Journal
STEROIDS
Volume 74, Issue 12, Pages 920-926Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.steroids.2009.06.007
Keywords
Testosterone; 5 alpha-Dihydrotestosterone; Liquid chromatography-electrospray ionization tandem mass spectrometry; Picolinyl derivatization; Prostatic disorder diagnosis
Funding
- Ministry of Education, Culture, Sports, Science and Technology of Japan
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We developed highly sensitive detection of testosterone (T) and 5 alpha-dihydrotestosterone (DHT) by liquid chromatography-electrospray ionization tandem mass spectrometry using high proton affinitive derivatization of 17 beta-hydroxyl group of T and DHT with picolinic acid, mobile phase consisting of MeCN-MeOH-H2O-formic acid and conventional octadecylsilica (ODS) column. Purification of the derivatives was carried out using solid-phase extraction with ODS cartridge. By this method, T and DHT were determined simultaneously with limits of quantification (LOQs) of 1 pg/0.2 ml in serum, and T and DHT with LOQs of 0.5 pg and 1 pg/3 mg in prostate tissue, respectively, under acceptable assay performance (intra-assay and inter-assay accuracy and precision). The present method provides reliable and reproducible results for quantification of T and DHT in small volumes of serum and prostate samples for diagnosis in prostatic disorders and male climacteric. (C) 2009 Elsevier Inc. All rights reserved.
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