Bitte vorab per E-Mail vereinbaren: s.pawlowski(at)dshs-koeln.de . Termine in Präsenz (Institutsgebäude II, Raum 313) oder WebEx möglich. Terminübersicht 28.07. von 14:00-16:00 Uhr vor Ort, ab 16 Uhr online 05.08. von 08:30-10:00 Uhr vor Ort, ab 10:00 Uhr online 02.09. von 09:00-10:30 Uhr 06.10. von 09:00-10:30 Uhr

Am 3. Juli 2026 wurde die Deutsche Sporthochschule Köln wieder zur Bühne für Ideen, spannende Persönlichkeiten und jede Menge Gründungsgeist. Beim großen Finale des Inkubator Programms „StarS-Kader“ präsentierten sieben Teams ihre Geschäftsideen vor Publikum und Jury. Am Ende überzeugte das Team nykraft und sicherte sich den Titel als bestes StarS-Kader Team 2026 und damit einen Preis in Höhe von 2.500 € (gesponsert durch CBH Rechtsanwälte) und Karten für die Loge der Besten in der Lanxess Arena (gesponsert durch die Sparkasse KölnBonn). Die Gewinner*innen des Abends: Platz 1: nykraft Platz 2: Fit-o-Saurus Platz 3: Sprintalyze Publikumspreis: Fit-o-Saurus Newcomer des Jahres: SwimSafe Gateway Alumni Startup des Jahres: AV8 Der Abend zeigte einmal mehr, wie viel Innovationskraft in den Bereichen Sport, Bewegung und Gesundheit stecken. Die Teams brachten unterschiedliche Themen auf die Bühne: von digitalem Coaching und Bewegungsanalyse über Gesundheitsbildung für Kinder bis hin zu Prothetik, Orientierung nach dem Schulabschluss und Männergesundheit. Anders als bei klassischen Pitch-Formaten mussten die Teams gleich drei Aufgaben meistern. Nach einem Imagevideo folgte der Live Pitch vor Publikum. In der Finalrunde „Roast my Startup“ wurde es dann besonders spannend: Die Jury konfrontierte die besten Teams mit möglichen Schwachstellen ihrer Ideen. In kurzer Zeit mussten sie zeigen, wie gut sie ihr Geschäftsmodell kennen und wie überzeugend sie auch unter Druck reagieren. Ein Highlight des Abends war das Panel „Athletes in Business“, powered by Athlete Capital. Jonas Hector, Luca Waldschmidt, Léa Krüger und Nelvie Tiafack sprachen darüber, wie sich Erfahrungen aus dem Spitzensport auf Unternehmertum übertragen lassen und welche Chancen entstehen, wenn Sportler*innen eigene Projekte aufbauen. Inspirierende Einblicke boten außerdem: Unternehmer und Big Wave Surfer Marc Ebinger, der über seinen Weg vom Unternehmensaufbau bis zu den größten Wellen der Welt sprach, Siegerin des StarS-Kader 2025 Janina Jaspers von wellplayd und Per Günther, der über seinen Werdegang vom Athlete Investor zum Athlete Founder sprach. Sie berichteten aus erster Hand von ihren Gründungserfahrungen und gaben den Teams und dem Publikum wertvolle Impulse für ihren weiteren Weg. Auch abseits der Bühne war viel los. Im Foyer kamen Gründer*innen, Studierende, Gäste aus Sport, Wissenschaft und Wirtschaft sowie Akteur*innen aus dem Kölner Startup Ökosystem miteinander ins Gespräch. Der Startup Market und die Newcomer Area machten sichtbar, wie vielfältig die Gründungsideen rund um die Spoho und die Gateway Hochschulen Köln sind. Das Finale wurde von den Gateway Hochschulen Köln ausgerichtet. Ziel des Programms ist es, Gründungsideen aus der Universität sichtbar zu machen, Teams auf ihrem Weg zu begleiten und Menschen aus Sport, Wissenschaft, Wirtschaft und Startup-Szene miteinander zu vernetzen. Ein besonderer Dank geht an CBH Rechtsanwälte , die Kreissparkasse Köln , die NRW.BANK und die Sparkasse KölnBonn , die mit ihrer Unterstützung die Preisgelder ermöglicht haben. Wir freuen uns, gemeinsam junge Gründungsteams auf ihrem Weg begleiten zu können. Die Fotos wurden von Manor Lux aufgenommen.

Ein weiteres Konzert findet am 13. Juli in der Evangelischen Kirche Köln Weiden statt. Die Veranstaltung in der Sporthochschule beginnt um 19:00 Uhr; das Konzert in Weiden geht um 20:00 Uhr los. Der Eintritt ist frei; Spenden sind willkommen.

Wissenschaftliche Studie zu Ecdysteron, Muskelaufbau und Kraftentwicklung Studienbeschreibung Mittels dieser wissenschaftlichen Studie der Deutschen Sporthochschule Köln möchten wir den Einfluss von Ecdysteron auf Muskelaufbau und Kraftentwicklung bei trainingserfahrenen Männern untersuchen. Anforderungsprofil trainingserfahrene Männer 18 bis 40 Jahre Vorteile Wir erfassen regelmäßig deine Kraftwerte, dein Blutbild sowie mittels Ultraschall und Körperanalyse deine Muskel- und Körperzusammensetzung. Als Teilnehmer erhältst du neben dem Ecdysteron-Supplement einen individuell abgestimmten Trainingsplan sowie die vollständige Auswertung deiner Kraft- und Körperwerte über den Studienverlauf. Ablauf / Ziel Die Intervention erstreckt sich über 12 Wochen mit einem wissenschaftlich ausgelegten, gruppengesteuerten Ganzkörpertraining mit der Langhantel, das dreimal wöchentlich stattfindet. Dabei ist das Ziel der Studie, die Wirkung von Ecdysteron auf Kraftwerte, Muskelaufbau und Körperzusammensetzung im Vergleich zu untersuchen. Anmeldung erfolgt über den QR-Code auf dem Flyer oder per E-Mail an Lynn.Starke(at)stud.dshs-koeln.de .

