Dr. Cowan, Head of the WADA-Accredited Drug Control Center at King’s College London, has provided the following update on his PCC funded research.
Project title: Developing a mass spectrometric method for PIIINP and other collagen bone biomarkers to detect rHGH administration.
Principal Investigator: Cowan, David, A
Name of Institution: King’s College London
Start Date of Project: August 17, 2015 (follow-on from 1st phase of overall project)
Summary:
Procollagen III amino-terminal propeptide (PIIINP), a biomarker for detecting growth hormone administration, is currently measured by immunoassays in the absence of international reference material. The main aim of this research is to demonstrate whether liquid chromatography tandem mass spectrometry can be used to quantify PIIINP in blood.
Post translational modifications of PIIINP are expected based on its topology, some of which have been identified for the first time by mass spectrometry of surrogate peptides, T1 and T5, obtained from trypsin digestion of intact bovine PIIINP. A basal blood PIIINP concentration of 5 ng/mL (117 pM) exists in humans; LC-MS methods using nano ESI have been developed with a LOD of 10 pM for both peptides. However, significant ion suppression of the peptides is observed in the presence human serum albumin (HSA) ≥20 µg/mL. To achieve these conditions in serum ≥99.7% HSA depletion is necessary. Appropriate separation of PIIINP from matrix to facilitate analysis by LC-MS methods are currently being investigated.
This research is scheduled for completion by December 31, 2017.
For more information on Dr. Cowan’s project, or other PCC sponsored research, please email Jenna Celmer at jcelmer@cleancompetition.org
