alexa Quantifying Therapeutic Peptides In Human Plasma By Mass Spectrometry: Our Experience Applied To Pharmacokinetic Studies In Clinical Trial | 69659
ISSN: 2157-7064

Journal of Chromatography & Separation Techniques
Open Access

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3rd International Conference and Exhibition on Advances in Chromatography & HPLC Techniques

Ania Cabrales Rico
Center for Genetic Engineering and Biotechnology, Cuba
Posters & Accepted Abstracts: J Chromatogr Sep Tech
DOI: 10.4172/2157-7064-C1-029
Abstract
Mass Spectrometry (MS) has impressive capabilities in terms of sensitivity, resolving power, mass accuracy and different scanmodes versatility. Either alone or in combination with liquid chromatography it is the analytical tool of choice for synthetic therapeutic peptide characterization. Nevertheless, for peptide quantitation in human plasma or other biological sample, the design of the Internal Standard (IS) and the optimization of the sample processing and LC-MS analysis are also key elements for a successful outcome. In consequence, all strategies involving the peptide quantitation in biological fluids are still a challenge and need to be tailored. We present here our recent experiences in the development and validation of customized bioanalytical methods applied to pharmacokinetic studies included in phase I clinical trials. For the absolute quantitation of these three therapeutic peptides, alternatives to the AQUA® methodology were used. However, the design of the IS, sample processing and mass spectrometry techniques were optimized case by case for CIGB-500, CIGB-300 and CIGB-814 candidates. IS for CIGB-500 and CIGB-814 were synthetic peptides labeled with stable isotopes (13C and/or 15N) in specific residues within the amino acids sequence, instead of IS for CIGB-300 that was a N-terminus acetylated peptide. Sample processing, mainly based on plasma proteins organic or acid precipitation was adapted according to the peptide recovery. In the particular case of CIGB-300, no liquid chromatography separation was needed before MS analysis by MALDI-TOF MS. For CIGB-500 it was applied LC-MS analysis with simultaneous ion monitoring (SIM) in full scan mode. For CIGB-814 it was used LC-MS analysis in single reaction monitoring mode (SRM). The three bioanalytical methods were fully validated and applied to pharmacokinetic (PK) analysis in a phase I clinical trials. It was possible to obtain PK profiles and main PK parameters for all of the assessed candidates.
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