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Chromatographie en phase liquide à interaction zwitterionique-hydrophile pour l'acide l-ascorbique 2-phosphate de magnésium, une matière première en thérapie cellulaire.

Le sel de 2-phosphate de magnésium (APMg) de l'acide l-ascorbique est un dérivé de la vitamine C fréquemment utilisé dans les milieux de culture cellulaire à des fins de recherche. Il est également utilisé comme matière première dans la fabrication des médicaments de thérapie innovante pour gènes, cellules et tissus (GMP) au sein du GMP. Cependant, les méthodes de qualité font actuellement défaut. Par conséquent, une méthode LC a été développée, basée sur la chromatographie en phase liquide à mode mixte à interaction hydrophile (HILIC) - échange d’ions (IE). La méthode finale consistait en un système isocratique avec une phase mobile de tampon KH2PO4 15 mM (pH 2,5 avec HCl), d’acétonitrile (30:70, v / v) sur une colonne HILT zwitterionique, contenant une fonctionnalité d’échange de cations incorporée à un ligand hydrophile et un anion de surface. groupe d'échange. Un débit de 0,4 mL / min et une détection UV à 240 nm ont été appliqués. La méthode de dosage des APMg a été validée, en obtenant une linéarité adéquate (R (2) = 0,999), une précision (RSD de 0,49%) et une précision (récupération globale de 100,4%). La méthode développée a été appliquée avec succès à cinq produits actuellement commercialisés provenant de différents fournisseurs, présentant différents profils d'impureté de substance apparentée. En utilisant la spectroscopie d'absorption atomique (AAS), le magnésium s'est lié à la phase stationnaire, nécessitant une forte phase mobile pour rincer la colonne. Enfin, les impuretés liées ont été identifiées à l'aide de MS / MS et de MS haute résolution, et se sont révélées être de l'acide ascorbique ainsi que des dérivés éthyliques, ce qui a été confirmé par RMN.


Source: https://www.ncbi.nlm.nih.gov/pubmed/30579234


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