Leffler C W, Desiderio D M, Wakelyn C E
Prostaglandins. 1981 Feb;21(2):227-41. doi: 10.1016/0090-6980(81)90140-4.
A method for isolating climax products of the arachidonic acid cascade from biological fluids is described which allows simultaneous measurement of PGs (PGE2, PGD2, 6-keto-PGF1 alpha, TXB2, 6-keto-PGE1, 6, 15-diketo-13,14-dihydro-PGF1 alpha) by electron capture detection of pentafluorobenzyloxime methyl ester trimethylsilyl (TMS) ether derivatives. PGs are adsorbed onto Amberlite XAD-2 from acidified solution and nonadhering substances removed by sequential administration of water, then petroleum ether. PGs are extracted into methanol. Following evaporation and reconstitution in water, the PGs are extracted into ethyl acetate at pH 3 and the ethyl acetate extracts are purified by lipidex column chromatography. Derivatization to pentafluorobenzyloxime methyl ester TMS ethers of PGs in the sample is followed by separation on either glass packed-columns or SCOT capillary columns, and detection by an electron capture detector. PGA2, added to the unpurified sample, is used as an internal standard for quantification. The methods have performed well on all biological fluids thus far examined. Examples of chromatographs from urine, Krebs-perfused lung effluents, and blood are shown.
描述了一种从生物流体中分离花生四烯酸级联反应终产物的方法，该方法可通过电子捕获检测五氟苄基肟甲酯三甲基硅烷基（TMS）醚衍生物同时测定前列腺素（PGs）（PGE2、PGD2、6-酮-PGF1α、TXB2、6-酮-PGE1、6,15-二酮-13,14-二氢-PGF1α）。PGs从酸化溶液中吸附到Amberlite XAD-2上，通过依次用水、然后石油醚处理除去非粘附物质。PGs用甲醇萃取。蒸发并在水中复溶后，PGs在pH 3时萃取到乙酸乙酯中，乙酸乙酯萃取物通过lipidex柱色谱法纯化。将样品中的PGs衍生化为五氟苄基肟甲酯TMS醚，然后在玻璃填充柱或SCOT毛细管柱上进行分离，并用电子捕获检测器检测。加入未纯化样品中的PGA2用作定量的内标。到目前为止，这些方法在所有检测的生物流体中表现良好。展示了来自尿液、Krebs灌注肺流出液和血液的色谱图示例。
