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Voltammetric Determination of 7-Dehydrocholesterol as a Biomarker of Smith-Lemli-Opitz Syndrom
Zárybnická, Adéla ; Schwarzová, Karolina (advisor) ; Nesměrák, Karel (referee)
The aim of this thesis was the development of a voltammetric method for the determination of 7-dehydrocholesterol as a biomarker of congenital disease Smith-Lemli-Opitz syndrome. The concentration of 7-dehydrocholesterol in plasma will increase hundred-fold to hundreds to thousands µmol L-1 in patients suffering from Smith-Lemli-Opitz syndrome. The determination of 7-dehydrocholesterol was carried out in artifical serum in this work, which was prepared to mimic the conditions of postnatal diagnosis of Smith-Lemli-Opitz syndrome. 7-dehydrocholestrol provides a well developed voltammetric signal at approximately +0.8 V vs. Ag/AgNO3 in acetonitrile. Differential pulse voltammetry with optimized parameters was used for the determination of 7-dehydrocholesterol. The presence of protein in artificial serum (human serum albumin) resulted in an unacceptably high detection limit of the method (178 µmol L-1 in artificial serum). Therefore, human serum albumin was eliminated from the artificial serum samples by precipitating it with acetonitrile and subsequently centrifuging the resulting suspension. At the same time, the ratio of aqueous and organic components in the studied medium was also adjusted with acetonitrile to achieve an optimal voltammetric response of 7-dehydrocholestrol. With regard to the...

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