采用高效液相色谱法和快原子轰击质谱法对大鼠脑中两种胆囊收缩素肽CCK - 4和CCK - 8s进行定量和鉴定。

Quantitation and identification of two cholecystokinin peptides, CCK-4 and CCK-8s, in rat brain by HPLC and fast atom bombardment mass spectrometry.

作者信息

Qureshi G A, Bednar I, Min Q, Södersten P, Silberring J, Nyberg F, Thörnwall M

机构信息

Department of Psychiatry, Karolinska Institute, Huddinge, Sweden.

出版信息

Biomed Chromatogr. 1993 Sep-Oct;7(5):251-5. doi: 10.1002/bmc.1130070503.

DOI:10.1002/bmc.1130070503

PMID:8305854

Abstract

A sensitive HPLC method has been described for quantitation of two cholecystokinin (CCK) peptides in discrete rat brain regions. Separation and quantitation was performed by the reversed-phase HPLC combined with electrochemical detection. Analytical recoveries of the tetrapeptide (CCK-4) and octapeptide-sulphate (CCK-8s) were 96% and 94%, respectively. The between assay coefficient of variation (CV) was less than 3% for both peptides. The within-assay CV was 4% and 6% for CCK-4 and CCK-8s and the detection limit was 2 and 10 pmol/mL, respectively. For identification of structures, the peptides were fractionated by semi-preparative HPLC using a novel SMART system for micropurification. The fractions were analysed by fast atom bombardment mass spectrometry (FAB MS) which confirmed the presence of both CCK-4 and CCK-8s in the rat brain tissue.

摘要

已描述了一种灵敏的高效液相色谱（HPLC）方法，用于定量大鼠离散脑区中的两种胆囊收缩素（CCK）肽。通过反相HPLC结合电化学检测进行分离和定量。四肽（CCK-4）和八肽硫酸盐（CCK-8s）的分析回收率分别为96%和94%。两种肽的测定间变异系数（CV）均小于3%。CCK-4和CCK-8s的测定内CV分别为4%和6%，检测限分别为2和10 pmol/mL。为了鉴定结构，使用新型SMART微纯化系统通过半制备HPLC对肽进行分级分离。通过快原子轰击质谱（FAB MS）分析这些级分，证实大鼠脑组织中同时存在CCK-4和CCK-8s。