Serum steroid profiling by isotopic dilution-liquid chromatography-mass spectrometry: Comparison with current immunoassays and reference intervals in healthy adults

被引：162
作者：
Fanelli, Flaminia [1 ,2 ]
Belluomo, Ilaria [1 ,2 ]
Di Lallo, Valentina D. [1 ,2 ]
Cuomo, Gaia [1 ,2 ]
De Iasio, Rosaria [3 ]
Baccini, Margherita [1 ,2 ]
Casadio, Elena [1 ,2 ]
Casetta, Bruno [4 ]
Vicennati, Valentina [1 ,2 ]
Gambineri, Alessandra [1 ,2 ]
Grossi, Gabriele [3 ]
Pasquali, Renato [1 ,2 ]
Pagotto, Uberto [1 ,2 ]
机构：
[1] Univ Bologna, S Orsola Malpighi Hosp, Endocrinol Unit, I-40138 Bologna, Italy
[2] Univ Bologna, S Orsola Malpighi Hosp, Ctr Appl Biomed Res, Dept Clin Med, I-40138 Bologna, Italy
[3] St Orsola Marcello Malpighi Hosp, Cent Lab, Bologna, Italy
[4] AB Sciex, I-20052 Monza, Italy
来源：
STEROIDS | 2011年 / 76卷 / 03期
关键词：
Tandem mass spectrometry; Immunoassay; Reference interval; ADRENAL-STEROIDS; GAS-CHROMATOGRAPHY; TESTOSTERONE; ASSAYS; QUANTIFICATION; RADIOIMMUNOASSAY; ANDROSTENEDIONE; PERFORMANCE; EXTRACTION; CORTISOL;
D O I：
10.1016/j.steroids.2010.11.005
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Background: The simultaneous, rapid and reliable measurement of a wide steroid panel is a powerful tool to unravel physiological and pathological hormone status. Clinical laboratories are currently dominated by high-throughput immunoassays, but these methods lack specificity due to cross-reactivity and matrix interferences. We developed and validated an isotopic dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) method for the simultaneous measurement of cortisol, corticosterone, 11deoxycortisol, androstenedione, deoxycorticosterone (DOC), testosterone, 17OHprogesterone, dehydroepiandrosterone (DHEA) and progesterone in serum, and compared it to routine immunoassays employed in our laboratory. We also established adult reference intervals in 416 healthy subjects. Methods: 0.9 ml of serum were spiked with labelled internal standards (IS) and extracted on C18 cartridges. Eluate was injected into a two-dimensional LC-system, purified in a perfusion column and separated on a C8 column during a 21 min gradient run. Analytes were revealed by atmospheric pressure chemical ionization (APCI) followed by multiple reaction monitoring (MRM) analysis. Results: Of the four immunoassays compared with the ID-LC-MS/MS method, only the results of ElecsysE170 for cortisol, testosterone in males and progesterone > 1 ng/ml were in agreement with ID-LC-MS/MS. ElecsysE170 for testosterone in females and progesterone <1 ng/ml, Immulite2000 for androstenedione, DSL-9000 for DHEA and 17OHP Bridge for 17OHprogesterone, respectively, showed poor agreement. Reference intervals and steroid age and fertility related fluctuations were established. Conclusion: Our ID-LC-MS/MS method proved to be reliable and sensitive in revealing steroid circulating concentrations in adults and in highlighting the limits of routine immunoassays at low concentrations. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码：244 / 253
页数：10
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