M. Raro et al.
steroids andmetabolites in human urine by liquid
chromatography-electrospray-tandem mass spectrometry. Anal.
Chem. 2008, 80, 1709.
In Fig. 3, the MS spectrum for MeTmet1 is shown as an exam-
ple. In this case, [M+H-2H2O]+ appears as the base peak of the
spectrum for the underivatized compound (Fig. 3g), whereas a
prominent [M+H-2TMSOH]+ was observed for the enol-TMS de-
rivative (Fig. 3h). In both cases, minor ions corresponding to
single losses of water or TMSOH were also observed.
[7] M. Thevis, H. Geyer, U. Mareck, W. Schänzer. Screening for unknown
steroids in human urine by liquid chromatography-tandem mass
spectrometry. J. Mass Spectrom. 2005, 40, 955.
[8] C. Gómez, O. J. Pozo, J. Marcos, J. Segura,R. Ventura. Alternative long-
term markers for the detection of methyltestosterone misuse.
Steroids 2013, 78, 44.
[9] O. J. Pozo, P. Van Eenoo, K. Deventer, F. T. Delbeke. Detection and
characterization of anabolic steroids in doping analysis by LC-MS.
TrAC Trends Anal. Chem. 2008, 27, 657.
Conclusions
[10] M. Galesio, R. Rial-Otero, J. Simal-Gándara, X. de la Torre, F. Botrè, J. L.
Capelo-Martínez. Improved ultrasonic-based sample treatment
for the screening of anabolic steroids by gas chromatogra-
phy/mass spectrometry. Rapid Commun. Mass Spectrom.
2010, 24, 2375.
[11] E. M. Brun, R. Puchades, Á. Maquieira. Analytical methods for anti-
doping control in sport: anabolic steroids with 4,9,11-triene structure
in urine. TrAC Trends in Anal. Chem. 2011, 30, 771.
[12] O. J. Pozo, P. Van Eenoo, K. Deventer. Development and validation of
qualitative screening method for the detection of exogenous ana-
bolic steroids in urine by liquid chromatography-tandem mass spec-
trometry. Anal. Bioanal. Chem. 2007, 389, 1209.
[13] O. J. Pozo, P. Van Eenoo, K. Deventer, F. T. Delbeke. Ionization of
anabolic steroids by adduct formation in liquid chromatography
electrospray mass spectrometry. J. Mass Spectrom. 2007, 42, 467.
[14] Y.-C. Ma, H.-Y. Kim. Determination of steroids by liquid chromatogra-
phy/Mass spectrometry. J. Am. Soc. Mass Spectrom. 1997, 8, 1010.
[15] E. C. Horning, D.I. Carroll, I. Dzidic, K. D. Haegele, S. Lin, C. U. Oertli, R.
N. Stillwell. Development and use of analytical systems based on
mass spectrometry. Clin. Chem. 1977, 23, 13.
[16] L. Hintikka, M. Haapala, T. Kuuranne, A. Leinonen, R. Kostiainen. Anal-
ysis of anabolic steroids in urine by gas chromatography-microchip
atmospheric pressure photoionization-mass spectrometry with chlo-
robenzene as dopant. J. Chromatogr. A 2013, 1312, 111.
[17] L. Hintikka, M. Haapala, S. Franssila, T. Kuuranne, A. Leinonen, R.
Kostiainen. Feasibility of gas chromatography-microchip atmo-
spheric pressure photoionization-mass spectrometry in analysis of
anabolic steroids. J. Chromatogr. A 2010, 1217, 8290.
The use of a new APCI interface coupled to GC–QTOF has been
investigated for the ionization of model AAS. The addition of wa-
ter in the interface promoted the proton transfer reaction pro-
ducing mainly protonated species, i.e. [M+H]+ and [M+H-H2O]+
(or [M+H-TMSOH]+ for TMS derivatives). A relationship between
ionization behavior and structure has been established. The main
conclusions are summarized in Table 6.
The low fragmentation observed in the APCI spectra (all ions
contained the intact steroid skeleton) would facilitate the selec-
tion of abundant and/or more specific precursor ions in tandem
MS experiments. Theoretically, this fact would lead to a neat sen-
sitivity and selectivity improvement in SRM-based methods. The
effect of this improvement in the control of the abuse of steroids
in sports should be investigated.
Furthermore, the knowledge gained regarding the behavior of
AAS ionization in the APCI source might be helpful in the identi-
fication and elucidation of unknown metabolites of steroids in
urine. Thus, based on the information depicted in Table 6, a
theoretical ion for a predicted metabolite can be postulated.
This can help in the target detection of potential metabolites
by GC–APCI–MS. Additionally, if an unknown steroid is detected
by untargeted methods, some valuable structural information
can be obtained based on the results showed in this work.
