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Novel direct dried blood spot-based approaches for forensic toxicology and clinical applications Nele SADONES 2017 Promoters: Prof. Dr. Christophe Stove & Prof. Dr. Willy Lambert Thesis submitted to obtain the degree of Doctor in Pharmaceutical Sciences II Copyright The author and the promoters give authorization to consult and copy parts of this thesis for personal use only. Any other use is limited by the laws of copyright, especially concerning the obligation to refer to the source whenever results are cited from this thesis. De auteur en promotoren geven de toelating dit proefschrift voor consultatie beschikbaar te stellen en delen ervan te kopiëren voor persoonlijk gebruik. Elk ander gebruik valt onder de beperkingen van het auteursrecht, in het bijzonder met betrekking tot de verplichting uitdrukkelijk de bron te vermelden bij het aanhalen van resultaten uit dit proefschrift. Ghent, 2017, The promoters, Prof. Dr. Christophe Stove Prof. Dr. Willy Lambert The author, Nele Sadones III IV Dankwoord Een woord van dank aan iedereen die rechtstreeks of onrechtstreeks heeft bijgedragen tot dit werk. Vooreerst wil ik mijn promotoren prof. Christophe Stove en prof. Willy Lambert bedanken. Zonder hen kon ik uiteraard nooit starten aan een doctoraat. Dank u wel om mij deze kans te geven! Jullie constructieve feedback heeft in belangrijke mate bijgedragen tot het realiseren van dit werk. Bedankt prof. Stove om het vele schrijfwerk telkens opnieuw kritisch te verbeteren. Gezien je overvolle agenda heb je hiervoor heel wat uren van je slaap opgeofferd. Prof. Lambert, bedankt voor de steun de voorbije jaren. Zowel op professioneel als op menselijk vlak ben je een voorbeeld. Bedankt aan alle collega’s waarmee ik heb samengewerkt. Dit werk is ook een deel van jullie. Jullie hebben er allemaal op een of andere manier voor gezorgd dat dit tot stand kwam. Ik kon steeds bij jullie terecht. In het bijzonder wil ik hierbij Jolien en Sofie bedanken om me te steunen! Jullie begrepen me met weinig woorden. Uitermate belangrijk aangezien ik niet zo’n grote spraakwaterval ben ;-) Ann-Sofie, bedankt om me in te leiden in de wereld van de GC. Filip, bedankt om de gasflessen te vervangen. Ook bij heel wat andere praktische problemen was een ingenieur soms iets handiger dan een apotheker ;-) Ook wil ik de thesisstudenten bedanken voor de vele analyses die ze uitgevoerd hebben en Annelies, sorry als ik jouw strakke planning moest doorbreken voor het uitvoeren van heel wat PCR-reacties. Iedereen die betrokken was bij de verschillende studies wil ik natuurlijk ook bedanken. Ann-Sofie, Michelle Wood, Dr. Archer, Dr. Dargan, Dr. Wood en prof. Neels wil ik bedanken voor hun bijdrage aan de GHB studie. Eveneens een woord van dank aan prof. Van Bortel en Elien voor de vlotte samenwerking en het opzetten van de gabapentine studie. Pieter en Sara, dank u wel, om mee te helpen bij het verzamelen van stalen. Prof. Delanghe wens ik te bedanken voor het delen van zijn haptoglobine kennis. Het 24 uurs-labo van UZ Gent verdient ook een speciaal woord van dank voor de vele bloedafnames en het uitvoeren van de hematocriet bepalingen. Jolien verdient natuurlijk nog een extra woord van dank. Zij was immers het creatieve brein die de kaft heeft ontworpen. Dikke merci Jolien! Zonder jou zou het een blanco kaft geweest zijn ;-) Mama, papa en Ben, 1000x bedankt! Geen woorden om te beschrijven hoe dankbaar ik jullie ben. V VI Table of contents List of abbreviations .......................................................................................................................................... XI Aim and outline of the thesis ............................................................................................................................. 1 Chapter 1: Analysis of abused substances using dried blood spots .................................................................. 11 1.1 Introduction ............................................................................................................................................ 13 1.2 Paper used and volume spotted .............................................................................................................. 16 1.3 Analytical techniques .............................................................................................................................. 17 1.4 Extraction................................................................................................................................................ 17 1.5 Lower limit of quantification (LLOQ)........................................................................................................ 19 1.6 Validation................................................................................................................................................ 20 1.7 Comparison with other biological matrices ............................................................................................. 21 1.8 Stability ................................................................................................................................................... 23 1.9 Applications ............................................................................................................................................ 23 1.9.1 Driving under the influence of drugs ................................................................................................ 23 1.9.2 Cannabinoids ................................................................................................................................... 24 1.9.3 Cocaine and metabolites ................................................................................................................. 25 1.9.4 Opiates ............................................................................................................................................ 27 1.9.5 Non-opiate opioids .......................................................................................................................... 29 1.9.6 Benzodiazepines and Z-drugs ........................................................................................................... 30 1.9.7 Amphetamine and analogues (methamphetamine, MDMA, MDA and MDEA) ................................. 33 1.9.8 Gamma-hydroxybutyric acid ............................................................................................................ 33 1.9.9 Ketamine and norketamine ............................................................................................................. 34 1.9.10 New psychoactive substances ........................................................................................................ 35 1.9.11 Alcohol markers ............................................................................................................................. 35 1.9.12 Cotinine ......................................................................................................................................... 37 1.10 Conclusion ............................................................................................................................................ 38 1.11 References ............................................................................................................................................ 40 Chapter 2: Correlation of GHB concentrations between vDBS and cDBS ......................................................... 49 2.1 Introduction ............................................................................................................................................ 51 2.2 Materials and methods ........................................................................................................................... 51 2.2.1 Samples ........................................................................................................................................... 51 2.2.2 Analytical procedures ...................................................................................................................... 52 2.2.3 Storage experiment ......................................................................................................................... 52 2.2.4 Data analysis .................................................................................................................................... 52 2.3 Results and discussion ............................................................................................................................. 53 2.4 Conclusion .............................................................................................................................................. 58 2.5 References .............................................................................................................................................. 59 Chapter 3: Microwave-assisted on-spot derivatization of polar low molecular weight compounds................ 61 3.1 Introduction ............................................................................................................................................ 63 3.2 Materials and methods ........................................................................................................................... 65 3.2.1 Chemicals and reagents ................................................................................................................... 65 3.2.2 Preparation of calibrators and quality control samples ...................................................................
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