Henden T, Strand H, Børde E, Semb A G, Larsen T S
Department of Medical Physiology, Institute of Medical Biology, University of Tromoø, Norway.
Prostaglandins Leukot Essent Fatty Acids. 1993 Nov;49(5):851-4. doi: 10.1016/0952-3278(93)90209-f.
Leukotrienes (LTs) are biologically active compounds derived from lipoxygenase catalyzed metabolism of arachidonic acid in mammalian tissues. The present report describes a simple method for extraction and isolation of dihydroxy-LTs; LTB4, LTB5 and the peptido-LTs; LTC4, LTD4 and LTE4 from human plasma, using a pretreatment cartridge which utilizes both hydrophobic and ion-exchange interactions. 5 ml acidified plasma or acetate buffer containing commercially available LT standards were passed through the cartridges under suction, and the absorbed LTs were subsequently eluted in a stepwise manner with acetate buffer containing increasing amounts of methanol. The eluted LTs were analyzed by reversed-phase high performance liquid chromatography (HPLC) on octadecylsilyl (ODS)-silica, using a Waters HPLC unit. Both with plasma and acetate buffer the present methodology resulted in good separation of the LTs with a total run-time of less than 32 min. Recovery of dihydroxy-LTs was approximately 80% (range 73-82%) both when the standards were dissolved in plasma and in acetate buffer. Recovery of the peptido-LTs was, however somewhat lower (47-50%). It should be noted that the present method has the advantage that exposure to chemicals of high toxicity is avoided.
白三烯（LTs）是哺乳动物组织中由脂氧合酶催化花生四烯酸代谢产生的生物活性化合物。本报告描述了一种从人血浆中提取和分离二羟基白三烯（LTB4、LTB5）和肽基白三烯（LTC4、LTD4和LTE4）的简单方法，该方法使用了一种利用疏水和离子交换相互作用的预处理柱。将5毫升酸化血浆或含有市售LT标准品的醋酸盐缓冲液在抽吸作用下通过柱子，随后用含有逐渐增加量甲醇的醋酸盐缓冲液逐步洗脱吸附的LTs。使用沃特斯高效液相色谱仪，通过反相高效液相色谱（HPLC）在十八烷基硅烷（ODS）硅胶上对洗脱的LTs进行分析。无论是血浆还是醋酸盐缓冲液，本方法都能实现LTs的良好分离，总运行时间不到32分钟。当标准品溶解在血浆和醋酸盐缓冲液中时，二羟基白三烯的回收率约为80%（范围为73 - 82%）。然而，肽基白三烯的回收率略低（47 - 50%）。应当指出，本方法的优点是避免了接触高毒性化学品。
