Related Experiment Video
Updated: Aug 13, 2026
High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
Published on: April 23, 2019
Highly sensitive simultaneous quantification of estrogenic tamoxifen metabolites and steroid hormones by LC-MS/MS
Janina Johänning1,2, Georg Heinkele1,2, Jana C Precht1,2
1Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology, Auerbachstr. 112, 70376, Stuttgart, Germany.
Abstract:
Tamoxifen is a mainstay in the treatment of estrogen receptor-positive breast cancer and is metabolized to more than 30 different compounds. Little is known about in vivo concentrations of estrogenic metabolites E-metabolite E, Z-metabolite E, and bisphenol and their relevance for tamoxifen efficacy. Therefore, we developed a highly sensitive HPLC-ESI-MS/MS quantification method for tamoxifen metabolites bisphenol, E-metabolite E, and Z-metabolite E as well as for the sex steroid hormones estradiol, estrone, testosterone, androstenedione, and progesterone. Plasma samples were subjected to protein precipitation followed by solid phase extraction. Upon derivatization with 3-[(N-succinimide-1-yl)oxycarbonyl]-1-methylpyridinium iodide, all analytes were separated on a sub-2-μm column with a gradient of acetonitrile in water with 0.1 % of formic acid. Analytes were detected on a triple-quadrupole mass spectrometer with positive electrospray ionization in the multiple reaction monitoring mode. Our method demonstrated high sensitivity, accuracy, and precision. The lower limits of quantification were 12, 8, and 25 pM for bisphenol, E-metabolite E, and Z-metabolite E, respectively, and 4 pM for estradiol and estrogen, 50 pM for testosterone and androstenedione, and 25 pM for progesterone. The method was applied to plasma samples of postmenopausal patients taken at baseline and under tamoxifen therapy. Graphical Abstract Sample preparation and derivatization for highly sensitive quantification of estrogenic tamoxifen metabolites and steroid hormones by HPLC-MS/MS.
Insights
A new HPLC-MS/MS method accurately quantifies tamoxifen metabolites and hormones in patient plasma. This aids understanding of how these compounds affect breast cancer treatment efficacy.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Endocrinology
Background:
- Tamoxifen is a key treatment for estrogen receptor-positive breast cancer.
- Tamoxifen is metabolized into over 30 compounds, but the in vivo relevance of estrogenic metabolites like bisphenol, E-metabolite E, and Z-metabolite E remains unclear.
- Understanding these metabolites is crucial for optimizing tamoxifen therapy.
Purpose of the Study:
- To develop a highly sensitive method for quantifying tamoxifen metabolites (bisphenol, E-metabolite E, Z-metabolite E) and key sex steroid hormones (estradiol, estrone, testosterone, androstenedione, progesterone).
- To apply this method to analyze plasma samples from postmenopausal patients undergoing tamoxifen therapy.
Main Methods:
- Developed a High-Performance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (HPLC-ESI-MS/MS) method.
- Utilized protein precipitation and solid-phase extraction for sample preparation.
- Employed derivatization with 3-[(N-succinimide-1-yl)oxycarbonyl]-1-methylpyridinium iodide.
- Separated analytes on a sub-2-μm column with a gradient elution and detected them using a triple-quadrupole mass spectrometer.
Main Results:
- Achieved high sensitivity, accuracy, and precision for all targeted analytes.
- Established lower limits of quantification as low as 4 pM for estradiol/estrone and 8 pM for E-metabolite E.
- Successfully applied the method to analyze patient plasma samples.
Conclusions:
- The developed HPLC-MS/MS method is suitable for sensitive quantification of tamoxifen metabolites and steroid hormones.
- This analytical tool can help elucidate the role of these compounds in tamoxifen efficacy for breast cancer treatment.
- Further research using this method may lead to personalized tamoxifen therapy strategies.
More Related Videos
07:37An Integrated Raman Spectroscopy and Mass Spectrometry Platform to Study Single-Cell Drug Uptake, Metabolism, and Effects
Published on: January 9, 2020
08:56Detection of Regulated Ergot Alkaloids in Food Matrices by Liquid Chromatography-Trapped Ion Mobility Spectrometry-Time-of-Flight Mass Spectrometry
Published on: November 22, 2024
