Quantitation of glutathione in urine and probing for further autism biomarkers utilizing both low and high resolution mass spectrometric and chromatographic techniques
Cruickshank, Charmion I. E.
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The search for biomarkers has increased enormously in the last decade due to the pressing need for early diagnosis of many diseases. The use of mass spectrometry has greatly aided detection and profiling, and more sensitive instrumentation enables concentrations in the femtomolar range to be detected. This dissertation will discuss one such biomarker, glutathione, from method development to its quantitation using mass spectrometry. Autism has come in the forefront in recent years due to its prevalence worldwide. It affects 1 in every 88 children in the United States, representing a 78 percent increase from 2002 to 2006, and a 600 percent increase in the past 20 years. A study published in 2011 reported that 1 in 38 children in South Korea are affected with these numbers growing. Glutathione is of crucial importance because of its role in oxidative stress in humans. Research by S Jill James has shown that plasma levels of total glutathione and the ratio of reduced to oxidized glutathione, were significantly decreased in autistic patients. Autistic plasma samples exhibited higher levels of oxidized glutathione compared to control samples hence justifying a possible biological test for early detection. Collecting urine is a relatively non-invasive technique. A comparison of controls versus autistic urine samples for glutathione levels provides insight into the oxidative stress theory. The use of a Thermo Advantage Ion Trap and a Bruker SolariX FT-ICR are used for mass spectrometric analysis coupled to liquid chromatography and nano-liquid chromatography respectively.