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Optimizacija protokola za določanje biooznačevalcev cirkadianega ritma melatonina in kortizola
ID Zevnik, Urša (Author), ID Rozman, Damjana (Mentor) More about this mentor... This link opens in a new window

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Abstract
Cirkadiani sistem usklajuje biološke procese z dnevnimi ritmičnimi spremembami v naravi. Ker so motnje cirkadianih ritmov povezane s številnimi boleznimi, je ključno, da jih lahko zanesljivo določamo. To običajno zahteva oceno dnevnega spreminjanja koncentracije glavnih cirkadianih biomarkerjev, melatonina in kortizola. Njuno koncentracijo v plazmi ali slini lahko izmerimo s tekočinsko kromatografijo, sklopljeno s tandemsko masno spektrometrijo (LC-MS/MS). V tem diplomskem delu smo sistematično optimizirali protokol za pripravo vzorcev za vnos v LC-MS/MS sistem. S poenostavljenim protokolom smo nato določili vpliv časovnega zamika na 24-urna ritma melatonina in kortizola. Pokazali smo, da se faza nihanja melatonina spremeni za čas, enak časovni razliki, nasprotno pa iz ritma kortizola zamika faze ni bilo mogoče natančno določiti.

Language:Slovenian
Keywords:cirkadiana ura, kortizol, melatonin, LC-MS/MS, časovni zamik (angl. jet lag)
Work type:Bachelor thesis/paper
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2024
PID:20.500.12556/RUL-159168 This link opens in a new window
Publication date in RUL:02.07.2024
Views:48
Downloads:5
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Secondary language

Language:English
Title:Protocol optimisation for the measurement of circadian biomarkers melatonin and cortisol
Abstract:
Circadian system coordinates biological processes with the daily rhythmic changes in nature. Since the disruption of the circadian rhythm is a likely player in the aetiology of numerous pathologies, it is crucial that we can reliably measure it. Typically, this involves assessing the daily fluctuations of key circadian biomarkers, namely melatonin and cortisol. Plasma or saliva samples are collected in regular time intervals and their concentrations can be quantified by Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). Here we optimised the LC-MS/MS sample preparation protocol, systematically altering one factor at a time. Simplified protocol was then used to investigate the effect of jet lag on the 24-hour rhythms of melatonin and cortisol. While melatonin exhibited a phase shift consistent with the time difference induced by jet lag, changes in cortisol levels did not enable precise phase shift calculations.

Keywords:circadian clock, cortisol, melatonin, LC-MS/MS, jet lag

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