高速 液體 Chromatography에 의한 Prostaglandin F_2α및 E_2定量에 관한 硏究 = A Study on the High Performance Liquid Chromatographic Determination of Prostaglandin F_2αand E_2

다국어 초록 (Multilingual Abstract)

Prostaglandin F_2α and E_2 are being applied for induction of labor, termination of first or second trimester pregnancies and postpartum atonic uterine bleeding.
Though various analytical methods for quantitation of prostaglandins were reported elsewhere, the development of a more sensitive, rapid and progressive method has long been desired.
This study was conducted to quantitate PGF_2α and PGE_2 simultaneously by high performance liquid chromatography after p-bromophenacylation of Sep-Pak C_18, cartridge extracts.
The results were as follows:
1. A high performance liquid chromatographic analysis were developed for the rapid, simultaneous separation and quantitative recovery of PGF_2α, PGE_2 as their p-bromophenacyl esters to improve chromatographic properties and to detect them at 254 nm.
The procedure involves p-bromophenacylation of PGF_2α and PGF_2 under N, N-diisopropyl ethylamine catalyst after rapid extraction using Sep-Pak C_18 cartridge, which is then chromatographed on μ Bondapak column (8 mm i.d. x 10 cm) using acetonitrile : water(1 : 1) as mobile phase.
2. HPLC peaks of PGF_2α and PGE_2 were observed at 10.8 min. and 12.3 min., respectively. Linear calibration plots of peak area versus PGF_2α and PGE_2 concentration were obtained over the range of 0.1-1.2㎍/㎕.
This method enables to determine simultaneously PGF_2α, and PGE_2, thus shortening the assay time in comparison with that of p-nitrophenacylation.
3. p-Bromophenacylation was complete in about 40 min. at room temperature, and aliquots of the reaction mixtures were capable to be analyzed directly by HPLC.
4. The recovery in the concentration of 20 ㎍-4 mg prostaglandin was 99% by single Sep-Pak C_18 cartridge(1 cm i.d. X 1 cm) extraction with 10 ml ethanol(over 70 % ). The amount of prostaglandin was measured by ultra-violet spectrophotometry and fluorometry. For the Sep-Pak C_18 extraction, the optimum pH range was acidic(pH 3. 0). The present method allowed to obtain a clean extract from various samples suitable for HPLC analysis.