M. THEVIS H. GEYER U. MARECK (EDITORS) RECENT ADVANCES IN DOPING ANALYSIS (32) Proceedings of the Manfred Donike Workshop 42nd Cologne Workshop on Dope Analysis 26th to 29th February 2024 SPORTVERLAG Strauß - Hellenthal 2024 RECENT ADVANCES IN DOPING ANALYSIS (32) 2 ISBN 978-3-86884-080-3 Bibliografische Information Der Deutschen Nationalbibliothek Die Deutsche Nationalbibliothek verzeichnet diese Publikation in der Deutschen Nationalbibliografie; detaillierte bibliografische Daten sind im Internet über abrufbar. Thevis, Mario; Geyer, Hans; Mareck, Ute (Eds.) Recent Advances in Doping Analysis (32). Proceedings of the Manfred Donike Workshop, 42nd Cologne Workshop on Dope Analysis, 26th to 29th February 2024/ [MDI e.V.] – 2024 Sportverlag Strauß. ISBN 978-3-86884-080-3 ©SPORTVERLAG Strauß Neuhaus 12 – 53940 Hellenthal Tel. +49 (0)2448/2470040 – Fax +49 (0)2448/9195610 E-mail: info@sportverlag-strauss.de www.sportverlag-strauss.de Satz: Autorensatz RECENT ADVANCES IN DOPING ANALYSIS (32) 3 ISBN 978-3-86884-080-3 MANFRED DONIKE WORKSHOP 2024 TABLE OF CONTENTS PAGE LECTURES Wang Y, Zhang L, Wang Z, Xing Y, Wang S, Shen L, Yang Z: Doping Control Laboratory operational strategy and outcome of sequential major competitions including the Hangzhou Asian Games in 2023 .....................................................15-20 Croft M, Edey C, Villanueva R, Murby EJ, Goebel C, Agon V: The use of dual column chromatographic separations for an efficient ITP ................................. 21-27 Al-Menhali A, El-Sayed I, El-Saftawy W, Al-Qahtani A, Al-Zeyara A, Aakel N, Kraiem S, Beotra A, Mohamad-Ali V, Al Maadheed M: Cobalt analysis in urine samples using ICP-MS: method validation and prevalence studies in athletes ........................................................................................................................ 28-32 Cutler C, Viljanto M, Hincks P, Taylor P, Habershon-Butcher J, Scarth J, van Eenoo P: Metabolism of SARMs and other doping substances available to purchase online as supplement products – a comparison between human and equine studies ............................... 33-40 Thilmany S, Thomas A, Vogel M, Scholl C, Thevis M: Influence of steroid receptor ligands on the neuronal proteome: A trigger for depression? .......................................................................................................................41-43 Sansoucy M, Soucy M, Morneau E, Ricard S, Daoust B, Naud J: In vitro SR9011 metabolism study using tandem mass spectrometry..........................................44-50 Barber A, Couture M, Ayotte C, Naud J: GC-C-IRMS confirmation of boldenone across sample matrices....................................................51-56 POSTER PRESENTATIONS Athanasiadou I, Gmeiner G: Occurrence of meldonium in Russian athletes’ population............................................................ 57-61 Stacchini C, de La Torre X, Meloni M, Terracciano F, Botrè F: Carbonic anhydrase isoforms and their relevance in anti-doping analysis: analytical strategies ............................................................................................................................... 62-66 RECENT ADVANCES IN DOPING ANALYSIS (32) 4 ISBN 978-3-86884-080-3 PAGE Kniess A, Vonaparti A, Voss S: Stability of small peptides under various storage conditions......................................................... 67-71 Wilairat P, Kongpatanakul S: Determination of LOD/LOI using 95% detection/identification rate: correct sigmoid function and regression method.............................................................................. 72-74 Jarek A, Wójtowicz-Zawadka M, Urbaniak-Zbyszynska A, Tarka M, Chajewska K, Pasik M, Kwiatkowska D: Influence of monitoring long-term metabolites on the detection window of the selected AASs in the last decade in the Polish Anti-Doping Laboratory........................................ 75-79 Kaliszewski P, Zamlynski M, Zalewska Z, Grucza K, Woznica M, Zamlynski J, Olejek A, Kwiatkowska D: Is pregnancy a confounding factor for hGH detection by using the biomarkers method? .................................................................................................................................................... 80-82 Grucza K, Wicka M, Drapala A, Konarski P, Bedecka M, Kaliszewski P, Stanczyk D, Kwiatkowska D: Monitoring of a potentially valuable metabolite of indapamide in routine doping control........................................................................................................................................................ 83-88 Mareck U, Fußhöller E, Geyer H, Görgens C, Guddat S, Petring S, Thevis M: Detection of the SARM S-23 in urine at low pg/mL levels after consumption of nutritional supplements contaminated with S-23 in the low ng/g range - a pilot study............89-92 Kraiem S, Al-Said I, Samsam W, Ali Saad K, Beotra A, Ali-Mohamed V, Al-Maadheed M: Detection of sibutramine and its metabolites in urine, serum and plasma by UHPLC MS: Case studies about unintentional doping with Hoodia gordonii ....................................................93-98 Cuervo D, Muñoz G: Doping-related drugs seized in 2023 during police operations in Spain: discussing statistics and new trends in traffic of doping substances ...........................................99-103 Mazzarino M, Khelifi S, Nayef Al Qasmi N, Ali Saad K, Kraiem S, Beotra A, Ali-Mohammed V, Al-Maadheed M Determination of dermorphin and 7 structural analogues in human urine by LC-HRMS for doping control purposes ..............................................................................................................104-109 Zhu S, Liu X, Zhang L, Wang S, Cheng Y: Potential risks of adverse analytical findings after consumption of metandienone contaminated beef from the market ...............................................................................................110-113 Ekström L, Suominen T: Longitudinal study of serum steroids in females ...........................................................................114-118 RECENT ADVANCES IN DOPING ANALYSIS (32) 5 ISBN 978-3-86884-080-3 PAGE Medojevic N, Grainger J, Brooker L, Agon V: Testicular cancer and the steroid profile: A case study ................................................................119-124 Pop A, Constantinescu G, Penu R, Toboc A, Stan C: Validation of 6-OXO and its metabolites confirmation method by GC-MS/MS and analysis of long-term storage samples ............................................................................................125-128 Sobolevsky T, Ahrens B: Carbon isotope ratio analysis of AICAr-derived ribose: a pilot study ........................................129-132 Sobolevsky T, Ahrens B: Formylation of steroids: an old method with a new outlook......................................................133-137 Williams E, van der Merwe M: Comprehensive determination of the carbon isotope ratios of endogenous steroids in human urine of the South African population by GC-C-IRMS......................................................138-143 Pettersson-Bohlin K, Buisson C, Marchand A, Pohanka A, Ericsson M: Metabolites found in human urine after Oxbryta (voxelotor) administration to sickle cell patients........................................................................................................................................... 144-147 Fußhöller G, Piper T, Sahu PL, Svambayev E, Thevis M, Geyer H Boldenone undecylenate residues in meat – pilot prevalence study and simulation of a contamination scenario with estimation of urinary boldenone and boldenone metabolite levels .................................................................................................................................148-152 Montes de Oca Porto R, Martinez Brito D, Terrero Serrano O, Rodríguez Fernández A, Correa Vidal T: Metabolism of 17 -methyl-19-nor-testosterone using gas chromatography coupled to mass spectrometry ..............................................................................................................................153-157 Soni A, Singh A, Singh S, Sahu PL: Investigation of triamterene excretion pattern: Expanding the detection window in sports doping ........................................................................................................................................158-161 Sahu PL, Handa T, Upadhyay A, Sahu K, Anandarajan T, Mahajan S, Bhawna V, Padder AH, Reddy DS, Ahmed Z, Ahmed QN: 17 -Hydroxy-androst-1,4,6-triene-3-one: Synthesis and characterization...............................162-165 Frelat C, Buisson C, Martinat N, Ericsson M: Automated extraction with positive pressure system for the GC-CIRMS sample preparation ...........................................................................................................................................166-169 Hülsemann F, Gougoulidis V, Blatt C, Thevis M: Purification process of commercial acetic anhydride for derivatization of steroids in GC/C/IRMS analysis.............................................................................................................................170-172 RECENT ADVANCES IN DOPING ANALYSIS (32) 6 ISBN 978-3-86884-080-3 PAGE Cristea C, Berghes B, Catana D, Radu M, Toboc A, Stan C: Liquid-liquid extraction approach for the identification of solriamfetol in doping control.................................................................................................................................................... 