Adrenal steroids are generated in the adrenal cortex and metabolized by various enzymes such as hydroxylases, dehydrogenases, and reductases. Determining the comprehensive metabolic signatures of adrenal steroids can provide insight into their metabolic functions and roles in the pathophysiology of adrenal diseases, including Cushing's syndrome (CS) and congenital adrenal hyperplasia (CAH). To this end, we developed an advanced quantitative profiling method of serum adrenal steroids with liquid chromatography-mass spectrometry (LC-MS) under molecular-specific scan modes. Twenty-seven steroids were separated on a 1.9-μm particle C18 column (50 × 2.1 mm) at a flow rate of 250 μL/min and quantified via triple-quadrupole MS with electrospray ionization. During validation, linearities ( r2) were higher than 0.940 with a limit of quantification of 0.1-5.0 ng/mL, and precision (coefficient of variation) and accuracy (%bias) of 3.7-14.3 % and 96.3-113.1 %, respectively. In contrast with the significantly increased serum levels of mineralocorticoids (P <  0.001), the present LC-MS assay revealed remarkably decreased levels of all glucocorticoids and androgens in a patient diagnosed with 17α-hydroxylase deficiency CAH (P <  0.001) compared to those of age- and sex-matched healthy and CS subjects. In the CAH patient, the metabolic ratios for 17α-hydroxylase were significantly decreased, whereas there was no reduction in the metabolic ratio of 17-hydroxyprogesterone to androstenedione, indicating 17,20-lyase activity. In particular, both pregnenolone and dehydroepiandrosterone sulfates, and their metabolic ratio, were identified as potential biomarkers for 17α-hydroxylase deficiency (all P <  0.001), which were also distinct from those of CS patients. The devised LC-MS assay clearly revealed the metabolic signatures of 17α-hydroxylase deficiency, as a rare phenotype of CAH, compared to both healthy and CS subjects, indicating its utility for screening adrenal diseases.

中文翻译：

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基于选择性LC-MRM / SIM-MS的肾上腺类固醇分析揭示了17α-羟化酶缺乏症的代谢特征。

肾上腺类固醇在肾上腺皮质中产生，并通过各种酶（例如羟化酶，脱氢酶和还原酶）代谢。确定肾上腺类固醇的全面代谢特征可以提供对它们的代谢功能和在包括库欣综合征（CS）和先天性肾上腺增生（CAH）在内的肾上腺疾病病理生理中的作用的了解。为此，我们开发了一种在分子特异性扫描模式下用液相色谱-质谱联用技术（LC-MS）对血清肾上腺类固醇进行定量分析的先进方法。在1.9μm的C18色谱柱（50×2.1 mm）上以250μL/ min的流速分离了27种类固醇，并通过三重四极杆质谱与电喷雾电离进行了定量。验证期间，线性度（r2）高于0.940，定量限为0。1-5.0 ng / mL，精密度（变异系数）和准确度（偏差百分比）分别为3.7-14.3％和96.3-113.1％。与盐皮质激素的血清水平显着升高（P <0.001）相比，本LC-MS分析显示，诊断为17α-羟化酶缺乏症CAH的患者中，所有糖皮质激素和雄激素的水平均显着降低（P <0.001）。年龄和性别相匹配的健康和CS受试者。在CAH患者中，17α-羟化酶的代谢率显着降低，而17-羟孕酮与雄烯二酮的代谢率没有降低，表明17,20-裂合酶活性。尤其是孕烯醇酮和去氢表雄酮硫酸盐，以及它们的代谢率，被确定为17α-羟化酶缺乏症的潜在生物标志物（所有P <0.001），这也与CS患者不同。与健康和CS受试者相比，设计的LC-MS测定法清楚地揭示了17α-羟化酶缺乏症的代谢特征，这是CAH的罕见表型，表明其可用于筛查肾上腺疾病。