A selective fluorescence reaction for peptides and chromatographic analysis
A selective fluorescence reaction for peptides and chromatographic analysis
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DOI：
10.1016/j.peptides.2007.11.014
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发表时间：
2008-03-01
期刊：
影响因子：
3
通讯作者：
Kai, Masaaki
中科院分区：
文献类型：
--
作者：
Kabashima, Tsutomu;Yu, Zhiqiang;Kai, Masaaki
提出了一种新的选择性荧光反应，用于反相液相色谱（RPLC）定量测定多肽。当肽在含有邻苯二酚、高碘酸钠和硼酸钠的中性水性介质（pH 7.0）中于120 ℃加热20分钟时，形成单一荧光产物。在反相柱上，以甲醇为流动相，硼酸钠为流动相（pH 7.0），梯度洗脱，Leu-Gly、Ala-Leu-Gly、Tyr-Gly-Gly-Phe-Leu和Leu-Leu-Leu四种多肽的荧光产物得到分离，荧光定量检测。测试肽的检测下限（SIN = 3）为0.5-1.0 pmol/注射体积（40 μ l）。此外，利用电喷雾飞行时间质谱（ESI-TOF/MS）对苯丙氨酸酰胺和Leu-Leu-Leu的荧光产物进行了鉴定，以确定其化学结构。(c)2007年爱思唯尔公司All rights reserved.
A novel and selective fluorescence reaction is proposed for the quantitative determination of peptides by reversed-phase liquid chromatography (RPLC). A single fluorescent product was formed when a peptide was heated at 120 degrees C for 20 min in a neutral aqueous medium (pH 7.0) containing catechol, sodium periodate, and sodium borate. The fluorescent products of four peptides such as Leu-Gly, Ala-Leu-Gly, Tyr-Gly-Gly-Phe-Leu, and Leu-Leu-Leu were easily separated on a reversed-phase column by gradient elution of methanol in a mobile phase containing sodium borate (pH 7.0), and then quantitatively detected by fluorimetry. The lower limits (SIN = 3) of the detection for the tested peptides were 0.5-1.0 pmol per an injection volume (40 mu l). In addition, the fluorescent products of phenylalanine amide and Leu-Leu-Leu were identified by electrospray ionization-time of flightmass spectrometry (ESI-TOF/MS) for the elucidation of their chemical structures. (c) 2007 Elsevier Inc. All rights reserved.