173-176 Liu Y, Zhang Y, Ma C, Wang Z, Dong T, Chang W: A comprehensive screening method for small peptides including BPC157: The crucial role of pH regulation in weak cation exchange SPE ..................................................177-182 Marchand A, Crepin N, Roulland I, Cerone H, Ericsson M: Evaluation of new kits in 96-well plate format for the detection of doping with human Growth Hormone................................................................................................................................. 183-187 Sakurai H, Akiyama K, Momobayashi A, Sato M, Okano M: Doping control analysis of darbepoetin alpha biosimilars...........................................................188-191 Mc Pherson Medina A, Fiallo Fernandez T, Martinez Brito D, Montes de Oca Porto R: Stability of CERA in urine. Influence of the use of Stemcell and MAIIA immuno- purification kits .................................................................................................................................... 192-195 He C, Shen L, He S, Gan J, Liu X, Sun G, Zhang L: Certain effects on the HBT result measured by flow cytometer .................................................196-200 Voss S, Schwenke D, Kniess A, Kirchbichler A, Robinson N: Comparison of K2EDTA and K3EDTA blood tubes for the parameters of WADAs ABP...........201-205 Donati F, de La Torre X, Botrè F: Variability of Serum and Plasma biomarkers associated to the Hematological module of the Athlete Biological Passport in elite athletes selected for doping control......................206-209 Harju L, Nieminen H, Siivonen A, Suominen T: Analysis of doping agents in dried blood spots (DBS) – method development of multiple classes screening..................................................................................................................................210-214 Chang W, Yan S, Yan X, Zhang Y, Liu Y, Yang S: Doping control analysis of BPC157 and dermorphin analogues in dried blood spots by SPE and UPLC-HRMS............................................................................................................................215-219 Mazzarino M, Salama S, Saleh A, Mahieddine S, Ali Saad K, Kraiem S, Beotra A, Ali-Mohammed V, Al-Maadheed M: Detection of steroid esters in dried blood spot by LC-MS/MS following microwave assisted derivatization ........................................................................................................................220-223 Yan X, Yang S, Zhang L, Wang Z, Yan S, Chang W: A study of UV/Visible spectra of hydroxylamine derivatized nandrolone esters on DBS by a computational and experimental approach...........................................................................224-228 RECENT ADVANCES IN DOPING ANALYSIS (32) 7 ISBN 978-3-86884-080-3 PAGE Stojanovic B, Pecher D, Geisendorfer T, Forsdahl G, Gmeiner G: DBS in doping analytics – opportunities and pitfalls.....................................................................229-233 Stojanovic B, Jeitler V, Leclerc M, Sylvestre A, Naud J, Gmeiner G: Interlaboratory comparison as the key to proof competency in DBS testing...........................234-237 Watanabe Y, Miyamoto A, Aoi A, Ota M, Sato M, Okano M: LC-MS/MS analysis of osilodrostat in urine and dried blood spots after administration ......238-242 Steen L, Mazzarino M, Deventer K, van Eenoo P: Preliminary experiments towards the quantitative analysis of ephedrines in dried blood micro sampling devices by liquid chromatography-mass spectrometry........................243-248 PRESENTATIONS ABSTRACTS - LECTURES Hernández Domínguez D, Coll Camenforte S, Bressan C, Ramírez Llanelis R, Monfort Mercader N, Ventura Alemany R: Impact of repeated oral administration of glucocorticoids on the urinary steroid profile............................................................................................................................................................ 249 Santos G, Carneiro G, Mendes J, Marques A, Padilha M, Pereira H: Bidimensional LC applied to antidoping analysis – update of small projects .................................. 250 Padilha M, Pereira H, Anselmo C, Estevam D, Meirelles JV, Brito B, Gomes M, Miranda B: Mosher strikes again: Alkylamine- and phenylalkylamine-like confirmation procedures based on -methoxy- -trifluoromethylphenylamide derivatives: a systematic study .........251-252 Nunes I, Mariana C, Oliveira F, Ana Carolina C, Anselmo C, Pereira H: Volumetric absorptive blood microsampling vs urine analysis after administration study in stimulant and diuretics. A pilot comparison .......................................................................... 253 Paßreiter A, Naumann N, Thomas A, Grogna N, Delahaut P, Thevis M The Duality of CRISPR in Anti-Doping Research Detection of sgRNA via SHERLOCK for CRISPR related gene doping control purposes.................. 254 Fedoruk M, Lewis L, Wright D, Marrapode M, Aranyosi A, Ahrens B, Sobolevskii T, Nicoli R, van Wagoner R, Eichner D Evaluating sweat as a novel complementary matrix to detect doping............................................. 255 Breuer J, Garzinsky A, Thomas A, Kliesch S, Fedoruk M, Geyer H, Thevis M: Multi-analyte LC-(HR)-MS/MS methods for the analysis of doping substance residues in human seminal fluid and case-related application ...................................................................256-257 RECENT ADVANCES IN DOPING ANALYSIS (32) 8 ISBN 978-3-86884-080-3 PAGE Heybroek M, Chaparala A, Monfredi A, Howa J, Nielsen B, Johnson H, Nair V: Evaluation of a proprietary recombinant -glucuronidase for suitability of use in manual and automated antidoping applications............................................................................258 Deventer K, Brutsaert A, de Wilde L, Roels K, van Eenoo P: Development of an online SPE-LC-MS/MS confirmation method for beta-blocking agents in urine .................................................................................................................................259 de La Torre X, Colamonici C, Martinez-Brito D, Montes de Oca Porto R, Parr MK, Lingyu L, Botrè F: Metabolism of clostebol by different administration routes.........................................................260 de La Torre X, Martinez-Brito D, Colamonici C, Curcio D, Botrè F: Detection of 6 -chloro-testosterone abuse by mass spectrometric techniques ..........................261 Garzinsky A, Fußhöller G, Hedeland M, Thevis M: Investigation of elimination profiles of microdosed dehydrochloromethyltestosterone (DHCMT) and methylclostebol by GC-MS/MS ................................................................................262 Peters I, Naumann N, Görgens C, Guddat S, Thevis M: Elucidation of the metabolism of dehydrochloromethyl-testosterone in human skin and liver using a co-culture microphysiological chip model...........................................................263 Wen H, Wagener F, Piper T, Neudörfl J, Thevis M, Schäfer M: Investigation structures of in vtro and in vivo metabolites of a novel 20-keto-steroid S42 by LC/GC-MS analysis and organic synthesis .................................................................... 