Use of UPLC-ESI-MS/MS to quantitate free amino acid concentrations in micro-samples of mammalian milk.
2013
Roucher, Véronique Ferchaud | Desnots, Emmanuelle | Nael, Charlotte | Agnoux, Aurore Martin | Alexandre-Gouabau, Marie Cécile | Darmaun, Dominique, D. | Boquien, Clair-Yves | Centre de Recherche en Nutrition Humaine | Physiopathologie des Adaptations Nutritionnelles (PhAN) ; Institut National de la Recherche Agronomique (INRA)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE) ; Université de Nantes (UN)-Université de Nantes (UN) | Direction de la Recherche Département Partenariats et Innovation (DHU 2020) | Centre Hospitalier Universitaire de Nantes = Nantes University Hospital (CHU Nantes)
Region Pays de la Loire (for the creation of the "Human milk" biocollection at Nantes University Hospital)
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Afficher plus [+] Moins [-]anglais. Although free amino acids (FAA) account for a small fraction of total nitrogen in mammalian milk, they are more abundant in human milk than in most formulas, and may serve as a readily available source of amino acids for protein synthesis, as well as fulfill specific physiologic roles. We used reversed phase Ultra Performance Liquid Chromatography (UPLC) coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS) technique for FAA profiling in milks from three species (human, rat and cow) with a simple and rapid sample preparation. The derivatization procedure chosen, combined with UPLC-ESI-MS/MS allowed the quantitation of 21 FAA using labeled amino acids (Internal Standards) over a 10 min run time in micro-samples of mammalian milk (50 μL). The low limit of quantitation was 0.05 pmol/μL for most FAA with good repeatability and reproducibility (mean CV of 5.1%). Higher levels of total FAA were found in human (3032 μM) and rat milk (3460 μM) than in bovine milk (240 μM), with wide differences in the abundances of specific FAA between species. This robust analytical method could be applied to monitor FAA profile in human breast milk, and open the way to individualized adjustment of FAA content for the nutritional management of infants.
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