Universität zu Köln

Thevis, Mario, Dib, Josef, Thomas, Andreas ORCID: 0000-0003-1199-0743, Hoeppner, Sebastian, Lagojda, Andreas, Kuehne, Dirk, Sander, Mark, Opfermann, Georg and Schaenzer, Wilhelm (2015). Complementing the characterization of in vivo generated N-glucuronic acid conjugates of stanozolol by collision cross section computation and analysis. Drug Test. Anal., 7 (11-12). S. 1050 - 1057. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

Detailed structural information on metabolites serving as target analytes in clinical, forensic, and sports drug testing programmes is of paramount importance to ensure unequivocal test results. In the present study, the utility of collision cross section (CCS) analysis by travelling wave ion mobility measurements to support drug metabolite characterization efforts was tested concerning recently identified glucuronic acid conjugates of the anabolic-androgenic steroid stanozolol. Employing travelling-wave ion mobility spectrometry/quadrupole-time-of-flight mass spectrometry, drift times of five synthetically derived and fully characterized steroid glucuronides were measured and subsequently correlated to respective CCSs as obtained in silico to form an analyte-tailored calibration curve. The CCSs were calculated by equilibrium structure minimization (density functional theory) using the programmes ORCA with the data set B3LYP/6-31G and MOBCAL utilizing the trajectory method (TM) with nitrogen as drift gas. Under identical experimental conditions, synthesized and/or urinary stanozolol-N and O-glucuronides were analyzed to provide complementary information on the location of glucuronidation. Finally, the obtained data were compared to CCS results generated by the system's internal algorithm based on a calibration employing a polyalanine analyte mixture. The CCSs Omega(N2) calculated for the five steroid glucuronide calibrants were found between 180 and 208 angstrom(2), thus largely covering the observed and computed CCSs for stanozolol-N1'-, stanozolol-N2'-, and stanozolol-O-glucuronide found at values between 195.1 and 212.4 angstrom(2). The obtained data corroborated the earlier suggested N- and O-glucuronidation of stanozolol, and demonstrate the exploit of ion mobility and CCS computation in structure characterization of phase-II metabolic products; however, despite reproducibly measurable differences in ion mobility of stanozolol-N1'-, N2'-, and O-glucuronides, the discriminatory power of the chosen CCS computation algorithm was found to be not appropriate to allow for accurate assignments of the two N-conjugated structures. Using polyalanine-based calibrations, significantly different absolute values were obtained for all CCSs, but due to a constant offset of approximately 45 angstrom(2) an excellent correlation (R-2=0.9997) between both approaches was observed. This suggests a substantially accelerated protocol when patterns of computed and polyalanine-based experimental data can be used for structure elucidations instead of creating individual analyte-specific calibration curves. Copyright (c) 2015 John Wiley & Sons, Ltd.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED Dib, Josef UNSPECIFIED UNSPECIFIED UNSPECIFIED Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Hoeppner, Sebastian UNSPECIFIED UNSPECIFIED UNSPECIFIED Lagojda, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuehne, Dirk UNSPECIFIED UNSPECIFIED UNSPECIFIED Sander, Mark UNSPECIFIED UNSPECIFIED UNSPECIFIED Opfermann, Georg UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaenzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-387669
DOI:	10.1002/dta.1907
Journal or Publication Title:	Drug Test. Anal.
Volume:	7
Number:	11-12
Page Range:	S. 1050 - 1057
Date:	2015
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1942-7611
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MOBILITY-MASS-SPECTROMETRY; ELECTRON-IMPACT FRAGMENTATION; ANABOLIC-STEROID METABOLITES; ION-MOBILITY; HUMAN URINE; ANDROGENIC STEROIDS; PROTEIN COMPLEXES; DRIFT GAS; IDENTIFICATION; METANDIENONE Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/38766