264-265 Möller T, Thevis M: Determination of the metabolic pathways of different Rycals in in-vitro samples for doping control purposes ..................................................................................................................266 Langer T, Nicoli R, Guillarme D, Rudaz S, Veuthey J, Kuuranne T, Musenga A: Supercritical fluid chromatography and endogenous steroids: Super possibilities and critical aspects ..................................................................................................................................267 Nair V, Hanelli T, Moore C, Eichner D: Evaluation of the Minimum Reporting Limit to determine incompetition use of stimulants .........................................................................................................................................268 Piper T, Fußhöller G, Thevis M: Investigations into the human metabolism of trestolone (7 -methyl-19-nortesto- sterone, MENT) – first results ..........................................................................................................269 Keiler AM, Thieme D, Krumbholz A, Bidlingmaier M, Hameder A, Stöver A, Graw M: Influence of ethanol and food intake on blood steroids ...............................................................270 RECENT ADVANCES IN DOPING ANALYSIS (32) 9 ISBN 978-3-86884-080-3 PAGE Honesova L, van Eenoo P, Polet M: Development of an LC-IRMS method for steroid analysis in urine Part 1: Evaluation of analytical columns suitable for high-temperature liquid chromatography - isotope ratio mass spectrometry analysis of anabolic steroids .................271-272 Honesova L, van Eenoo P, Polet M: Development of an LC-IRMS method for steroid analysis in urine Part 2: Evaluation of analytical columns suitable for high-temperature liquid chromatography - isotope ratio mass spectrometry analysis of anabolic steroids .................273-274 Coll S, Shiomoura S, Alechaga E, Bressan C, Monfort N, Ventura R, Okano M: Detection of oral testosterone undecanoate administration in Asian population: steroid profile, IRMS markers and sulfate metabolites....................................................................... 275 Piper T, Thevis M: Improving the determination of carbon isotope ratios of endogenous steroids found in human serum............................................................................................................................................... 276 Colpaert T, Risseeuw M, van Eenoo P, Deventer K: Updated method for screening and confirmation of small peptides in urine using dilute- and-shoot methods combined with direct urinary injection into (nano) LC-HRMS ................277-278 Walpurgis K, Thomas A, Sato M, Okano M, Geyer H, Thevis M: Mass spectrometric detection of the GH analogue Somatrogon from doping control serum samples ............................................................................................................................................ 279 Miller G, Goodrum J, Crouch A, Eichner D: Comparing detectability of somapacitan-beco (Sogroya®), a longacting GH drug, and somatropin following controlled administrations in healthy volunteers......................................... 280 Miller G, Goodrum J, Peek K, Eichner D: Comparing serum-based doping control analytes in traditional venous serum and capillary serum collected with the Tasso+ SST device ......................................................................... 281 Requena-Tutusaus L, Coll S, Bressan C, Anselmo I, Bergés R, D'onofrio G, Ventura R: Impact of repeated glucocorticoid oral administration on the hematological passport............... 282 Terracciano F, de La Torre X, Donati F, Botrè F: Characterization of red blood cells changes during storage by atomic force microscope................................................................................................................................................... 283 Al-Jaber M, Al-Nesf M, Manickam C, Sastry KS, Rammal S, Mohd S, Cherif A, Beotra A, Al-Maadheed M, Mohamed-Ali V: Investigation of the sensitivity of STR analysis for the in-vivo detection of homologous blood transfusion........................................................................................................................................ 284 RECENT ADVANCES IN DOPING ANALYSIS (32) 10 ISBN 978-3-86884-080-3 PAGE Reichel C, Lorenc B, Scheiblhofer V, Gmeiner G, Thevis M: Data from an EPO theta administration study and Updated protocol for high sensitivity direct immunoprecipitation of epoetins from urine .....................................................................285-286 Zhou X, He S, Niu C, Liu X, Martin L, Wu D: Development of a confirmation method for recombinant EPO using reverse-normal immunopurification coupled with Western Blotting ....................................................................287-288 Naumann N, Do C, Vollmert C, Krajina M, Thevis M: Development and characterization of a multiplex panel for the simultaneous testing of 7 gene doping targets using Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry.....................................................................................................................................289 Albertsdóttir AD, van Gansbeke W, van Eenoo P, Polet M: Comparison of the detection time of sulfated metabolites of anabolic steroids using four analytical techniques: update with 17 -methyltestosterone.............................................209-291 PRESENTATIONS ABSTRACTS - POSTERS Görgens C, Möller T, Guddat S, Thomas A, Svambayev E, Geyer H, Thevis M: Urinary metabolites indicative of the administration of hypoxen monitored by liquid chromatography – high resolution/accurate mass tandem mass spectrometry ............................292 Krug O, Thomas A, Tobergte L, Sahu P, Thevis M: Quantification of emoxypine, emoxypine sulfate, and emoxypine glucuronide in cow's milk and human urine ................................................................................................................................293 Wicka M, Drapala A, Wojtkowiak K, Grucza K, Konarski P, Bedecka M, Kwiatkowska D: Elimination profile of semaglutide and its metabolite in human urine ....................................294-295 Wicka M, Drapala A, Wojtkowiak K, Grucza K, Konarski P, Bedecka M, Kwiatkowska D Identification of compounds used in therapy for type 2 diabetes as potential doping substances with particular emphasis on glucagonlike peptide-1 (GLP-1) analogues .............296-297 Keiler AM, Derwand D, Zierau O, Thieme D: Effects of 5 -hydroxy-laxogenin in the orchiectomized rat model ...................................................298 Loungyot T, Thangboonjit W, Prompittayayut P, Yaikwawong M, Songsaeng R, Panan S, Thitamadee S, Kongpatanakul S: Transcriptomic profiling of clenbuterol-induced human myotubes..................................................299 RECENT ADVANCES IN DOPING ANALYSIS (32) 11 ISBN 978-3-86884-080-3 PAGE Charlebois A, Naud J, Ayotte C: Replacing the paper documentation with electronic operation trails, including the chain of custody of samples, processed samples, reagents, and reference materials...................300 Park H, Cho Y, Park S, Chin A, Xu Y, Jeon M, Jung S, Kim M, Son J: Enhancing the accuracy of intelligent doping diagnosis through nonlabeling machine learning optimization .................................................................................................................................301 Fedoruk M, Lewis L, Sobolevskii T, Ahrens B, Nicoli R, Thevis M: Cases of contamination resuling from athlete exposure to pet medications..................................302 He G, Zhu S, Liu X, Lu J, Wang Z, Zhang L, Li J: A multi-technique based analytical platform for characterization and identification of a newly emerged steroid 6 -chlorotestosterone and its in-vivo epimeric metabolite.....................303 Krug O, Naumann N, Geyer H, Thomas A, Piper T, Thevis M: Black market products suspected to contain doping relevant ingredients - report for 2022-2023.....................................................................................................................................................304 Miyamoto A, Watanabe Y, Ota M, Sato M, Okano M: Comparison of doping control analysis of testosterone enanthate in Asian males .......................305 Piper T, Fußhöller G, Thevis M: Employing 11-ketotestosterone as target analyte for adrenosterone (11OXO) administrations in doping controls – reference population derived thresholds.............................306 Martinez-Brito D, Leogrande P, Jardines Garcia D, Botrè F, de La Torre X: Stability of 6-chloro-testosterone by LCMS and GCMS........................................................................307 Sobolevsky T, Fedoruk M, Dellanna F, Geyer H, Ahrens B, Thevis M: Long-term excretion of roxadustat in urine ...........................................................................................308 Alhalabi H, Garzinsky AM, Thomas A, Thevis M: Characterization of a LC-MS method for differentiation between doping and contamination scenarios concerning the anabolic agent SARM S-23 ...............................................309 Walpurgis K, Gäde A, Thomas A, Gochard S, Delahaut P, Thevis M: Detection of