Analysis of aspartyl peptide degradation products by high-performance liquid chromatography and high-performance liquid chromatography-mass spectrometry.
Analysis of aspartyl peptide degradation products by high-performance liquid chromatography and high-performance liquid chromatography-mass spectrometry.
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通过高效液相色谱和高效液相色谱-质谱法分析天冬氨酰肽降解产物。
DOI：
10.1016/j.chroma.2003.09.026
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
G. Scriba
中科院分区：
文献类型：
--
作者：
Silvia De Boni;C. Oberthür;M. Hamburger;G. Scriba
建立了反相高效液相色谱法分析模型肽Phe-Asp-GlyNH 2和Gly-Asp-PheNH 2在pH 2和10条件下孵育后的降解产物。大多数化合物可以用含0.1%三氟乙酸的乙腈水溶液梯度分离。在磷酸盐缓冲液（pH 5.0）中，使用乙腈梯度实现异构体对l-Phe-α-l-Asp-GlyNH 2/l-Phe-β-l-Asp-GlyNH 2和l-Phe-α-d-Asp-GlyOH/l-Phe-β-d-Asp-GlyOH的分离。在酸性条件下，主要降解途径是裂解肽骨架酰胺键，产生二肽和氨基酸，C-末端脱酰胺以及形成succidinimyl肽。在碱性pH值下，观察到C-末端酰胺的脱酰胺以及Asp的异构化和伴随的对映异构化。通过参比物质和在线电喷雾质谱法鉴别峰。结果进行了比较，以前开发的毛细管电泳方法。不能通过毛细管电泳分离的肽的非对映异构体对通过HPLC分离，而与毛细管电泳相反，通过HPLC并不总是能够实现α-和β-Asp肽的相应对的分离，这说明这两种技术在肽分析中可以互补。
A reversed-phase HPLC method for the analysis of degradation products of the model aspartyl tripeptides Phe-Asp-GlyNH2and Gly-Asp-PheNH2after incubation at pH 2 and 10 was developed. Most of the compounds could be separated with a gradient of acetonitrile in water containing 0.1% trifluoroacetic acid. Resolution of the isomeric pairs l-Phe-α-l-Asp-GlyNH2/l-Phe-β-l-Asp-GlyNH2and l-Phe-α-d-Asp-GlyOH/l-Phe-β-d-Asp-GlyOH was achieved with a gradient of acetonitrile in phosphate buffer, pH 5.0. Under acidic conditions the major degradation pathway was cleavage of the peptide backbone amide bonds yielding dipeptides and amino acids, C-terminal deamidation as well as formation of succidinimyl peptides. At alkaline pH both deamidation of the C-terminal amide as well as isomerization and concomitant enantiomerization of Asp were observed. The peaks were identified both by reference substances and by online electrospray mass spectrometry. The results were compared to a previous developed capillary electrophoresis method. Diastereomeric pairs of peptides that could not be separated by capillary electrophoresis were resolved by HPLC while the separation of corresponding pairs of α- and β-Asp peptides was not always achieved by HPLC in contrast to capillary electrophoresis illustrating that both techniques can be complimentary in peptide analysis.
DOI：
10.1016/s0021-9258(18)53661-9
发表时间：
1993-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. Roher;J. Lowenson;S. Clarke;C. Wolkow;Rong Wang;R. Cotter;I. Reardon;H. Zurcher-Neely;R. Heinrikson;M. Ball
通讯作者：
A. Roher;J. Lowenson;S. Clarke;C. Wolkow;Rong Wang;R. Cotter;I. Reardon;H. Zurcher-Neely;R. Heinrikson;M. Ball
影响因子：
2.9
作者：
JOHNSON, BA;ASWAD, DW
通讯作者：
ASWAD, DW