Sensitive simultaneous quantitation of testosterone and estradiol in serum by LC–MS/MS without derivatization and comparison with the CDC HoSt program Journal Article


Authors: Schofield, R. C.; Mendu, D. R.; Ramanathan, L. V.; Pessin, M. S.; Carlow, D. C.
Article Title: Sensitive simultaneous quantitation of testosterone and estradiol in serum by LC–MS/MS without derivatization and comparison with the CDC HoSt program
Abstract: Background Very sensitive measurements of serum estrogens and testosterone are important in adult and pediatric endocrinology and immunoassays are known to lack the required performance at very low levels. Our aim was to develop a sensitive HPLC–MS/MS assay for both estradiol (E2) and testosterone (Te) in serum without the need for chemical derivatization and using commercially available calibrators. Methods Serum samples were prepared by the addition of internal standards followed by extraction using hexane:ethyl acetate. Chromatographic separation was achieved using a C18 column and mass spectrometry was performed in both positive and negative ion modes. Results The lower limits of quantitation (LLOQs) of E2 and Te were 5 pg/mL and 1 ng/dL, respectively. The analytical measurement range (AMR) for E2 was 5–600 pg/mL and 1–1,170 ng/dL for Te. Assay accuracy was determined both by comparison with a LC–MS/MS method performed at a national laboratory and the CDC HoSt program. Comparison with samples analyzed by both methods showed excellent correlation. Within-day (N = 10) and between-day (N = 20) CVs at concentrations spanning the AMR were less than 7% for both analytes. Conclusion We have developed an accurate and highly sensitive assay to measure E2 and Te levels in serum by HPLC–MS/MS without chemical derivatization and using commercially available calibrators. © 2017 Elsevier B.V.
Keywords: mass spectrometry; body fluids; high performance liquid chromatography; estradiol; liquid chromatography; testosterone; spectrometry; endocrinology; chromatography; negative ions; method correlation; cdc host; nist srm 971; chemical derivatization; chromatographic separations; lower limits of quantitations; sensitive measurement
Journal Title: Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
Volume: 1048
ISSN: 1570-0232
Publisher: Elsevier Science, Inc.  
Date Published: 2017-03-24
Start Page: 70
End Page: 76
Language: English
DOI: 10.1016/j.jchromb.2017.02.006
PROVIDER: scopus
PUBMED: 28222335
PMCID: PMC5379634
DOI/URL:
Notes: Article -- Export Date: 2 March 2017 -- Source: Scopus
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  1. Dean Christian Carlow
    33 Carlow