Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

HPLC separation of derivatized amino acids in mead

Autoři: Klikarová Jitka | Česlová Lenka | Fischer Jan

Rok: 2017

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	HPLC separation of derivatized amino acids in mead	Mead is an alcoholic beverage produced by fermentation of honey solution using cultivated wine yeast. Honey is the source of many biologically active compounds, which come into mead during production. In our study, we are focused on determination of free amino acids that are minor but important ingredients in both honey and mead. The main amino acid presented in honey and mead is proline, other amino acids are contained in small amount. The concentration of proline is used for evaluation of authenticity of honey and can be used as a marker of falsification of mead. However, the final amino acid profile could provide information about geographical and botanical origin of honey and thus mead. The primary problem in determination of amino acids using HPLC is absence of chromophore or fluorophore, which is necessary for their detection. Therefore, several derivatization agents have to be used for spectrophotometric or fluorescent detection. We have focused on two pre-column derivatization agents and those are phenyl isothiocyanate (PITC) and dansyl chloride (DNS). Separation of PITC derivatives was performed by reversed-phase HPLC on Zorbax SB-Aq column with spectrophotometric detection at 254 nm and DNS derivatives on Ascentis Express C18 column with fluorometric detection (λEX = 330 nm λEM = 530 nm). Both derivatization procedures have been applied to real samples of mead from which the amino acid had to be firstly extracted by different cation-exchange tubes.	mead, HPLC, fluorescence, derivatization