Automated extraction and HPLC resolution of lipoxygenase and cyclooxygenase products utilizing high pressure column switching.

An automatable column switching technique for extraction of cyclooxygenase and lipoxygenase products has been developed. The extraction column is a 0.4 cm X 3 cm stainless steel column slurry packed with Polygosil (C18, 25-40 mu particle diameter). Supernatants of incubations terminated by the addition of 1 volume MeOH, bath sonicated and centrifuged were pumped onto the extraction column. The non-adherent polar material was eluted to waste with up to five times the sample volume with a high polarity mobile phase. Following this, the extraction column was then eluted to an analytical column with a mobile phase of decreased polarity.