Chromatographie

Pour obtenir le produit pur d’oxilofrine l’échantillon correspondant à la formulation 16 a été purifié dans les conditions génériques explicitées ci-après :

3.5.4.1.1 Mise au point analytique

Pour mettre au point les conditions analytiques 3 gélules de la formulation 16 ont été dissoutes dans 6 mL d’un mélange acétonitrile-H2O (80-20 v-v). Nous avons travaillé sur une UPLC Waters Acquity muni d’un double mode de détection PDA MS simple quadripôle (mass spectrometry).

La colonne utilisée est une colonne Acquity BEH (Bridged Ethylene Hybrid) en phase inverse C18, avec une granulométrie de 1,7 µm et des dimensions de (2,1*100) mm.

137 Un mélange binaire homogène de solvant MeOH (+0.1% HCOOH) [A] et H2O (+0,1 % HCOOH) [B] a été utilisé dans les conditions d’élution suivante avec un volume d’injection de 5 µL et un débit de 0,6 mL/min à une température d’analyse de 40°C.

Temps (min) % [A] MeOH (+0,1% HCOOH) % [B] H2O (+0,1 % HCOOH)

0 5 95

2 5 95

6 100 0

7,5 100 0

8 5 95

Les analyses ont été menées avec une détection UV à 254 nm et une détection de masse en electrospray positif avec une tension de cône fixée à 20V.

3.5.4.1.2 Conditions de purification

Les conditions de chromatographie préparative ont été adaptées des conditions analytiques. Les analyses ont été menées sur un système d’auto-purification waters à pompe binaire 245 muni d’un collecteur de fraction 2767, équipé cette fois uniquement d’un détecteur UV. La colonne utilisée est une XBridge en phase inverse C18 avec une granulométrie de 5 µm et de dimension (19*150) mm possédant la même chimie de colonne que la BEH utilisée en analytique. Le même mélange binaire de solvant que précédemment [A] et [B] a été utilisé selon les conditions d’élution présentés ci-après avec un volume d’injection de 1 mL et débit de 20,5 mL/min.

Temps (min) % [A] MeOH (+0.1% HCOOH) % [B] H2O (+0.1 % HCOOH)

0 5 95

2 5 95

12 100 0

14 100 0

14,5 5 95

138

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