Beaming steviol glycoside analysis into the next dimension.

Area of Science:

• Analytical Chemistry
• Chromatography
• Food Science

Background:

• Steviol glycosides are natural sweeteners with complex separation challenges.
• Accurate quantification and identification are crucial for food and pharmaceutical industries.

Purpose of the Study:

• To evaluate state-of-the-art reversed-phase (RP) columns for UHPLC separation of steviol glycosides.
• To identify columns providing optimal resolution and retention for key compounds.
• To develop a robust and efficient method for steviol glycoside analysis.

Main Methods:

• Testing nine different RP columns for UHPLC.
• Assessing resolution of critical peak pairs (rebaudioside A and stevioside).
• Evaluating retention times of rebaudioside D using UV and MS compatible, buffer-free eluents.

Main Results:

• Three columns achieved a resolution of 2 or more for the critical peak pair.
• Two columns provided sufficient retention of rebaudioside D (retention factors 0.98 and 1.24).
• Separation of nine steviol glycosides was achieved within 11 minutes.

Conclusions:

• The study identified optimal UHPLC columns for steviol glycoside separation.
• A rapid, buffer-free method compatible with UV and MS detection was developed.
• The presented method is proposed for adoption as a new official analytical method.