extracellular hemoglobin from Arenicola marina in doping control plasma and serum samples by means of LC-HRMS/MS: In vivo studies.........................................................310 Martinez-Brito D, Leogrande P, de La Torre X, Botrè F: Detection of thyroid hormones in urine by LC-MS/MS. An investigation on high- performance athletes.................................................................................................................................311 Martinez-Brito D, Leogrande P, de La Torre X, Botrè F: Preliminary results on thyroid hormones metabolism. A case study................................................312 RECENT ADVANCES IN DOPING ANALYSIS (32) 12 ISBN 978-3-86884-080-3 PAGE Korsmeier L, Krombholz S, Thevis M: Analysis of the metabolic profile, bioavailability, and elimination behavior of the selective androgen receptor modulators LGD-4033, RAD-140, and S-23 after trans- dermal application for doping control purposes............................................................................313-314 Sahu PL, Samandram R, Matsa R, Jat S, Pullapanthula R, Murty U, Handa T, Raj A, Sahu K, Anandarajan T, Kumar P: Synthesis and characterization of molidustat glucuronide .................................................................315 Sahu PL, Matsa R, Samandram R, Pawar S, Pullapanthula R, Murty U, Handa T, Sahu K, Anandarajan T, Kumar P: Synthesis and characterization of YK-11 metabolite (M1) ..................................................................316 Sahu PL, Handa T, Upadhyay A, Bhardwaj A, T. A, Sarkar AR, Mahajan S, Bhawna V, Padder AH, Nandi U, Ahmed QN, Reddy DS, Ahmed Z: Synthesis and characterization of Growth Hormone Releasing Peptide metabolite: GHRP-1 ..... 317 Almeida CVP, Neng NR, Nogueira JMF, Ruivo J: Application of a bar adsorptive microextraction based methodology for doping control of alkylamine stimulants in urine matrices .....................................................................................318-319 Kim KH, Cho EB, Jeon M, Son J: Multiresidue LC-MS screening by using solid-phase extraction and optimized enzymatic hydrolysis in sports doping .......................................................................................................................320 Pfeffer S, Gmeiner G, Forsdahl G: Methylation: a supplementary derivatization technique for the analysis of phase II metabolites of endogenous anabolic androgenic steroids ..........................................................321-323 Heo S, Cho Y, Kim J, Sung C, Lee KM, Kim HJ, Jung H, Son J: Developing innovative markers to enhance the detection windows for small peptide hormones .....................................................................................................................................................324 Niu C, Liu X, Zhou X, He S: Study on internal standards applicable to the detection of VAR-EPO in doping control ..............325 Fiallo Fernandez T, Mc Pherson Medina A, Martinez Brito D, Montes de Oca Porto R: Detection of ERAs in urine by purification kits from MAIIA Diagnostics ..........................................326 Sung C, Kim M, Yi J, Jeong Y, Hwang NS, Park HH: Validation study of the high-throughput gene doping assay (HiGDA) platform for exogenous EPO gene doping in human blood .......................................................................................327 RECENT ADVANCES IN DOPING ANALYSIS (32) 13 ISBN 978-3-86884-080-3 PAGE Akiyama K, Momobayashi A, Okano M: Direct detection of CRISPR-Cas9 ribonucleoprotein gene doping using RNA immuno- precipitation and quantitative PCR .........................................................................................................328 Teixeira P, Levy R, Muniz S, Azevedo A, Barros T, Pereira H: Gel card kit (ID-system, BioRad) – An alternative to homologous blood transfusion detection? ....................................................................................................................................................329 Couture M, Barber A, Naud J: Detection of testosterone esters in blood by LC-MS/MS ....................................................................330 Somay Selbes Y, Menderes Y, Akin S, Ekinci YE, Ilicepinar ÖF, Nemutlu E, Demirel HA: Do plasma volume markers reflect changes in the athlete's biological passport? .........................331 Almeida Oliveira J, Loria F, Mumenthaler C, Kuuranne T, Leuenberger N: Comparison between standard hematological parameters and blood doping bio-markers in dried blood spots within the athlete population of Swiss Sport Integrity (SSI) ..........................332 Langer T, Musenga A, Ericsson M, Gmeiner G, Harju L, Mongongu C, Pecher D, Stojanovic B, Suominen T, Tsivou M, Kuuranne T, Nicoli R: Analysis of testosterone esters in serum and DBS samples – results from an inter- laboratory study..........................................................................................................................................333 Lee J, Jeong TY, Kang M, Kim HJ, Kim KH, Son J, Lee KM: substances in dried blood spots (Tasso-M20).................................................................................................................334 Min H, Hwang J, Lee H, Son J, Lee HJ: Simultaneous detection method of muscle developing monoclonal antibodies using LC-MS/MS from dried blood spots ..........................................................................................................335 Aoi A, Miyamoto A, Ota M, Momobayashi A, Sato M, Okano M: Detection of plasma volume expander hydroxyethyl starch in DBS and urine and impact on the blood parameters and blood steroid profiles ...........................................................................336 Boschetti G , Langer T , Nicoli R , Mumenthaler C , Medana C , Kuuranne T , Musenga A: Development of a qualitative screening method for small peptides in dried blood spots ..............................................................................................................................................................337 MDI PAGE Akiyama K, Momobayashi A, Okano M: Direct detection of CRISPR-Cas9 ribonucleoprotein gene doping using RNA immuno- precipitation and quantitative PCR ooo... ccececscsesececsssesesecasscsesecasssseseasassesesecassesesacassnseeeeaes 328 Teixeira P, Levy R, Muniz S, Azevedo A, Barros T, Pereira H: Gel card kit (ID-system, BioRad) — An alternative to homologous blood transfusion COteCtiOn? oo... eee ecccccceccesesessessesesescesescsesesnssucscacansnsusueasacecscssesasacensnsnsusasacenensesasasasensnsesisaseasensnseeesasetentees 329 Couture M, Barber A, Naud J: Detection of testosterone esters in blood by LC-MS/MS ............ ccc ccccceceeteteeeeeeeeeeeeeeees 330 Somay Selbes Y, Menderes Y, Akin S, Ekinci YE, llicepinar OF, Nemutlu E, Demirel HA: Do plasma volume markers reflect changes in the athlete's biological passport?....................... 331 Almeida Oliveira J, Loria F, Mumenthaler C, Kuuranne T, Leuenberger N: Comparison between standard hematological parameters and blood doping bio-markers in dried blood spots within the athlete population of Swiss Sport Integrity (SSI)............0..00.... 332 Langer T, Musenga A, Ericsson M, Gmeiner G, Harju L, Mongongu C, Pecher D, Stojanovic B, Suominen T, Tsivou M, Kuuranne T, Nicoli R: Analysis of testosterone esters in serum and DBS samples — results from an inter- Vablooratory Study... cece cecceeeeeeeeseseeeeseseseseseeeeseaeseseeeeeeaeeeeeeeeeeaeasaeseseeesesasaseeseseeasaseneeeeeeeaseseneeeeeeaeatees 333 Lee J, Jeong TY, Kang M, Kim HJ, Kim KH, Son J, Lee KM: UHPLC-MS/MS method for simultaneous screening of 257 prohibited substances in Cried blood spots (TaSSO-M20)............cccccccccscscscscscscscscscsesesesesesesescsssessssseseseasseasseseseseseseseaeseseeeseseeeeeees 334 Min H, Hwang J, Lee H, Son J, Lee HJ: Simultaneous detection method of muscle developing monoclonal antibodies using LC-MS/MS from dried DIOOd Spots 00.0... ccc cccccecsseseeecscseseseeecscsesesesasscsesesacassesesacasseseeecassesesacatasseetenes 335 Aoi A, Miyamoto A, Ota M, Momobayashi A, Sato M, Okano M: Detection of plasma volume expander hydroxyethyl starch in DBS and urine and impact on the blood parameters and blood steroid profiles... ccc ccscsesecscseseeecscseseseeecssssseteeeeasees 336 Boschetti G , Langer T , Nicoli R, Mumenthaler C , Medana C , Kuuranne T , Musenga A: Development of a qualitative screening method for small peptides in dried blood cS 0 0) ee cee eee 337 RECENT ADVANCES IN DOPING ANALYSIS (32) 13 ISBN 978-3-86884-080-3 RECENT ADVANCES IN DOPING ANALYSIS (32) 14 ISBN 978-3-86884-080-3 SCIENTIFIC