Razvoj i validacija metode za kvantifikaciju biološki aktivnih tvari UHPLC tehnikom

master's thesis

• Title (english): Development and validation of UHPLC method for the quantitative determination of biologically active compounds
• Author: Ines Topalović
• Mentor: Sandra Babić (mentor)
• Committee member: Sandra Babić (predsjednik povjerenstva)
• Committee member: Irena Škorić (član povjerenstva)
• Committee member: Martina Biošić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2019-07-09, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Prirodni spojevi koji najčešće nemaju nutritivnu vrijednost, ali utječu na zdravlje, nazivaju se biološki aktivne tvari. U središtu ovoga istraživanja bili su fitoestrogeni (daidzin, daidzein, genistin, genistein, ononin, sisotrin, formononetin i biohanin A), biološki aktivne tvari sličnih strukturnih i/ili funkcionalnih svojstava kao ženski spolni hormoni estrogeni. Fitoestrogeni se najčešće ekstrahiraju iz soje i livadne djeteline, a na tržištu se mogu naći u obliku dodataka za prehranu koji se koriste za menopauzalne tegobe. Cilj ovoga rada bio je razviti i validirati metodu za kvantifikaciju ranije navedenih osam molekula fitoestrogena UHPLC tehnikom. UHPLC tehnika korištena je kako bi se postiglo kraće vrijeme trajanja analize. Za razvoj metode korišten je eksperimentalni dizajn kako bi se skratilo vrijeme razvoja, uštedile kemikalije te pronašli najbolji uvjeti za razdvajanje. Validacijom metode potvrđeno je da je dobivena metoda prihvatljiva za određivanje ranije navedenih molekula fitoestrogena jer su sve ispitivane izvedbene značajke unutar granica prihvatljivosti. Dodatno je provedena i analiza sedamnaest komercijalnih uzoraka, dodataka prehrani u kojima je određen sadržaj fitoestrogena te su dobiveni rezultati uspoređeni s deklaracijama koje navode proizvođači kako bi se ispitala njihova ispravnost.

Abstract (english)

Natural compounds that usually do not have nutritional value but somehow affect human health are called biologically active compounds. At the center of this study were phytoestrogens (daidzin, daidzein, genistin, genistein, ononin, sissotrin, formononetin, biochanin A), biologically active compounds of similar structural and/or functional properties as female sex hormones estrogens. Phytoestrogens are most often extracted from soy and meadow clover and can be found on the market in the form of dietary supplements used for menopausal symptoms. The aim of this paper was to develop and validate the UHPLC method for quantitative determination of the eight previously mentioned phytoestrogens. UHPLC technique was used to achieve a shorter duration of the analysis. The experimental design was used to shorten method development time, save chemicals, and find the best separation conditions. The validation of the method confirmed that the obtained method was acceptable for the determination of previously mentioned phytoestrogen molecules as all tested performance features were within the limits of acceptability. Additionally, an analysis of seventeen commercial samples (dietary supplements) was carried out in which the content of phytoestrogen was determined. The results were compared with the declarations cited by the manufacturers to ascertain their correctness.

• Keywords: biološki aktivne tvari; fitoestrogeni; eksperimentalni dizajn; validacija; UHPLC
• Keywords (english): biologically active compounds; phytoestrogens; experimental design; validation; UHPLC
• Language: croatian
• URN:NBN: urn:nbn:hr:149:561529
• Study programme: Title: Materials Science and Engineering - Graduate study; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka kemijskog inženjerstva (magistar/magistra inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2021-07-09
• Created on: 2022-02-27 18:24:50