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Prostaglandin extraction

VD Valeria Di Dato
RB Roberta Barbarinaldi
AA Alberto Amato
FC Federica Di Costanzo
CF Carolina Fontanarosa
AP Anna Perna
AA Angela Amoresano
FE Francesco Esposito
AC Adele Cutignano
AI Adrianna Ianora
GR Giovanna Romano
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One µg of Prostaglandin E2-d4 (CAYMAN CHEMICAL, Michigan, USA) was added as internal standard to the media (100–200 mL) recovered from the cell culture, from day 3 to day 10, through a filtration step on 1.2 µm nitrocellulose filters (Millipore RAWP04700). Culture media were loaded onto pre-packed CHROMABOND® HR-X cartridges (500 mg/6 mL) previously activated with methanol (12 mL) and milliQ water (12 mL). After a preliminary desalting step with 12 mL of milliQ water, collection of the organic components was achieved by elution with 16 mL of methanol followed by 16 mL of methanol/dichloromethane (1:1). The two organic fractions were combined and dried under reduced pressure for LC-MS/MS analysis.

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