Development and Validation of a Stability-Indicating HPLC Method for the Simultaneous Determination of trans-Resveratrol and cis-Resveratrol in an Injectable Solution.

Esmeralda Mota-Lugo,Elvia A Morales-Hipólito,Iris A Blanco-Alcántara,Mariana Dolores-Hernández,Hugo Cuatecontzi-Flores,Raquel López-Arellano

doi:10.1155/2021/8402157

Abstract

trans-Resveratrol, a phytochemical compound with antioxidant power and various therapeutic effects, such as cardioprotective, chemopreventive, and neuroprotective, among others, has disadvantages of poor solubility and limited stability, creating difficulties for the development of new strategies for its quantification. This study developed and validated an analytical stability method for trans-resveratrol by high-pressure liquid chromatography with photodiode-array detection (HPLC-PDA), which allowed its quantification in the presence of its degradation products. The quantification of trans-resveratrol occurred at a retention time of 2.6 min, with ammonium formate (10 mM, pH = 4)/acetonitrile, 70/30 v/v, as mobile phase. The validation met the ICH Q2 criteria of specificity, method linearity (2.8–4.2 μg/ml), precision and accuracy, robustness, quantification limit (0.176 μg/ml), and detection (0.058 μg/ml). As degradation products, cis-resveratrol was observed at 3.9 min, which could be resveratrone in 3.2 min and five unidentified products in 0.7, 1.0, 1.4, 1.8, and 5 min. Some solutions subjected to temperature stress of 40 and 60°C, UV light, and acidic and basic hydrolysis exhibited colour changes. An analytical method was obtained by HPLC-PDA, which allowed quantifying the stability of trans-resveratrol in a fast and specific manner in the presence of its degradation products.

Highlights

Laboratory of Pharmaceutical Development Tests, Multidisciplinary Research Unit, Faculty of Higher Education Cuautitlan, National Autonomous University of Mexico, Carr
Academic Editor: Cecilia Cagliero trans-Resveratrol, a phytochemical compound with antioxidant power and various therapeutic effects, such as cardioprotective, chemopreventive, and neuroprotective, among others, has disadvantages of poor solubility and limited stability, creating difficulties for the development of new strategies for its quantification. is study developed and validated an analytical stability method for trans-resveratrol by high-pressure liquid chromatography with photodiode-array detection (HPLC-photodiode-array detector (PDA)), which allowed its quantification in the presence of its degradation products. e quantification of trans-resveratrol occurred at a retention time of 2.6 min, with ammonium formate (10 mM, pH 4)/acetonitrile, 70/30 v/v, as mobile phase. e validation met the International Conference on Harmonisation (ICH) Q2 criteria of specificity, method linearity (2.8–4.2 μg/ml), precision and accuracy, robustness, quantification limit (0.176 μg/ml), and detection (0.058 μg/ml)
Some studies have indicated that t-RSV had limited stability under the influence of light, basic pH levels, and temperature, which could cause isomerization to cisresveratrol (c-RSV) or t-RSV degradation. e c-RSV (Figure 1(b)) is not found naturally; it can be obtained from trans-resveratrol by exposure to sunlight, or by ultraviolet (UV) radiation at wavelengths of 254 nm or 365 nm [22,23,24,25,26]

Summary

Introduction

Laboratory of Pharmaceutical Development Tests, Multidisciplinary Research Unit, Faculty of Higher Education Cuautitlan, National Autonomous University of Mexico, Carr. Is study developed and validated an analytical stability method for trans-resveratrol by high-pressure liquid chromatography with photodiode-array detection (HPLC-PDA), which allowed its quantification in the presence of its degradation products. Introduction trans-resveratrol (3,5,4′-trihydroxystilbene; t-RSV) is a polyphenolic phytoalexin mainly found in the skin of grapes and in at least 72 species of plants, peanuts, berries, cocoa, and almonds (Figure 1(a)) It has been found in food products such as wine, chocolate, grape juice, and peanut butter [1]. E c-RSV (Figure 1(b)) is not found naturally; it can be obtained from trans-resveratrol by exposure to sunlight, or by ultraviolet (UV) radiation at wavelengths of 254 nm or 365 nm [22,23,24,25,26] There are very few studies that addressed the quantification of t-RSV in the face of its degradation products generated by forced degradation [25, 39], and most of them have focused on photolytic studies with UV light [22,23,24, 26, 40]

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Conclusion