Derivatization enhances analysis of estrogens and their bioactive metabolites in human plasma by liquid chromatography tandem mass spectrometry
| العنوان: | Derivatization enhances analysis of estrogens and their bioactive metabolites in human plasma by liquid chromatography tandem mass spectrometry |
|---|---|
| المؤلفون: | Nina, Denver, Shazia, Khan, Ioannis, Stasinopoulos, Colin, Church, Natalie Zm, Homer, Margaret R, MacLean, Ruth, Andrew |
| المصدر: | Analytica Chimica Acta |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | LC-MS, liquid chromatography-mass spectrometry, Estrogen metabolites, cps, counts per second, IS, internal standards, CID, collision-induced dissociation, RSD, relative standard deviation, Article, LLOQ/ULOQs, lower/upper limits of quantitation, Limit of Detection, Tandem Mass Spectrometry, LODs, limits of detection, PPZ, 1-(5-fluoro-2,4-dinitrophenyl)-4-methylpiperazine, MeOE1/2, methoxyestr-one/adiol, Humans, RME, relative mean error, Methylpiperazine, Piperazine, FA, formic acid, ESI, electrospray ionization, MPPZ, methyl-PPZ, Solid Phase Extraction, LC-MS/MS, liquid chromatography tandem mass spectrometry, SNR, signal/noise, OHE1/2, hydroxyestr-one/adiol, Estrogens, Derivatization, Estrogen, E1, estrone, SPE, solid phase extraction, Liquid chromatography tandem mass spectrometry, Linear Models, PAH, pulmonary arterial hypertension, SD, standard deviation, Blood Chemical Analysis, Chromatography, Liquid, MRM, multiple reaction monitoring |
| الوصف: | Estrogens regulate many diverse biological processes in health and disease. They circulate at a wide range of concentrations in females generating several active metabolites (hydroxy and methoxyestrogens). The metabolites are assumed to be present in much lower levels and are thought to contribute to diseases such as pulmonary arterial hypertension (PAH). Estrogen metabolites are challenging to quantify in plasma and currently available immunoassays are non-specific. Here we have developed and validated a novel assay to simultaneously quantify parent estrogens and their metabolites by mass spectrometry (MS). Estrogens were extracted from human plasma using solid phase extraction and derivatized using 1-(5-fluoro-2, 4-dinitrophenyl)-4-methylpiperazine (PPZ) before quaternization by methylation (“MPPZ”). MPPZ derivatives were separated and quantified by liquid chromatography tandem MS (LC-MS/MS) in positive electrospray ionization mode, using a QTrap 6500 + coupled to a Shimadzu Nexera X2. Separation was achieved using an ACE Excel 2 C18-PFP column (2 μm, 2.1 mm × 150 mm). The limits of quantification (LOQ) were 0.43–2.17 pg on column with a linear range from 2 or 10 - 2000 pg mL-1. Intra and inter-day precision and accuracy were acceptable ( Graphical abstract Image 1 Highlights • LC-MS/MS method for simultaneous quantification of estrogens and their bioactive metabolites in human plasma. • High recoveries and reduced matrix effects using MCX-SPE® extraction cartridges followed by MPPZ derivatization. • Generation of stable derivatives allowing quantification of estrogens in low endogenous levels. |
| تدمد: | 1873-4324 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=pmid________::6c42c2feaf6bb5dd75f9640cfc27c83eTest https://pubmed.ncbi.nlm.nih.gov/30712596Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.pmid..........6c42c2feaf6bb5dd75f9640cfc27c83e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 18734324 |
|---|