A high-performance liquid chromatography electrospray tandem mass spectrometry analysis of cortisol and metabolites in placental perfusate
A high-performance liquid chromatography electrospray tandem mass spectrometry analysis of cortisol and metabolites in placental perfusate
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DOI：
10.1006/abio.1997.2074
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发表时间：
1997-05-01
影响因子：
2.9
通讯作者：
Pond, SM
中科院分区：
文献类型：
--
作者：
Dodds, HM;Taylor, PJ;Pond, SM
采用反相高效液相色谱-电喷雾-串联质谱法（HPLC-ESI-MS/MS）定量测定皮质醇、可的松、20 α -和β -二氢皮质醇、20 α -和β -二氢可的松、四氢皮质醇和四氢可的松。该技术用于分析分离的人胎盘小叶的灌注液中皮质醇及其代谢物。用C18固相萃取筒从灌注培养基中制备分析物。分析的内标为6 α -甲基强的松龙。色谱柱为Novapak C18，室温下，流动相为53%甲醇和47% 10 mM甲酸铵缓冲液（pH 4.0），流速为80 μ L/min。采用PE-SCIEX API III三重四极杆质谱仪进行质谱检测。在负离子和正离子模式下使用离子喷雾（气动辅助电喷雾）界面。测定结果在100-2000 μ g/L范围内呈线性。仪器检出限为50 pg，在400和800 μ g/L时测定不精密度小于等于10%（总变异系数）。20 -二氢化可的松的准确度在83.2%到102.6%之间。1000 μ g/L可的松的回收率为91.3%，四氢皮质醇的回收率为109.7%。(C) 1997学术出版社。
A reversed-phase high-performance liquid chromatography-electrospray-tandem mass spectrometry assay (HPLC-ESI-MS/MS) was developed to quantitate cortisol, cortisone, 20 alpha- and beta-dihydrocortisol, 20 alpha- and beta-dihydrocortisone, tetrahydrocortisol, and tetrahydrocortisone. The technique was used to analyze perfusate from the isolated human placental lobule for cortisol and its metabolites. Analytes were prepared from the perfusion medium using C18 solid-phase extraction cartridges. The internal standard for the analyses was 6 alpha-methylprednisolone. Chromatography was performed on a Novapak C18 column at ambient temperature using 53% methanol and 47% 10 mM ammonium formate buffer (pH 4.0) as mobile phase, at a flow rate of 80 mu L/min. A PE-SCIEX API III triple quadrupole instrument was used for mass spectrometric detection. An ionspray (pneumatically assisted electrospray) interface was used in negative and positive ionization mode. The assay was linear over the range 100-2000 mu g/L for each analyte. The instrumental limit of detection was 50 pg. Assay imprecision at 400 and 800 mu g/L was less than or equal to 10% (total coefficient of variation). Accuracy ranged between 83.2% for 20 beta-dihydrocortisone to 102.6% for cortisone. Recovery of 1000 mu g/L analyte ranged from 91.3% for cortisone to 109.7% for tetrahydrocortisol. (C) 1997 Academic Press.