Senior Mass Spectroscopist.
Androgenic biomarker profiling in human matrices and cell culture samples using high throughput, electrospray tandem mass spectrometry
Article first published online: 21 FEB 2014
© 2014 Wiley Periodicals, Inc.
Volume 74, Issue 7, pages 722–731, May 2014
How to Cite
Wilton, J. H., Titus, M. A., Efstathiou, E., Fetterly, G. J. and Mohler, J. L. (2014), Androgenic biomarker profiling in human matrices and cell culture samples using high throughput, electrospray tandem mass spectrometry. Prostate, 74: 722–731. doi: 10.1002/pros.22792
Conflicts of interest: No potential conflicts of interest were disclosed.
- Issue published online: 15 APR 2014
- Article first published online: 21 FEB 2014
- Manuscript Accepted: 27 JAN 2014
- Manuscript Received: 2 OCT 2013
- Department of Defense. Grant Number: DAMD 17-03-2-0052
- Department of Defense Prostate Cancer Research Program. Grant Numbers: W81XWH-10-1-0273, W81XWH-10-1-1000
- PCF Challenge Award
- National Cancer Institute. Grant Number: NCI-CCSG-CA016056
- androgen axis;
- prostate cancer;
- steroid 5α-reductase;
A high throughput, high pressure liquid chromatographic (HPLC) method with triple quadrupole mass spectral detection (LC/MS/MS) was validated for the measurement of 5 endogenous androgens in human plasma and serum and applied to various in vivo and in vitro study samples to pursue a better understanding of the interrelationship of the androgen axis, intracrine metabolism, and castration-recurrent prostate cancer (CaP).
A Shimadzu HPLC system interfaced with a Sciex QTRAP 5500 mass spectrometer with electrospray ionization was used with inline column-switching. Samples were liquid/liquid extracted and chromatographed on a Luna C18(2) column at 60°C with a biphasic gradient using a 15-min run time.
The method was validated for five androgens in human plasma and serum, and applied to four sets of samples. Plasma (n = 188) and bone marrow aspirate (n = 129) samples from patients with CaP, who received abiraterone acetate plus prednisone for up to 945 days (135 weeks), had undetectable androgens after 8 weeks of treatment. Plasma dehydroepiandrosterone (DHEA) concentrations were higher in African Americans than Caucasian Americans with newly diagnosed CaP. Analysis of prostate tumor tissue homogenates demonstrated reproducible testosterone (T) and dihydrotestosterone (DHT) concentrations with a minimal sample size of ∼1.0–2.0 mg of tissue. Finally, cell pellet and media samples from the LNCaP C4-2 cell line showed conversion of T to DHT.
The proposed LC/MS/MS method was validated for quantitation of five endogenous androgens in human plasma and serum, and effectively profiles androgens in clinical specimens and cell culture samples. Prostate 74:722–731, 2014. © 2014 Wiley Periodicals, Inc.