| Polar Derivatization of 5α-Dihydrotestosterone and Sensitive Analysis by Semimicro-LC/ESI-MS |
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a) Pharmaceuticals Group, Nippon Kayaku Co., Ltd. (3-31-12 Shimo, Kita-ku, Tokyo 115-8588, Japan) |
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| Polar derivatization method for electrospray ionization mass spectrometry (ESI-MS) was developed and applied to the sensitive analysis of 5α-dihydrotestosterone (DHT) in prostate and plasma. Rat prostate was dissolved with alkaline solution and DHT in the prostate was extracted by solid phase extraction, and derivatized to the N-methylpyridinium-DHT as a polar derivative. The N-methylpyridinium derivative of DHT was purified by solid-phase extraction using Bond Elut C18, and determined by a semimicro-LC/ESI-MS with selected reaction monitoring (SRM). Linear responses of the N-methylpyridinium derivative of DHT were observed for analyte concentration from 5 to 100 pg/tube. DHT values were 12.0±3.8 ng/g tissue in rat prostates, 171.7±127.2 pg/mL in rat plasma and 286.2±123.5 pg/mL in human plasma. | ||
| Key words: Polar derivatization, LC/ESI-MS, Dihydrotestosterone, Prostate, Plasma | ||
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