Determination of Sarcosine in Human Urine by Derivatization and Fluorimetric Liquid Chromatography
| Autor: | Jian-De Li, 李建德 |
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| Rok vydání: | 2012 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 100 Sarcosine, an N-methyl derivative of glycine, is reported to be a potential biomarker for prostate cancer with conflicting results. To accurately assess the diagnostic efficiency of sarcosine for prostate cancer, there is a need of a sensitive and reliable analytical method to determine sarcosine. A high performance liquid chromatography-fluorescence detection (HPLC-FLU) method was studied for the analysis of urinary sarcosine at trace levels. Pre-column derivatization of sarcosine with a novel fluorescent reagent levofloxacin acyl chloride (L-OAC) was performed, and the resulting derivative was highly responsive to a fluorescence detector (excitation at 290 nm and emission at 460 nm). The derivatized sarcosine was separated on a hexyl-phenyl column with the mobile phase of sodium acetate buffer solution (50 mM, pH 3.8) and tetrahydrofuran by gradient elution. The linear range of sarcosine in water was over 0.5-20 μM, and the limit of detection (LOD) was 0.1 μM (S/N = 3). Intra-day and inter-day precision (relative standard deviation, RSD) and accuracy (relative error, RE) were all less than 4.8%, indicating this method was precise and accurate. This simple, rapid, sensitive and eco-friendly method has been successfully applied to the determination of sarcosine in urine samples from healthy male volunteers, which proved this HPLC-FLU technique is feasible for clinical and biochemical research. Application of this method to study the correlation between sarcosine and prostate cancer is in progression. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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