利用新的化學方法增進醣類分子在質譜儀中的偵測效率

張瑜玲

doi:10.6342/NTU.2009.03343

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利用新的化學方法增進醣類分子在質譜儀中的偵測效率

Novel chemical methods to improve detection efficiency of saccharides in mass spectrometry

張瑜玲 , Masters　　Advisor：陳仲瑄　　

英文 DOI： 10.6342/NTU.2009.03343

基質輔助雷射脫附游離質譜儀；電噴灑游離質譜儀；離子液體；醣類偵測極限；醣類的修飾； MALDI-MS ； ESI-MS ； ionic liquid (IL) ； detection limit of oligosaccharides ； glycopeptides ； 2-amino-1-naphthalensulfonic acid ； 7-amino-1,3-naphthalendisulfonic acid

Abstract │ Reference (51) │ Times Cited (1) │ Altmetrics

Abstract 〈TOP〉

在質譜儀的應用上，基質輔助雷射脫附游離法及電噴灑游離法是現階段常用來研究蛋白質或核酸等生物分子的分析工具。此外，由於生物科技的快速發展，醣類分子在生物體內的重要性也逐漸受到重視。研究發現，細胞與細胞間的辨識作用，蛋白質功能的正確與否，以及許多與代謝相關的疾病，都受其表面修飾之醣分子所影響。發展快速且正確分析醣類分子的方法越見重要性。但由於醣類本身質子吸附能力較蛋白質或核酸等生物分子來的低，導致醣類在質譜儀中離子化效率較差，增加偵測醣類分子的困難性。

為了增進醣類分子在質譜儀中的偵測效率，透過還原胺化的方式，於寡醣分子Maltoheptose (DP7)上修飾胜肽片段Bradykinin，利用質子吸附能力較高的胜肽片段提高醣類的游離效率。發現在基質輔助雷射脫附游離質譜儀中可增強訊號強度，同時偵測極限可由原先 54.6±14.9 fmol降低至 1.2±0.6 fmol；而在電噴灑質譜儀中則是可增強訊號強度最多8倍。

另一方面，也是利用還原胺化的方式，在寡醣分子DP7上修飾2-amino-1-naphthalensulfonic acid (ANS)或7-amino-1,3-naphthalendisulfonic acid (A13S)。在基質輔助雷射脫附游離質譜儀之負電模式下，藉由ANS / A13S易失去氫離子而帶負電的特性，使原先在負電下無法偵測之醣類分子，偵測極限可達 3.9±1.6 fmol / 2.7±0.9 fmol，此實驗數據優於前人所發表之60fmol；而在電噴灑質譜儀中，DP7-ANS與DP7-A13S之訊號強度最多可為DP7之9倍與13倍。

此外，研究發現，以離子液體DHBB (2,5-dihydroxybenzoic acid butylamine)為基質輔助雷射脫附游離法之基質使用，可偵測大的醣類分子。

將此觀念延伸至電噴灑游離質譜儀中，使用DHBB或CHCA-B (α-cyano-4-hydroxycinnamic acids butylamine)兩種離子液體進行分析多醣分子Pullulans (PL6K / 10K)之實驗。發現加入離子液體後，可改善醣類分子之訊雜比並增強訊號強度。分析圖譜可知，在正電模式下偵測時，正電荷由離子液體中之[butylamine+H]+所提供；同理在負電模式下，帶負電的[DHB-H]-或[CHCA-H]-也會吸附並增加醣類分子之游離效率；且電荷之來源完全由離子液體所提供，簡化醣類分子因鈉離子或鉀離子吸附而帶電的可能性，幫助回推多醣分子之分子量。

本實驗透過兩種不同的策略：針對醣類分子修飾胜肽片段或是特定基團，及將離子液體應用於電噴灑質譜儀中，皆可增加醣類在質譜儀的偵測效率，降低偵測極限或增強訊號強度，改善訊雜比。

Parallel Abstract 〈TOP〉

Matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) were successfully developed as the soft ionization techniques for biomolecules, such as proteins and nucleotides. The benefit of MALDI-mass spectrometry (MS) for analysis of polymers is predominantly from the formation of singly charged molecular ions that lead to simple mass spectra for polymer distribution analysis. In view of the increasing biological importance of saccharides associated with cell-cell recognition, protein targeting and metabolic diseases, oligosaccharides and their glycoconjugate research attracted more attention lately. But the ionization efficiency of saccharides is low compared with other biomolecules due to the low proton affinity of carbohydrates. In order to detect oligosaccharides, we utilized the reductive amination processes to produce glycopeptides (DP7-BK) and derivatives of oligosaccharides with 2-amino-1-naphthalensulfonic acid (DP7-ANS) or 7-amino-1, 3-naphthalendisulfonic acid (DP7-A13S). In our study, modifications were found to improve the detection limit of MALDI mass spectrometry.

In MALDI-MS, the detection limit of DP7-BK 1~9 is 1.2±0.6 fmol which is better than 54.6±14.9 fmol of DP7. The detection limits of DP7-ANS and DP7-A13S are 3.9±1.6 fmol and 2.7±0.9 fmol in negative ion mode. In ESI-MS, the signal intensity of DP7-BK 1~9 is 8 times higher than that of DP7 and the signal intensities of DP7-ANS and DP7-A13S in negative ion mode are 9 times and 13 times higher than that of DP7, respectively.

Ionic liquid (IL) has been used as the matrix to increase ionization efficiency of polysaccharides in MALDI-MS. In this work, we applied IL in ESI-MS. The ionic liquid used are 2, 5-dihydroxybenzoic acid butylamine (DHBB) and α-cyano-4-hydroxycinnamic acids butylamine (CHCA-B). The signal to noise ratios (S / N) of polysaccharides PL6K / 10K are better in positive ion mode. Ion attachments from ionic liquid to polysaccharides are observed. This is the first time that addition of ionic compounds allows us to predict ionic adducts in ESI-MS.

Reference ( 51 ) 〈TOP〉

(3) Pfeffer, S. R.; Rothman, J. E. Ann. Rev. Biochem. 1987, 56, 829-852.
連結：
(4) David J, H. Rapid Commun. Mass Spectrom. 1993, 7, 614-619.
連結：
(10) Hoffmann, E. d.; Stroobant, V. Mass Spectrometry : principles and applications (2nd), 2001.
連結：
(12) Karas, M.; Hillenkamp, F. Anal. Chem. 1988, 60, 2299-2301.
連結：
(13) Paul, G.; University of Bristol 2004.
連結：

Times Cited (1) 〈TOP〉

王雪華(2011)。以水溶液電漿分解纖維素之製程研究。臺灣大學化學工程學研究所學位論文。2011。1-157。

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