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Updated: Jun 21, 2026
Improved UPLC-UV Method for the Quantification of Vitamin C in Lettuce Varieties (Lactuca sativa L.) and Crop Wild Relatives (Lactuca spp.)
Published on: June 30, 2020
A Green UPLC-UV Method for L-Ascorbic Acid Determination Based on a Biodegradable Chelating Agent and Synergistic
Wen Gu1, Zuyan Fang1, Bingwen Lin1
1Herbalife (China) Health Products Ltd., 197 Su Hong Xi Road, Suzhou Industrial Park, Suzhou, Jiangsu Province, 215021, China.
Background:
Ascorbic acid (Vitamin C) is the most widely consumed dietary supplement, which necessitates extensive quantitative testing of these products for batch release and post-market surveillance. However, most methods for ascorbic acid determination rely on hazardous reagents. It is therefore essential to adopt a green analytical chemistry (GAC) approach to evaluate and improve these methods, thereby mitigating their adverse environmental impact of analytical activities.
Objective:
This study aimed to develop a green ultra-performance liquid chromatography (UPLC) method for determining L-ascorbic acid in different types of dietary supplements, using biodegradable reagents and minimizing organic solvent consumption.
Method:
A phosphate solution (pH 3.0) containing 5 mmol/L oxalic acid (OA) was used for sample preparation. A 2-µL aliquot of the ascorbic acid extract was injected into a UPLC system. Separation was performed on an AX-C18 column (2.1 mm i.d. × 100 mm, 1.9 µm) with a mobile phase of 0.4% (v/v) phosphoric acid in water. Isocratic elution was carried out at a flow rate of 0.2 mL/min for 3 minutes. Ascorbic acid was detected at 245 nm and quantitated against standard solutions of L-ascorbic acid.
Results:
OA demonstrated effectiveness in stabilizing ascorbic acid comparable to that of conventional chelators, while offering superior environmental sustainability. When coupled with a UPLC system, this method achieved a low limit of quantitation (0.02 μg/mL) and excellent recoveries (98.3%-101.2%). Analytical Eco-Scale and AGREE metrics were used to evaluate the greenness of this method, indicating its excellent environmental performance.
Conclusions:
This newly developed method demonstrated robust performance across diverse sample matrices, providing an eco-friendly and highly efficient approach for determining vitamin C in dietary supplements and health foods fortified with L-ascorbic acid.
Highlights:
Minimized organic solvent usage, faster turnaround time, and improved environmental compatibility make this a precise, efficient, and green method for vitamin C determination.
