Improvement in isolation and identification of food-derived peptides in human plasma based on precolumn derivatization of peptides with phenyl isothiocyanate
Improvement in isolation and identification of food-derived peptides in human plasma based on precolumn derivatization of peptides with phenyl isothiocyanate
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DOI：
10.1021/jf060531s
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发表时间：
2006-07-26
影响因子：
6.1
通讯作者：
Sato, Kenji
中科院分区：
文献类型：
--
作者：
Aito-Inoue, Misako;Ohtsuki, Kozo;Sato, Kenji
为了分离和检测血液中的食物源性肽，建立了一种基于异硫氰酸苯酯（PITC）衍生化肽的方法。这种方法允许通过反相（RP）HPLC分离和特异性检测亲水性肽。为了快速捕获和澄清人血浆中的肽，使用填充有强阳离子交换剂的微型旋转柱（5 mm x 5 mm）进行固相萃取。通过尺寸排阻色谱法（SEC）进一步分级分离澄清的肽级分。SEC级分中的肽用PITC衍生化，并通过使用乙酸铵缓冲液或三氟乙酸系统的RP-HPLC拆分衍生物。基于Edman降解的自动肽序列分析仪，通过修改程序，可以直接分析解析的衍生物。该方法成功地分离和鉴定了人血浆中的一些合成肽和食源性肽。
For the isolation and detection of food-derived peptides in blood, an approach based on the derivatization of peptides with phenyl isothiocyanate (PITC) was developed. This approach allows hydrophilic peptides to be resolved and specifically detected by reversed-phase (RP) HPLC. For the rapid capturing and clarification of peptides in human plasma, solid-phase extraction by using a mini spin column (5 mm x 5 mm) packed with a strong cation exchanger was used. The clarified peptide fraction was further fractionated by size-exclusion chromatography (SEC). The peptides in the SEC fractions were derivatized with PITC, and the derivatives were resolved by RP-HPLC by using an ammonium acetate buffer or a trifluoroacetic acid system. An automatic peptide sequencer based on Edman degradation with a modified program can directly analyze the resolved derivatives. Some synthetic peptides and food-derived peptides in human plasma were successfully isolated and identified by this approach.