QUANTIFICATION OF THE MAJOR URINARY METABOLITE OF PROSTAGLANDIN-D2 BY A STABLE ISOTOPE-DILUTION MASS-SPECTROMETRIC ASSAY
QUANTIFICATION OF THE MAJOR URINARY METABOLITE OF PROSTAGLANDIN-D2 BY A STABLE ISOTOPE-DILUTION MASS-SPECTROMETRIC ASSAY
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DOI：
10.1016/0003-2697(91)90054-w
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发表时间：
1991-02-15
影响因子：
2.9
通讯作者：
ROBERTS, LJ
中科院分区：
文献类型：
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作者：
MORROW, JD;PRAKASH, C;ROBERTS, LJ
前列腺素 D2 (PGD2) 已被发现是许多人类疾病的重要病理生理介质。因此，评估 PGD2 内源性产生的方法具有相当大的临床价值。为了实现这一目标，我们开发了一种通过气相色谱/负离子化学电离质谱法定量 PGD2、9α,11β-二羟基-15-oxo-2,3,18,19-tetranorprost-5-ene-1,20-dioic Acid 的主要尿代谢物的方法。该代谢物经过化学合成并转化为 18 O4 标记的衍生物，用作内标。利用该分子的下侧链在酸性 pH 下进行环化形成半缩酮、γ-内酯并通过甲肟化进行解环的能力，将新颖的衍生化和纯化程序纳入测定中。测定精度为±7%，准确度为96%。灵敏度下限约为 50 pg。 18 名正常成人中该代谢物的尿排泄正常水平为 1.08 ± 0.72 ng/mg 肌酐（平均值 ± 2SD）。在前列腺素 D2 释放量增加的临床情况下，发现代谢物的尿排泄量显着升高。因此，该测定提供了一种敏感且准确的方法来评估前列腺素 D2 的内源性产生，作为探索前列腺素 D2 在人类疾病中的病理生理学作用的手段。
Prostaglandin D2(PGD2) has been found to be an important pathophysiological mediator in a number of human disorders. Thus a means to assess the endogenous production of PGD2is of considerable clinical value. To accomplish this goal, we developed a method for the quantification of the major urinary metabolite of PGD2, 9α, 11β-dihydroxy-15-oxo-2,3,18,19-tetranorprost-5-ene-1,20-dioic acid, by gas chromatography/negative ion chemical ionization mass spectrometry. This metabolite was chemically synthesized and converted to an18O4-labeled derivative for use as an internal standard. Novel derivatization and purification procedures were incorporated in the assay taking advantage of the ability of the lower side chain of this molecule to undergo cyclization at acidic pH to form a hemiketal, γ-lactone, and uncyclization with methoximation. Precision of the assay is ±7% and accuracy is 96%. The lower limit of sensitivity is approximately 50 pg. Normal levels for the urinary excretion of this metabolite in 18 normal adults was found to be 1.08 ± 0.72 ng/mg creatinine (mean ± 2SD). Substantial elevations in the urinary excretion of the metabolite were found in clinical situations in which prostaglandin D2has been shown to be released in increased quantities. Thus, this assay provides a sensitive and accurate method to assess endogenous production of prostaglandin D2as a means to explore the pathophysiological role of prostaglandin D2in human disease.