PhD: Optimization of analytical methods for control of biogenic amines and their determination in Macedonian wines

Krste Tashev
Institute of Chemistry, Faculty of Natural Sciences and Mathematics, Ss. Cyril and Methodius University, Skopje, Macedonia
December, 2016


Biogenic amines are organic nitrogen compounds with low molecular weight whose presence plays a vital role in oenology. They can be found in the grape must, but they can also be formed during alcoholic and malolactic fermentation, as well as during wine aging. Their presence at high levels in wine can lead to health problems. The most important biogenic amines in wine are tryptamine, putrescine, histamine, phenylethylamine, tyramine, cadaverine, spermine, and spermidine.
Two analytical methods for determination of the main biogenic amines in red and white wines have been optimized and validated. One method is based on pre-column derivatization with dansyl chloride followed by reversed phase-high performance liquid chromatography with diode array detection (RP-HPLC/DAD) and the other one uses direct determination with ultra-performance liquid chromatography (UPLC) coupled with triple quadruple mass spectrometry (TQ/MS). Both methods demonstrated excellent linearity, recovery, repeatability and reproducibility and were applied for analysis of commercial white and red wines from Macedonian origin. The content of the individual biogenic amines was below the maximal allowed concentration in all tested wines.
Also a RP-HPLC method for quantitative determination of tartaric, malic, shikimic, lactic, citric and succinic acids in wines has been developed and validated after sample clean up with solid-phase extraction (SPE). Validation parameters such as linearity, limit of detection, limit of quantification, precision and recovery after clean-up of the wine samples through C18 SPE columns, were found satisfactory. The validated method was applied for analysis of white and red wines from Macedonian origin.
Experimental wines from the grape variety Vranec, have been produced, applying six different types of commercial malolactic bacteria from different producers. Biogenic amines and organic acids in these experimental wines were analyzed with the developed HPLC/DAD and UPLC-TQ/MS method. Sensory evaluation of the wine has also been performed in order to evaluate the impact of the different strains of malolactic bacteria on the wine aroma profile. The results have demonstrated positive impact of the controlled malolactic fermentation with selected bacteria on the content of biogenic amines during the fermentation process and aging of wine as well as on the sensory characteristics of the produced wines.

Keywords: wine; biogenic amines; organic acids; derivatization; HPLC/DAD; UPLC-TQ/MS