Accurate quantification of impurities in pure peptide material - angiotensin I: Comparison of calibration requirements and method performance characteristics of liquid chromatography coupled to hybrid tandem mass spectrometry and linear ion trap high-resolution mass spectrometry.

摘要: Rationale The prohormone angiotensin I (ANG I) [amino acid sequence: DRVYIHPFHL] and other structurally related peptide hormones play an essential role in the regulation of water electrolyte balance human body as well blood pressure. ANG is a biomarker for hypertension diabetes. Therefore, well-characterized pure reference materials comparable SI-traceable analytical characterization methods are required to establish measurement systems (RMS) laboratory medicine. Methods Two based on liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization have been developed validated in-house. Both high-resolution MS (hrMS) hybrid-tandem MS/MS were used identification quantification major impurities I. The quantified by use external calibrations original impurity standards. Mass fraction values corresponding expanded uncertainties calculated. Results Five degradation products detected 'pure' material. peptides (2-10) [RVYIHPFHL], II [DRVYIHPF] three isomers [DRVYLHPFHL, DRVYIHPFHI DRVYLHPFHI] identified mass calculated that range from 0.66 4.86 mg/g. Conclusions The material obtained both LC/hrMS LC/MS/MS excellent agreement. This study emphasizes importance spectrometric techniques application approaches value uncertainty assignment substance peptides. Copyright © 2015 John Wiley & Sons, Ltd.