Separation and identification of neuropeptide Y, two of its fragments and their degradation products using capillary electrophoresis-mass spectrometry.
This paper describes the development of an analytical method for the separation and identification of neuropeptide Y (NPY) and two important NPY fragments by capillary electrophoresis (CE) and mass spectrometry (MS). A satisfactory separation and the highest sensitivity were obtained with formic acid at high concentrations (250 mM, pH 2.75). The addition of 25 or 50 mM triethylamine (TEA) improved the separation. When applying full scan CE-MS, the separated peptides could be detected and identified using the spectra of each peak. The use of TEA as an additive to the formic acid slightly decreased the sensitivity but was compensated by the improved efficiency. The best compromise for optimal separation and MS detection was found to be 50 mM formic acid to which 50 mM TEA was added. CE-MS could be used for identification of the decomposition products of NPY. Decomposition products with one amino acid difference, which could not be distinguished with CE-UV, could be distinguished with CE-MS.
Ensing
K
K
Groningen Institute for Drug Studies, Department of Analytical Chemistry and Toxicology, The Netherlands. k.ensing@farm.rug.nl
de Boer
T
T
Schreuder
N
N
de Zeeuw
R A
RA
D000595:Amino Acid Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D005021:Ethylamines / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000737:chemistry / Q000302:isolation & purification*; D011487:Protein Conformation; D012680:Sensitivity and Specificity
D005021:Ethylamines; D007202:Indicators and Reagents; D009478:Neuropeptide Y; C016162:triethylamine