BOARD X. de la Torre, Rome (Italy) F. Donati, Rome (Italy) H. Geyer, Cologne (Germany) G. Gmeiner, Seibersdorf (Austria) R. Kempkes, Cologne (Germany) U. Mareck, Cologne (Germany) M. Mazzarino, Rome (Italy) G. Miller, Salt Lake City (USA) M. Okano, Tokyo (Japan) T. Piper, Cologne (Germany) C. Reichel, Seibersdorf (Austria) S. Rzeppa, Dresden (Germany) W. Schänzer, Cologne (Germany) M. Thevis, Cologne (Germany) A. Thomas, Cologne (Germany) P. van Eenoo, Ghent (Belgium) R. Ventura, Barcelona (Spain) S. Voss, Dresden (Germany) 15 MDI ex Lecture salt MANFRED DONIKE WORKSHOP 2024 Wang Y, Zhang L, Wang Z, Xing Y, Wang S, Shen L, Yang Z Doping Control Laboratory operational strategy and outcome of sequential major competitions including the Hangzhou Asian Games in 2023 Beijing Anti-Doping Laboratory, Beijing Sport University, Beijing, People’s Republic of China Abstract World Anti-Doping Agency (WADA) accredited Beijing Anti-Doping Laboratory (Beijing lab) analyzes 18,000 to 20,000 doping control samples annually from year 2018 to 2022. Affected by the COVID-19 pandemic, several competitions of recent 2 years were postponed to year 2023. This includes the Chengdu 2021 FISU World University Games, the 2022 Hangzhou Asian Games and Guangxi Student/Youth Games of China. As a result, a total of 26267 samples were analyzed in year 2023 by Beijing lab, which has a roughly 30% increase than past years. With all out-of-competition samples included, Beijing lab analyzed 12174 samples from July to November, 2023, with an average reporting Turnaround Time (TAT) of 6.9 days, which is tremendously shorter than the 20 days requirement proposed in WADA's International Standards for Laboratories (ISL). Among all these analysis missions, Hangzhou Asian Games is the most demanding event where 1665 samples were analyzed with an average TAT of 2.1 days, in which eighteen (18) Adverse Analytical Findings (AAFs) and seven (7) Atypical Findings (ATFs) were reported. Compared to the anti-doping project for the Beijing 2022 Winter Olympics, an optimized setup of about 50% workforce were employed during this period. This presentation summarizes details of the laboratory ’s operational strategy along with discussion of several highlighted quality assurance measures. Introduction Doping control is a fundamental aspect of maintaining the integrity of sports competitions worldwide. With the increasing frequency of major sporting events, doping control laboratories face significant challenges in ensuring timely, accurate, and reliable test procedures. The unique challenges posed by hosting multiple high-level events in a short timeframe highlights the need for robust, proactive, and adaptable risk management frameworks. This paper explores the risks associated with Beijing Anti- Doping Laboratory during sequential major competitions, including the Hangzhou Asian Games in 2023, and outlines a comprehensive risk assessment and management framework. Experimental The daily number of samples tested in routine analysis of Beijing Anti-Doping Laboratory is about 75, and the reporting Turn-round Time (TAT) is usually 20 days. From July to November 2023, the larger number of samples coupled with fast TAT requirements, and the unexpectedly high rate of AAFs and ATFs have posed significant challenges for the laboratory [1]. RECENT ADVANCES IN DOPING ANALYSIS (32) 15 ISBN 978-3-86884-080-3 16 MDI 1 Lecture of MANFRED DONIKE WORKSHOP 2024 Timea Number of | TAT Requirments Samples 2021 Chengdu FISU Jul 25 to Aug 679 48 hours for standard menu, World University 10 72 hours for others Games 2022 Hangzhou Asian Sepi7teOct 2055 48 hours for standard menu, Games 10 72 hours for others 2022 Hangzhou Asian Oct20to Oct 465 43 hours for standard menu, Para-Games 29 72 hours for others Guangxi Student/Youth Oct23toNov 1679 5 days for all menu Games of China i8 China Anti-Doping AugiSteOct3 4449 3-5 days for all menu Agency's Pre-Game Screening Missions Table 1. Number of samples and TAT requirements for sequential major competitions Reselt Date recelwee! Fret Subrdason Lest Uipdete Subetences 19-norandrosterone > Long/mL 25-S0p-20 27-Sep-23 lostebal C/CARMS confirmed exogenous origin of Ts 3 rine Badminton 25-Sep-23] 30-Sep-23] jerythropotetin (EPO) cod _‘(eycling 15-Sep-23] 30-Sep-24 erythropoietin (EPO) Ririne ==» Eyecling 27-509-25 20-5ep24 4-Octe2 mpuigaceaRaiek rine Athbeties 28-Sep-23] 2-Cret-23 19-norandrosterone, GC/C/IRMS negative WWrine Cycling 28-Sep-23] 30-Sep-23 idenone at 0.6 ng/ml, < LOO of GC/CHAMS [Urine Handball 29-Sep-23) 1-Oct-23| 4-Oct-23)GC/(C/IRMS confirmed exogenous origin of Ts ood fAthbetics 29-Sep-23) 3-Oet-2 panercen (dEPO) rythropoletin (EPO) lrine Athletics 2-Sep-23) 29-Sep-28 3-Oet-2adarbepoetin (dEPO) rine urash a Sep-23] 4-Oet-23, darbepoetin (dEPO) [Blood Kasing 30-Sep-23 B-Cet-23] methoxy polyethylene glycol-apoetin beta (CERA} Urine thiatics 1-Oct-23) 0 S-Oect-23] larnfetamine . ; id rachlorome shestostanane Urine Weightlifting 1-024 4-Oct-23 aepldalaes ans rine PVeightlifting 32-Oct-23] S-Oct-23) drostanclone ood fWrestling uring [eabaddi rine Boxing 23-Oct-23] #-Cet-23 23-Oct-23] B-Cet-23 3-Chct-23] &-Cet-23] rythrapoietin (EPO) 15-norandrosterone, GC/C/IAMS negative Kit 1 Ratio < DL. Further Testi recommended Bl alalele| & [B[slela) & feiss] & [ele lUrine Handball $-Oct-23] er hropoletin (EPO) jurine Cycling ATE 4-Oct-23] 9-Oct- 24] idenone at 0.9 ng/mL, < LOW of GC/CARMS [Blood Athbeties ATF a-Oiet= 23] 14-Oet-23 @ak signal for rEPO, other sample from the same athlete reported AGF rine Sport Climbing AF 5-Oect-23 10-Det-24 brinzolamide ring resting JAF 5-Oct-23] T-Oct-23 19-norandrosterone rine Basketball AF 5-Oct-23] £-Oct-23 metandienone rine Basketball |AAF 7T-Oct-23] 10-Ciet-231 Carboxy-THC greater than the Decision Limit Urine jAthieties JAF TeOhct-23]° | 10-Cet- 23} dehydrochloremethyl-testosterone Table 2. AAFs and ATFs in Hangzhou Asian Games Despite limited staffing and instrumentation, the laboratory completed the test tasks for sequential major competitions, including the Hangzhou Asian Games in 2023. The reporting time during the competitions fulfilled the fast TAT requirements. The reporting time of samples unrelated to the competitions was within 20 days, which is compliant with International Standard for Laboratories (ISL) of the World Anti- Doping Program [2]. After all competitions, some reporting time exceeded 20 days due to the reduction of staff and instruments, but it was still within the reporting time agreed by each Testing Authority (TA). RECENT ADVANCES IN DOPING ANALYSIS (32) 16 ISBN 978-3-86884-080-3 17 MDI Gana Lecture alt MANFRED DONIKE WORKSHOP 2024 Workload (Urine) Distribution and Report Turnaround Time - July to November, 2023 400 IC Sampler 0OC Samples mic TAT (dais) == (C0 TAT (ednys} 4 20 f 200 f 150. + Figure 1. Workload (Urine) distribution and report TAT Results and Discussion To ensure the proper operation during sequential major competitions, the laboratory has conducted a comprehensive risk assessment and management. The maximum daily sample number is calculated based on TAT and the length of single injection for mass spectroscopy (MS) test methods, while batch size optimization versus cost efficiency is undertaken for biochemistry methods, considering the crucial instrumentation and consumables with short shelf-life [3]. In cases where instruments are insufficient, the option to lease or borrow is considered. Sufficient workforce is allocated for sample reception and aliquoting, data reviewing, and specialized biochemistry test methods like polyacrylamide gel electrophoresis (PAGE), Western Blot, and hormone isoform analysis. The support from peer World Anti-Doping Agency (WADA) labs of Tokyo and Seoul, staff from Shanghai candidate lab, and China Anti-Doping Agency (CHINADA) food and nutrition lab is actively sought to augment the workforce capabilities. University student volunteers are offered enough time for training, ensuring they are prepared to contribute effectively to the workforce. To prevent staff depletion due to illnesses like COVID-19 and flu, sufficient staff is recruited for rotation schedules, and quarantine measures are minimized to maintain uninterrupted operations. Instrument stability is maintained through on-site support from Agilent and Thermofisher engineer, who built an temparory inventory of instrument parts, and verifying instrument status at end of shift after the first sample injection. To mitigate tube swapping errors, deuterated internal standards (ISTD) are introduced during sample aliquoting, and one-tube-at-a-time handling is enforced during critical steps, with additional supervisory quality