The use of solid phase extraction for fractionation of lipids in cheese matrix

This bachelor thesis deals with Solid Phase Extraction, as a method for isolation of free fatty acids from cheese matrix to be subsequently determined using gas chromatography with FID detection. The theoretical part provides information about milk fat composition and related reactions being a source of flavour compounds, free fatty acids including. The main objective of the theoretical research is to summarize alternatives to determining fatty acids in cheese, focusing on current advancement and use of Solid Phase Extraction for this purpose. The practical part discovers proper technique and conditions for fractionation of lipids by Solid Phase Extraction, using aminopropyl bonded phase columns. Agilent SampliQ Amino columns were utilized to separate lipid mixture and different lipid classes were eluted using suitable organic solvents. Composition and purity of lipid extracts were verified combining thin layer chromatography and gas chromatography. Before being determined by gas chromatography, free fatty acids were converted into methylesters, based on ČSN EN ISO 12966-2. Besides discussing proper experimental conditions, the assesment also comes with solving optimal volumes of sample and eluting solvents to apply. The results of this thesis are considered to be useful base for subsequent experimental optimization and following integration of validated method into process of determining free fatty acids