Abstract
Three different high performance liquid chromatography columns were accessed for phenolic compounds (PC) separation in the hydrophilic fraction of extra virgin olive oil (EVOO). Two fully porous C18 bonded silica phases and one partially porous biphenyl column were used. Biphenyl column allowed for an increase of more than 30% in peak capacity (nc), higher selectivity (α) (1.045), and improved retention (k), with a reduction of 22.1% in the retention time. The higher resolution (Rs) was obtained by using the biphenyl column, with a fair separation of oleuropein aglycone isomers (OAI) and a good identification of caffeic acid (CA). Tyrosol (T), hydroxytyrosol (HT), and dihydroxyphenyl glycol (DHPG) were also well separated and identified. Moreover, the method using a biphenyl column was fully validated according to the requirements for new methods. For all parameters, the method applying the biphenyl column proved to be a reliable, accurate, and robust tool for separation, identification, and quantification of the main PCs in EVOOs.
Highlights
Extra virgin olive oil (EVOO), which represents the primary source of fat intake in the Mediterranean diet, is well known for its health benefits, such as the protection of low density lipoprotein particles from oxidative damage, the maintenance of normal blood high density lipoprotein cholesterol concentrations, the maintenance of normal blood pressure, and its anti-inflammatory properties, among many others [1]
Phenolic compounds (PC) are among these health-promoters that are present in EVOO [2,3,4,5,6], which were highlighted by the European Food Safety Authority in 2012 with a health claim for virgin olive oil PCs, which contribute to the protection of blood lipids from oxidative stress [7]
The use of C18 columns for olive oil PCs separation, identification, and quantification has been extensively reported in the literature [19,20], but little is known about the use of the biphenyl phases on the analyses of these specific EVOO compounds
Summary
Extra virgin olive oil (EVOO), which represents the primary source of fat intake in the Mediterranean diet, is well known for its health benefits, such as the protection of low density lipoprotein particles from oxidative damage, the maintenance of normal blood high density lipoprotein cholesterol concentrations, the maintenance of normal blood pressure, and its anti-inflammatory properties, among many others [1]. Withthime cpornosveecudtievfefilcoisesnescitehsa,ttahree dimespcroirbteadnacbeoovfe.fFaisntaelrlys,etphaerUaVtiosnpsechtraadofablsoothbceoemnptoauknindgs (pλrmeaxceatdence in re2c3e7ntanydea2r7s9.nHm)igahreerveflryowsimrailtaers, wreitdhuthcee UthVesrpuenctrtuimmer,epbourtteadlsion ltihteeraetfufirceifeonrcOie[s2,7d].ue to resistance of mLaisCshtArroassnpfshofererr.athlleOowncheedrwofmoaraytthotegorsaoeppvhaeirrcactoisoemnpeaarntadhtiicosonnpsoreofqbutlehenemt iiddiseennttthiiffriioceaudtgiochnomtohfpesoixuinnPcdCossr, pinbootrhtahetiEoKVniOnoeOtfeexnxotarnanpcdtorous partic(Fleigsusruer3ro),uwnhdieledobnylyafothuirnoputoorof usisxsPhCesllw(ceorerei-dsehnetlilfipedarbtiycluessi)n,gptrhoeviSdpihnegriasorrebdcuocleudmpn,awthitfhorboatnhalytes to traOvAeIlsineltuotitnhgeasttaatsiionnglaerpyepakh.aCseo,mwpohuenndcsosmucphaarsedO taondfuitlslydeproivroatuivseps aarreticclloesse,lwy aitshsoinciaatnedacwciethptable backpheraelstshubreen[e2f3it]s., and their correct identification and quantification is of high importance. Both T and HT are secoiridoid derivative compounds that are present in the olive oil polar fraction, the second one (HT) being extensively studied and well-associated with cardio-protective, antiinflammatory and platelet aggregation, and antimicrobial and anticancer activities, among others. The results revealed an improved peak capacity, retention factor, selectivity, peak asymmetry and overall resolution when using the biphenyl core-shell column, and this was the method that was fully validated according to the requirements for the new methods
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