Saccharin: analysis

Sample preparation for saccharin analysis by HPLC can be performed as indicated in Section I.C. Methods for extraction have also been described for desserts and sweets containing food thickeners (38) soy sauce, orange juice, and yogurt (60) chewing gum (17,39), and different food samples (42). Aminobenzoic acid, theophyllin, sodium fumarate, and adenine sulfate have been used as internal standards (17,31,39,44). 

Thin Layer Chromatography Systems for Saccharin Analysis... 

Zemplen helped his students in many ways. I remember an occasion in the difficult postwar period. The production of the famous Hungarian salami, interrupted by the war, was just in the process of being restarted for export. The manufacturer wanted a supportive analysis from the well-known professor. Zemplen asked for a suitable sample of some hundreds of kilograms, on which the whole institute lived for weeks. When it was gone he rightly could offer an opinion that the product was quite satisfactory. After the war, grain alcohol was for a long time the only available and widely used laboratory solvent, and, not unexpectedly, it also found other uses. Later, when it was denatured to prevent human consumption, we devised clever ways for its purification. The lab also manufactured saccharine, which was... 

Cosgrove K., Carroll M. Effects of bremazocine on self-administration of smoked cocaine base and orally delivered ethanol, phencyclidine, saccharin, and food in rhesus monkeys a behavioral economic analysis. J. Pharmacol. Exp. Ther. 301 Jun 2002, 2002. 

Sweeteners can be roughly divided into two groups bulk and intense sweeteners. Prodolliet (1996) and Gloria (2000) reviewed thoroughly the analysis and properties of intense sweeteners acesulfame-K, alitame, cyclamate, aspartame, glycyrrhizin, neohesperidin DC, saccharin, stevioside, sucralose and thaumatin. They are generally used in low calorie products such as diet... 

There is a recent trend towards simultaneous CE separations of several classes of food additives. This has so far been applied to soft drinks and preserved fruits, but could also be used for other food products. An MEKC method was published (Lin et al., 2000) for simultaneous separation of intense sweeteners (dulcin, aspartame, saccharin and acesulfame K) and some preservatives (sorbic and benzoic acids, sodium dehydroacetate, methyl-, ethyl-, propyl- and isopropyl- p-hydroxybenzoates) in preserved fruits. Ion pair extraction and SPE cleanup were used prior to CE analysis. The average recovery of these various additives was 90% with good within-laboratory reproducibility of results. Another procedure was described by Frazier et al. (2000b) for separation of intense sweeteners, preservatives and colours as well as caffeine and caramel in soft drinks. Using the MEKC mode, separation was obtained in 15 min. The aqueous phase was 20 mM carbonate buffer at pH 9.5 and the micellar phase was 62 mM sodium dodecyl sulphate. A diode array detector was used for quantification in the range 190-600 nm, and limits of quantification of 0.01 mg/1 per analyte were reported. The authors observed that their procedure requires further validation for quantitative analysis. 

This technique has been established for many years particularly for water, soil and feeding-stuff analysis, where a large number of analyses are required for quality control or monitoring purposes. A number of applications have been published for food additives including aspartame (Fatibello et al., 1999), citric acid (Prodromidis et al., 1997), chloride, nitrite and nitrate (Ferreira et al., 1996), cyclamates (Cabero et al., 1999), sulphites (Huang et al., 1999 AOAC Int, 2000), and carbonate, sulphite and acetate (Shi et al., 1996). Yebra-Biumm (2000) reviewed the determination of artificial sweeteners (saccharin, aspartame and cyclamate) by flow injection. 

For the analysis and separation of benzoic acid, caffeine, aspartame, and saccharin in dietetic soft drinks, a HPLC system consisting of a Varian MCH-5N-CAP 150 x 4.6 mm column and a variable wave length UV/VIS detector was recommended [32]. The mobile phase is a gradient, beginning with 90% 0.01 M KH2PO4 (pH = 2) and methanol, and ending in 25 minutes with 60 % buffer / 40 % methanol. 

Herrmann et al. (24) used ion-pair chromatography for the determination of cyclamate. The efficiency of LiChrosorb RP-18 and Hypersil MOS 3 with a mobile phase of 5 mM tetrabutylammonium p-toluenesulfonate, pH 3.5, mixed with 12% methanol for the separation of cyclamate from other sweeteners was investigated. With the first column, cyclamate separated from saccharin, but the second was the recommended column for the analysis of cyclamate, saccharin, aspartame, and dulcin in a single run. 

Various methods have been developed for the simultaneous determination of several sweeteners in a single run. Most of the methods described in the literature and summarized in Table 3 have been developed for the separation of three sweeteners, especially for saccharin, acesulfame-K, and aspartame. Herrmann et al. (24), Veerabhadrarao et al. (27), and Hausch (66) developed methods for the simultaneous determination of four sweeteners. Prodolliet and Bruelhart (33) and Wu et al. (47) separated five sweeteners. The most comprehensive method is the one developed by Lawrence and Charbonneau (16), which allows the simultaneous analysis of seven sweeteners. With the increased number of sweeteners available and their use being approved for use in specified food products and beverages by different countries, methods capable of separating several sweeteners simultaneously are still needed. 

TA Biemer. Analysis of saccharin, acesulfame-K and sodium cyclamate by high performance ion chromatography. J Chromatogr 463(2) 463-468, 1989. 

H Terada, Y Sakabe. Studies on the analysis of food additives by high-performance liquid chromatography. V. Simultaneous determination of preservatives and saccharin in foods by ion pair chromatography. J Chromatogr 346(5) 333-340, 1985. 

