| 英文摘要 |
Blood sampling by the dried blood spot (DBS) technique has become commonly applied innewborn screening. It is often used for analysis of small molecules, such as metabolites.Recently, DBS sampling has been applied for quantification of post-translational proteinmodifications. Glyoxal and methylglyoxal are two simple oxoaldehydes released fromglycated proteins in the Maillard reaction. They are widely distributed in the environment(e.g. cigarette smoke) and found in foods and beverages. Glyoxal and methylglyoxal areshown to react with biomolecules including DNA and proteins. In this laboratory, wepreviously identified the sites of modification by these two oxoaldehydes in human hemoglobinand found that the extents of modification at certain sites of lysine and arginineresidues are significantly higher in type 2 diabetes mellitus patients than in nondiabeticindividuals. In this study, we examine the stability of these modifications of hemoglobinstored on DBS cards at room temperature or 4 _C in the ambient air. After hemoglobin wasextracted from the DBS cards, it was digested by trypsin and analyzed by nanoflow liquidchromatography coupled with nanospray ionization tandem mass spectrometry. The resultsshow that the extents of all these PTMs are stable within 14 and 21 days when storedon DBS at room temperature and at 4 _C, respectively. Extraction of globin from DBS cardsis mostly advantageous for hemolytic blood samples. This assay is sensitive as only aquarter of a DBS card containing ca. 12 mL of blood is required. Thus, it is practically usefulto measure the extents of glyoxal- and methylglyoxal-induced hemoglobin modificationsfrom DBS cards. |
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