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LC-MS/MS ANALYSIS FOR PEPTIDES USING HUMAN BRADYKININ AND TWO OF ITS FRAGMENTS AS MODEL MOLECULES

VALENTIN ION, SILVIA IMRE*, ANCA-GABRIELA CARJE, ALINA BALINT, DANIELA-LUCIA MUNTEAN

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Peptide analysis in the context of targeted or discovery proteomics applications assisted by liquid chromatography coupled with mass spectrometry gained increasingly more interest in the last ten years. In any of the proteomics applications cases, an efficient separation of the analysed peptides alongside the chromatographic column is foreseen. That is why this article aimed offer a better insight of peptide chromatographic and mass spectrometric behaviour. As model molecules, human bradykinin and two of its fragments, Bradykinin 1-8 and Bradykinin 1-7 were selected. A reversed phase-mode liquid chromatography-triple quadrupole-mass spectrometry analytical method was developed, investigating the influence of the elution gradient slope of the mobile phase organic component and chromatographic column temperature influence, over the method selectivity α. This two studied components proved to be critical in respect with method selectivity and analytes separations. As a step forward, a fast and simple sample preparation method was developed for the analysis of the three peptides in human urine, overall sample preparation-liquid chromatographic-QQQ method complying with general bioanalytical validation parameters and thresholds.