PMID- 26331928
OWN - NLM
STAT- MEDLINE
DCOM- 20160711
LR  - 20150903
IS  - 1097-0231 (Electronic)
IS  - 0951-4198 (Linking)
VI  - 29
IP  - 19
DP  - 2015 Oct 15
TI  - High-temperature high-performance liquid chromatography on a porous graphitized 
      carbon column coupled to an Orbitrap mass spectrometer with atmospheric pressure 
      photoionization for screening exogenous anabolic steroids in human urine.
PG  - 1779-88
LID - 10.1002/rcm.7278 [doi]
AB  - RATIONALE: The presence in a urinary matrix of a large number of endogenous 
      steroids and corticosteroids with similar structures can hamper the detection of 
      specific exogenous steroids using liquid chromatography/mass spectrometry (LC/MS) 
      with reversed-phase columns. Therefore, the development of LC/MS methods using 
      alternative columns is of great interest. Porous graphitized carbon is a unique 
      stationary phase for high-performance liquid chromatography (HPLC), with 
      properties differing from traditional silica-based and polymeric stationary 
      phases. METHODS: The new method involves enzymatic hydrolysis, liquid-liquid 
      extraction, and determination by high-temperature HPLC/Orbitrap mass spectrometry 
      (HTLC/Orbitrap MS) with atmospheric pressure photoionization (APPI). To achieve 
      APPI of doping substances, the mobile phase consisted of 0.1% CF3COOH (A) and a 
      mixture of acetonitrile/2-propanol (25:75 v/v), containing 0.1% CF3COOH (B), 
      which was used as an effective proton source. RESULTS: A screening method for the 
      detection of 57 exogenous steroids has been developed. The method was validated 
      by spiking 10 different blank urine samples at different concentration levels. 
      Validation parameters included limit of detection (LOD), selectivity, ion 
      suppression, extraction recovery, and repeatability. All studied compounds had an 
      LOD lower than the minimum required performance level. Of the 57 steroids 
      studied, 55 showed recovery better than 70%. For all of the analytes, the 
      relative retention times proved to be stable between days, with relative standard 
      deviations (RSDs) smaller than 0.3%. In addition, the interday RSDs of the peak 
      area ratios ranged between 0.7% and 14.5%. CONCLUSIONS: The proposed method 
      matches the basic requirements of all methods used to analyze drugs or 
      metabolites in an antidoping laboratory, i.e., sensitivity, selectivity, and 
      specificity. The acquisition of full-scan mass spectra with accurate masses can 
      be a valuable tool in the retrospective evaluation of analyzed samples for 
      anabolic steroids recently added to the prohibited list.
CI  - Copyright (c) 2015 John Wiley & Sons, Ltd.
FAU - Virus, Edward D
AU  - Virus ED
AD  - Institute of General Pathology and Pathophysiology, Baltiskaya 8, 125315, Moscow, 
      Russia.
FAU - Luzyanin, Boris P
AU  - Luzyanin BP
AD  - Institute of General Pathology and Pathophysiology, Baltiskaya 8, 125315, Moscow, 
      Russia.
FAU - Ivanov, Alexander V
AU  - Ivanov AV
AD  - Institute of General Pathology and Pathophysiology, Baltiskaya 8, 125315, Moscow, 
      Russia.
FAU - Kubatiev, Aslan A
AU  - Kubatiev AA
AD  - Institute of General Pathology and Pathophysiology, Baltiskaya 8, 125315, Moscow, 
      Russia.
LA  - eng
PT  - Journal Article
PL  - England
TA  - Rapid Commun Mass Spectrom
JT  - Rapid communications in mass spectrometry : RCM
JID - 8802365
RN  - 0 (Anabolic Agents)
RN  - 7782-42-5 (Graphite)
SB  - IM
MH  - Anabolic Agents/*urine
MH  - Chromatography, High Pressure Liquid/methods
MH  - Doping in Sports
MH  - Graphite/chemistry
MH  - Hot Temperature
MH  - Humans
MH  - Male
MH  - Mass Spectrometry/*methods
MH  - Porosity
EDAT- 2015/09/04 06:00
MHDA- 2016/07/12 06:00
CRDT- 2015/09/03 06:00
PHST- 2015/02/10 00:00 [received]
PHST- 2015/07/02 00:00 [revised]
PHST- 2015/07/12 00:00 [accepted]
PHST- 2015/09/03 06:00 [entrez]
PHST- 2015/09/04 06:00 [pubmed]
PHST- 2016/07/12 06:00 [medline]
AID - 10.1002/rcm.7278 [doi]
PST - ppublish
SO  - Rapid Commun Mass Spectrom. 2015 Oct 15;29(19):1779-88. doi: 10.1002/rcm.7278.