Abstract
The objective of this work was the evaluation of different conditions for the selected free amino acids extraction from cheese samples, followed by capillary isotachophoresis analysis. Parameters of the extraction: concentration and type of extraction reagents (solvents: methanol, ethanol and mineral acids: perchloric acid, trichloroacetic acid, sulphuric acid and hydrochloric acid), time and temperature, and isotachophoretic separation conditions (voltage, time) were studied. Repeatability, reproducibility and stability of this method at different conditions are discussed. The best yield of free amino acids was obtained after triple extraction (30min each process) with 0.1M hydrochloric acid at 50°C. Histidine, lysine, arginine, ornithine, tyrosine, and phenylalanine were determined using calibration curves and the standard additions method in different types of cheeses. The highest level of amino acids was determined in semi-hard cheese (2626.3mg100g−1 – calibration curve, and 2679.2mg100g−1 – standard additions method), whereas the lowest were found in sheep cheese (426.3mg100g−1 and 455.7mg100g−1, respectively), and Gouda type cheese samples (441.5mg100g−1 and 472.8mg100g−1, respectively).
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