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Quantitation and identification of two cholecystokinin peptides, CCK‐4 and CCK‐8s, in rat brain by HPLC and fast atom bombardment mass spectrometry
Author(s) -
Qureshi G. Ali,
Bednar Ivan,
Min Qian,
Södersten Per,
Silberring Jerzy,
Nyberg Fred,
Thörnwall Madeleine
Publication year - 1993
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130070503
Subject(s) - chemistry , cholecystokinin , fast atom bombardment , chromatography , mass spectrometry , high performance liquid chromatography , tetrapeptide , detection limit , peptide , quantitative analysis (chemistry) , biochemistry , receptor
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.