Novel Techniques for Mass Spectrometric Analysis of Biomolecules
Quinn, Kevin D.
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A three dimensional quadrupole ion trap was used to obtain low resolution mass spectra of Infasurf® ® , calf lung surfactant extract (CLSE), and surfactant protein B (SP-B) utilizing both online electrospray ionization and static nano electrospray ionization. A separation technique utilizing C8 packing medium was modified to be performed on an ion trap with the use of a 2.1x 100mm C8 column with a flow rated of 200uL/min. The method successfully separated SP-B from the CLSE matrix though the use of chloroform/methanol, and methanol/water solvents for direct mass spectrometric analysis. The Collision Induced Dissociation (CID) fragmentation patterns of stercobilin, urobilin, and bilirubin were probed using a quadrupole ion trap at various relative fragmentation energies. Stercobilin's and urobilin's carboxylic acid groups were methylated to form a methyl ester at room temperature. CID mass spectra were obtained of the methyl esters that were formed at various fragmentation energies. A fragmentation pathway was determined for stercobilin and further supported by information obtained from the methyl ester of stercobilin. Fragmentation data was analyzed for urobilin and bilirubin, but no definitive pathway was determined dictating the need for higher resolution data or higher degrees of tandem mass spectrometry, MS 3 . Paper Spray Ionization is an atmospheric pressure ionization technique that utilizes an offline electro-osmotic flow to generate ions off a paper medium. This technique can be performed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometer by modifying the existing nano-spray source. High resolution paper spray spectra were obtained for both organic and biological samples to demonstrate the benefit of linking the technique with a high resolution mass analyzer. Error values in the range 0.23 to 2.75ppm were obtained for Calf Lung Surfactant extract with broadband resolving power as high as 61637 utilizing an external calibration standard.