Razvoj in validacija metode za določanje urolitina A

Kuhar, Maša (author), Kralj Cigić, Irena (mentor)

Kostanjev tanin iz vrste pravega kostanja (lat. Castanea sativa) spada v skupino taninov, ki jih imenujemo elagitanini. Sesalci lahko iz omenjenih taninov v črevesju po bakterijski poti tvorijo posebne metabolite, urolitine. Urolitini imajo odlične lastnosti, med drugim naj bi imeli antioksidativne, protivnetne in antikancerogene učinke, prehajajo lahko v kri in naprej do različnih organov, nekaj pa se jih lahko izloči skupaj z blatom in urinom. V diplomskem delu sem predstavila razvoj in validacijo metode za določanje urolitina A. Cilj naloge je bil preveriti uporabnost tekočinske kromatografije visoke ločljivosti za določitev urolitina A. Določanje urolitina A sem izvedla s tekočinsko kromatografijo visoke ločljivosti (HPLC) z uporabo UV-VIS detekcije. Separacija je bila izvedena na C18 koloni, kot mobilni fazi pa sem uporabila 100 % acetonitril in 0,1 % raztopino trifluoroocetne kisline v vodi, s pretokom 1 mL/min. Uporabljeni standard je predstavljal urolitin A, topilo zanj pa je bila 50 % raztopina metanola v vodi. Z validacijo sem preverjala linearnost metode, mejo zaznave, mejo določitve, ponovljivost in stabilnost. Ugotovila sem, da je metoda linearna v koncentracijskem območju med 0,4 mg/L in 25 mg/L. Mejo zaznave sem določila kot dvakratnik šuma, mejo določitve pa kot desetkratnik šuma. Ugotovila sem, da je metoda ponovljiva, stabilnost raztopin standarda pa je odvisna od pogojev shranjevanja.

• Language: Slovenian
• Keywords: urolitin A, validacija, HPLC
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2022
• PID: 20.500.12556/RUL-139589
• COBISS.SI-ID: 126443779
• Publication date in RUL: 05.09.2022

Secondary language

• Language: English
• Title: Development and Validation of the Method for the Determination of Urolithin A

Abstract

Chestnut tannin from the true chestnut species (lat. Castanea sativa) belongs to the group of tannins called ellagitannins. From these tannins, mammals form special metabolites, urolithins, in the intestines via bacterial pathways. Urolithins have excellent properties, including antioxidant, anti-inflammatory and anti-carcinogenic effects, they can also pass into the blood and continue to various organs, and some of them can be excreted together with feces and urine. In the thesis, the development and validation of a method for the determination of urolithin A were presented. The aim of the thesis was to verify the applicability of high-resolution liquid chromatography for the determination of urolithin A. The determination of urolithin A was performed by high-performance liquid chromatography (HPLC) using UV-VIS detection. The separation was carried out on a C18 column, and as mobile phase 100% acetonitrile and a 0.1% solution of trifluoroacetic acid in water were used, with a flow rate of 1 mL/min. The standard used was urolithin A, and its solvent was a 50% solution of methanol in water. With the validation, the linearity of the method, the limit of detection, the limit of quantitation, reproducibility and stability were checked. It was found that the method is linear in the concentration range between 0.4 mg/L and 25 mg/L. The limit of detection was set as two times multiple of the noise, and the limit of quantitation as ten times the noise. It was found that the method is reproducible, and the stability of the standard solutions depends on the storage conditions.

• Keywords: urolithin A, validation, HPLC