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  • Determination of eight artificial sweeteners and common Stevia rebaudiana glycosides in non-alcoholic and alcoholic beverages by reversed-phase liquid chromatography coupled with tandem mass spectrometry.

Determination of eight artificial sweeteners and common Stevia rebaudiana glycosides in non-alcoholic and alcoholic beverages by reversed-phase liquid chromatography coupled with tandem mass spectrometry.

Analytical and bioanalytical chemistry (2014-12-05)
Paweł Kubica, Jacek Namieśnik, Andrzej Wasik
ABSTRACT

The method for the determination of acesulfame-K, saccharine, cyclamate, aspartame, sucralose, alitame, neohesperidin dihydrochalcone, neotame and five common steviol glycosides (rebaudioside A, rebaudioside C, steviol, steviolbioside and stevioside) in soft and alcoholic beverages was developed using high-performance liquid chromatography and tandem mass spectrometry with electrospray ionisation (HPLC-ESI-MS/MS). To the best of our knowledge, this is the first work that presents an HPLC-ESI-MS/MS method which allows for the simultaneous determination of all EU-authorised high-potency sweeteners (thaumatin being the only exception) in one analytical run. The minimalistic sample preparation procedure consisted of only two operations; dilution and centrifugation. Linearity, limits of detection and quantitation, repeatability, and trueness of the method were evaluated. The obtained recoveries at three tested concentration levels varied from 97.0 to 105.7%, with relative standard deviations lower than 4.1%. The proposed method was successfully applied for the determination of sweeteners in 24 samples of different soft and alcoholic drinks.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Neohesperidin, ≥90%
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Asp-Phe methyl ester, ≥98%
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Acetonitrile, puriss. p.a., ACS reagent, ≥99.5% (GC)
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Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 5 mm × 8 in.
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Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
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Methanol-12C, 99.95 atom % 12C
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Acetone solution, certified reference material, 2000 μg/mL in methanol: water (9:1)
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Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%
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Acetonitrile, Preparateur, ≥99.9% (GC), One-time steel-plastic (SP) drum
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Mettler-Toledo Calibration substance ME 51143091, Saccharin, traceable to primary standards (LGC)
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Aspartame, European Pharmacopoeia (EP) Reference Standard
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Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
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Rebaudioside C, analytical standard
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Aspartame, Pharmaceutical Secondary Standard; Certified Reference Material
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Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard
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Aspartame, United States Pharmacopeia (USP) Reference Standard
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Acetonitrile, electronic grade, 99.999% trace metals basis
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Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
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Acetonitrile, for HPLC, for UV, ≥99.9% (GC)
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Acetonitrile, HPLC Plus, ≥99.9%
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Acetonitrile, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5% (GC)
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