diplomsko delo
Lara Vraničar (Author), Mitja Kolar (Mentor)

Abstract

V diplomski nalogi sem predstavila razvoj in validacijo metode za določanje resorcinola. V Taninu Sevnica iz lesa pravega kostanja pridobivajo kostanjev ekstrakt, ki se uporablja kot naravno sredstvo za preprečevanje diareje pri živalih. Kostanjev ekstrakt je bogat z elagitanini, iz katerih sesalci v debelem črevesju razvijajo urolitine. Urolitini imajo zelo ugodne učinke na telo, saj podaljšujejo življenjsko dobo, preprečujejo raka, povečajo mišično moč, preprečujejo srčne motnje in še veliko drugega. Kot surovina za sintezo urolitina A se uporablja resorcinol. Resorcinol je bela kristalinična snov, po strukturi je 1,3-benzodiol in je uporaben za veliko različnih namenov. Razvila sem metodo za določitev resorcinola s tekočinsko kromatografijo visoke ločljivosti. Standard, ki sem ga uporabila, je bil resorcinol, ki sem ga raztopila v 60 % destilirane vode in 40 % metanola. Uporabila sem kolono Kinetex 2,6 qm C18 100A 100 x 4,6 mm, mobilna faza je sestavljena iz 60 % vode z 0,1 % $H_3PO_4$ in 40 % metanola z 0,1 % $H_3PO_4$, ki ima pretok 0,5 mL/min in izokratsko elucijo. Uporabljena valovna dolžina je bila 280 nm. Metoda poteka 10 min, retencijski čas resorcinola sem določila pri 2,7 min. Metodo sem tudi preizkusila na vzorcu resorcinola in filtratu sinteze urolitina A, ki vsebuje nezreagiran resorcinol. Ko sem metodo razvila, sem validirala. Pri validaciji sem preverjala stabilnost in ugotovila, da je resorcinol stabilen vsaj 15 dni na svetlobi, temi ali v hladilniku. Določila sem, da je LOD pri koncentraciji 0,05 μg/mL, LOQ pa pri koncentraciji 0,2 μg/mL. Preverjala sem tudi ponovljivost metode in ponovljivost injiciranja terugotovila, da je ponovljivost dobra. Določila sem še linearno območje, ki je od 0,2186 μg/mL do 4,9183 μg/mL in ima $R^2$ enak 0,9999.

Keywords

kostanjev les;urolitin A;resorcinol;HPLC;validacija metode;diplomske naloge;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [L. Vraničar]
UDC: 543.544.5HPLC(043.2)
COBISS: 173447171 Link will open in a new window
Views: 64
Downloads: 8
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Other data

Secondary language: English
Secondary title: Development and validation of the method for the determination of resorcinol
Secondary abstract: In this thesis, I present the development and validation of a method for determining resorcinol. Tanin Sevnica extracts chestnut extract from chestnut wood, which is used as a natural remedy for preventing diarrhea in animals. Chestnut extract is rich in ellagitannins, which mammals convert into urolithins in the large intestine. Urolithins have numerous beneficial effects on the body, including extending lifespan, preventing cancer, increasing muscle strength, preventing heart disorders, and many others. Resorcinol is used as a raw material for the synthesis of urolithin A. Resorcinol is a white crystalline substance, structurally known as 1,3-benzenediol, and is used for various purposes. I developed a method for resorcinol determination using high-performance liquid chromatography (HPLC). The standard used was resorcinol dissolved in a mixture of 60% distilled water and 40% methanol. I used a Kinetex 2.6 μm C18 100A 100x4.6 mm column with a mobile phase composed of 60% water with 0,1% $H_3PO_4$ and 40% methanol with 0,1% $H_3PO_4$, flowing at a rate of 0.5 mL/min with an isocratic elution. The detection wavelength was set at 280 nm. The method had a total runtime of 10 min, with a retention time of 2.7 min for resorcinol. I also tested the method on a sample of resorcinol and the urolithin A synthesis filtrate, which contains unreacted resorcinol. When I developed the method, I performed validation. Stability testing showed that resorcinol remained stable for at least 15 days under light, dark, and refrigeration conditions. The limit of detection (LOD) was determined to be 0.05 μg/mL, and the limit of quantification (LOQ) was found to be 0.2 μg/mL. I also assessed method repeatability and injection repeatability, both of which demonstrated good results. Additionally, I established the linear range, which is from 0,2186 μg/mL to 4,9183 μg/mL and has an $R^2$ value of 0.9999.
Secondary keywords: resorcinol;urolithin A;HPLC;validation;Tekočinska kromatografija;Univerzitetna in visokošolska dela;
Type (COBISS): Bachelor thesis/paper
Study programme: 1000374
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, VSŠ Kemijska tehnologija
Pages: 33 str.
ID: 20034103