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Determination of Melatonin and Its Metabolites in Biological Fluids and Eggs Using High-Performance Liquid Chromatography with Fluorescence and Quadrupole-Orbitrap High-Resolution Mass Spectrometry

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Abstract

Melatonin is a hormone found in animals, plants, fungi, and bacteria, which can be used as a sleep aid and in the treatment of sleep disorders. Here, a rapid and accurate method was developed for the determination of melatonin in plasma, milk, and egg using high-performance liquid chromatography with fluorimetric detection (HPLC-FLD) and ultra high performance liquid chromatography–quadrupole-orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Sample was extracted with methanol and cleaned by passing through hydrophilic–lipophilic balanced (HLB) solid phase extraction cartridge. The method validation results showed that recovery for melatonin was between 80.3 and 105.1 % with relative standard deviations less than 9.8 %. UHPLC-Q-Orbitrap HRMS was employed as a further means of confirmation to assure accuracy of the results. Limits of detection for melatonin were 0.13 ng g−1 and 0.14 ng mL−1 for solid and liquid samples, respectively. Thus, the proposed method can be applied to detect melatonin in milk, plasma, and egg samples. Meanwhile, the metabolites of melatonin were identified using Q-Orbitrap HRMS. The approach identified five metabolites; among them, a novel metabolite, named mono-oxygenation-demethylation-melatonin, was found in plasma.

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Acknowledgments

The financial support from the National Basic Research Program of China (973 Program, 2013CB127305), the Special Fund for Agro Scientific Research in the Public Interest (201203088), and the National Natural Science Foundation of China (31472126) are gratefully acknowledged.

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Conflict of Interest

All the authors declare that they have no conflict of interest in the present work.

Ethical Approval

In this study, all applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

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Not applicable

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Authors and Affiliations

  1. State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, 100193, People’s Republic of China

    Bingjie Yin, Tingting Li, Tong Dang & Pingli He

  2. State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, People’s Republic of China

    Zhen Li

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  1. Bingjie Yin
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  2. Tingting Li
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  3. Zhen Li
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  4. Tong Dang
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  5. Pingli He
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Correspondence to Pingli He.

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Yin, B., Li, T., Li, Z. et al. Determination of Melatonin and Its Metabolites in Biological Fluids and Eggs Using High-Performance Liquid Chromatography with Fluorescence and Quadrupole-Orbitrap High-Resolution Mass Spectrometry. Food Anal. Methods 9, 1142–1149 (2016). https://doi.org/10.1007/s12161-015-0288-2

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  • DOI: https://doi.org/10.1007/s12161-015-0288-2

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