Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7626904 | Journal of Pharmaceutical and Biomedical Analysis | 2018 | 8 Pages |
Abstract
An LC-MS/MS method was developed and validated to analyze simultaneously estrogens (estradiol, E2; estrone, E1), androgens (testosterone, T; androstenedione, A4; dehydroepiandrosterone, DHEA), progestagens (17a-hydroxypregnenolone, 17OHP5; 17a-hydroxyprogesterone, 17OHP4; progesterone, P4), glucocorticoids (cortisol, F; cortisone E; corticosterone, B; 11-deoxycortisol, S; 21-hydroxyprogesterone, 21OHP4), and mineralocorticoids (aldosterone, A) from 150â¯Î¼l of human serum, plasma, or endometrium and endometriotic tissue homogenates. Samples spiked with isotope-labeled steroids as internal standards were extracted with toluene prior to LC-MS/MS analysis. The chromatographic separation of underivatized steroids was achieved on a biphenyl column with 0.2â¯mM NH4F as the eluent additive and a water-methanol gradient to improve E2 and E1 ionization. Method validation was performed with human plasma samples, and analysis of certified E2, T, F, and P4 reference serums (BCR-576, ERM-DA346, ERM-DA192, ERM-DA347), as well as homogenates of endometrium and endometriotic tissue. A total of 27 steroids were included in the method development to ensure the specificity of the method. After validation, the method was found suitable for quantitative analysis of 11 steroids: E2 (6.7â¯pM-13â¯nM), E1 (1.3â¯pM-6.6â¯nM),â¯T (3.3â¯pM-13â¯nM), A4 (13â¯pM-33â¯nM), 17OHP5 (32â¯pM-65â¯nM), 17OHP4 (33 pM-13â¯nM), F (33â¯pM-133â¯nM), E (13â¯pM-130â¯nM), B (33â¯pM-134â¯nM), S (13â¯pM-129â¯nM), and A (32â¯pM-32â¯nM). In addition, DHEA (333â¯pM-32â¯nM), P4 (13â¯pM-13â¯nM) and 21OHP4 (13â¯pM-13â¯nM) can be analyzed semiquantitatively.
Keywords
DHT17OHP421-hydroxyprogesteroneDHEA11-DeoxycortisoldehydroepiandrosteroneMTBEACNLLERSDCIDLLOQESILC–MSaldosteroneEndometriumMatrix effectCollision-induced dissociationinternal standardLiquid-liquid extractionEstradiolSteroidEstroneAcetonitrilerelative standard deviationandrostenedioneEndometriosiscollision energyTissuetert-butyl methyl ethertestosteronepregnenoloneDihydrotestosteroneRetention timeSerumlower limit of quantitationProgesteronequality controlcortisolcortisoneCorticosteroneelectrospray ionization
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Merja R. Häkkinen, Taija Heinosalo, Niina Saarinen, Tero Linnanen, Raimo Voutilainen, Timo Lakka, Jarmo Jääskeläinen, Matti Poutanen, Seppo Auriola,