Abstract
Melatonin (MLT) was involved in regulating various stages of plant growth and development. However, due to the low concentration and complex matrixes of plant, the analysis of MLT is a challenging task. In this study, we developed a rapid and efficient method with simplified sample preparation by employing UPLC coupled with a high resolution Orbitrap mass spectrometry, and stable isotope-labeled MLT (MLT-d4) was first used as internal standard in the developed analytical method. In the developed method, we used one-step liquid–liquid extraction to purify the crude extracts both from shoot and root of rice for the analysis, which remarkably simplify the sample preparation process. The method exhibits high specificity and recovery yield (>96.4%). Good linearities were obtained for MLT ranging from 0.01 to 20 ng/ mL with determination coefficient (R2) of 0.9991. The limit of detection for MLT was 0.03 pg. Reproducibility of the method was evaluated by intra-day and inter-day measurements and the results showed that relative standard deviations were less than 7.2%. Moreover, MLT quantification was accomplished by using only 100 mg fresh plant tissues. Additionally, the established method was successfully applied to investigate the spatiotemporal distributions of MLT in rice under cadmium (Cd) stress condition. We found that the content of MLT in shoot and root of rice increased under Cd stress, suggesting that MLT would play a crucial role in modulating the responses to Cd stress in different plant tissues.
Highlights
Studies on melatonin (N-acetyl-5-methoxytryptamine, MLT) in plants have attracted great attention of researchers in recent years
We successfully investigated the dynamic distributions of MLT in response to Cd stress in shoot and root of rice
High-resolution and accurate MS would benefit for simplifying procedures for sample preparation procedure and reducing analysis time, so an UPLC-high resolution Orbitrap MS was applied to the analysis of MLT in our study
Summary
Studies on melatonin (N-acetyl-5-methoxytryptamine, MLT) in plants have attracted great attention of researchers in recent years. LC coupled with MS has been proved as a powerful method for MLT analysis because of its high sensitivity, reproducibility and accuracy (Cao et al, 2006; Hernández-Ruiz and Arnao, 2008; Huang and Mazza, 2011b; Gomez et al, 2012, 2013). Multi reaction monitoring (MRM) (Gomez et al, 2012, 2013; Kocadali et al, 2014) and selective reaction monitoring (SRM) modes (Huang and Mazza, 2011b) have been used previously to the quantification of MLT They have been reported to be powerful tools to improve the sensitivity and specificity of analytes. We successfully investigated the dynamic distributions of MLT in response to Cd stress in shoot and root of rice
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