Simultaneous Determination of Nonnutritive Sweeteners in Foods by HPLC/ESI-MS

Abstract

Nonnutritive sweeteners are the low calorie substances used to replace sugar and other caloric ones. Determination of these sweeteners in foods is important to ensure consistency in product quality. In this study, seven artificial (aspartame, saccharin, acesulfame-K, neotame, sucralose, cyclamate, and alitame) and one natural sweetener (stevioside) were simultaneously determined in different foods using high performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometric detection (ESI-MS). The target compounds were quantified using a selective ionization recording (SIR) at m/z 178, 397, 377, 293, 641, 312, 162, and 182 to cyclamate, sucralose, neotame, aspartame, stevioside, alitame, acesulfame-K, and saccharin, respectively, with warfarin sodium (SIR m/z 307) being used as an internal standard. The correlation coefficient of the calibration curve was better than 0.998 (n = 6), in the range of 0.05 to 5.00 microg/mL for cyclamate, 0.30 to 30.0 microg/mL for sucralose, 0.10 to 10.0 microg/mL for neotame, 0.20 to 20.0 microg/mL for aspartame, 0.50 to 15.0 microg/mL for stevioside, 0.08 to 8.00 microg/mL for alitame, 0.10 to 15.0 microg/mL for acesulfame-K, and 0.05 to 5.00 microg/mL for saccharin. The limits of detection (LODs) were below 0.10 microg/mL, whereas the limits of quantification (LOQs) were below 0.30 microg/mL. It is concluded that the method has merits such as high sensitivity, specificity, and simplicity versus the those of the other methods reported in the literature.

Recent Analytical Methods for the Analysis of Sweeteners in Food: A Regulatory Perspective

“…Liquid-liquid extraction (LLE) is sometimes used as a simple, low-cost method to prepare samples prior to instrumental analyses [5]. LLE involves addition of an organic solvent to the food in liquid form.…”

“…Solid-liquid extraction (SLE) is the process of partitioning target analytes from a solid state into a solvent prior to dilution and filtration. Solid fi samples can be homogenized, vortexed and centrifuged to separate the supernatant [5,22,26].…”

“…Yang and Chen [5] used LLE and SLE to extract sweeteners from a water/ methanol solution (50:50, v/v). Beverages were degassed when necessary and solid samples were homogenized and extracted.…”

“…Generally, it is important to have internal standards for quantitation to account for possible ion suppression from matrix interferences in the complex composition of foods [5]. Although it is ideal to have isotopically labeled standards for MS detection methods for each compound being analyzed, these are sometimes unavailable and cost prohibitive.…”

“…Neohesperidine dihydrochalcone (NHDC) is a semi-synthetic sweetener, while stevioside (STV) and rebaudioside (REB) A are natural sweeteners [4]. The list of allowable sweeteners varies among nations worldwide [5]. Th For example, CYC and NHDC are not approved for use as food additives by the US Food and Drug Administration (FDA) but are authorized in the European Union (EU) [6].…”