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Validacija HPLC metode za določanje organskih kislin in sladkorjev v vinu
Kristjan Mesar, 2022, undergraduate thesis

Abstract: V tem diplomskem delu smo obravnavali proces validacije dveh analiznih metod za določanje kakovosti vina. In sicer smo določali organske kisline kot so vinska kislina, jabolčna kislina, mlečna kislina, citronska kislina, ocetna kislina, in monosaharide glukozo in fruktozo. Analiza teh parametrov je pomembna pri nadzoru procesa pridelave vina kot tudi spremljanju kakovosti vina. V diplomski nalogi smo pregledali glavne metode za določanje kislin in sladkorjev v grozdju in vinu, v samem poskusu pa smo uporabili kromatografsko tehniko na osnovi tekočinske kromatografijo visoke ločljivosti v kombinaciji z detekcijo v ultravijoličnem (UV) za organske kisline in z detekcijo lomnega količnika (RID) za monosaharidne sladkorje. HPLC-UV/RID sistem omogoča enostavno pripravo vzorca na analizo (samo filtracija, mogoče redčitev, če je potrebna), hitro analizo in omogoča zadovoljive meje detekcije za potrebe določevanja omenjenih parametrov. Obe metodi smo opisali z različnimi parametri, kot je ponovljivost, linearnost, izkoristek v štirih različnih matriksih (dve beli vini, eno rdeče in eno jabolčno vino – cider), meja detekcije in meja kvantifikacije. Rezultate kvantifikacije skupnih sladkorjev in organskih kislin v vzorcih vin smo primerjali z rezultati akreditiranega laboratorija kjer je bilo to mogoče in ugotovili, da kvantifikacija z metodo standardnega dodatka omogoča primerljive rezultate z akreditiranim laboratorijem, predvsem za parameter skupni fermentirani sladkorji (glukoza in fruktoza). Vsebnost skupnih kislin določenih kromatografsko je bila nekoliko manjša od parametra skupne titrabilne kisline, kar je najverjetneje posledica manjšega števila posameznih kislin, ki smo jih vključili v ta parameter, pa tudi drugačne metode določanja kislin.
Keywords: vino, cider, HPLC, UV, RID, organske kisline, sladkorji, validacija
Published in RUNG: 02.08.2022; Views: 1804; Downloads: 59
.pdf Full text (1,10 MB)

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Identification of sweet cherry anthocyanins and hydroxycinnamic acids using HPLC coupled with DAD and MS detector
Polonca Trebše, Branka Mozetič Vodopivec, 2004, original scientific article

Abstract: In absence of standards, HPLC coupled with DAD offers identification of polyphenols by scanning UV-Vis spectra of individual components, which spectral characteristics are unique, but not selective. At the same time HPLC-DAD determination methods of polyphenols differ in mobile phase solutions resulting in DAD scanned spectra deviation between different studies, aggravating the precise identification based on agreement to UV-Vis data from literature. Mass spectrometry (MS) detection with molar weight determination of the individual components in the sample enables more precise identification of compounds eluted from the column. Sweet cherry Petrovka polyphenols were separated on C18 Hypersil PEP 300 column (250 x 4.6 mm, 5 [mu]m) using gradient solvent mixture consisting of methanol, water and formic acid. MS and UV-Vis spectra of eluted anthocyanins and hydroxycinnamic acid were recorded. HPLC-MS analyses were performed using a LCQ [sup]TM ion trap, Finnigan, MAT mass spectrometer by atmospheric pressure chemical ionisation (ACPI). Molecular and fragmented ion masses of sweet cherry hydroxycinnamic acids and anthocyanins were determined and with UV-Vis spectra, in the range of 190-600 nm, used for identification of compounds. Electro spray mass spectrum of two hydroxycinnamic acids produced ions with m/z ratios of 353.0 and 337.0, which corresponded to molecular weights of neochlorogenic acid and 3'-p-coumarylquinic acid. The molecular weights of 5 anthocyanins corresponded to cyanidin-3-glucoside (449.0), cyanidin-3-rutinoside (595.1), peonidin-3-glucoside (463.1), pelargonidin-3-rutinoside (579.1) and peonidin-3-rlltinoside (609.2).
Keywords: češnje, antociani, hidroksicimetne kisline, HPLC-DAD/MS
Published in RUNG: 10.07.2015; Views: 5856; Downloads: 23
URL Link to full text

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Lipids in freshwater ecosystems
Tadej Mezek, 2009, doctoral dissertation

Keywords: sladkovodni ekosistemi, bentoški nevretenčarji, biomarkerji, lipidi, maščobne kisline, disertacije
Published in RUNG: 15.10.2013; Views: 5499; Downloads: 431
URL Link to full text
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