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采用毛细管电泳-激光诱导荧光检测法测定人体体液中缓激肽相关肽。

Assay of bradykinin-related peptides in human body fluids using capillary electrophoresis with laser-induced fluorescence detection.

作者信息

Chen Ying, Xu Liangjun, Lin Jinming, Chen Guonan

机构信息

Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety, Fuzhou University, Fuzhou, Fujian, PR China.

出版信息

Electrophoresis. 2008 Mar;29(6):1302-7. doi: 10.1002/elps.200700594.

Abstract

A CE with LIF detection was developed for separation and determination of bradykinin (BK)-related peptides, such as BK, kallidin (Kal), and neurokinin A (NKA). BK-related peptides were derivatized with FITC prior to CE-LIF analysis. Sodium borate 10 mmol/L at pH 9.5 was selected as derivatization media in order to get the high efficiency. Three peptides were baseline-separated within 10 min by using 110 mmol/L sodium borate-sodium hydroxide solution at pH 10.0 as the running buffer. Concentration detection limits (S/N = 3) for BK, Kal, and NKA were 0.08, 0.5, and 0.2 nmol/L, respectively. Meanwhile we have also developed a simple, quick, and sensitive large-volume sample stacking (LVSS) technique for CE-LIF detection of BK, Kal, and NKA. By using this stacking technique, the detection limits (S/N = 3) for BK, Kal, and NKA were 0.02, 0.05, and 0.04 nmol/L, respectively. This method has been applied to the assay of human saliva and cerebrospinal fluid with satisfactory results.

摘要

开发了一种用于分离和测定缓激肽(BK)相关肽(如BK、胰激肽(Kal)和神经激肽A(NKA))的毛细管电泳-激光诱导荧光检测(CE-LIF)方法。在进行CE-LIF分析之前,BK相关肽用异硫氰酸荧光素(FITC)进行衍生化。选择pH 9.5的10 mmol/L硼酸钠作为衍生化介质以获得高效衍生化效果。以pH 10.0的110 mmol/L硼酸钠-氢氧化钠溶液作为运行缓冲液,三种肽在10分钟内实现了基线分离。BK、Kal和NKA的浓度检测限(信噪比S/N = 3)分别为0.08、0.5和0.2 nmol/L。同时,我们还开发了一种简单、快速且灵敏的大体积样品堆积(LVSS)技术用于CE-LIF检测BK、Kal和NKA。通过使用这种堆积技术,BK、Kal和NKA的检测限(S/N = 3)分别为0.02、0.05和0.04nmol/L。该方法已应用于人体唾液和脑脊液的检测,结果令人满意。

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