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摘要
We have reported previously a method for the detection of sugars via in-situ derivatization with phenylboronic acid PhB(OH)(2) using reactive desorption electrospray ionization (DESI, Chen et al, Chem. Commun. (2006) 597-599). The present study describes an improved method that employs modified phenylboronic acids including 3-nitrophenylboronic acid and N-methyl-4-pyridineboronic acid iodide. In contrast to using PhB(OH)(2), enhanced sensitivity of using 3-nitrophenylboronic acid was observed due to the stabilization of the resulting boronate ester anion by the electron-withdrawing nitro group and the limit of detections (LODs) for glucose in water using 3-nitrophenylbornic acid and phenylboronic acid were determined to be 0.11 mM and 0.40mM, respectively. In the case of N-methyl-4-pyridineboronic acid iodide, the corresponding LOD is 6.9 mu M and the higher sensitivity obtained is attributed to the efficient ionization of both the reactive DESI reagent and reaction product since the precursor acid with a quaternary ammonium group is pre-charged. In this case, additional important features are found: (i) unlike using phenylboronic acid or 3-nitrophenylbornic acid, the experiment, performed in the positive ion mode, is applicable to neutral and acidic saccharide solutions, facilitating the analysis of biological fluids without the need to adjust pH: (ii) simply by changing the spray solvent from water to acetonitrile, the method can be used for direct glucose analyses of both urine and serum samples via online desalting, due to the low solubility of salts of these biofluids in the sprayed organic solvent; (iii) in comparison with other sugar derivatizing reagents such as the Girard's reagent T, the N-methyl-4-pyridineboronic acid iodide shows higher reactivity in the reactive DESI; and (iv) the ions of saccharide DESI reaction products undergo extensive ring or glycosidic bond cleavage upon CID, a feature that might be useful in the structure elucidation of saccharides. In addition, a variant sample introduction protocol using pipette tips for saccharide solutions was also demonstrated in the reactive DESI experiments, allowing the analysis of multiple samples of a small volume (e.g., 200 nL) in a short period of time (e.g., five samples in 4 min). The method reported in this study with improved sensitivity and high selectivity along with online desalting and high throughput capability could find useful applications in saccharide analysis in complicated biological samples.
- 出版日期2010-1-15
