[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-PGF1 alpha, TXB2, PGF2 alpha, PGE2 and 13,14-dihydro PGF2 alpha 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 min 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.