Optimizacija uvjeta detekcije MEKC-MS/MS metode za istovremeno određivanje šest lijekova u terapiji raka dojke

master's thesis

• Title (english): Optimization of MEKC-MS/MS detection conditions for the simultaneous determination of six drugs in breast cancer therapy
• Author: Petra Stjepanović
• Mentor: Miranda Sertić (mentor)
• Committee member: Miranda Sertić (predsjednik povjerenstva)
• Committee member: Maja Ortner Hadžiabdić (član povjerenstva)
• Committee member: Dubravka Vitali Čepo (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2024-06-20, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Rak dojke najčešća je zloćudna bolest kod žena. U današnje vrijeme mortalitet mu opada zbog novih terapijskih
metoda, unatoč trajnom porastu incidencije. Kako bi se osiguralo pravilno doziranje terapije koje će osigurati najbolju
učinkovitost lijeka te njegove nuspojave svesti na minimum, važno je pratiti koncentracije lijekova u plazmi. Kod
ispitivanja bioloških uzoraka, važno je da metoda bude osjetljiva kako bi mogla detektirati analite niskih koncentracija
te ih također učinkovito razdvojiti od interferencija koje su također prisutne. U ovome radu, analiti su bili šest lijekova
za rak dojke: ribociklib, abemaciklib i palbociklib kao CDK4/6 inhibitori, anastrozol i letrozol kao nesteroidni
inhibitori aromataze te fulvestrant kao kompetitivni antagonist ER. U literaturi još nije opisana uporaba MEKCMS/MS u analizi ovih lijekova.
Cilj ovoga rada bio je odrediti optimalne MS/MS uvjete kako bismo osigurali zadovoljavajuću osjetljivost metode i
kvantifikaciju ovih lijekova. Optimizirani uvjeti bili su: temperatura (75- 250 °C) i protok pomoćnog plina (3-11
L/min), protok (2-12 μL/min) i sastav pomoćne tekućine - prikladno organsko otapalo (metanol, izopropanol, etanol,
acetonitril), njegov udio (50-80%, v/v) i sadržaj mravlje kiseline (0-0,5%, v/v). Također su optimizirani napon
mlaznice (0-2000 V) i napon kapilare (2000-4000 V). Parametri plina u ESI izvoru koji su optimizirani uključuju
temperaturu (200-350 °C), protok (5-11 L/min) i tlak plina za raspršivanje (6-14 psi). Dobiveni su sljedeći rezultati:
optimalna temperatura pomoćnog plina je 250°C, a njegov protok 5 L/min. Kao najbolje organsko otapalo u sastavu
pomoćne tekućine pokazao se metanol i to u udjelu od 70%. Optimalni udio mravlje kiseline u pomoćnoj tekućini je
0,2%, a protok pomoćne tekućine 1 mL/min. Optimalni napon mlaznice je 2000 V, a napon kapilare 2500 V.
Parametri plina u ESI izvoru su: temperatura 350 °C, protok 9 L/min i tlak plina za raspršivanje 12 psi.

Abstract (english)

Breast cancer is the most common malignant disease in women. Nowadays, its mortality rate is decreasing due to new
therapeutic methods, despite a continuous increase in incidence. To ensure proper dosing of therapy that will
maximize drug efficacy and minimize side effects, it is important to monitor drug concentrations in plasma. When
analyzing biological samples, it is essential for the method to be sensitive enough to detect analytes at low
concentrations and effectively distinguish them from interfering substances that are also present. In this study, the
analytes were six breast cancer drugs: ribociclib, abemaciclib, and palbociclib as CDK4/6 inhibitors, anastrozole and
letrozole as non-steroidal aromatase inhibitors and fulvestrant as a competitive ER antagonist. The use of MEKCMS/MS in the analysis of these drugs has not yet been described in the literature.
The aim of this study was to determine the optimal MS/MS conditions to ensure satisfactory method sensibility and
quantification of these drugs. The optimized conditions were: temperature (75-250 °C) and sheath gas flow (3-11
L/min), flow rate (2-12 μL/min) and sheath liquid composition - suitable organic solvent (methanol, isopropanol,
ethanol, acetonitrile), its proportion (50-80%, v/v) and formic acid content (0-0.5%, v/v). Nozzle voltage (0-2000 V)
and capillary voltage (2000-4000 V) were also optimized. The ESI source gas parameters which were optimized are
temperature (200-350 °C), flow (5-11 L/min) and nebulizing gas pressure (6-14 psi). The following results were
obtained: the optimal sheath gas temperature is 250°C, with a flow rate of 5 L/min. Methanol at a concentration of
70% proved to be the best organic solvent in the composition of the sheath liquid. The optimal formic acid content in
the sheath liquid is 0.2%, with a sheath liquid flow rate of 1 mL/min. The optimal nozzle voltage is 2000 V, while the
capillary voltage is 2500 V. The ESI source gas parameters are: temperature 350°C, flow rate 9 L/min, and nebulizing
gas pressure 12 psi.

• Keywords: rak dojke; spektrometrija masa; micelarna elektrokinetička spektrometrija
• Keywords (english): breast cancer; mass spectrometry; micellar electrokinetic spectrometry
• Language: croatian
• URN:NBN: urn:nbn:hr:163:883689
• Study programme: Title: Pharmacy - integrated undergraduate and graduate - university study; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni magistar farmacije (sveučilišni magistar farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-09-27 10:26:57