title: Liquid Chromatography-High Resolution Mass Spectrometry for Peptide Drug Quality Control.
abstract: A liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed using three peptide drugs: salmon calcitonin, bivalirudin, and exenatide as model systems to assess the suitability of this approach for monitoring peptide drug product quality. Calcitonin and its related impurities displayed linear responses over the range from 0.1 to 10 μM (R (2) values for calcitonin salmon, Glu(14)-calcitonin, and acetyl-calcitonin were 0.995, 0.996, and 0.993, respectively). Intra-assay precision in terms of relative standard deviation (%RSD) was less than 10% at all tested concentrations. The accuracy of the method was greater than 85% as measured by spiking 0.1, 0.3, and 1% of Glu(14)-calcitonin and acetyl-calcitonin into a stock calcitonin solution. Limits of detection for calcitonin, Glu(14)-calcitonin, and acetyl-calcitonin were 0.02, 0.03, and 0.04 μM, respectively, indicating that an impurity present at less than 0.1% (0.1 μM) of the drug product API concentration (107 μM) could be detected. Method validation studies analyzing bivalirudin and exenatide drug products exhibited similar results to calcitonin salmon in regard to high selectivity, sensitivity, precision, and linearity. Added benefits of using LC-HRMS-based methods are the ability to also determine amino acid composition, confirm peptide sequence, and quantify impurities, even when they are co-eluting, within a single experiment. LC-HRMS represents a promising approach for the quality control of peptides including the measurement of any peptide-related impurities. While the development work performed here is focus on peptide drug products, the principles could be adapted to peptide drug substance.
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, Center for Drug Evaluation and Research, Food and Drug Administration, 645 S. Newstead Avenue., St Louis, Missouri, 63110, USA.
authors.forename: Kui
authors.initials: K
authors.lastname: Zeng
authors.forename: Ilan
authors.initials: I
authors.lastname: Geerlof-Vidavisky
authors.forename: Ashley
authors.initials: A
authors.lastname: Gucinski
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Jiang
authors.forename: Michael T
authors.initials: MT
authors.lastname: Boyne
mesh_terms: D000595:Amino Acid Sequence; D002116:Calcitonin / Q000032:analysis / Q000592:standards; D002853:Chromatography, Liquid / Q000379:methods*; D000077270:Exenatide; D006629:Hirudins / Q000032:analysis / Q000592:standards; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D010446:Peptide Fragments / Q000032:analysis / Q000592:standards; D010455:Peptides / Q000032:analysis* / Q000592:standards; D011786:Quality Control; D011994:Recombinant Proteins / Q000032:analysis / Q000592:standards; D014688:Venoms / Q000032:analysis
chemical_list: D006629:Hirudins; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D014688:Venoms; C028815:salmon calcitonin; D002116:Calcitonin; D000077270:Exenatide; C074619:bivalirudin