Comparison of isotope pattern deconvolution and calibration curve quantification methods for the determination of estrone and 17β-estradiol in human serum
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Other documents of the author: Pitarch-Motellón, Jorge; Fabregat-Cabello, Neus; Le Goff, C.; Roig-Navarro, Antoni F.; Sancho, Juan V; Cavalier, E.
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Show full item recordcomunitat-uji-handle:10234/9
comunitat-uji-handle2:10234/33596
comunitat-uji-handle3:10234/33597
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Title
Comparison of isotope pattern deconvolution and calibration curve quantification methods for the determination of estrone and 17β-estradiol in human serumAuthor (s)
Date
2019-04Publisher
ElsevierBibliographic citation
PITARCH-MOTELLÓN, J., et al. Comparison of isotope pattern deconvolution and calibration curve quantification methods for the determination of estrone and 17β-estradiol in human serum. Journal of pharmaceutical and biomedical analysis, 2019, 171: 164-170.Type
info:eu-repo/semantics/articlePublisher version
https://www.sciencedirect.com/science/article/pii/S0731708519303255Version
info:eu-repo/semantics/acceptedVersionSubject
Abstract
A Liquid Chromatography coupled to tandem mass spectrometry (LC–MS/MS) based method have been developed for the determination of the main estrogen compounds –estrone (E1) and 17β-estradiol (E2)– in human serum. Two ... [+]
A Liquid Chromatography coupled to tandem mass spectrometry (LC–MS/MS) based method have been developed for the determination of the main estrogen compounds –estrone (E1) and 17β-estradiol (E2)– in human serum. Two isotope dilution mass spectrometry (IDMS) quantification procedures have been used: a classical calibration curve-based method were compared to a recently developed isotope pattern deconvolution (IPD) method. IPD is based on isotopic abundance measurements and multiple linear regression. Validation was performed in terms of intra-assay repeatability (n = 5), inter-assay reproducibility (n = 9) and accuracy using spiked steroid-free serum at 5 concentration levels and 3 certified reference materials. Both methodologies meet EMEA requirements yielding recoveries between 79–106% and coefficient of variations of 1.7–8.3% along all experiments. Limits of quantification as low as 5 ng/L were achieved. 40 real samples were analysed for comparison purposes showing a great correlation between calibration and IPD concentration values. Real samples were also quantified by routine immunoassay analysis, which showed a significant proportional bias of 2.55 for E1 and good correlation for E2. While methods were considered suitable for routine or countercheck analysis within the context of hospital’s needs, IPD has demonstrated to be faster and cost saving. [-]
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