Praćenje metabolizma anaboličko-androgenih steroida

master's thesis

• Title (english): Metabolism monitoring of anabolic-androgenic steroids
• Author: Dino Đurinek
• Mentor: Mirza Bojić (mentor)
• Committee member: Mirza Bojić
• Committee member: Željan Maleš
• Committee member: Renata Jurišić Grubešić
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medicinal chemistry); Zagreb
• Defense date and country: 2015-05-29, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

6β-hidroksilacija testosterona predstavlja marker reakciju enzima CYP3A4/5. Ova reakcija
korištena je kao ogledni primjer praćenja najznačajnije skupine nedozvoljenih tvari u sportu -
anaboličko-androgenih steroida.
U svrhu praćenja 6β-hidroksilacije testosterona primjenjen je vezani sustav tekućinske
kromatografije visoke djelotovornosti s detektorom s diodnim nizom. Za provedbu reakcije
neophodan je koenzim citokroma P450 odnosno NADPH, te drugi enzimi koji sudjeluju u
prijenosu elektrona kao što su NADPH reduktaza i citokrom b5. Kako su citokromi P450
vezani na membrane endoplazmatskog retikuluma i eksprimirani u jetri idealan sustav za
praćenje metaboličke aktivnosti su humani jetreni mikrosomi.
Rezultati analize prikazuju se u obliku ostatne aktivnosti u odnosu na pozitivnu kontrolu, a
kao negativna kontrola primjenjuje se marker inhibitor CYP3A4/5 na primjer ketokonazol.
HPLC spregnut s DAD predstavlja osnovni analitički sustav za praćenje anaboličkoandrogenih
steroida koji se nadograđuje sa spektrometrijom masa i nadopunjuje s plinskom
kromatografijom.

Abstract (english)

6β-hydoxylation of testosterone represents a marker reaction of CYP3A4/5 enzyme. This
reaction was used as an example for monitoring the most significant group of prohibited
substances in sports – anabolic-androgenic steroids.
High prformance liquid chromatography with diode array detector was used for monitoring
6β-hydoxylation. In order to perform the reaction, it is essential to have a coenzyme of
cytochrome P450 - NADPH, and other enzymes involved in electron transfer; NADPHreductase
and cytochrome b5. Human liver microsomes are ideal system for monitoring the
metabolic activity because cytochrom P450 enzymes are linked to membranes of
endoplasmatic reticulum and are expressed in liver.
The results of analysis are usually presented as residual activity in comparison to positive
control. Ketoconazole, as a marker inhibitor of CYP3A4/5, was used as a negative control.
High performance liquid chromatography with diode array detector represents a basic
analytical system for monitoring anabolic-androgenic steroids accompanied with mass
spectrometry and gas chromatography.

• Keywords: steroidi; CYP3A4/5; testosteron; 6β-hidroksitestosteron; biotransformacija; detekcija
• Keywords (english): steroids; CYP3A4/5; enzyme; testosterone; 6β-hydroxytestosterone; biotransformation; detection
• Language: croatian
• URN:NBN: urn:nbn:hr:163:974142
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2015-12-15 14:57:58