Abstract
This study provides a fast, accurate and reproducible method for L-ascorbic acid (L-AA) determination in milk samples by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). A small volume of a low toxicity organic solvent (ethanol) was used for degreasing and deproteinization steps. Ethylenediaminetetraacetic acid disodium salt (EDTA) and formic acid were used as stabilizing agents. The method was successfully validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, and inter/intra-day precision and applied in raw and processed milk samples. Good linearity (r2 > 0.9915) and low LOD and LOQ, 1.5 and 5.0 µg L-1, respectively, were obtained. The recoveries for 500 and 1000 µg L-1 spikes were higher than 90% and the precision values expressed in terms of relative standard deviation (RSD) were ≤ 6.8%. For the first time, the determination of L-AA in a 500 µg L-1 concentration range was reported for milk samples.
Highlights
Milk has been the most consumed food throughout the world
L-Ascorbic acid (L-AA) is the biologically active form of vitamin C, the general term used to name a set of compounds that exhibit the biological activity of L-AA, such as L-dehydroascorbic acid (L-DHA) and other oxidation products, which can be readily converted
A fast, low cost, low oxidative, accurate and reproducible methodology based on ultra-high-performance liquid chromatography (UHPLC)‐MS/MS was developed, validated and successfully applied for the analysis of L-AA in different milk samples
Summary
Milk has been the most consumed food throughout the world. In 2015, global per capita dairy consumption was estimated at 111.3 kg. Milk has great economic and social relevance, as the dairy industry employs millions of people and is an important source of income in the global economic scenario.[1,2] Nutritionally, milk is an important source of macro and micronutrients, e.g., proteins, lipids, carbohydrates, vitamins and minerals, as well as bioactive compounds such as antioxidants. Among the antioxidants that can be found in milk samples, vitamin C is one of the most important for the proper functioning of an organism, and diet is the only source of this vitamin for humans.[3]. L-Ascorbic acid (L-AA) is the biologically active form of vitamin C, the general term used to name a set of compounds that exhibit the biological activity of L-AA, such as L-dehydroascorbic acid (L-DHA) and other oxidation products, which can be readily converted
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