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An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables. Evaluation of degradation rate during storage

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Abstract

This study provides a versatile validated method to determine the total vitamin C content, as the sum of the contents of L-ascorbic acid (L-AA) and dehydroascorbic acid (DHAA), in several fruits and vegetables and its degradability with storage time. Seven horticultural crops from two different origins were analyzed using an ultra-high-performance liquid chromatographic–photodiode array (UHPLC-PDA) system, equipped with a new trifunctional high strength silica (100% silica particle) analytical column (100 mm × 2.1 mm, 1.7 μm particle size) using 0.1% (v/v) formic acid as mobile phase, in isocratic mode. This new stationary phase, specially designed for polar compounds, overcomes the problems normally encountered in HPLC and is suitable for the analysis of large batches of samples without L-AA degradation. In addition, it proves to be an excellent alternative to conventional C18 columns for the determination of L-AA in fruits and vegetables. The method was fully validated in terms of linearity, detection (LOD) and quantification (LOQ) limits, accuracy, and inter/intra-day precision. Validation experiments revealed very good recovery rate of 96.6 ± 4.4% for L-AA and 103.1 ± 4.8 % for total vitamin C, good linearity with r 2-values >0.999 within the established concentration range, excellent repeatability (0.5%), and reproducibility (1.6%) values. The LOD of the method was 22 ng/mL whereas the LOQ was 67 ng/mL. It was possible to demonstrate that L-AA and DHAA concentrations in the different horticulture products varied oppositely with time of storage not always affecting the total amount of vitamin C during shelf-life. Locally produced fruits have higher concentrations of vitamin C, compared with imported ones, but vegetables showed the opposite trend. Moreover, this UHPLC-PDA methodology proves to be an improved, simple, and fast approach for determining the total content of vitamin C in various food commodities, with high sensitivity, selectivity, and resolving power within 3 min of run analysis.

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Acknowledgments

The authors show their gratitude to Sonae MC for supplying the fruits and vegetables samples used in this study. This research was supported by Fundação para a Ciência e a Tecnologia (FCT) with funds from the Portuguese Government (Project PEst-OE/QUI/UI0674/2011).

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  1. Centro de Química da Madeira (CQM), Centro de Ciências Exactas e da Engenharia da Universidade da Madeira, Campus Universitário da Penteada, 9000-390, Funchal, Portugal

    Vítor Spínola, Berta Mendes, José S. Câmara & Paula C. Castilho

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  1. Vítor Spínola
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  2. Berta Mendes
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  3. José S. Câmara
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  4. Paula C. Castilho
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Correspondence to Paula C. Castilho.

Additional information

Published in the special issue Euroanalysis XVI (The European Conference on Analytical Chemistry) with guest editor Slavica Ražić.

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Spínola, V., Mendes, B., Câmara, J.S. et al. An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables. Evaluation of degradation rate during storage. Anal Bioanal Chem 403, 1049–1058 (2012). https://doi.org/10.1007/s00216-011-5668-x

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  • DOI: https://doi.org/10.1007/s00216-011-5668-x

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