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Determination of peptide hormones of brain and intestine by CE with ESI‐MS detection
Author(s) -
Xia Shifei,
Zhang Lan,
Tong Ping,
Lu Minghua,
Liu Wei,
Chen Guonan
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600756
Subject(s) - chemistry , formic acid , ammonium formate , chromatography , neurotensin , peptide , electrolyte , mass spectrometry , analytical chemistry (journal) , biochemistry , neuropeptide , receptor , electrode
Abstract A new method for the determination of the peptide hormones of brain and intestine based on CE coupling with a DAD and ESI‐MS was established. Several electrophoretic and ESI‐MS parameters were investigated in detail, such as electrolyte nature and concentration, organic solvent and sheath liquid compositions, nebulization gas pressure and the ESI capillary voltage. Optimized conditions were achieved with 25 mM formic acid‐ammonium formate (pH 2.9) as the optimal electrolyte, 2 mM formic acid in 80% methanol in water as the sheath liquid, and 20 kV applied voltage. Under the optimized conditions, four protonated peptides were separated by CE and selectively detected by a quadrupole mass spectrometer with a sheath flow ESI interface. LODs for the four peptides (neurotensin hexapeptide, neurotensin, cholecystokinin tetrapeptide, and pentagastrin) were in the range of 0.10–0.60 µmol/L at an S/N of 3. The RSDs ( n = 8) of the method were 0.70–1.5% for migration times and 1.6–6.1% for peak areas. This method is simple, rapid, and selective compared with RIA and ELISA techniques, and has been applied to the analysis of rat hypothalamus tissue.