Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

Optimization of HPLC separation of amino acids presented in mead

Autoři: Klikarová Jitka | Šebková Jana | Česlová Lenka | Fischer Jan

Rok: 2017

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Optimization of HPLC separation of amino acids presented in 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 fluorescence detection. We have focused on two pre-column derivatization agents and those are phenyl isothiocyanate (PITC) and dansyl chloride (DNS). The separation of both types derivatives were performed by reversed-phase HPLC. In case of PITC derivatives, the final separation was performed on Zorbax SB-Aq column with gradient elution of mobile phase consisted of 70mM ammonium acetate (pH 6.55) and 60% aqueous acetonitrile. The Ascentis Express C18 column and mixture of acetonitrile with water (+1% acetic acid and 0.1 % triethylamine) was used for separation of DNS derivatives. The spectrophotometric detection at 254 nm was employed for PITC derivatives and fluorescence detection (λEX = 330 nm λEM = 530 nm) for DNS derivatives. Further, both derivatization procedures have been applied to real samples of mead from which the amino acid had to be primarily extracted by cation-exchange tubes. The main amino acid presented in 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.	phenyl isothiocyanate, dansyl chloride, HPLC, amino acids