Daniel V C, Minton T A, Brown N J, Nadeau J H, Morrow J D
Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232-6602.
J Chromatogr B Biomed Appl. 1994 Mar 4;653(2):117-22. doi: 10.1016/0378-4347(93)e0432-p.
Endogenous prostacyclin production is best assessed by the measurement of its excreted metabolites, of which a major one is 2,3-dinor-6-ketoprostaglandin F1 alpha (2,3-dinor-6-keto-PGF1 alpha). Gas chromatographic-mass spectrometric (GC-MS) assays have been developed for this compound but are cumbersome and time-consuming. We now report a modified assay for the measurement of 2,3-dinor-6-keto-PGF1 alpha employing GC-MS in which sample preparation time is markedly shortened by replacing a number of extraction steps with reversed-phase column extraction and by modifying derivatization procedures. Precision of the assay is +/- 5% and the accuracy is 98%. The lower limit of detection in urine is approximately 15 pg/mg creatinine. Normal urinary levels of this metabolite were found to be 141 +/- 54 pg/mg creatinine (mean +/- S.D.). Urinary excretion of 2,3-dinor-6-keto-PGF1 alpha is markedly altered in situations associated with abnormalities of prostacyclin generation when quantified using this assay. Thus, this assay provides a sensitive and accurate method to assess endogenous prostacyclin production and to further explore the role of this compound in human health and disease.
内源性前列环素的产生情况最好通过对其排泄代谢产物的测量来评估，其中主要的一种代谢产物是2,3-二去甲-6-酮前列环素F1α（2,3-二去甲-6-酮-PGF1α）。已开发出用于该化合物的气相色谱-质谱（GC-MS）分析法，但操作繁琐且耗时。我们现在报告一种采用GC-MS测量2,3-二去甲-6-酮-PGF1α的改良分析法，其中通过用反相柱萃取替代多个萃取步骤并修改衍生化程序，显著缩短了样品制备时间。该分析法的精密度为±5%，准确度为98%。尿液中的检测下限约为15 pg/mg肌酐。发现这种代谢产物的正常尿液水平为141±54 pg/mg肌酐（平均值±标准差）。当使用该分析法进行定量时，在与前列环素生成异常相关的情况下，2,3-二去甲-6-酮-PGF1α的尿排泄量会发生显著变化。因此，该分析法提供了一种灵敏且准确的方法来评估内源性前列环素的产生情况，并进一步探索该化合物在人类健康和疾病中的作用。
