Abstract
A high-performance liquid chromatographic method with evaporative light scattering detection (HPLC-ELSD) was optimized for the simultaneous determination of non-caloric sweeteners—NCS (acesulfame-K, aspartame, cyclamic acid, neotame, saccharin, and sucralose) in non-alcoholic beverages. The procedure involved a simple dilute-and-shoot extraction of the six NCS with a formic acid/triethylamine/water solution and filtration (0.22 μm) followed by an HPLC–ELSD analysis. Validation steps included regression of the analytical curve, recovery, repeatability, and intermediate precision (relative standard deviation—RSD and RSDip, respectively), and limits of quantification and detection (LOQ and LOD, respectively). The method allowed lower consumption of solvents in a 35-min run. The figures of merit were satisfactory for all sweeteners (r2 ≥ 0.99; recovery from 99.0 to 101%; RSD and RSDip both < 1.0; LOQ ≤ 4.0 μg/mL; LOD ≤ 1.33 μg/mL). The method was used in the analysis of NCS in 64 non-alcoholic beverages from the market. NCS were not detected in 19 samples that did not declare their use. Higher concentrations of aspartame were found in carbonated soft drinks (7.97 mg/100 mL) and powdered drinks (22.46 mg/100 mL), while nectars had higher levels of acesulfame-K (8.69 mg/100 mL) and sports drinks only had sucralose (1.66 mg/100 mL). The concentrations found were 54.1 up to 194.0% of declared levels. The estimated daily intake of NCS in non-alcoholic beverages by the Brazilian population indicated that men, adolescents, residents of urban areas, and inhabitants of the southern region are the most widely exposed to NCS, but the risk is characterized as low.
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Abbreviations
- Anvisa:
-
National Health Surveillance Agency
- ADI:
-
Acceptable daily intake
- EDI:
-
Estimated daily intake
- HPLC–ELSD:
-
High performance liquid chromatography–Evaporative light scattering detection
- JECFA:
-
Joint FAO/WHO Expert Committee on Food Additives
- LOQ:
-
Limit of quantification
- LOD:
-
Limit of detection
- NCS:
-
Non-caloric sweeteners
- RQ:
-
Risk quotient
- RSD:
-
Relative standard deviation
- RSDip:
-
Relative standard deviation to intermediate precision
- SPE:
-
Solid phase extraction
- WHO:
-
World Health Organization
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The authors acknowledge Coordenação de Pessoal de Nível Superior—CAPES, Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq (Brasília, DF, Brazil), and Fundação de Amparo a Pesquisa do Estado de Minas Gerais—FAPEMIG (Belo Horizonte, MG, Brazil) for the financial support.
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RCSd Sousa: conception, material preparation, data collection and analysis, writing; K Costa: material preparation, data collection; MdF Gomides: material preparation, data collection; MRR Cunha: material preparation, data collection; MdO Almeida: material preparation, data collection; FB Custódio: material preparation, data collection and analysis; and MBA Gloria: conception, material preparation, data collection and analysis, writing, reviewing; funding acquisition. All authors reviewed the manuscript.
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Roberto Cesar Santos de Sousa declares that he has no conflict of interest. Maria de Fatima Gomides declares that she has no conflict of interest. Karen Costa declares that she has no conflict of interest. Mariem R. Ribeiro Cunha declares that she has no conflict of interest. Mariana de Oliveira Almeida declares that she has no conflict of interest. Flavia B. Custódio declares that she has no conflict of interest. Maria Beatriz A. Gloria declares that she has no conflict of interest.
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de Sousa, R.C.S., de Fatima Gomides, M., Costa, K. et al. Optimization and Validation of an Analytical Method for the Determination of Sweeteners in Beverages by HPLC-ELSD. Food Anal. Methods 17, 207–225 (2024). https://doi.org/10.1007/s12161-023-02562-w
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DOI: https://doi.org/10.1007/s12161-023-02562-w