Fitohemijski markeri botaničkog i geografskog porekla meda

Abstract

Predmet istraživanja ove doktorske disertacije bio je određivanje polifenolnog i šećernog profila meda i njihove veze sa botaničkim i geografskim poreklom. Postoji veliki broj eksperimentalnih metoda kombinovanih sa hemometrijskim tehnikama, već opisanih u literaturi, koje se bave utvrđivanjem autentičnosti meda, odnosno njegovog botaničkog i geografskog porekla analizom specifičnih fitohemikalija. Pronalaženje hemijskih biomarkera, odnosno fitohemikalija karakterističnih za pojedine vrste meda je predmet interesovanja značajnog broja istraživača. U literaturi su kao potencijalni biomarkeri botaničkog i geografskog porekla meda opisani niži šećeri i njihovi karakteristični odnosi, aminokiseline, isparlјive supstance, minerali, polifenoli i druge supstance. Do sada, kvantitativni sadržaj polifenola i šećera u kombinaciji sa analizom glavnih komponenata nije korišćen za procenu botaničkog i geografskog porekla meda. U cilјu identifikacije polifenolnih jedinjenja u medu korišćena je ultra-visoko efikasna tečna hromatografija sa hibridnim masenim detektorom visoke rezolucije koji kombinuje linarni trap-kvadrupol i orbitrap maseni analizator (UHPLC-LTQ OrbiTrap XL), dok je za kvantifikaciju polifenola korišćen UHPLC sa ultravioletnim detektorom sa više dioda (DAD) i masenim detektorom sa tri analizatora - trostruki kvadrupol (QQQ, triple quadrupol, UHPLC-DAD MS/MS). Profil šećera određen je visokoefikasnom anjonskom hromatografijom sa elektrohemijskom detekcijom (HPAECPAD). Tečno-masena hromatografija se pokazala kao brza i efikasna metoda za određivanje velikog broja polifenola u medu, dok je jonska hromatografija bila veoma pogodna metoda za određivanje šećernog profila meda. Odabrane su dve botaničke vrste meda: lipov med sa dva lokaliteta (Fruška Gora i Istočna Srbija) i med od žalfije (kadulјe) sa područja Hrvatske (Primorsko-goranska regija). Kvantitativni podaci (sadržaj polifenola i šećera) dobijeni analizom ovih medova upotreblјeni su za utvrđivanje veze između određenih hemijskih karakteristika meda i njegovog botaničkog, odnosno geografskog porekla...

The research topic of this doctoral thesis was determination of polyphenolic and sugar profiles of honey and their relations with its botanical and geographical origin. There are a large number of experimental methods combined with multivariate analysis techniques, already described in the literature, engaged in determining the authenticity of honey, its botanical and geographic origin by analyzing specific phytochemical constituents. Finding phytochemicals characteristic for particular species of honey is the subject of interest of a significant number of researchers. Potential biomarkers of botanical and geographical origin of honey can be sugars, amino acids, volatiles, minerals, antioxidants and others, as well as their characteristic relations. Until now, the quantitative content of polyphenols and sugars combined with principal component analysis was not used to estimate the botanical and geographical origin of honey. Identification of the polyphenolic compounds from honey was achived using ultra-high performance liquid chromatography coupled with a hybrid mass spectrometer which consists of a linear ion trap and an Orbitrap mass analyzer (UHPLC-LTQ Orbitrap XL), while the quantification of the them was done by UHPLC with DAD (doide array detection) and triple quadrupole mass spectrometry (UHPLC-DAD MS/MS) detection. Sugar profile was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Liquid chromatography-mass spectrometry technique are proved to be a quick and efficient method for the determination of a large number of polyphenols in honey, while anion exchange chromatography was very suitable method for the determination of the sugars in honey. In order to isolate polyphenols from honey samples solid phase exraction (SPE) was used. Analysis of individual sugar was performed from aqueous solution of honey sample. For this invastigation, two botanical varieties of honey were selected: linden honey from two locations (Fruška Gora and Eastern Serbia) and sage honey from Croatia (Primorsko-goranska region). Quantitative data (polyphenols and sugars) obtained by the analysis of these honeys were used to establish the relationship between certain chemical properties of honey and its botanical or geographical origin...