Abstract
A new method using ultra high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) methodology was developed for the determination of ascorbic acid (AA) and dehydroascorbic acid (DHAA) contents in liquid and solid vegetable samples. The advantages of this method are speed, high sensitivity and practical application. In accordance with these advantages, the present method allows the simultaneous determination of AA and DHAA without previous reduction/derivatization of DHAA and without the use of internal standards in the samples. This is of high interest in routine analysis, providing a simpler sample preparation, as well as enhanced accuracy and robustness. Its validation included selectivity, sensitivity and linearity, precision and accuracy, matrix effect, and recovery. The results showed high selectivity and sensitivity, with calibration curves ranging from 10 to 500 ng mL−1 and from 50 to 500 ng mL−1 for AA and DHAA, respectively. Appropriate dilutions for each sample are necessary to avoid the matrix effect with accepted recoveries.
Highlights
Vitamin C is one of the most important water-soluble vitamins for health
The Recommended Dietary Allowance (RDA) of vitamin C is 90 and 75 mg per day, on average, for adult men and women, respectively [2], specific pathophysiological states can modulate the requirements of this vitamin [3]
The validation of this method was performed according to the International Guidelines on Validation of Analytical Procedures (ICH) [19] and the FDA Guidance for Bioanalytical Method
Summary
Vitamin C is one of the most important water-soluble vitamins for health. It is unable to be synthesized in humans and is involved in many biochemical functions, such as neutralization of free-radicals, the absorption of iron at the gastrointestinal level, and the synthesis and protection from oxidation of collagen, catecholamines, cholesterol, amino acids and some peptide hormones as an essential enzyme cofactor [1]. The Recommended Dietary Allowance (RDA) of vitamin C is 90 and 75 mg per day, on average, for adult men and women, respectively [2], specific pathophysiological states can modulate the requirements of this vitamin [3]. From a technological point of view, vitamin C could be used as an antioxidant preservative in food and pharmaceutical industries, contributing to protecting manufactured products from spoilage [5]. The methodology for vitamin C analysis should involve the quantification of L-ascorbic acid (AA), and of its oxidation product, the dehydroascorbic acid (DHAA), as this compound exhibits equivalent biological activity to AA [6,7]. The evaluation of only AA may lead to an underestimation of the biological value of a given food matrix as a source of this vitamin [8]. Among the Molecules 2019, 24, 1632; doi:10.3390/molecules24081632 www.mdpi.com/journal/molecules
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