2537 Bioanalysis (2015) 7(19), 2537–2556 ISSN 1757-6180
Review
part of
10.4155/bio.15.172 © 2015 Future Science Ltd
Derivatization is one of the most important steps during sample preparation in
doping control analysis. Its main purpose is the enhancement of chromatographic
separation and mass spectrometric detection of analytes in the full range of laboratory
doping control activities. Its application is shown to broaden the detectable range
of compounds, even in LC–MS analysis, where derivatization is not a prerequisite.
The impact of derivatization initiates from the stage of the metabolic studies of
doping agents up to the discovery of doping markers, by inclusion of the screening
and confirmation procedures of prohibited substances in athlete’s urine samples.
Derivatization renders an unlimited number of opportunities to advanced analyte
detection.
Since the introduction of chromatographic
methods in the determination of xenobiot-
ics in biological luids – a ield related to vet-
erinary drug analysis, forensic science and
doping control analysis – and together
with the implementation of chromatographic
methods coupled with different types of
mass spectrometers for the unquestionable
analyte identiication in routine analysis,
derivatization procedures have played a
central and critical role.
The term derivatization is used to describe
the procedures in which chemical groups
(e.g., protons from protic sites) are replaced
by groups that modify physical and chemical
properties of analytes, like molecular formula,
thermal stability, volatility, polarity, chro-
matographic behavior, light absorption, ion-
ization ef iciency, mass spectra fragmentation,
protection of labile groups, etc, thus resulting
in more sensitive and robust detection. For a
successful derivatization, a single and stable
derivative should be formed for each com-
pound and the derivatization reaction should
be simply and rapidly performed under mild
conditions with a high and reproducible yield.
Excellent handbooks and recent reviews
are available on the general advantages of
derivatization procedures [1–6] .
Hundreds of chemically different drugs
are included in the List of Prohibited Sub-
stances [7] issued by the World Anti-Doping
Agency (WADA). The Prohibited List is
revised annually and comprises substances
grouped according to their pharmacological
action. The demanding task of doping con-
trol laboratories is to develop analytical
methods able to screen a large number of
substances, minimizing by the same token,
sample preparation and costs.
Primarily, doping control analysis requires
the use of chromatographic methods [8–10] ,
particularly GC–MS and LC–MS. Applica-
tion of GC–MS and LC–MS for screening
purposes is complementary, depending on
the properties of the compounds like vola-
tility, polarity, molecular weight, ionization
properties etc.
GC–MS has played major role in the
doping control analysis for several decades,
despite the increasing use of LC systems in
doping control laboratories. The screening
beneits from GC–MS (e.g., ionization, cap-
illary column chromatographic resolution,
etc.) are signiicant, especially for the low
ng/ml detection of anabolic androgenic
steroids (AAS), since LC–MS ionization
for AAS lacking at least a conjugated keto-
Current status and recent advantages in
derivatization procedures in human doping
control
Ioanna Athanasiadou
1,2
,
Polyxeni Kiousi
3
, Nasia
Kioukia-Fougia
3
, Emmanouil
Lyris
3
& Yiannis S Angelis*
,3
1
Laboratory of Biopharmaceutics &
Pharmacokinetics, Faculty of Pharmacy,
National & Kapodistrian University of
Athens, Greece
2
Anti-Doping Lab Qatar, Sports City
Road, Aspire Zone, P.O. Box 27775,
Doha, Qatar
3
Doping Control Laboratory of Athens,
Olympic Athletic Center of Athens
(OAKA), 15123 Maroussi, Greece
*Author for correspondence:
Tel.: +30 210 686 8412
Fax: +30 210 683 4021
y_angelis@yahoo.gr
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