Ανάπτυξη και Επικύρωση Μεθόδου Ανίχνευσης Απαγορευμένων Ουσιών με Υγροχρωματογραφία-Φασματομετρία Μαζών και Αναλυτή Χρόνου Πτήσης

Postgraduate Thesis uoadl:1315901 631 Read counter

Unit:
Κατεύθυνση Χημική Ανάλυση-Έλεγχος Ποιότητας
Library of the School of Science
Deposit date:
2011-12-19
Year:
2011
Author:
Μπουρμποπούλου Αικατερίνη
Supervisors info:
Κουππάρης Μιχαήλ Καθηγητής ΕΚΠΑ Επιβλέπων, Θωμαίδης Νικόλαος Επίκουρος Καθηγητής ΕΚΠΑ, Οικονόμου Αναστάσιος Επίκουρος Καθηγητής ΕΚΠΑ
Original Title:
Ανάπτυξη και Επικύρωση Μεθόδου Ανίχνευσης Απαγορευμένων Ουσιών με Υγροχρωματογραφία-Φασματομετρία Μαζών και Αναλυτή Χρόνου Πτήσης
Languages:
Greek
Summary:
This Master Thesis presents the development and validation of a screening
method detecting prohibited substances with Liquid Chromatography- Mass
Spectrometry and the use of a Time of Flight mass analyzer coupled with a
quadrupole (LC-QTOF). Analytes were ionized with an electrospray ionization,
performed in the positive mode [(+)ESI].
During the first stage of this Thesis, nearly all of the prohibited
substances were analyzed, using standard solutions, in order to determine their
chromatographic characteristics: the retention time, the relative retention
time (in relation to the internal standard) and the molecular adduct ions of
the highest abundance. In addition, co-eluted substances and isomers were
examined.
The second stage of this study was the development of the sample
preparation procedure. Different parameters were examined, including the type
of extracting solvent, the adjustment of the pH before the extraction, the
salting-out conditions and the reconstitution solvent. The chosen conditions
were those providing the optimum analytical results in respect to the
resolution, the sensitivity and the stability of the retention time for the
majority of the substances.
Especially for the detection of boldenone, methyldienolone and
methyltrienolone, a coupled mass spectrometry (MS/MS) along with the single
mass spectrometry (MS) were used, due to matrix interferences for these
substances.
The developed method -using ethyl acetate as the extracting solvent- , was
validated according to the current International Standard for Laboratories
(ISL). 251 substances and metabolites were analyzed, including anabolics,
glucocorticosteroids, stimulants, narcotics, diuretics, b-blockers, b2-agonists
etc. Validation parameters included chromatographic characteristics,
identification capability, limit of detection, specificity, carryover
contamination, matrix interferences, mass accuracy, extraction recovery and
matrix effect.
In order to show the efficiency of the proposed method -in addition to the
validation experiments- a study on the detection of metabolites in excretion
urine was conducted. 40 different metabolites of 18 parent substances were
detected. Some of those metabolites -such as the sulpho-conjugated compounds-
were detected for the first time in urine, with the use of ethyl acetate as the
extracting solvent.
As a result, the proposed method was considered suitable for the detection
of 227 prohibited substances and metabolites.
Keywords:
Liquid Chromatography, Mass Spectrometry, Time of Flight analyzer, Ethyl acetate, Doping control
Index:
Yes
Number of index pages:
xi-xvii, 131-135
Contains images:
Yes
Number of references:
61
Number of pages:
xvii, 168
File:
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