Clinical Development Laboratory, Merck Research Laboratories, Rahway, NJ 07065, United States.
J Pharm Biomed Anal. 2011 Nov 1;56(3):600-3. doi: 10.1016/j.jpba.2011.06.019. Epub 2011 Jun 29.
A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed for the quantitation of 6-keto PGF(1α) in human urine and plasma. Prostacyclin (PGI(2)) is a locally acting prostanoid, which mediates vasorelaxation and inhibition of platelet aggregation. 6-Keto PGF(1α) is the most-immediate metabolite of PGI(2). Samples were spiked with an internal standard (6-keto PGF(1α)-d(4)), purified by immuno-affinity chromatography and selected reaction monitoring (SRM) was performed. Analytical validation of the 6-keto PGF(1α) assay was performed in urine. This included an assessment of assay precision, recovery, stability, sensitivity and linearity. Urinary 6-keto PGF(1α) concentrations were also correlated to urinary 2,3-dinor-6-keto PGF(1α) (PGIM) concentrations using urine samples collected from 16 healthy volunteers. The mean concentration of 6-keto PGF(1α) in urine (mean ± SD) was 92 ± 51 pg/ml or 168 ± 91 pg/mg creatinine. Overall, there was a statistically significant correlation between urinary 6-keto PGF(1α) and PGIM (r(2)=0.55, p ≤ 0.001; slope=2.7; y-intercept=130). However, PGIM was approximately 3-fold more abundant than 6-keto PGF(1α) in urine. In addition, 6-keto PGF(1α) concentrations were measured in EDTA plasma samples obtained from 7 healthy donors. The mean concentration of 6-keto PGF(1α) in plasma was 1.9 ± 0.8 pg/ml (± SD).
建立了一种灵敏的液相色谱/串联质谱（LC/MS/MS）法，用于定量人尿液和血浆中的 6-酮前列腺素 F1α（6-keto PGF(1α)）。前列腺素（PGI(2)）是一种局部作用的前列腺素，介导血管舒张和血小板聚集抑制。6-酮前列腺素 F1α（6-keto PGF(1α)）是 PGI(2)的最直接代谢产物。样品用内标（6-酮前列腺素 F1α-d(4)）进行了标记，通过免疫亲和色谱进行了纯化，并进行了选择反应监测（SRM）。在尿液中对 6-酮前列腺素 F1α 测定进行了分析验证。这包括评估测定的精密度、回收率、稳定性、灵敏度和线性。使用从 16 名健康志愿者收集的尿液样本，将尿液中的 6-酮前列腺素 F1α 浓度与尿液中的 2,3-二去甲-6-酮前列腺素 F1α（PGIM）浓度相关联。尿液中 6-酮前列腺素 F1α 的平均浓度（平均值±SD）为 92±51pg/ml 或 168±91pg/mg 肌酐。总体而言，尿液中 6-酮前列腺素 F1α 与 PGIM 之间存在统计学显著相关性（r(2)=0.55，p≤0.001；斜率=2.7；y 轴截距=130）。然而，PGIM 在尿液中的含量约为 6-酮前列腺素 F1α 的 3 倍。此外，还测量了来自 7 名健康供体的 EDTA 血浆样品中的 6-酮前列腺素 F1α 浓度。血浆中 6-酮前列腺素 F1α 的平均浓度为 1.9±0.8pg/ml（±SD）。
