Analytical Quality by Design-Assisted Eco-Friendly RP-HPLC Method for the Simultaneous Estimation of Artificial Sweeteners in Commercial Food Samples Utilizing a Green Ultrasound-Assisted Extraction Technique: Greenness, Blueness, and Whiteness Appraisal.

Background: Acesulfame K (E950) and saccharin Na (E954) are commonly utilized synthetic sweeteners that are added to various processed food products to improve the sweet flavor. Environmentally friendly technology must be prioritized when evaluating the artificial sweeteners, as excessive consumption of these sweeteners presents serious health hazards.

Objective: The main aim of this study was to develop an analytical quality by design-aided eco-friendly RP-HPLC technique that can detect both acesulfame K and saccharin Na simultaneously, incorporating green analytical chemistry (GAC) principles and white analytical chemistry (WAC), using the ultrasound-assisted extraction (UAE) technique on commercial food samples.

Methods: The usage of ethanol was in accordance with eco-friendly ideals due to its ease of use, speed, and lack of environmental impact. Rotatable central composite design (rCCD) was used for method optimization. A mobile phase consisting of an ethanol-1% aqueous acetic acid (1 + 1, by volume) mixture was used and the separation was carried out on a Zorbax column (SB-C18, 150 × 4.6 mm, 5 µm) at a flow rate of 1 mL/min and a detection wavelength of 217 nm.

Results: Acesulfame K had a retention time of 1.134 min and saccharin Na of 2.134 min. Acesulfame K and saccharin Na recovery rates varied betweeen different commercially available food samples, ranging from 65 to 102% and 75 to 101%, respectively.

Conclusion: At the defined operating point, the developed procedure displays conformity with the previously defined requirements for linearity, accuracy, sensitivity, and repeatability. The most accurate assessments of greenness were produced by the Green Analytical Procedure Index (GAPI), Analytical Eco Scale (AES), and Analytical GREEnness metrics (AGREE) tools. Results from the Red-Green-Blue 12 (RGB 12) algorithm for whiteness and Blue Applicability Grade Index (BAGI) for blueness indicate that the method is very practical, cost-effective, and environmentally friendly.

Highlights: The results of this study could pave the way for more eco-friendly and effective AQbD methods to be used in the future for evaluating various sweeteners using green solvents.