Greenness assessment and validation of HPLC method for simultaneous determination of Resveratrol and Vitamin E in dietary supplements

Background There is an increasing interest in the use of a combination of trans-resveratrol and vitamin E in dietary supplements. Determination of the content of both components is essential for confirmation of the quality of the product. Objective To establish the applicability and ensure the greenness of the previously developed high-throughput HPLC/UV method for the simultaneous determination of trans-resveratrol and alpha-tocopherol acetate (vitamin E) in dietary supplements. Methods Separation was performed on RP C8 Select B chromatographic column, using acetonitrile and water in the mobile phase, with gradient elution. Full method validation was performed in accordance with ICH Q2(R1). The greenness of the method was assessed using the analytical eco-scale (AES) methodology and the analytical greenness metric (AGREE). Results The method is selective, linear, precise and accurate over defined concentration ranges (185—369 µg/mL of trans-resveratrol and 37—75 µg/mL of alpha-tocopherol acetate), and has a suitable sensitivity (limits of detection and quantification are 7.7 µg/mL and 23.3 µg/mL for resveratrol and 2.6 µg/mL and 7.8 µg/mL for tocopherol acetate, respectively). The obtained analytical eco-scale score of 77 and the pale green AGREE pictogram with an overall score of 0.61 confirm the method’s greenness. Conclusion The sensitivity and selectivity of the method, its short analysis time (7 minutes), the low negative environmental impact, and simple sample preparation make the method readily applicable to in-line quality control procedures. Highlights A method for simultaneously analysing vitamin E and resveratrol in dietary supplements is presented. The method is rapid, includes a simple sample preparation procedure, and has a low environmental impact.