Application of Triple Quadrupole Tandem Mass Spectrometry to the Bioanalysis of Collision-Induced Dissociation-Resistant Cyclic Peptides – Ultra-Sensitive Quantification of the Somatostatin-Analog Pasireotide Utilizing UHPLC-MS/MS
Posted by Adam Awdish on
Innovative Grade US Origin Canine Beagle Single Donor Plasma from Innovative Research was used in the following study:
Max Sauter, Philipp Uhl, Jürgen Burhenne, Walter E. Haefeli
Journal of Pharmaceutical and Biomedical Analysis
February 5, 2021
Peptides are short chains of amino acids that are highly specific and are typically not rejected by host bodies, making them promising candidates in therapeutic drug development. Typical peptides are unfortunately vulnerable to enzymatic hydrolysis, and therefore sequences of peptides are commonly cyclized to increase their stability and target affinity. In fact, there are currently over 40 different cyclic peptides that are already used clinically, and more still that are currently in clinical trials to determine their safety and effectiveness.
The current standard for peptide bioanalysis involves using LC-MS/MS methods, despite their various potential shortcomings in accuracy resulting from broad isotopic patterns and abundance of product ions resulting from collision-induced dissociation (CID), among other things. Despite the analytic shortcomings of LC-MS/MS methods, analysis via triple quadrupole (TQ) MS/MS remains challenging due to poor CID efficiency and the need for at least two consecutive dissociation reactions at the peptide backbone.
Previous works have sought to remedy the CID-resistance of cyclic peptides that hinders bioanalysis using post-extraction reduction methods. Unfortunately, this method proves difficult to use with amide-cyclized peptides due to their stability against proteolytic degradation. Attempts to work around the necessity of CID have investigated monitoring precursors either directly or as a pseudo-transitions in MS/MS instruments, however both methods fall short in terms of bioanalytic sensitivity due to matrix interference. Some studies, however, have observed that high-resolution MS may be a viable method for circumventing the need for precursor fragmentation by eliminating isobaric interferences caused by CID.
In this study, researchers sought to further investigate the CID characteristics of Pasireotide and developed an ultra-sensitive UHPLC-MS/MS assay that boasted a significantly lower limit of quantification (LLOQ) than other quantification methods. Their assay was validated according to EMA and FDA guidelines using 100L of plasma. They were able to achieve the highest sensitivity seen thus far in TQMS/MS quantification of samples with an accuracy of 90.8% and a precision of 4%.
Related products available from Innovative Research also include:
Innovative Grade US Origin Canine Beagle Plasma