Presenter: Dr. Anas El-Aneed, Associate Professor
College of Pharmacy and Nutrition at the University of Saskatchewan
Full Talk title: Targeted mass spectrometric methods for the identification of asthma biomarkers in human urine and the determination of the cellular fate of nanoparticle gene delivery agents.
Dr. El-Aneed is an Associate Professor at the College of Pharmacy and Nutrition, University of Saskatchewan. He obtained his B.Sc. in Pharmacy and Pharmaceutical Chemistry in 1997 from Tishreen University, Syria. He then completed a M.Sc. in 2003 in Pharmacy and Ph.D. in 2007 in Biochemistry from Memorial University of Newfoundland. He was awarded the Governor General Gold medal for his Ph.D. thesis. He worked as Pharmacy Research Specialist at the Newfoundland and Labrador Center for Health Information 2006-2007 and joined the University of Saskatchewan in January 2008. In 2012, he completed an MBA degree from the University of Saskatchewan. Dr. El-Aneed’s main area of research is focused on the use of different mass spectrometry platforms for the qualitative and quantitative analysis of small organic compounds, with recent emphasis on diagnostic urine metabolites and gene delivery agents. He is currently the co-chair of the Saskatchewan Mass Spectrometry User Group. In addition to basic science, Dr. El-Aneed is engaged in applied health research, currently investigating the educational needs for community pharmacists concerning substance abuse and addiction.
Abstract: High pressure liquid chromatography- tandem mass spectrometry (HPLC-MS/MS) methods are developed to quantify 50 highly polar, low molecular-weight urine metabolites that were suggested as potential biomarkers for respiratory illnesses, such as asthma and COPD. Targeted metabolites were classified based on common functional groups into amines and phenols (group-1), carboxylic acids (group-2), and miscellaneous (group-3). Groups 1 and 2 contain 42 metabolites and were analyzed based on the differential isotope labeling (DIL) approach using C12/C13 labeled derivatizing reagents. Dansyl chloride (DNS-Cl) and dimethylaminophenacyl bromide reagent (DmPA) were selected for the derivatization of groups 1 and 2, respectively. The derivatization allows for the successful separation of the metabolites on a C18 column using gradient elution, while enhancing the MS response during positive electrospray ionization. Despite complexity, the developed methods were successfully validated according to the FDA guidelines. Preliminary clinical data showed separation of COPD and asthma patients based on their quantitative metabolomic profiles.
Similarly, we developed various fully validated MS-based quantitative methods to monitor the fate of two leading gemini surfactants within cells. Gemini surfactants are used as gene delivery agents and we aimed at understanding the varying toxicities among various structures, based on their cellular fate. It was shown that the two structures followed the same pattern in terms of uptake and release/degradation. However, our preliminary data regarding the subcellular localization showed varying patterns which may provide new insights into the efficiency and toxicity of gemini surfactant as nanoparticle gene delivery agents.
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