Optimizacija HPLC-DAD-MS metode za analizu sterigmatocistina i 5-metoksisterigmatocistina

master's thesis

• Title (english): Optimization of HPLC-DAD-MS method for the analysis of sterigmatocystin and 5-methoxysterigmatocystin
• Author: Monika Popčević
• Mentor: Miranda Sertić (mentor)
• Committee member: Miranda Sertić (predsjednik povjerenstva)
• Committee member: Maja Šegvić Klarić (član povjerenstva)
• Committee member: Anita Hafner (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2017-07-11, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Mikotoksini kao toksični sekundarni metaboliti plijesni predstavljaju vrlo značajne kontaminante uzrokujući mikotoksikoze kod ljudi i životinja. Osim ingestijom kontaminirane hrane, vrlo značajan put unosa je i inhalacijom konidija s mikotoksinima. Kod sterigmatocistina ovaj je put izrazito važan jer se njegov glavni proizvođač plijesan Aspergillus versicolor vrlo često nalazi u prostorijama koje su oštećene vlagom. Međunarodna agencija za istraživanje raka smješta ga u skupinu 2B karcinogena. Svoj mutageni učinak ostvaruje kovalentnim vezanjem na molekulu DNA nakon metaboličke aktivacije. Njemu sličan metabolit 5-metoksisterigmatocistin također je važan jer ima slične učinke.
Zbog svoje visoke osjetljivost i selektivnosti, za metodu analize odabrana je tekućinska kromatografija visoke djelotvornosti povezana s masenom spektrometrijom. Cilj rada bio je optimizirati uvjete za istovremenu analizu sterigmatocistina i njemu sličnog metabolita 5-metoksisterigmatocistina. Tako je odabran gradijentni program eluacije s najboljim razlučivanjem i najkraćim vremenom analize. Uz pomoć masenog detektora, utvrđeni su karakteristični ioni za pojedini analit na temelju kojih se vršila njihova strukturna karakterizacija te potvrda identiteta u uzorcima. Koristeći kromatogram izoliranih iona pojedinog analita moguća je precizna kvantifikacija analita usprkos tome što u kromatogramu ukupnog toka iona njihovi pikovi nisu bili potpuno razdvojeni. Provjerom utjecaja matriksa na rezultate analize, utvrđeno je da on nije značajan. Predložena HPLC-DAD-MS metoda prikladna je za istovremenu analizu sterigmatocistina i 5-metoksisterigmatocistina.

Abstract (english)

Mycotoxins are toxic secondary metabolites of molds, which represent important contaminants because they can cause mycotoxicosis in humans and animals. They enter human body not only by ingestion of contaminated food, but also by inhalation of conidia containing mycotoxins. For sterigmatocystin this pathway is very important since its main producer, mold Aspergillus versicolor, is often found in water-damaged rooms. International Agency for Research on Cancer has categorized it as group 2B carcinogen. After metabolic activation it covalently binds the DNA molecule thus expressing its mutagenic effect. Sterigmatocystin-related metabolite 5-methoxysterigmatocystin is also important because it possesses similar properties.
Due to its high sensitivity and selectivity, the technique of choice was high performance liquid chromatography mass spectrometry. The aim of this thesis was to optimize the conditions for the simultaneous analysis of sterigmatocystin and related metabolite 5-methoxysterigmatocystin. The gradient elution program with high resolution and shortest analysis time was chosen. The characteristic ions for the analytes were determined using the MS detector and they were utilized for structural characterization and identity confirmation in samples. Using extracted ion current chromatogram of each analyte precise quantification is possible, although the peaks in total ion current chromatogram were not completely separated. The influence of the sample matrix was tested and the results showed no significant effect. The proposed HPLC-DAD-MS method is appropriate for simultaneous analysis of sterigmatocystin and 5-methoxysterigmatocystin.

• Keywords: sterigmatocistin; 5-metoksistergmatocistin; LC-MS
• Keywords (english): sterigmatocystin; 5-methoxysterigmatocystin; LC-MS
• Language: croatian
• URN:NBN: urn:nbn:hr:163:677623
• 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: 2017-11-22 13:27:02