20.500.12556/DKUM-11045 RAZVOJ IN VALIDACIJA HPLC-UV METODE ZA SOČASNO DOLOČANJE IZBRANIH HIDROKSCIMETNIH KISLIN V VZORCU VINA DEVELOPMENT AND VALIDATION OF HPLC-UV METHOD FOR DETERMINATION OF CHOSEN HYDROXY CINNAMIC ACIDS IN WINE SAMPLE Preučevali smo vsebnost p-kumarne in ferulne kisline v vzorcih vin. Metoda temelji na analizi HPLC z UV zaznavo. Uporabili smo izokratsko elucijo. Na osnovi topnosti standardov p-kumarne in ferulne kisline smo izbrali mobilno fazo meÅ¡anice vode in metanola (82:18) z 2 % ocetne kisline. Kot interni standard smo uporabili o-kumarno kislino. Kromatografske pogoje smo optimirali, da bi dobili najboljÅ¡o ločitev preiskovanih spojin. Spojine smo identificirali glede na retencijski čas in s pomočjo podatkov iz literature. Standardne raztopine smo pripravlili v koncentracijskem območju od 1 do 10 μg/ml in od 0,1 do 1 μg/ml. Rezultate smo statistično ovrednotili v skladu z zahtevami validacije (delovno območje, linearnost, ponovljivost, točnost, meja določanja LOQ, mejo zaznave LOD). Vzorce rdečega vina in belega vina smo pripravili z ekstrakcijo, s centrifugiranjem in s filtriranjem. Iz rezultatov je razvidno, da vzorec rdečega vina vsebuje viÅ¡je koncentracije preiskovanih spojin kot vzorec belega vina. Dokazali smo, da je predlagana analizna metoda za določevanje p-kumarne in ferulne kisline z uporabo HPLC primerna, natančna in točna. The content of p-coumaric acid and ferulic acid in wine samples was studied. The method is based on HPLC analysis with UV-detection. Isocratic elution was used. Based on the solubility of standard substances, namely p-coumaric acid and ferulic acid, the mixture of water and methanol (82:18) with 2 % acetic acid was chosen as the most suitable mobile phase. As an internal standard o-coumaric acid was used. Chromatographic conditions were optimized to perform best separation of examined substances. Compounds were identified according to retention times of individual substances and data from literature. Standard solutions were prepared in concentration range from 1 to 10 μg/ml and from 0,1 to 1 μg/ml. The content of p-coumaric acid and ferulic acid was evaluated from calibration curves of standard solutions. Results were statistically evaluated according to the validation requirements (working range, linearity, repeatability, accuracy, limit of quantitation LOQ, limit of detection LOD). Samples of red and white wines were prepared with extraction, centrifugation and filtration. From results it is evident that a sample of red wine contains higher concentrations of investigated compounds then a sample of white vine. We have demonstrated that the proposed analytical method using HPLC analytical procedure is suitable for determination of p-coumaric and ferulic acid in wine samples as it is precise and accurate. Fenolne spojine rdeče vino belo vino validacija hidroksicimetne kisline p-kumarna kislina ferulna kislina o-kumarna kislina tekočinska kromatografija. Phenolic compounds red wine white wine validation hidroxycinnamic acids p-coumaric acid ferulic acid o-coumaric acid liquid chromatography. true true false [M. Lovrec] Slovenski jezik Angleški jezik Diplomsko delo 2009-06-23 10:35:46 2009-12-22 09:40:41 2022-04-11 23:26:23 2022-04-17 03:27:33 2009 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 543.544.5:547.633.4:663.2(043.2) 13326358 URN:SI:UM:DK:PFOV1U9K VS_Lovrec_Mateja_2009.pdf VS_Lovrec_Mateja_2009.pdf 1 60D2B20960BDDAE406BE262DCB651B50 b3dcd67258c1111c1db3d748cfcd960bd2d0d43340700977f4e02069b35ce357 01ada1d1-7c03-11eb-bb7a-00155d0001ca 20.500.12556/dkum/18bd269d-d022-47bc-9f25-6fca10ad8594 https://dk.um.si/Dokument.php?lang=slv&id=9549 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0