Coker S J, Clarke B, Zeitlin I J
J Pharmacol Methods. 1982 May;7(3):207-17. doi: 10.1016/0160-5402(82)90037-7.
Radioimmunoassay techniques are described that enable the quantification of the local release of prostaglandin (PG) E2, PGF2 alpha, TxB2 (the metabolite of thromboxane A2), and 6-keto PGF1 alpha (a metabolite of prostacyclin) from various organs in vivo. Plasma samples were acidified and the prostanoids extracted with ethyl acetate, prior to assay. The mean recovery of tritiated internal standards was 60% for 6-keto PGF1 alpha and 84% for the other prostanoids. Sample extracts were redissolved in phosphate-buffered saline and the appropriate tritiated antigen and specific antibody were added. After overnight incubation the antibody-bound and free prostanoid were separated using dextran-coated charcoal. The intraassay variation and the interassay variation were similar for all prostanoids; the mean values being 8.5% and 13.2%, respectively. There were variations in the sensitivity of the radioimmunoassay for each prostanoid. Using 250 microliter plasma samples the detection limit for PGE2 was 75 pg/ml, PGF2 alpha 50 pg/ml, TxB2 25 pg/ml, and 6-keto PGF1 alpha 125 pg/ml. These assays have been used to determine prostanoid concentrations in rats, cats, dogs, and humans.
本文描述了放射免疫分析技术，该技术能够对体内各种器官中前列腺素（PG）E2、PGF2α、TxB2（血栓素A2的代谢产物）和6-酮-PGF1α（前列环素的代谢产物）的局部释放进行定量。在测定之前，将血浆样品酸化并用乙酸乙酯提取前列腺素。6-酮-PGF1α的氚化内标平均回收率为60%，其他前列腺素为84%。将样品提取物重新溶解于磷酸盐缓冲盐水中，并加入适当的氚化抗原和特异性抗体。过夜孵育后，使用葡聚糖包被的活性炭分离结合抗体的和游离的前列腺素。所有前列腺素的批内变异和批间变异相似；平均值分别为8.5%和13.2%。每种前列腺素的放射免疫分析灵敏度存在差异。使用250微升血浆样品时，PGE2的检测限为75皮克/毫升，PGF2α为50皮克/毫升，TxB2为25皮克/毫升，6-酮-PGF1α为125皮克/毫升。这些分析方法已用于测定大鼠、猫、狗和人类体内的前列腺素浓度。
