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Simultaneous determination of multiple androgens in mice organs with liquid chromatography tandem mass spectrometry
Shu Fang Soh1, Xiaoxing Yin2, Jiaquan Sun3
1Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 209 Tongshan Road, Xuzhou 221004, PR China; Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, 5 Lower Kent Ridge Road, 119074 Singapore, Singapore.
Abstract:
Prostate cancer (PCa) is the most commonly diagnosed cancer in men worldwide. It is essentially dependent on potent androgens, such as testosterone (T) and dihydrotestosterone (DHT). The precursors of T and DHT, which includes androstenedione (A4) and dihydroepiandrosterone (DHEA), and also the metabolites of DHT, 5α-androstane-3α,17β-diol (3α-Diol) and 5α-androstane-3β,17β-diol (3β-Diol) are able to affect the development of PCa. Therefore, it is important to simultaneously determine all these key androgens. This study aims to develop and validate an LC-MS/MS quantification method to simultaneously detect and quantify the six related androgens, including T, DHT, A4, DHEA, 3α-Diol, and 3β-Diol in limited sample volume. The sample preparation involved liquid extraction with methyl tert-butyl ether (MTBE), following by chemical derivatisation with hydroxylamine. The limits of quantitation for T, DHT, A4, and DHEA were 0.05nM and 3α-Diol and 3β-Diol were 0.5nM with S/N ratio of at least 5:1 by using 100μL samples.
Insights
This study developed a new method to measure six key androgens, including testosterone and dihydrotestosterone, crucial for prostate cancer (PCa) development. This allows for precise analysis even with small sample volumes.
Area of Science:
- Endocrinology
- Analytical Chemistry
- Oncology
Background:
- Prostate cancer (PCa) is a prevalent cancer in men globally.
- PCa growth is significantly influenced by androgens like testosterone (T) and dihydrotestosterone (DHT).
- Androgen precursors (androstenedione, dihydroepiandrosterone) and DHT metabolites (3α-Diol, 3β-Diol) also impact PCa development.
Purpose of the Study:
- To develop and validate a simultaneous LC-MS/MS method for quantifying six key androgens.
- To enable androgen measurement in limited sample volumes, crucial for PCa research.
- To improve the understanding of androgen roles in prostate cancer.
Main Methods:
- Liquid extraction using methyl tert-butyl ether (MTBE).
- Chemical derivatization with hydroxylamine.
- Simultaneous detection and quantification using LC-MS/MS.
Main Results:
- Successful simultaneous quantification of testosterone (T), dihydrotestosterone (DHT), androstenedione (A4), dihydroepiandrosterone (DHEA), 3α-Diol, and 3β-Diol.
- Achieved limits of quantification of 0.05nM for T, DHT, A4, and DHEA.
- Achieved limits of quantification of 0.5nM for 3α-Diol and 3β-Diol using 100μL samples.
Conclusions:
- A validated LC-MS/MS method allows for the simultaneous determination of critical androgens and their related compounds.
- This method is effective for analyzing small sample volumes, facilitating PCa research.
- Accurate quantification of these androgens is vital for understanding and potentially targeting PCa development.
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