3. FOOD ChEMISTRy & bIOTEChNOLOGy 3.1. Lectures

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Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology P99 ThE IMPACT OF FERMENTATION PROCESS ON AMINO ACID PROFILE OF GRÜNER VELTLINER whITE wINE LEnKA ŠťAVíKOVá and MIROSLAV HTKA Department of Food Chemistry and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 612 00 Brno, Czech Republic, stavikova@fch.vutbr.cz Introduction Amino acids represent 30–40 % of total wine nitrogen. It is generally accepted, that they may act as nutrients for both yeast and malolactic bacteria during the wine fermentation. In the yeasts’ growing phase, they can be either partially or totaly metabolised, excreted by living yeasts at the end of the fermentation or released by proteolysis during the autolysis of the dead yeasts. In addition, amino acids can serve as substrate for the production of aroma compounds 3–5 or biogenic amines 6,7 in wine. On the contrary, their production by enzymatic degradation of the grape proteins was described 1,2 . There is no doubt, that their content in wines is dependent on the fertilization and climatic conditions, duration of skin maceration in the must, field treatments and on other factors 2,6–8 . Recently, amino acids’ content in wines is studied extensively for several reasons, e. g, due to the differentiation of Champagnes and sparkling wines 2 , the determination of geographic origin, variety and vintage 2,7,8 , the assessment of substances with enological interest (e.g., of aroma compounds) 3–5 , or toxicological interest (biogenic amines etc.) 6,7,9,10 , the monitoring of nitrogen metabolism during the fermentation 5,11 or their changes during the ageing of wines 12,13 . The determination of amino acid in wines is usually employed by HPLC with precolumn derivatization using o-phthaldialdehyde (OPA), fluorenylmethyl-chloroformate (FMOC) or 6-aminoquinolyl-n-hydroxysuc-cinimidyl carbamate (AQC). 2,4,6–8,10–14 Ion-exchange chromatography with post-column derivatization with ninhydrin reagent 1,9,15 or comprehensive two-dimensional gas chromatography 16 was successfully applied, as well. In this contribution, the impact of spontaneous and inoculated fermentations on amino acids profile of Grüner Veltliner (Vitis vinifera) white wine is presented, applying an ion exchange chromatography with post-column derivatization with ninhydrin and spectrophotometric detection system. Experimental W i n e S a m p l e Grüner Veltliner (Veltlínské zelené) white wine variety from Velké Pavlovice sub-region (South Moravia region, Czech Republic) was investigated. Wine samples were prepared by two different technological processes. In the first one, the spontaneous fermentation was used, whereas in the second, wine was prepared by inoculated fermentation. The s800 sampling procedure was performed every other day continuously, starting from cider up to the final young wine. C h r o m a t o g r a p h i c C o n d i t i o n s Respective wine sample was analysed to free amino acids (FAA) content after its filtration and dilution in Li-citric buffer (pH 2.2), employing the AAA T339 amino acid analyser (Mikrotechna, Prague, Czech Republic) equipped with Ostion LG AnB ion-exchange resin column. Amino acids separation was achiewed by stepwise gradient elution using the Li-citric buffer system. Post-column derivatization with ninhydrin reagent and spectrophotometric measurement was used for their determination. Results Concentration profile of the following 36 compounds potentially present in wine samples under study was investigated: cysteic acid (CystAC), tautine (Tau), phosphoethanolamine (PE), urea, aspartic acid (Asp), hydroxyproline (Hpro), threonine (Thr), serine (Ser), asparagine (Asn), glutamic acid (Glu), glutamine (Gln), α-aminoadipic acid (α- AAD), proline (Pro), glycine (Gly), alanine (Ala), citrulline (Cit), α-aminobutyric acid (α-ABA), valine (Val), cysteine (Cys), methionine (Met), cystathionine (Cysne), isoleucine (Ile), leucine (Leu), tyrosine (Tyr), phenylalanine (Phe), αalanine (α-ALA), β-aminobutyric acid (β-ABA), γ-aminobutyric acid (GABA), ethanolamine (EA), ammonium (nH 3 ), ornithine (Orn), lysine (Lys), histidine (His), 1-methyl histidine (1-met His), 3-methyl histidine (3-met His), arginine (Arg). Results obtained showed, that total FAA concentration in wine samples prepared by spontaneous fermentation ranged from 302.30 mg dm –3 up to 1,578.25 mg dm –3 , whereas in those prepared by inoculated fermentation, from 289.64 mg dm –3 up to 1,371.41 mg dm –3 . In the sample prepared by a spontaneous fermentation, the total concentration of free amino acids slowly increased during the first three days from 1,299.75 g dm –3 to the maximum of 1,371.47 mg dm –3 , then in next six days of fermen- Fig. 1. Total free amino acids content monitored during 24 days in the wine samples prepared by spontaneous (�) and inoculated (�) fermentation processes
Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology Fig. 2. The content of individual amino acids in a) cider, b) semi – fermented product, c) young wine prepared by spontaneous (black fill) and inoculated (gray fill) fermentation s801
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3. FOOD ChEMISTRy & bIOTEChNOLOGy 3.1. Lectures

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