductivity. Herrmann et al. (24) used indirect photometry for the detection of cyclamate at 267 nm against a UV-absorbing mobile-phase component, p-toluenesulphonate. Biemer (17) and Wu et al. (47) used a conductivity detector for the determination of cyclamate. According to Biemer (17) the use of this detector offers distinct advantages, since compounds coeluting with cyclamate may not exhibit an electrochemical response and, hence, not appear in the chromatogram. 

Since HtS dissolved in water is very corrosive to carbon 4 steel, a comprehensive corrosion-control program is being conducted. In the field, each well is treated once per month by displacing inhibitor down to the perfora-. tions with stock tank oil. Corrosion coupons in the flow-lines are inspected every 6 months Little corrosion has been detected in the field. In the plants, corrosion in-hibitor is added daily to the gas-sweetening solvent, the salt water system, and the stabilizer overhead. Inhibitor is -Jj also added to bulk chemicals as received. Numerous corrosion coupons and probes are installed in each facility and are pulled for inspection every 1 to 3 months Corrosion rates have been low (less than I mil/year) asY result of the inhibitor injection program. 

These may contain levels of sugars or artificial sweeteners, coloring and flavoring agents, which make the HPLC determination more complex. Many of these components will be detected by HPLC and may interfere with the components that are being determined. The container should be stored such that the contents are in contact with the closme system. 

Liquid chromatography Four LC systems are in general use for analysis of bulk sweeteners and sugar alcohols (all systems require a sample with sugars concentration from <1 to 10% and the detection of separated peaks is usually by differential refracto-metry) ... 

RPLC with MS detection was used for the analysis of seven artificial sweeteners (aspartame, saccharin, acesulfame-K, neotame, sucralose, cyclamate, and alitame) and one natural sweetener (stevioside). Samples were extracted using methanokwater and injected without any cleanup into the LC—MS system. Separation is carried out using a Cis column and gradient elution. Sweeteners were quantified using selective-ionization recording (SIR) at m/z 178, 397, 377, 293, 641, 312, 162, and 182 for cyclamate, sucralose, neotame, aspartame, stevioside, alitame, acesulfame-K, and saccharin, respectively, with a warfarin sodium m/z = 307) used as an internal standard [24]. For a detailed discussion of other analytical methods to determine artificial sweeteners, refer to [25]. 

Some sweeteners (aspartame, cyclamate, saccharin, and acesulfame K) were determined by CE-SIA with contactless conductivity detection (Stojkovic et al., 2013). The analyses were carried out in an aqueous running buffer consisting of 150 mM 2-(cyclo-hexylamino)ethanesulfonic acid and 400 mM tris(hydroxymethyl)aminomethane at pH 9.1 in order to render all analytes in the fully deprotonated anionic form. The four compounds were determined successfully in food samples the experimental set-up and typical analysis results are illustrated in Figure 2.9. Another SIA system combined with solenoid valves was used to automate an enzymatic method for the determination of aspartame in commercial sweetener tablets. The method involves the enzymatic conversion of aspartame to hydrogen peroxide by the chymotrypsin-alcohol oxidase system, followed by the use of 2,2-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ARTS) as electron donor for peroxidase. Chymotrypsin and alcohol oxidase enzymes were immobilized on activated porous silica beads (Pena et al., 2004). 

A monolithic minicolumn was incorporated in a FIA manifold for the simultaneous analysis of eight analytes, including sweeteners (aspartame, acesulfame-K, saccharin) and a few preservatives and antioxidants (Garcia-Jimenez et al., 2007). The singlechannel FIA system with a short monolithic C18 column, which allowed the separation of analytes according to their retention time, was used for quantification by measuring the intrinsic UV absorption of the analytes. The system was applied to the detection in different foodstuffs and the results obtained were in agreement with a reference FiPLC method. 

A flow injection system coupled to a monolithic column has been described for the simultaneous determination of antioxidants (PG and BHA), sweeteners (potassium acesulfame, sodium saccharin, and aspartame), and preservatives (methylparaben, eth-ylparaben, propylparaben, and butylparaben), using photometric detection [56]. The monolithic column used as separation system was a 5 mm commercial precolumn of silica Cjg. The mixture was separated in only 400 s with resolution factors greater than 1.1 in all cases. Detection was accomplished by means of a DAD at the respective wavelength of each compound. The detection limit obtained for PG was 0.02 pg/mL. The method was applied to the analysis of food and cosmetic samples and the results were compared with those obtained using a conventional LC method. 

No methods could be found in the literature for the individual flow analysis determination of acesulfame-K. The first flow analysis method for acesulfame-K was proposed by Nikolelis et al. 2001 [72]. This method allowed the electrochemical flow injection monitoring and analysis of mixtures of acesulfame-K, cyclamate, and saccharin using stabilized systems of filter-supported bilayer lipid membranes (BLMs). Detection consisted of t time-dependent appearance of a transient ion current peak in which the time-dependence could be used to distinguish the presence of different artificial sweeteners, and the peak magnitude was related to the concentration of the artificial sweetener. The BLM-based system is able to monitor each artificial sweetener in mixtures. The apparatus for the formation of stabilized BLMs is shown in Figure 24.4. The method also offers response times of less than 1 min, which are the fastest times reported for any similar... 

Cabero et al. [80] developed a method based on the conversion of cyclamate to cyclo-hexylamine and the subsequent reaction with l,2-naphthoquinone-4-sulfonate, yielding a spectrophotometrically active derivative, which is detected at 480 nm thus, other sweeteners, such as saccharin or aspartame, do not interfere in these determinations. The hydrolysis step is performed batchwise by treatment of cyclamate with hydrogen peroxide and hydrochloric acid, while the cyclohexylamine derivatization is carried out in the flow injection system (Figure 24.9). Rocha et al. [81] reported a flow system based on multicommutation for fast and clean determination of cyclamate. The procedure exploits the reaction of cyclamate with nitrite in an acidic medium and the spectrophotometric determination of the excess of nitrite by iodometry. The flow system was designed with a set of solenoid micropumps to minimize reagent consumption and waste generation (Figure 24.10). 

Only two flow analysis methods have been published for multianalyte determination including cyclamate as analyte. Both methods determine cyclamate with other artificial sweeteners. One of them used stabilized systems of filter-supported BLMs in a FIA manifold [72], and the other is based on CE with contactless conductivity detection employing a sequential injection manifold based on a syringe pump [77]. 

Saccharin is also determined with FIA methodologies, which allow multianalyte determination. These methodologies include methods cited previously for acesulfame-K and/or cyclamate a method based on electrochemical detection of these three artificial sweeteners using stabilized systems of filter-supported BLMs [72] online dialysis for sample pretreatment prior to the simultaneous determination of saccharin and caffeine, benzoic acid, and sorbic acid by HPLC-FID [76] and molecular spectroscopic methods in the UV region based on the transient retention of analytes on solid phases, silica Cjg [74], and quaternary amine ion exchanger [75]. 

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