Measurement of steroid synthesis in zona glomerulosa cells by liquid chromatography-electrospray ionization-mass spectrometry: inhibition by nitric oxide.
Measurement of steroid synthesis in zona glomerulosa cells by liquid chromatography-electrospray ionization-mass spectrometry: inhibition by nitric oxide.
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通过液相色谱-电喷雾电离-质谱法测量球状带细胞中的类固醇合成：一氧化氮的抑制。
DOI：
10.1016/j.ab.2004.11.025
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Campbell,WilliamB
中科院分区：
文献类型：
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作者：
Nithipatikom,Kasem;Holmes,BlytheB;Isbell,MarilynA;Hanke,CraigJ;Gomez-Sanchez,CelsoE;Campbell,WilliamB
建立了液相色谱-电喷雾电离-质谱法同时测定牛肾上腺皮质球细胞中醛固酮、皮质酮、皮质醇、脱氧皮质酮、去甲烯醇酮和孕酮浓度的方法。甾体类化合物采用液-液萃取法提取，反相C18柱分离，电喷雾电离，正离子模式单四极杆质谱检测。所有类固醇形成高丰度的钠加合物。研究了影响钠加合物形成和信号的因素。使用选择离子监测的检测限（S/N=3）对于这些类固醇为2 pg，对于去甲烯醇酮为10 pg。DETA NONOate是一种一氧化氮供体，以浓度依赖性方式抑制ZG细胞中这些类固醇的基础、血管紧张素II刺激和25-羟基胆固醇刺激合成。该技术证明了在肾上腺ZG细胞中醛固酮合成的每个酶促步骤和类固醇生成酶的活性中确定单个类固醇的能力。
A liquid chromatography–electrospray ionization–mass spectrometry method was developed to simultaneously determine the concentrations of aldosterone, corticosterone, cortisol, deoxycorticosterone, pregnenolone, and progesterone in bovine adrenal zona glomerulosa (ZG) cells. Steroids were extracted by liquid–liquid extraction, separated on a reverse-phase C18column, ionized by electrospray, and detected by single-quadrupole mass spectrometry in a positive ion mode. All steroids formed sodium adducts at high abundance. Factors affecting the formation and signal of sodium adducts were investigated. The limits of detection (S/N=3) using selected ion monitoring are 2pg for these steroids and 10pg for pregnenolone. DETA NONOate, a nitric oxide donor, inhibited the basal, angiotensin-II-stimulated, and 25-hydroxycholesterol-stimulated syntheses of these steroids in ZG cells in a concentration-dependent manner. The technique demonstrates the ability to determine the individual steroid in each enzymatic step of aldosterone synthesis and the activity of steroidogenic enzymes in adrenal ZG cells.