PMID- 22818945
OWN - NLM
STAT- MEDLINE
DCOM- 20121105
LR  - 20161125
IS  - 1873-376X (Electronic)
IS  - 1570-0232 (Linking)
VI  - 902
DP  - 2012 Aug 1
TI  - Validation of a sequential extraction and liquid chromatography-tandem mass 
      spectrometric method for determination of dihydrotestosterone, androstanediol and 
      androstanediol-glucuronide in prostate tissues.
PG  - 84-95
LID - 10.1016/j.jchromb.2012.06.031 [doi]
AB  - Androgens are key mediators of prostate development and function, a role that 
      extends to the development of prostate diseases such as benign prostatic 
      hyperplasia (BPH) and prostate cancer. In prostate, DHT is the major androgen and 
      reduction and glucuronidation are the major metabolic pathways for DHT 
      elimination. A streamlined method for quantitation of dihydrotestosterone (DHT), 
      5alpha-androstan-3alpha,17beta-diol (3alpha-diol), and 3alpha-diol glucuronide (diol-gluc) was 
      established and validated for use with archived prostate tissue specimens to 
      facilitate examination of the roles of the underlying metabolism. This involved a 
      sequential 70/30 hexane/ethyl acetate (hex/EtOAc) extraction of steroids, 
      followed by an ethyl acetate extraction for diol-gluc. Derivatization of the 
      hex/EtOAc fraction with2-fluoro-1-methylpyridinium p-toluene-4-sulfonate (FMP) 
      was used to enhance sensitivity for hydroxyl steroids and liquid 
      chromatography-tandem mass spectrometry (LC-MS/MS) was utilized for analysis of 
      both fractions. The method was validated with calibration standards followed by 
      recovery assessment from spiked samples of BPH and normal prostate. Lower limits 
      of quantitation (LLOQ) were 50 pg/g, 20 pg/g and 100 pg/g for DHT, 3alpha-diol and 
      diol-gluc, respectively for extracts from 50mg equivalents of tissue. Prepared 
      samples were stable for up to three weeks at 4  degrees C and 37  degrees C. The method provides 
      excellent sensitivity and selectivity for determination of tissue levels of DHT, 
      3alpha-diol, and diol-gluc. Furthermore, this protocol can easily be extended to 
      other hydroxyl steroids, is relatively straightforward to perform and is an 
      effective tool for assessing steroid levels in archived clinical prostate 
      samples.
CI  - Crown Copyright (c) 2012. Published by Elsevier B.V. All rights reserved.
FAU - Adomat, Hans H
AU  - Adomat HH
AD  - The Prostate Centre, Vancouver General Hospital, Vancouver, British Columbia, 
      Canada. hadomat@prostatecentre.com
FAU - Bains, Onkar S
AU  - Bains OS
FAU - Lubieniecka, Joanna M
AU  - Lubieniecka JM
FAU - Gleave, Martin E
AU  - Gleave ME
FAU - Guns, Emma S
AU  - Guns ES
FAU - Grigliatti, Thomas A
AU  - Grigliatti TA
FAU - Reid, Ronald E
AU  - Reid RE
FAU - Riggs, K Wayne
AU  - Riggs KW
LA  - eng
PT  - Journal Article
DEP - 20120701
PL  - Netherlands
TA  - J Chromatogr B Analyt Technol Biomed Life Sci
JT  - Journal of chromatography. B, Analytical technologies in the biomedical and life 
      sciences
JID - 101139554
RN  - 0 (Benzenesulfonates)
RN  - 08J2K08A3Y (Dihydrotestosterone)
RN  - 25126-76-5 (Androstane-3,17-diol)
RN  - 27195-25-1 (androstane-3,17-diol glucuronide)
RN  - QGV5ZG5741 (4-toluenesulfonic acid)
SB  - IM
MH  - Androstane-3,17-diol/*analogs & derivatives/analysis/chemistry
MH  - Benzenesulfonates/chemistry
MH  - Chromatography, Liquid/*methods
MH  - Dihydrotestosterone/*analysis
MH  - Drug Stability
MH  - Humans
MH  - Male
MH  - Prostate/*chemistry
MH  - Prostatic Hyperplasia/metabolism
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Tandem Mass Spectrometry/*methods
EDAT- 2012/07/24 06:00
MHDA- 2012/11/06 06:00
CRDT- 2012/07/24 06:00
PHST- 2012/01/04 00:00 [received]
PHST- 2012/06/19 00:00 [revised]
PHST- 2012/06/24 00:00 [accepted]
PHST- 2012/07/24 06:00 [entrez]
PHST- 2012/07/24 06:00 [pubmed]
PHST- 2012/11/06 06:00 [medline]
AID - S1570-0232(12)00376-5 [pii]
AID - 10.1016/j.jchromb.2012.06.031 [doi]
PST - ppublish
SO  - J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Aug 1;902:84-95. doi: 
      10.1016/j.jchromb.2012.06.031. Epub 2012 Jul 1.