|Opis:||One of objectives of this master's thesis was to simplify the pharmacopoeia analytical procedure for determination of medium-chain triglycerides used in pharmaceutical industry. The procedures in EU and US pharmacopeia’s are very complex, time-consuming and they requires a lot of laboratory inventory, which can be found in the laboratory for organic synthesis, rather than in a classical analytical laboratory. With this master's thesis we developed and validated the simplified and user-friendly analytical procedure for determination of fatty acids in samples of beechnuts and edible dormouse with derivatization procedure of sample esterification. We also performed a comparison of all three analytical procedures to prove the equivalence of the optimized analytical procedure with existing pharmacopoeia’s procedures.
In the literature we found claims where authors reported that, the fat of domestic pigs, which were fed with beechnuts, showed a significant drop in melting point similar to that of edible dormouse. At the same time, it's generally known that edible dormouse fat, with exception of other mammals, is liquid. Therefore, we assumed that the specific physical properties of the fat are caused by beechnut's oil, which is mostly the food of edible dormice. We analysed two samples: first, edible dormouse fat and second, beechnut's oil. With the comparison of the fatty acids composition between edible dormouse fat and beechnut's oil, we wanted to prove the correlation of fatty acids composition between them. In the case of a good matching in fatty acids composition, we prove the effect of beechnuts in the diet of edible dormice and therefore prove the cause for special properties of edible dormouse fat.
The first analysis was made on gas chromatograph with mass detector using the analytical procedure of European pharmacopoeia. This analysis was used for identification of fatty acids of edible dormouse fat and beechnut oil. Then we performed analysis by GC-FID with analytical procedure of American pharmacopoeia (USP29, <401> Fats and fixed oils) and then with European pharmacopoeia (PhEur 9.0 Composition of fatty acids by GC) analytical procedure. Then the analytical procedure was modified, simplified and validated. With optimization of the analytical procedure we shorten the preparation of standards and samples and achieved significantly simplified sample preparation as well. For the preparation of standards in the optimized analytical procedure, we adopted PhEur standard preparation and at the same time we changed chromatographic conditions (temperature program and gradient program for mobile phase flow was added) in order to improve resolution between peaks and shorten the time of chromatographic run and analysis in general.
In the validation process we performed tests for: stability of the standard solutions and sample, linearity, precision of the system and the method, determination of detection and quantitation limit and selectivity.
We found out that there is a sufficient similarity between the beechnut oil and the dormouse fatty acids composition, and concluded that the consumption of beechnuts causes, the special fatty acids composition of dormouse fat, which consequently causes the special physical properties of dormouse fat.|