PLs séparés par CCM sont transméthylés en présence de 500?L du mélange toluène/MeOH (1 :1) et 500 ?L du mélange BF 3 /MeOH 14%. Le tube est fermé sous azote et mis dans le vortex. Il est chauffé à 100°C pendant 90min. La réaction est arrêtée en plongeant les tubes dans la glace et en ajoutant 1,5 mL de carbonate de potassium (K 2 CO 3 ) à 10% afin de stopper la réaction ,
on ajoute 500 ?L de toluène/EtOH, on ferme sous azote ,
100 ?L EtOH, 1 mL tampon borate de sodium 0.1 M + CaCl 2 (6 .10-3 M) pH 7.4 et 1 mL diéthyl éther. Le tube est fermé sous azote, mis dans le vortex et mis à soniquer durant 5 minutes. Ensuite, 2 ?L PLA 2 (coupe les AG dans la position 2) sont ajoutés et laissés sous agitation à 37°C (bain marie) pendant une nuit. Ensuite la réaction est arrêtée en plongeant les tubes dans la glace ,
extrait de cholestérol total (évaporé sous azote) est repris dans un faible volume CHCl 3 /EtOH Après le vortex, tout le volume est transféré dans un flacon conique (lavage 3 fois avec acétone ,
EtOH est ajouté, une évaporation sous azote est effectué, s'en suit un ajout de 100 ?L N,O-Bis (triméthylsilyl) trifluoroacétamide (BSTFA) ,
Une deuxième évaporation avec 10 ml d'éthanol absolu permet l'élimination de l'eau résiduelle ,
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