HPLC DETERMINATION OF PROSTAGLANDINS WITH SMALL-BORE ODS COLUMN

Several prostaglandins can be separated and identified with 2.6mm small bore ODS column-HPLC at 204 nm. The mobile phase used was 26% acetonitrile at the flow rate of 0.3 ml/min. The PGs studied were 6-keto-PGF_1a, TXB₂, PGF_2a, PGE₂ and 13,14-dihydro PGF_2a with retention times of 3.3, 5.6, 7.8, 8.8 and 11 minutes, respectively.Rabbit gastric mucosa was homogenized, incubated with ¹⁴C-AA (33 nmol)for 20 rain and then extracted with ethyl acetate. The organic phase was dried under nitrogen, the residue was dissolved in 25% acetonitrile and injected into the HPLC system. Fractions of effluent were collected every 20 seconds and the radioactivity was measured. The results show that the peaks of radioactivity coincided well with those of the PG standard samples.When indomethacin was added to the incubation system, the radioactivity counts decreased significantly.