assurance (QA) staff provided. A dual reviewer system is implemented for both data reviewing and LIMS reporting is reinforced to ensure accuracy. RECENT ADVANCES IN DOPING ANALYSIS (32) 17 ISBN 978-3-86884-080-3 18 MDI Gee Lecture cate MANFRED DONIKE WORKSHOP 2024 Risk Resolution « Calculation: l nctearmentation Rc Critical > » MS methods: maximum daily sample number / TAT eee ears __ /length of single injection » Biochemistry methods: batch size vs cost efficiency * Lease or borrow instrument - Calculation: Workload » Sample Reception and Aliquoting | Capacity > MS method: data reviewing va Biochemistry: PAGE and Western Blot, CG/LH, GH Isoforms, HBT — Avoid multitasking as much as possible Students — enough time for training Support from peer WADA labs Shanghai candidate lab CHINADA’s food and nutrition lab No quarantine Recruit sufficient staff for rotation Risk Resolution * On-site Agilent and Thermofisher engineer Instrument Stability * Set up inventory of instrument parts in the facility « End of shift after first injection of actual sample . Deuterated ISTD at sample aliquoting ; « Handle one tube only at a time during capping, Random Tube swonpiie | centrifugation, put on injector, etc human | + Extra supervisory QA staff at Sample Prep. section mistakes Peak misreading in ITP for r Fully independent two reviewers + LIMS reporting review | MS method eee es Planaine: 0g _ = Don't perform data reviewing late at night or overnight!! _ Workforce Staff depletion - COVID and flu? te * Early communication Paperless DCF into LIMS glithes On-site support + Review the in-house validated Scope and RM inventory Rare Findings against WADA Testing Figures and 10 report of previous | _ Asian Games Table 3. Risk assessment and resolution In addition, extra Internal Quality Assurance Scheme (iQAS) planning is conducted to account for unforeseen circumstances and ensure that the operations remain robust. Late-night or overnight data reviewing was not assgined to prevent peak misreading in ITP for MS methods, expect for some of the Confirmation Procedure of rare cases. For smooth paperless Doping Control Form (DCF) integration into in-house Laboratory Information Management System (LIMS), early communication with the Sample Collection Authority, namely China Anti-Doping Agency, and on-site support from LIMS engineer are ensured to streamline operations further. For rare findings, a retrospective analysis is conducted by reviewing the in-house validated scope and Reference Material (RM) inventory against WADA testing figures report and independent observers report from previous events like the Asian Games [4,5]. This helps identify patterns and prepare for future occurrences. RECENT ADVANCES IN DOPING ANALYSIS (32) 18 ISBN 978-3-86884-080-3 19 MDI Gee Lecture cate MANFRED DONIKE WORKSHOP 2024 Section Substances) Matrix Preparation Events OP? Small peptidesiHexarelin (2 ng/mL) rine (Spiked ERAS PQ BRA (1 ILL) rine (Spiked DES Testosterone Cypionate (10 ngymL) OBS Spiked FISU Lc ETG (10 ug/ml) rine (Spiked FISU Gc Oxabolone (5 ng/mL) Urine Splked FISU Lc Fluocortelone (60 ng/ml) Urine [Spiked FISU hCG hCG (10 IU/L) rine — Spiked |FISU LC HES (300 ug/mi) Urine [Spiked GH InGH isoforms (rec; pit 2.17) Berum [Spiked GCEIRMS |15-NA (4 ng/mL) Urine Spiked IRMS CP Lc SWO?42 Sulfoxide (4 rg} Liriree Spiked 6c Danazol (2.5 ng/ml) rine [Spiked (OG LC Cratetamide (100ng/mL} lUrine [Spiked [OCA _|Re-ITP ERAS CERA (5 pg/ml) rine = Spiked KOH A [Small peptidesiGHRP-6 (2 ng/ml} Urine [Spiked OCA Lc Fenbutrazate (100 ng/mL) rine = Spikmed OCA, Gc [Methylstenbolone (2.5 ng/ml} Urine [Spiked OCA, Le Higenamine (20 ng/mt) Wrine (Spiked LARC Gc eranal (5 ng/ml) rine Spiked APC hcG nGG (10-1 L) rine [Spiked Le Higenamine (20 ng/mL) rine (Spiked Gc Mesterolone Met2(1o-Methyl-Sa-androstane-3a,17B-diol) (Sng/ml) [Urine [Spiked Lc [Torasemice (40 ng/ml) rine [Spiked Small peptidesPaforelin (2 ng/ml) ring = (Spiked ERAS EPO BRP (1 IU/L) Urine [Spiked LC Trimvetazicine Livin Excretion LC Dexamethasone ri ree [Exeretian Le WOT42 rine [Excretion Table 4. iQAS planned and analyzed Conclusions During sequential major competitions, a comprehensive risk assessment and management is necessary for a doping control laboratory to ensure that risks are identified promptly, analyzed thoroughly, and addressed effectively through proactive measures and continuous monitoring. This not only safeguards the quality and integrity of laboratory tests but also fosters a safe and productive work environment. References 1. World Anti-Doping Agency. 2022 Asian Games IO Report. Montreal (2024) https://www.wada- ama.org/sites/default/files/2024-05/asian_games_io_report_final_revised.pdf (access date 17.04.2024) 2. World Anti-Doping Agency. International Standard for Laboratories. International Standard, Montreal (2021) https://www.wada-ama.org/sites/default/files/resources/files/is|_ 2021.pdf (access date 01.01.2021) 3. Wang Y, Zhang L, Xing Y. (2022) Operation of the anti-doping laboratory for the Beijing 2022 Olympic and Paralympic Winter Games. Drug Test Anal. 14(11-12), 1853-1863 4. World Anti-Doping Agency. 2021 Testing Figures Report. Montreal (2023) https://www.wada- ama.org/sites/default/files/2023-01/2021_anti-doping_testing_figures_en.pdf (access date 17.01.2021) 5. World Anti-Doping Agency. 2018 Asian Games IO Report. Montreal (2019) https://www.wada- ama.org/sites/default/files/resources/files/wada_io_report_-_asian_games_- final_draft_20122018 - _clean.pdf (access date 05.02.2019) Acknowledgements We would like to express our genuine gratitude to World Anti-Doping Agency and and International Testing Agency for their prompt communication and consultation; to all the experts from other WADA accredited laboratories of Tokyo lab and Seoul lab, Shanghai candidate lab and CHINADA food and RECENT ADVANCES IN DOPING ANALYSIS (32) 19 ISBN 978-3-86884-080-3 20 MDI Gana Lecture alt MANFRED DONIKE WORKSHOP 2024 nutrition lab, who came or remotely supported us with extensive expertise; to all the student volunteers from Beijing Sport University, and to all the on-site engineers from Agilent and Thermofisher for their dedicated work during the Games. RECENT ADVANCES IN DOPING ANALYSIS (32) 20 ISBN 978-3-86884-080-3 21 MDI ex Lecture salt MANFRED DONIKE WORKSHOP 2024 Croft M, Edey C, Villanueva R, Murby EJ, Goebel C, Agon V The use of dual column chromatographic separations for an efficient ITP NMI, Australian Sports Drug Testing Laboratory (ASDTL), Sydney, Australia Abstract An efficient approach to the initial testing procedure for urine that employs two separate dual column chromatographic separations on GC-MSMS and LC-MSMS instruments has been developed. The dual column LC-MSMS procedure traps analytes from a single injection of a urine extract and uses column switching to allow their concurrent separation on HILIC and reverse phase columns with independent gradients. The effluent from the two columns is combined via a T-Piece prior to entry into the mass spectrometer and the acquisition of a single chromatogram. Compounds eluting from one column are interspersed with those eluting from the second column in the chromatogram, minimising run time. The dual column GC-MSMS screening procedure incorporates an alternative selectivity for separation of analytes using sequential injections of a sample on two different GC columns on the one instrument. This has improved efficiency by reducing sample handling and streamlining data processing. The use of larger internal diameter columns improved peak shape for compounds at high concentrations while also reducing retention shifts due to co-extractives. Calibrators prepared in urine matrix were used to improve the quantification of the steroid profile. The dual column ITP was validated in accordance with WADA requirements. Introduction The wide range of chemical properties of substances on the WADA prohibited list precludes the use of selective clean-up techniques when screening urine, requiring the specificity of mass spectrometry for detection. Multiple different chromatographic separations are still required, using both LC and GC, to reduce interferences from co-extractives and satisfy ever increasing WADA sensitivity requirements. The ability to perform different chromatographic separations from one injection via column switching and the use of different column chemistries increases the range of analytes that can be analysed in a single injection [1]. This improves efficiency by reducing sample handling, instrument run time and data processing and review. A single-injection, dual-column LC-MSMS method was developed in which compounds that are not well retained on a