Razvoj in validacija HPLC metode za določanje resveratrola

Vukadinović, Nataša (author), Zupančič, Tatjana (mentor)

Resveratrol je eden od manj znanih, vendar zelo močnih antioksidantov. Poleg tega ima še protivnetne lastnosti ter deluje estrogeno, antiagregacijsko in antikoagulativno. Najdemo ga v različnih rastlinah, največ v grozdju in jagodičevju. S procesom fermentacije in maceracije se ga nekaj ekstrahira iz grozdja v vino. V tem diplomskem delu sem uporabila metodo tekočinske kromatografije visoke ločljivosti. Prednost te tehnike je visoka ločljivost, občutljivost, ponovljivost, možnost avtomatizacije in široka možnost uporabe. Preizkusila sem dve HPLC analizni metodi za določitev vsebnosti resveratrola. Obe sem preizkušala na istem HPLC sistemu ter uporabila isto kolono, Gemini NX 150x4,6 mm - 5µm delci z ustrezno predkolono. Ker pri nobeni metodi nisem dobila ustrezne ločbe resveratrola od matriksa v vzorcu vina, sem modificirala HPLC analizno metodo. Spreminjala sem delež metanola v topilu, delež acetonitrila v mobilni fazi ter temperaturo kolone. Kolono je bilo treba zaradi obrabljenosti zamenjati z novejšo. Ko je bila dosežena ustrezna ločba resveratrola, sem metodo validirala po kriterijih validacije. Z validirano metodo sem določila resveratrol v različnih vzorcih slovenskih vin, v vzorcu prehranskega dopolnila in v soku aronije. Med rdečimi vini je imel najvišjo vsebnost resveratrola teran, nato cviček ter najnižjo rumeni muškat. V soku aronije resveratrola ni bilo zaznati.

• Language: Slovenian
• Keywords: resveratrol, vino, HPLC, validacija
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2019
• PID: 20.500.12556/RUL-109670
• COBISS.SI-ID: 1538326467
• Publication date in RUL: 06.09.2019

Secondary language

• Language: English
• Title: Development and validation of HPLC method for determination of resveratrol

Abstract

Resveratrol is a very strong antioxidant but not very known. In addition, it has estrogenic, antiaggregation and anticoagulant properties. It is found in various plants, mostly in berries. Because of the fermentation and maceration processes some of it can be found in wine. In this diploma thesis the high performance liquid chromatography method was used. The advantage of this technique is high resolution, sensitivity, repeatability, possibility of automation and wide range of use. Two HPLC methods were tested for the resveratrol determination. Methods were tested on same HPLC system, same column, Gemini NX 150x4.6 mm - 5µm particles with the appropriate precolumn. Proper separation of resveratrol form matrix in samples of wine, was not obtained by any method, so the HPLC method was modified. The proportion of methanol in solvent, proportion of acetonitrile in mobile phase and temperature of column were changed. Due to triteness, the column had to be replaced with a new one. When an appropriate resolution of resveratrol from matrix was reached, the method was validated according to the validation criteria. After the validation, resveratrol was determined in various samples of Slovenian wines, in a dietary supplement and in aronia juice. The highest content of resveratrol in red wines was determined in teran, followed by cviček and the lowest in yellow muscat. Resveratrol was not detected in aronia juice.

• Keywords: resveratrol, wine, HPLC, validation