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使用毛细管电泳-激光诱导荧光检测法分离和测定肌肽相关肽。

Separation and determination of carnosine-related peptides using capillary electrophoresis with laser-induced fluorescence detection.

作者信息

Huang Ying, Duan Jianping, Chen Hongqing, Chen Min, Chen Guonan

机构信息

Department of Chemistry, Fuzhou University, Fujian, China.

出版信息

Electrophoresis. 2005 Feb;26(3):593-9. doi: 10.1002/elps.200406130.

Abstract

A capillary electrophoresis (CE) method with laser-induced fluorescence (LIF) detection was developed for the separation and detection of carnosine-related peptides (carnosine, anserine, and homocarnosine). A sensitive and fluorogenic regent, 3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde (CBQCA) was selected as a precapillary labeling reagent for imidazole dipeptides to form isoindole derivatives. The optimized molar ratio between CBQCA and peptide was found to be 75:1, and 50 mmol/L borate buffer (pH 9.2) was used for the derivatization in order to achieve good efficiency. Three imidazole dipeptides were baseline-separated within 20 min by using 112 mmol/L sodium borate (pH 10.4-10.8) as running buffer. Concentration detection limits (signal-to-noise ratios) for carnosine, anserine, and homocarnosine were 4.73, 4.37, and 3.94 nmol/L, respectively. This method has been applied to the analysis of human cerebrospinal fluid (CSF) and meat dry powder of pig and sheep. Recoveries were in the range of 82.9-104.8% for homocarnosine in CSF. For carnosine and anserine, the recoveries are 98.3% and 80.2% in meat dry powder of pig and 111.2% and 112.8% in meat dry powder of sheep, respectively.

摘要

建立了一种采用激光诱导荧光(LIF)检测的毛细管电泳(CE)方法,用于分离和检测肌肽相关肽(肌肽、鹅肌肽和高肌肽)。选择一种灵敏的荧光试剂3-(4-羧基苯甲酰基)喹啉-2-甲醛(CBQCA)作为咪唑二肽的毛细管前标记试剂,以形成异吲哚衍生物。发现CBQCA与肽之间的最佳摩尔比为75:1,为了获得良好的效率,使用50 mmol/L硼酸盐缓冲液(pH 9.2)进行衍生化。以112 mmol/L硼酸钠(pH 10.4 - 10.8)作为运行缓冲液,三种咪唑二肽在20分钟内实现了基线分离。肌肽、鹅肌肽和高肌肽的浓度检测限(信噪比)分别为4.73、4.37和3.94 nmol/L。该方法已应用于人体脑脊液(CSF)以及猪和羊的肉干粉分析。CSF中高肌肽的回收率在82.9 - 104.8%范围内。对于肌肽和鹅肌肽,在猪的肉干粉中的回收率分别为98.3%和80.2%,在羊的肉干粉中的回收率分别为111.2%和112.8%。

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