reverse phase column were transferred to a HILIC column prior to concurrent elution of all compounds from both columns using independent gradients. The dual column set up for GC-MSMS combined the laboratory’s previously validated methodology, where samples were injected on two separate instruments (GC-MSMS and GC-QTOF), onto a single instrument to improve efficiency. Technical modifications were incorporated into the method setup to address limitations in the previous methodology. A second more polar column (DB-35MS) was installed to provide alternative separation for selected compounds. Larger internal diameter columns were used to reduce the occurrence of retention RECENT ADVANCES IN DOPING ANALYSIS (32) 21 ISBN 978-3-86884-080-3 22 MDI 1 Lecture of MANFRED DONIKE WORKSHOP 2024 shifts in sample extracts with high levels of co-extractives. Calibrators prepared in urine matrix were used to improve the quantification of the steroid profile. Experimental Sample hydrolysis and solid phase extraction was fully automated (Janus, PerkinElmer) with the extract split into two portions for LC and GC analysis. Dried extracts were either reconstituted in buffer for LC- MSMS or TMSI derivatised for GC-MSMS analysis. A multipoint calibration prepared separately in urine matrix was used for quantification of the markers of the Steroid Profile. Single injection dual column LC-MSMS analysis was performed using an Agilent 1290 Infinity II System interfaced to an Agilent 6495 TQ-MS with Jet Stream source. Chromatographic separation utilised a C18- trap (Zorbax Eclipse XDB-C18 in-line cartridge, 2.1 x 15 mm, 1.8 ym) fitted with pre-column filter and two analytical columns with different chemistries: a HILIC (Poroshell HILIC-Z, 3.0 x 50 mm, 2.7 um, 30°C) and a C18 (Zorbax EclipsePlus C18, 2.1 x 50 mm, 1.8 um, 45°C). Two 10-port switching valves controlled by MassHunter software allowed selection of dual column mode and directed eluent flow from the trap. The system included both a binary and a quaternary pump operating independent gradients. Initial Position: Second Position: Loud sample te CES trap and HILIC column Transier to C18 Coleman and continue AIL ution BaiART Fa wl | Burry 00 Bingay aro a0 001 204 570 3.0 250 Ceaternany 0.00 O45 99.85 asa Cuaternare 2.01 10.00 0 0 0.350 Cofummn Selection Volver {05¥) , lex Cube Voter (FCW), Anotosampier (4,5 Figure 1. LC-MSMS dual column valve configuration showing initial and second position RECENT ADVANCES IN DOPING ANALYSIS (32) 22 ISBN 978-3-86884-080-3 23 MDI Ge Lecture cate MANFRED DONIKE WORKSHOP 2024 In the initial position, the sample (5 UL) is loaded onto the trap at 61 uL/min with a mobile phase containing 3% acetonitrile. Compounds that are trapped on the C18 in-line cartridge are later separated on the C18 column. Compounds that are not trapped proceed to a T-piece where the mobile phase is diluted with acetonitrile for retention and separation on the HILIC column. In the second position the two columns are eluted with independent gradients. Effluent from the two columns is combined prior to the mass spectrometer. Total flow to the MS is maintained at 600 uL/min throughout. A total of 782 MRM transitions were monitored using dMRM in positive and negative ion modes. Hardware and valve configuration are shown in Figure 1, mobile phases, gradient programs and valve switching events in Figure 2. Temp: Hivbax bolipie ABDC Binary Furmp Quaternary Pomp Column: Zorbax Eckpse Plus C18, tamp 45 °C Column: Poroshiall 120 HILIC-2 +. lamp 30°C Gradient program: Grodlent program: Time Flow Time Flow (rin) (rl tnuba) Salad wie (min) (mitmin) an me 0.) O 051 57.00 3.0 oo O439 O14 59.65 2.00 0.061 97.0 3.0 1.0 1s 0.15 99.08 2.01 O FA aF.0 40 161 0.438 10.00 S00 2410 O25) 65.0 15.0 2.00 049 10.00 0.00 6. 0.250 67.0 43.0 204 0.350 70.00 B0.00 B10 Oo #0 ath. o BO aH 0450 710.00 4) 00 So o350 oo 100 6.60 O50 24,50 75.50 1440 Oo 250 | Tih O F400 a-a50 640 3h 00 140 O20 57.6 3.0 10.98 0.350 6400 36.00 17.410 0750 97.0 38 Ve 0350 10.00 20.00 1? 40 Oo 0f4 oro Af Vi 40 03451 10.00 S00 17.10 0.350 10.00 3,00 17.40 539 O15 99.05 At 10 mh ieennten formate, O00 8 (ee) boat netic in Ae Ar 0 1% fv) acedic occ in 20 Ba: 6.2% formic acd In acetonttrite B: 100% acetonitrile Valve swiich evienis: Time (min) Walve awiteh Flow fron the bhiary and quetienny pongpe: combined poe ty netic bes 0.00 Initial position mass spectrometer, Runilme 22 min 2.00 Second position 17.45 Initial postion Figure 2. LC-MSMS method parameters, at initial conditions the composition of the mobile phase at the HIILIC column is 90% ACN Dual column GC-MSMS analysis was performed using an Agilent 8890/7010B GC-MSMS with PAL autosampler. An HP Ultral column (25 m x 0.32 mm x 0.17 um) was fitted to the front inlet and DB35-MS (25 m x 0.32 mm x 0.25 um) to the back. A short piece of narrow bore HP Ultral (2 m x 0.2 mm x 0.33 um) was used as a restrictor in front of each column, connected using an Agilent Ultimate Union. The two columns were connected to the mass spectrometer by a T-Piece (Trajan 3-port MCD) and deactivated fused silica (0.5 m x 0.32 mm). Sample injections alternated between front and back columns within the sequence. The hardware set-up and temperature programs are shown in Figure 3. Helium was used as carrier gas at 1.2 mL/min constant flow on the active column. To avoid excess flow to the mass spectrometer the flow was reduced on the inactive column to 0.6 psi constant pressure. Splitless injection (1 uL) was used for both columns, the inlet was set at 250°C, transfer line at 300°C and ion source at 230°C. Data acquisition used dynamic MRM mode. RECENT ADVANCES IN DOPING ANALYSIS (32) 23 ISBN 978-3-86884-080-3 24 MDI eae Lecture Falty MANFRED DONIKE WORKSHOP 2024 PFrent Column: HP Ua 1 foe eater 0 1m Peo-column- restlcion: 1 Whe), ime 2 ome Be Back jeg a Crees program: i prdene 26 ob eons G Ramp Penny Hole | {Cimin} (i fenin] | oF itera O*¢ | m6 is ted L if io a 0 i | ia 285 a Freed inet SS oo u20 a) : ty CHR Pa Back Column: OB 35-5 Soe 2 0 Sen 2 Oo 25 yn Pro-column rewiricion 1 Litre ), 2 mai Fm 2 0) pe wiih oP ra [tte Earl depiard wir a ‘(Cheee program: Purdene 24 min Ramp Terre Hokl | [Cini] ic) tein | nag oe itera 4d 184 a £ #55 a if 234 q i Pet) d | x aa i Figure 3. Dual column GC-MSMS hardware setup and method parameters Results and Discussion Dual Column LC-MSMS Chromatograms showing the concurrent elution of compounds from the HILIC and C18 columns are provided in Figure 4. Compounds separated on the HILIC column are broader while those separated on the C18 column are concentrated in the trap and are eluted as a narrow peak. The C18 separation is superior with narrow peaks and very stable retention times (RT) so it is the preferred column. The advantage of the HILIC column is the ability to separate highly polar analytes that are not well retained by C18. Urinary salts that may interfere with early eluting compounds are directed to the HILIC column where they are strongly retained. Switching the C18 trap effluent from the HILIC column to the C18 column at 2 minutes sends only the least retained compounds to the HILIC column. HILIC peak shape and resolution can be improved by the addition of a buffer such as ammonium acetate however the formation of precipitate in the presence of a high percentage of acetonitrile can lead to LC flow path blockages. Dynamic MRM was used to monitor 782 MRM transitions with generally two transitions per compound. The maximum number of concurrent MRMs was 102 with a minimum dwell time of 0.82 ms which resulted in at least 7-8 points across the sharpest peaks. To maximise dwell time MRM windows were kept as narrow as possible, typically 0.5 min on the C18 column, requiring very stable and reproducible RTs. Although desirable, the software did not allow assignment of higher dwell times to compounds with very low MRPLs. The LC-MSMS dual column procedure was validated in accordance with WADA Technical note on Analytical Method Validation [2]. A total of 363 compounds were assessed, 9 of these were eluted on the HILIC column, 340 substances met the validation criteria. Between instrument RT reproducibility was very good on the C18 column, generally

Es finden zwei Aufführungen im Musischen Forum (MuFo) statt: um 18:00 und um 20:00 Uhr. Eintritt: 4 € (Studierende), 5 € (Gäste) Das ITB-Lehrteam freut sich auf euer/Ihr Kommen!

Die Veranstaltung findet am 22. Juli in Hörsaal 1 statt. Los geht es um 19:30 Uhr. Eintritt: 5€ Karten: Der Vorverkauf findet am 09.07., 14.07. und 16.07. statt sowie Sonntag, 19.07. (während der Generalprobe) an der Mensa, jeweils von 11.00 bis 13.00 Uhr. Das ITB-Lehrteam freut sich auf euer/Ihr Kommen!