A Hannisdal. Analysis of acesulfame-K, saccharin and preservatives in beverages and jams by HPLC. Z Lebensm Unters Forsch 194(6) 517-519, 1992. 

Veerabhadrarao et al. (76) used reverse-phase HPLC for the determination of some food additives (acesulfame, saccharine, BA, p-hydroxybenzoic acid). The samples (beverages, tomato sauce) were diluted and then separated on a /rBondapak CJg column with methanol/acetic acid/water (20 5 75) or (35 5 60) as mobile phases. The determination was done at 254 nm. Recoveries varied from 98 to 106% for direct analysis and from 91.6 to 101.8% for extraction of samples (76). 

A paired-ion, reversed-phase high-performance liquid chromatographic method was developed for the simultaneous determination of sweeteners (dulcin, saccharin-Na, and acesulfame-K), preservatives (sodium dehydroacetate, SA, salicyclic acid, BA, succinic acid, methyl-para-hydroxybenzoic acid, ethyl-para-hydroxybenzoic acid, n-propyl-para-hydroxybenzoic acid, n-butyl-para-hydroxybenzoic acid, and isobutyl-para-hydroxybenzoic acid), and antioxidants (3-tertiary-butyl-4-hydroxyanisole and tertiary-butyl-hydroquinone). A mobile phase of acetonitrile-50 ml aqueous tr-hydroxyisobutyric acid solution (pH 4.5) (2.2 3.4 or 2.4 3.6, v/v) containing 2.5 mM hexadecyltrimethylammonium bromide and a Clg column with a flow rate of 1.0 ml/min and detection at 233 nm were used. This method was found to be very reproducible detection limits ranged from 0.15 to 3.00 p,g. The retention factor (k) of each additive could be affected by the concentrations of hexadecyltrimethylammonium bromide and a-hydroxyisobu-tyric acid and the pH and ratio of mobile phase. The presence of additives in dried roast beef and sugared fruit was determined. The method is suitable for routine analysis of additives in food samples (81). 

Synergism occurs with fructose (Hyvonen el al., 1978), aspartame, cycla-mate (Bakal, 1987) and sucralose (Tate Lyle Pic, 1986). Negative synergy (i.e. suppression) occurs with acesulfame K blends. Analysis of saccharin is usually done using HPLC (Halm Gilikson, 1987) or spectrophotometric methods (Ramappa Nayak, 1983). 

Two methods have been published which were designed to analyse a range of sweeteners and preservatives in one run. The fust method, published in German by Hagenauer-Hener et al. (1990), describes the analysis of aspartame, acesulfame K, saccharin, caffeine, sorbic acid and benzoic acid in soft drinks and foods. The method relies on a similar system to that given above but with a less complex solvent system (Figure 10.5). The solvent system has been modified to include a gradient portion to elute the preservatives more quickly. 

Three spectrophotometric procedures are given in the AOAC compendium of methods (960.22, 962.13 and 967.11) for the analysis of caffeine, all of which have an extraction stage followed by a quantification procedure. There is also an HPLC method, discussed earlier, which was designed to measure saccharin, benzoic acid and caffeine at the same time (AOAC, 978.08). Again, the HPLC method, EN 12856 1999 (Anon, 1999a), can be used for the analysis of caffeine, but this analyte was not included in the collaborative study. 

Egan, H., Kirk, R.S. and Sawyer, R. (1990a) Analysis of saccharin, in Pearson s Chemical Analysis of Foods, 8th edn, Longman, Harlow, p. 215. 

As a result of decisions of International Congresses of Applied Chemistry, among them that held at Paris (1900) and of the International Commission for Standard Methods of Sugar Analysis, the specific gravity of saccharine solutions should be determined at 20° C. and referred to water at 40 (sp. gr. at 20°/4°), i.e., it should indicate the weight of a true c.c. of solution at 200. Use is, however, largely made in practice of the sp. gr. at i745°/i7-50 and sometimes at i5°/i5°. 

Analysis of refined sugars is mostly reduced to the determination of the saccharose by direct polarisation. In some cases the ash and moisture are also determined, and in rare instances other determinations may be necessary, such as that of the reducing sugars (invert sugar and, perhaps, glucose or lactose added as adulterant) or raifinose a test for saccharin is occasionally required. 

To ascertain the quantity of sugar (saccharose) used in the preparation of the preserved fruit, from the total sugar (invert sugar x 0 95 plus saccharose) contained in the sample must be deducted the natural sugar (also calculated as saccharose) due to the quantity of fruit in the sample. For this purpose it is, of course, necessary to know the saccharine contents of different fruits, so that an analysis of fruit of the same quality preserved in water in the same conditions as in the syrup must be made. 

IR and Raman spectra of ofloxacin and levofloxacin were measured and evaluated [06SA(A)159]. Saccharin salt of ofloxacin was prepared and characterized by FTIR, X-ray powder diffraction, thermal analysis, and and 13C NMR spectroscopy in solution and in solid state (09JPS3788). Nondestructive discrimination between levofloxacin and ofloxacin containing tablets was achieved by using diffusion reflectance NIR spectroscopy (08PHA628). 

Polarographic methods of analysis have been applied to samples of foods containing saccharin (1+1-1+1+ ). In a procedure (1+1+) saccharin is extracted into organic solvents in an acidic medium. Further purification is achieved by column chromatography. The residue obtained is dissolved in 0.1 N NaOH and an aliquot is polarographed in a supporting electrolyte of 0.1 N HC1, 0.1 N KC1 and 0.1 Bu N Br. 

Thin layer chromatography has been used in qualitative and quantitative analysis of saccharin, when present in artificial sweetening agents, beverages, food and pharmaceuticals. Several systems have been used and are listed in table 5-... 

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