A Novel and Sensitive LC/MS/MS Method for Quantitation of Pyochelin in Human Sputum Samples from Cystic Fibrosis Patients

Abstract

Pyoverdines and pyochelin are siderophore compounds secreted by Pseudomonas aeruginosa present in cystic fibrosis affected patients. The available literature on the quantification of pyoverdines and pyochelin in sputa of cystic fibrosis patients is based on detecting the fluorescence properties of the targets detecting pyoverdines and pyochelin without differentiating the types by chromagraphic separation. Within these studies pyochelin was always far less detected than pyoverdines. This is in contrast to gene expression data and suggets the pyochelin fluorescence detection method lacks sensitivity which results in the molecule being non-detectable. In this report, we describe a novel and sensitive LC/MS/MS method for quantitation of pyochelin in human sputum from cystic fibrosis affected patients with Pseudomonas aeruginosa (culture positive) infection. Using a validated bioanalytical method, 26 sputum samples from cystic fibrosis patients from a biobank were analyzed. Data showed the LC/MS/MS method met bioanalytical validation acceptance criteria and results of pyochelin were consistent with reported results of Pseudomonas aeruginosa concentration in cystic fibrosis patients. This new approach for quantitation of pyochelin in human sputum is more sensitive, reproducible and easier to reliably measure the biomarker pyochelin in cystic fibrosis patients. Monitoring pyoverdines and pyochelin in the sputum samples from cystic fibrosis patients may provide better analytical tools for cystic fibrosis new drug development. DOI: 10.29011/2576-9588.100035