This work deals with development and optimization of a method for separation and detection of pro- tamines using capillary zone electrophoresis. The developed method uses a fused silica capillary with inner diameter of 50 µm and effective length of 41,5 cm. Driving voltage is 30 kV. Background electrolyte is aque- ous solution of 45 mmol dm−3 phosphoric acid. Analyte is detected spectrophotometrically at wavelength of 200 nm. Sample is injected hydrodynamically. The method allows for determination of protamines in the concentration range between 11 µg ml−1 to 1000 µg ml−1 , limit of detection is 4 µg ml−1 . Viablity of the method has been verified with a real sample of NPH insulin injection. For sufficiently sensitive detection of protamines in NPH insuline it is necessary to prepare the sample in acidic environment. For the pur- poses of this work the sample was acidified by addition of background electrolyte into the sample so that the background electrolyte concentration in the resulting solution of the sample was 18 mmol dm−3 . The main advantage of the method is the rapid analysis, migration time of the analyte is about 2 min. Disadvantage of the method in comparison to alternate methods using CZE or RP-HPLC is the inability to separate individ- ual protamine peptides. This disadvantage...
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:351753 |
| Date | January 2015 |
| Creators | Malý, Michal |
| Contributors | Křížek, Tomáš, Kubíčková, Anna |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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