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Andrology. 2015 May;3(3):496-505. doi: 10.1111/andr.12027. Epub 2015 Apr 8.

Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance ((1) H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS).

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Faculty of Medicine, Human Genetics Research Group, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain.
Biochemistry and Molecular Genetics Service, Hospital Clinic, Barcelona, Spain.
Biology of Reproduction and Stem Cell Group, CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
PhD Program in Experimental Biology and Biomedicine (PDBEB), Center for Neuroscience and Cell Biology, Coimbra, Portugal.
Institute for Interdisciplinary Research (IIIUC), University of Coimbra, Coimbra, Portugal.
Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Institut d'Investigació Sanitària Pere Virgili (IISPV) and Universitat Rovira i Virgili (URV), Tarragona, Spain.
Clinic Institute of Gynaecology, Obstetrics and Neonatology, Hospital Clinic, Barcelona, Spain.
Department of Life Sciences, University of Coimbra, Coimbra, Portugal.


The objective of this study was to contribute to the first comprehensive metabolomic characterization of the human sperm cell through the application of two untargeted platforms based on proton nuclear magnetic resonance ((1) H-NMR) spectroscopy and gas chromatography coupled to mass spectrometry (GC-MS). Using these two complementary strategies, we were able to identify a total of 69 metabolites, of which 42 were identified using NMR, 27 using GC-MS and 4 by both techniques. The identity of some of these metabolites was further confirmed by two-dimensional (1) H-(1) H homonuclear correlation spectroscopy (COSY) and (1) H-(13) C heteronuclear single-quantum correlation (HSQC) spectroscopy. Most of the metabolites identified are reported here for the first time in mature human spermatozoa. The relationship between the metabolites identified and the previously reported sperm proteome was also explored. Interestingly, overrepresented pathways included not only the metabolism of carbohydrates, but also of lipids and lipoproteins. Of note, a large number of the metabolites identified belonged to the amino acids, peptides and analogues super class. The identification of this initial set of metabolites represents an important first step to further study their function in male gamete physiology and to explore potential reasons for dysfunction in future studies. We also demonstrate that the application of NMR and MS provides complementary results, thus constituting a promising strategy towards the completion of the human sperm cell metabolome.


gas chromatography mass spectrometry (GC-MS); human sperm; metabolites; metabolome; pathways analysis; proton Nuclear Magnetic Resonance (1H-NMR) Spectroscopy

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