AN INVESTIGATION INTO THE VERSATILITY OF A TITANIUM:SAPPHIRE REGENERATIVE AMPLIFIER LASER SYSTEM FOR AMBIENT MASS SPECTROMETRY

Abstract

This dissertation details an investigation into the use of laser pulses from a titanium:sapphire regenerative amplifier laser system to vaporize analytes in ambient air for mass spectral analysis. The laser system was modified to operate in one of two distinct modes. In femtosecond (fs) mode the laser produced 2.5 mJ, ~60 fs laser pulses centered at 800 nm. In nanosecond (ns) mode the laser produced 2.4 mJ, ~10 ns laser pulses centered at 800 nm. Using appropriate optical components the laser pulse energy was attenuated to achieve pulses varying from 0.15 mJ to 2.0 mJ. Laser pulses were used to vaporize liquid and solid samples on different substrates. The laser vaporized material was captured and ionized by an electrospray source and then detected via a mass spectrometer instrument. It was discovered that samples on glass substrate could be vaporized by fs laser pulses, but not by ns laser pulses. Samples on metal substrate were successfully vaporized by both fs and ns laser pulses. Low energy ns laser pulses were less efficient than fs laser pulses of the same energy for vaporizing off metal substrate. A comparison of vaporization from aluminum, copper and stainless steel substrates revealed limited vaporization from copper by ns laser pulses. The electrospray ionization (ESI) mass spectral response of wet and dry proteins on stainless steel was similar for both fs and ns laser pulses. Experiments to test the capabilities of ns laser electrospray mass spectrometry (ns-LEMS) revealed that sample vaporization was limited to analysis on metal surfaces. This dissertation details methods for femtosecond laser electrospray ionization (fs-LEMS) to be used to quantify non-covalent protein-ligand interactions. Hen egg white lysozyme (HEWL) and N,N’,N”-triacetylchitotriose (NAG3) interactions were quantified via dissociation constant (Kd) measurements. The Kd for HEWL and N,N’,N”,N”’-tetraacetylchitotetraose (NAG4) were also measured. This dissertation also reports a miniaturized flowing atmospheric pressure afterglow (micro-FAPA) for use as an alternative ionization source of fs-laser vaporized analytes. Loratadine pills were vaporized and reacted with the gas stream from the micro-FAPA source to generate ions which were then detected by a mass analyzer. The ions detected varied in distribution as a response to the distance the sample was vaporized from the ion source. Complexed samples were tested and molecular assignments were difficult due to the numerous pathways for ion formation. The use of an ion filter to decrease the energy imparted on sample molecules during the ionization process of the micro-FAPA is also reported.