Research On Biological Monitoring Of Exposure To Sulfur Mustard For Glutathione Adducts

Surfur mustard [bis-(2-chloroethyl)sulfide, SM] was called as"king of toxic agents", it has been widely used as a powerful chemical weapon in World War I, World War II, and Iran-Iraq War. Recently, it also could be used as one of the threat tool for terrorism. The analysis of biological samples to access exposure to sulfur mustard is critical for responding to a terrorist attack, accidental release, or military use of SM. The terrorism threat of SM is severe; therefore, developing a fast and accurate detection method for SM plays an important role in emergence treatments. The research on the analysis of SM in biomedical samples is still a hot topic for military analysts.SM belongs to a kind of vesicant agent, the skin, respiratory, oral and mucous membrane contact are all the exposure approaches for SM. As a bifunctional alkylating agent, it reacts rapidly with nucleophiles under physiological conditions. These reactions in the human body include the alkylation of DNA, proteins and glutathione. About 50% of SM reacts with glutathione and then products urinary metabolites. Urine samples from individuals without exposure to SM have no SBMTE detected. This compound can be applied to demonstrate exposure to SM. In this dissertation, the gas chromatography-mass spectrometric method (GC-MS) and other modern techniques were used to quantitatively determine the glutathione adduct. This dissertation consists of five chapters.Chapter 1 was the preface of the dissertation. The toxicology and metabolism of SM in human body were described. The detection of glutathione is so important that can help to verify SM exposure. The objective and contents of this dissertation are then presented.In chapter 2, 1,1'-sulfonylbis[methylthio]ethane(SBMTE) was synthesized.purified and identified. It was synthesized by the oxidized reactions of SM. SBMTE was isolated and purified by column chromatography and recrystallisation, its purity was up to 98.9% by GC-MS analysis. It was identified by MS and 1H-NMR.Their spectra were full consistent with the structure of chemical. In chapter 3, a simple GC-FPD and GC-FID method was developed to measure SBMTE in PBS buffer and urine samples. The the linear calibration curve was ranged from 0.25 to 10μg/mL(FID) and the limit of detection(LOD) was 10ng/mL(FPD). It can be used as a method for the retrospective detection of sulfur mustard. However, there is a requirement to enhance the LOD.In chapter 4, a simple and accurate GC-MS method was developed to measure SBMTE in PBS buffer which is used as simulated urine samples. Solid-phase extraction(SPE) method was chosen as the sample preconcentration method after comparing with liquid-liquid extraction method. The selectivity of different SPE collumns and the efficiency of different elution solvents were investigated. The loaded pH and steps of extraction were also optimized. The the linear calibration curve was ranged from 10to 1000ng/mL and the limit of detection(LOD) was 5ng/mL.In chapter 5, GC-MS method was developed to determine SBMTE in urine samples. Different procedures to remove proteins were carried out, since biological samples always contain proteins which has more or less interference on the detection method. The recovery of those complicated removal procedures was unsatisfied and of time-consuming. Therefore, choosing other internal standards, and optimizing MS conditions were finally adopted in this chapter to avoid of the interference from urine samples. After the progress of concentration, the the linear calibration curve was ranged from 1 to 100ng/mL and the limit of detection(LOD) was 0.5ng/mL.