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High-Performance Thin-Layer Chromatography Analysis of Saccharin in Foods and Beverages

Mohd Idris, Seema Srivastava, Tulsidas R. Baggi, and Sudhir Kumar Shukla*

Key Words
Saccharin HPTLC Foods Beverages

Summary
A simple, accurate, and inexpensive high-performance thin-layer chromatography (HPTLC) method has been established for analysis of saccharin in foodstuffs, for example cola drinks, lemon juices, betel nut powder, mouth fresheners, ice candy, and tabletop sweeteners. Chromatography was performed on silica-gel 60 F254 plates with chloroformmethanolacetic acid 64:35:1 or acetoneisopropanolacetic acid 60:39:1 as mobile phases. The bands corresponding to the saccharin were scanned in absorbance mode at 230 nm. The calibration plot showed peak area was a linear function of concentration over the range 2501250 ng L1. The relationship between peak area and the amount of saccharin was evaluated by linear regression analysis. The limits of detection and quantification of saccharin were 40 and 130 ng, respectively. Mean recovery from spiked samples was 102.3% for cola drinks and 98.8% for lemon juices. Relative standard deviation (RSD) for cola drinks, lemon juices, ice candy, mouth freshener, betel nut powders, and tabletop sweeteners were 2.1, 4.2, 3.4, 3.0, 4.9, and 4.1%, respectively. The procedure was validated for analysis of saccharin in these food products.

Figure 1 The structure of saccharin.

mately 80% as saccharin and the rest as hydrolysis products. Although saccharin causes urinary bladder cancer in laboratory rats, its ban was rescinded after a public outcry. In 1984, the World Health Organization suggested an intake limit of 2.5 mg day1 kg1 b.w. Saccharin is used in a variety of beverages and foods, for example soft drinks, fruit juice drinks, processed fruit, chewing gum, betel nut powder (consumed extensively in India), confectionary, gelatine desserts, juices, jams, toppings, sauces, and dressings. It is suitable for cooking and baking. It is also used in cosmetics, pharmaceutical products, and other non-food applications, for example nickel-plating brightener, animal food sweetener, etc. The analysis of saccharin in beverages, food products, and pharmaceutical preparations has economic and social relevance for both health and legal reasons. Saccharin in foodstuffs has been analyzed by visible spectrophotometry [2], UV spectrophotometry [3], potentiometry [47], polarography [8, 9], non aqueous titrimetry [10], flowinjection analysis [1113], IR spectroscopy [14], FT-Raman spectroscopy [15], gas chromatography [1618], high-performance liquid chromatography [1926], and ion chromatography [2733]. Because most of the methods outlined above are poorly selective and because of the need for analysis of this sweetener in mixtures with other additives, for example preservatives and antioxidants, separative methods are most commonly used. Planar chromatography, especially thin-layer chromatography (TLC), has been widely used for identification of this intense
DOI: 10.1556/JPC.23.2010.5.7

1 Introduction
Saccharin (1,2-benzisothiazol-3(2H)-one-1,1-dioxide; Figure 1), used as the sodium or calcium salt, is a non-glucose, low-calorie product with a very sweet taste. Its sweetness is between 200 and 700 times that of a 2% sucrose solution and it is the least expensive sweetener per sweetness equivalent. Saccharin is characterized by a rapid onset, short persistence, and bitter and metallic aftertaste [1]. To reduce or eliminate this aftertaste via a synergistic effect, saccharin is usually blended with other sweeteners, for example cyclamate, aspartame, or sucralose. Saccharin provides no energy because it is not metabolized fully by humans, but rather is rapidly excreted via the kidneys, approxi-

M. Idris, S. Srivastava, and S.K. Shukla, Central Forensic Science Laboratory, Directorate of Forensic Science, Ministry of Home Affairs, Government of India, Ramanthapur, Hyderabad-500 013, India; T.R. Baggi, Forensic Science Unit, Department of Chemistry, University College of Science, Osmania University, Hyderabad-500 007, India. E-mail: drskshukla@gmail.com

Journal of Planar Chromatography 23 (2010) 5, 339342 0933-4173/$ 20.00 Akadmiai Kiad, Budapest

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