用细口径反相柱HPLC分离多种前列腺素

HPLC与放射性同位素联用,可同时分离鉴定多种花生四烯酸代谢产物。目前国外一般是使用普通口径(4.6 mm)的反相柱,以30%左右乙腈为流动相,流速为1～3 ml/min。这种HPLC方法乙腈用量较大,费用太贵,不适合国内实验室常规操作。据报道,细口径

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), TXB_2, PGF_(2a), PGE_2 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 ~(14)C-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.