| Summary: | 碩士 === 高雄醫學大學 === 藥學系碩士班 === 105 === One capillary electrophoresis (CE) method had been developed for simultaneous determination of protein-bound uremic toxins in the serum of hemodialysis patients. The other method, chemical sensor, had also been developed for determination of iron chelating medicine in the plasma of β-thalassemia patients.
The first part of the thesis is determination of four uremic toxins in the serum of chronic kidney disease (CKD) patients, including indole-3-acetic acid (IAA), para-cresol sulfate (pCS), 3-indoxyl sulfate (3-INS), and hippuric acid (HA), by field-amplified sample stacking-sweeping-micellar electrokinetic chromatography (FASS-sweeping-MEKC). Chemometric experimental design was used to determine the optimal condition for analysis. The p value of the concentration of phosphate buffer and SDS and pH value of phosphate buffer were less than 0.05, indicating that the factors affected the separation in statistical significance. In a further discussion, the optimal condition was set at 100.00 mM NaH2PO¬4 (pH 4.0) containing 150.00 mM SDS and 7.50% MeOH. The separation could be finished within 15 minutes. The calibration curves were established by standard addition method. The limit of detection (LOD) of IAA, pCS, 3-INS and HA were 0.08, 0.28, 0.34, and 0.25 μg/mL, respectively. The relative standard deviation (RSD) was less than 14.15% and the relative error (RE) was less than 13.71%. The proposed method could be successfully applied in quantitative determination of the four uremic toxins in the serum of 100 hemodialysis patients.
The second part of the thesis is determination of iron chelating medicine in the plasma of β-thalassemia patients by fluorescent polymer dots (Pdots). The PFBT (poly(fluorene-alt-benzothiadiazole)) polymer dots functionalized with PSMA (poly(styrene-co-maleic anhydride)) polymer was synthesized by reprecipitation. The optimal condition for pretreatment of plasma samples was deproteinized with acetonitrile and salting-out with ammonium sulfate. The optimal condition for analysis is using 50.00 μg/mL copper ions to quench the fluorescence of the Pdots which diluted with 10.00 mM HEPES buffer (pH 6.0) and the fluorescence was recovered by adding the pretreated plasma samples containing deferiprone (DFP). The pretreatment time of plasma samples was only 10 minutes and the reaction time was less than 1 minutes, indicating that the proposed method could be applied in the rapidly screening plasma samples from β-thalassemia patients. The LOD was 2.50 μg/mL and the linear range was 5.00-30.00 μg/mL. The RSD and RE were lower than 12.23% and 14.82%, respectively. The specificity of the method was good. The proposed method could be successfully applied in quantitative determination of the DFP in the plasma of 2 β-thalassemia patients. The specificity test was ongoing.
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