Skrinska V, Lucas F V
Prostaglandins. 1981 Sep;22(3):365-75. doi: 10.1016/0090-6980(81)90099-x.
A method was development for the isolation of prostacyclin (PGI2) from whole blood in a fraction suitable for high pressure liquid chromatography (HPLC) separation of PGI2 and 6-keto-prostaglandin F1 alpha (6-K-PGF1 alpha). Prostacyclin was stabilized in whole blood by rapidly raising the pH to 10 with Na2CO3 and cooling the samples to 0 degree C. Under these conditions, 2.9% hydrolysis was observed after 20 min. Reverse phase extraction columns were used to directly extract both PGI2 and 6-K-PGF1 alpha from the alkaline plasma with recoveries of greater than 95% using an acetonitrile/2mM Na2B4O7, pH 10, 40/60 elution solvent mixture. An additional 1.7% hydrolysis was found during the column extraction procedure. Final separation of PGI2 and 6-K-PGF1 alpha was performed with HPLC using an alkaline solvent system. This method is capable of rapidly and efficiently extracting and separating PGI2 and 6-K-PGF1 alpha from whole blood or plasma. It introduces less than 5% hydrolysis of PGI2, thus providing a means of applying highly sensitive 6-K-PGF1 alpha assays to the determination of PGI2 levels in physiological samples.
已开发出一种从全血中分离前列环素（PGI2）的方法，所得组分适用于对PGI2和6-酮-前列腺素F1α（6-K-PGF1α）进行高压液相色谱（HPLC）分离。通过用Na2CO3迅速将pH值提高到10并将样品冷却至0℃，可使前列环素在全血中稳定。在这些条件下，20分钟后观察到2.9%的水解。使用反相萃取柱从碱性血浆中直接萃取PGI2和6-K-PGF1α，使用乙腈/2mM Na2B4O7（pH 10，40/60）洗脱溶剂混合物时回收率大于95%。在柱萃取过程中发现额外有1.7%的水解。PGI2和6-K-PGF1α的最终分离采用HPLC，使用碱性溶剂系统。该方法能够快速有效地从全血或血浆中萃取和分离PGI2和6-K-PGF1α。它使PGI2的水解率低于5%，从而为将高灵敏度的6-K-PGF1α测定法应用于生理样品中PGI2水平的测定提供了一种方法。
