ANALYSIS OF LEUKOTRIENES, PROSTAGLANDINS, AND OTHER OXYGENATED METABOLITES OF ARACHIDONIC-ACID BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
ANALYSIS OF LEUKOTRIENES, PROSTAGLANDINS, AND OTHER OXYGENATED METABOLITES OF ARACHIDONIC-ACID BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
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DOI：
10.1016/0003-2697(84)90137-4
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发表时间：
1984-01-01
影响因子：
2.9
通讯作者：
ELING, TE
中科院分区：
文献类型：
--
作者：
HENKE, DC;KOUZAN, S;ELING, TE
开发了高效液相色谱[HPLC]方法，其分离白三烯（LT）、羟基脂肪酸（HETE）、前列腺素（PG）、前列环素的稳定代谢物（6-酮-PGF 1 α），血栓素A_2（TXB_2）、12-羟基十七碳三烯酸（HHT）和花生四烯酸（AA）的稳定代谢产物。介绍了两种反相柱的分离方法。一种方法使用径向压缩系统，另一种方法使用传统的钢柱。两种系统均采用甲醇和缓冲水作为溶剂。径向压缩系统需要60分钟来分离AA代谢物，而常规系统需要100分钟。两种方法都提供了6-酮-PGF 1 α的良好分离和回收，TXB2、PGE2、PGF2 α、PGD 2、LTC 4、LTB 4、LTD 4、LTE 4、HHT、15-、12-和5-HETE;和AA。5S，12 S-二羟基-6-反式、8-顺式、10-反式、14-顺式-二十碳四烯酸（5S，12 S-diHETE）是LTB 4的立体异构体，与LTB 4共洗脱。为了确定该方法对生物系统的适用性，在2种模型（豚鼠肺微粒体和大鼠肺泡巨噬细胞）中研究了AA代谢。两种HPLC系统均证明了2种生物系统中类二十烷酸的良好回收率和分离度。一个简单的蒸发技术HPLC，样品制备，避免使用色谱和其他耗时的方法，也被描述。
High-performance liquid chromatography [HPLC] procedures were developed which separate leukotrienes (LT), hydroxy-fatty acids (HETE), prostaglandins (PG), the stable metabolite of prostacyclin (6-keto-PGF1.alpha.), the stable metabolite of thromboxane A2 (TXB2), 12-hydroxyheptadecatrienoic acid (HHT) and arachidonic acid (AA). Two methods employing revere-phase columns are described. One method uses a radial compression system, the other a conventional steel column. Both systems employ methanol and buffered water as solvents. The radial compression system requires 60 min for separation of the AA metabolites, while the conventional system requires 100 min. Both methods provide good separation and recovery of 6-keto-PGF1.alpha., TXB2, PGE2, PGF2.alpha., PGD2, LTC4, LTB4, LTD4, LTE4, HHT, 15-, 12- and 5-HETE; and AA. The 5S,12S-dihydroxy-6-trans, 8-cis, 10-trans, 14-cis-eicosatetraenoic acid (5S,12S-diHETE), a stereoisomer of LTB4, coelutes with LTB4. To determine the applicability of the methods to biologic systems, AA metabolism was studied in 2 models, guinea pig lung microsomes and rat alveolar macrophages. Both HPLC systems demonstrated good recovery and resolution of eicosanoids from the 2 biological systems. A simple evaporation technique for HPLC, sample preparations, which avoids the use of chromatographic and other time-consuming methodology, is also described.