Development of a Novel, Sensitive, Selective, and Fast Methodology to Determine Malondialdehyde in Leaves of Melon Plants by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Melisa E Yonny,Soledad Cerutti,Ariel Rodríguez Torressi,Mónica A Nazareno

doi:10.1155/2017/4327954

Abstract

Early production of melon plant (Cucumis melo) is carried out using tunnels structures, where extreme temperatures lead to high reactive oxygen species production and, hence, oxidative stress. Malondialdehyde (MDA) is a recognized biomarker of the advanced oxidative status in a biological system. Thus a reliable, sensitive, simple, selective, and rapid separative strategy based on ultra-high-performance liquid chromatography coupled to positive electrospray-tandem mass spectrometry (UPLC-(+)ESI-MS/MS) was developed for the first time to measure MDA, without derivatization, in leaves of melon plants exposed to stress conditions. The detection and quantitation limits were 0.02 μg·L−1 and 0.08 μg·L−1, respectively, which was demonstrated to be better than the methodologies currently reported in the literature. The accuracy values were between 96% and 104%. The precision intraday and interday values were 2.7% and 3.8%, respectively. The optimized methodology was applied to monitoring of changes in MDA levels between control and exposed to thermal stress conditions melon leaves samples. Important preliminary conclusions were obtained. Besides, a comparison between MDA levels in melon leaves quantified by the proposed method and the traditional thiobarbituric acid reactive species (TBARS) approach was undertaken. The MDA determination by TBARS could lead to unrealistic conclusions regarding the oxidative stress status in plants.

Highlights

Harsh environmental conditions such as drought, salinity, and extreme temperatures can delay growth and development of plants, inflict lethal injuries to the plant structure, and reduce crop yield
Gradient and isocratic elution conditions were evaluated and the results showed that isocratic elution using a mixture of water/Acetonitrile high performance liquid chromatography (HPLC) (ACN) in a 70 : 30 proportion was optimum for MDA release from the column
This result is in agreement with Syslovaet al. [25], who found that MDA elution occurs in an isocratic section of the chromatographic run and when the proportion of aqueous component of the mobile phase is higher than the organic component

Summary

Introduction

Harsh environmental conditions such as drought, salinity, and extreme temperatures can delay growth and development of plants, inflict lethal injuries to the plant structure, and reduce crop yield These abiotic stress factors cause the overproduction and accumulation of reactive oxygen species (ROS) [1]. Limitation of CO2 uptake, caused by stress-induced stomatal closure, favors photorespiratory production of H2O2 in the peroxisome and production of superoxide and H2O2 or singlet oxygen by the overreduced photosynthetic electron transport chain [2] In this conditions, when the ROS production exceeds the intrinsic antioxidant defense mechanisms in a biological system, oxidative stress is produced causing damage to cell molecules and, affecting the normal cell functions and cell death [3]. These stresses produce plants damage in many ways: plant growth, membrane integrity, pigment

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