Analysis of B-type natriuretic peptide impurities using label-free data-independent acquisition mass spectrometry technology

摘要: Objectives Synthetic B-type natriuretic peptide (BNP) is employed in most clinical testing platforms as a raw material of calibrator. Characterization impurities with structures similar (BNPstrimp compounds) to that BNP reasonable way decrease measurement errors and improve drug safety. Methods A novel quantitative method targeted towards BNPstrimp compounds was developed. First, the samples were separated identified using ultra-performance liquid chromatography, coupled high-resolution mass spectrometry (MS). To evaluate biological activity further, immunoaffinity investigated western blot (WB) assays. Second, label-free data-independent acquisition (DIA) MS approach developed, internal standard (ISP) hydrolyzed. Absolute quantification performed an isotope dilution (ID-MS) approach. Third, precision C-peptide reference material. Results Seventeen synthetic BNP, 10 them successfully sequenced. The immunoassay results indicated deaminated, oxidized, isomerized exhibited weaker than intact BNP1-32. fraction solid ISP1-16, quantified by ID-MS, 853.5 (±17.8) mg/g. Validation developed effective accurate for quantitation well-separated impurities. Conclusions easy perform, it suitable parallel low-abundance when they good separation chromatography. comparable traceable. This promising tool product quality safety assessment.