A rapid and sensitive detection of D-Aspartic acid in Crystallin by chiral derivatized liquid chromatography mass spectrometry
A rapid and sensitive detection of D-Aspartic acid in Crystallin by chiral derivatized liquid chromatography mass spectrometry
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DOI：
10.1016/j.chroma.2016.07.021
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发表时间：
2016-10-07
影响因子：
4.1
通讯作者：
Toyo'oka, Toshimasa
中科院分区：
文献类型：
--
作者：
Mizuno, Hajime;Miyazaki, Yasuto;Toyo'oka, Toshimasa
建立了测定多肽和蛋白质中D-天冬氨酸（D-Asp）及其D/L比值的方法。该方法通过手性衍生化和ADME柱分离相结合，实现了D/L手性肽对的良好分离。此外，一种阳离子衍生试剂，DBD-Py-NCS，增加了ESI-MS/MS检测的灵敏度。为了确认全面的肽分析，分析了合成的α-晶体蛋白胰蛋白酶肽，其中包括D-Asp残基。包括肽非对映体的5对D/L-Asp得到了很好的分离。峰分离度均大于1.5，结果重复性好（RSD
A method for the determination of D-Aspartic acid (D-Asp) and its D/L ratio in peptides and proteins has been developed. This method was carried out with good separation of the D/L chiral peptide pairs by combination of a chiral derivatization and an ADME column separation. Furthermore, a cationic derivatization reagent, DBD-Py-NCS, increased the sensitivity of the ESI-MS/MS detection. To confirm the comprehensive peptide analysis, synthesized alpha-Crystallin tryptic peptides, which included D-Asp residues, were analyzed. The 5 pairs of D/L-Asp that included peptide diastereomers were well separated. Their peak resolutions were more than 1.5 and the results were reproducible (RSD