[18] C. N. McEwen,R. G. McKay. A Combination Atmospheric Pressure
LC/MS:GC/MS Ion Source: Advantages of Dual AP-LC/MS:GC/MS
Instrumentation. J. Am. Soc. Mass Spectrom. 2005, 16, 1730.
[19] R. Schiewek, M. Lorenz, R. Giese, K. Brockmann, T. Benter, S. Gäb, O. J.
Schmitz. Development of a multipurpose ion source for LC-MS and
GC-API MS. Anal. Bioanal. Chem. 2008, 392, 87.
[20] A. Carrasco-Pancorbo, E. Nevedomskaya, T. Arthen-Engeland, T. Zey,
G. Zurek, C. Baessmann, A. M. Deelder, O. A. Mayboroda. Gas chro-
matography/Atmospheric Pressure Chemical Ionization-time of
flight mass spectrometry: Analytical validation and applicability to
metabolic profiling. Anal. Chem. 2009, 81,10071.
[21] T. Portolés, J. V. Sancho, F. Hernández, A. Newton, P. Hancock. Poten-
tial of atmospheric pressure chemical ionization source in GC-QTOF
MS for pesticide residue analysis. J. Mass Spectrom. 2010, 45, 926.
[22] T. Portolés, L. Cherta, J. Beltran, F. Hernández. Improved gas
chromatography-tandem mass spectrometry determination of pesti-
cide residues making use of atmospheric pressure chemical ioniza-
tion. J. Chromatogr. A 2012, 1260, 183.
[23] T. Portolés, J. G. J. Mol, J. V. Sancho, F. Hernández. Advantages of
atmospheric pressure chemical ionization in gas chromatography
tandem mass spectrometry: Pyrethroid insecticides as a case study.
Anal. Chem. 2012, 84, 9802.
[24] T. Bristowm M. Harrison, M. Sims. The application of gas chromatog-
raphy/atmospheric pressure chemical ionisation time-of-flight mass
spectrometry to impurity identification in Pharmaceutical Develop-
ment. Rapid Commun. Mass Spectrom. 2010, 24, 1673.
[25] R. García-Villalba, T. Pacchiariotta, A. Carrasco-Pancorbo, A. Segura-
Carretero, A. Fernández-Gutiérrez, A. M. Deelder, O. A. Mayboroda.
Gas chromatography-atmospheric pressure chemical ionization-
time of flight mass spectrometry for profiling of phenolic com-
pounds in extra virgin olive oil. J. Chromatogr. A 2011, 1218, 959.
[26] C. Domeño, E. Canellas, P. Alfaro, A. Rodríguez-Lafuente, C. Nerin.
Atmospheric pressure gas chromatography with quadrupole time
of flight mass spectrometry for simultaneous detection and
quantification of polycyclic aromatic hydrocarons and nitro-
Acknowledgements
The authors acknowledge the financial support of the Ministry of
Education and Science, Spain, in the project DEP2011-28573-C02-
01/02. The authors from University Jaume I also acknowledge the
support from Generalitat Valenciana (Research Group of Excel-
lence Prometeo/2009/054). The authors from IMIM acknowledge
the support from Generalitat de Catalunya (Consell Català de
l’Esport and DIUE 2009SGR4929).
M. Raro is also grateful to the Ministry of Education and
Science for her predoctoral grant.
References
[1] P. Hemmersbach. History of mass spectrometry at the Olympic
Games. J. Mass Spectrom. 2008, 43, 839.
[3] C. Gómez, A. Fabregat, O. J. Pozo, J. Marcos, J. Segura, R. Ventura.
Analytical strategies based on mass spectrometric techniques for the
study of steroid metabolism. TrAC. Trends Anal. Chem. 2014, 53, 106.
[4] P. Van Eenoo, W. Van Gansbeke, N. De Brabanter, K. Deventer, F. T.
Delbeke. A fast, comprehensive screening method for doping
agents in urine by gas chromatography-triple quadrupole mass
spectrometry. J. Chromatogr. A 2011, 1218, 3306.
[5] M. A. Delgadillo, L. Garrostas, O. J. Pozo, R. Ventura, B. Velasco, J.
Segura, J. Marcos. Sensitive and robust method for anabolic agents
in human urine by gas chromatography-triple quadrupole mass
spectrometry. J. Chromatogr. B 2012, 897, 85.
[6] O. J. Pozo, K. Deventer, P. Van Eenoo, F. Delbeke. Efficient ap-
proach for the comprehensive detection of unknown anabolic
wileyonlinelibrary.com/journal/jms
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J. Mass Spectrom. 2014, 49, 509–521