Date of Award
Open Access Thesis
Master of Science (MS)
Darrin L. Smith
A direct analysis in real time mass spectrometry method was developed and validated for the analysis and quantitation of sugars that would be generated from pretreated and hydrolyzed switchgrass. This research aspect can be divided into two sections:
i) Literature Review: A review is presented on the status of energy security and how biomass and biofuels can be utilized as a source of transportation fuel. To this end, biomass (specifically switchgrass) can be broken down by pretreatment methods and then enzymatically hydrolyzed to simple sugars. These sugars can assist with algae growth that can eventually be converted into biofuels. Direct Analysis in Real Time Mass Spectrometry (DART-MS), an ambient mass spectrometric method described in this study, can readily analyze these generated simple sugars.
ii) Optimization of DART-MS for the quantitation of glucose: Being the first study to use DART-MS to quantify sugars, the DART-MS instrumental parameters, such as gas heater temperature, helium pressure, linear rail speed, distance of DART source from the mass spectrometer orifice, and the grid voltage were varied to determine the optimal ionization conditions for sugar standards. Reproducibility experiments were performed to determine the robustness of the method. For quantification experiments, a dynamic linear range was developed using sugar standards in matrix-free solvents with the use of internal standards.
iii) Validation of the DART-MS method for quantitation of six-carbon sugars in saccharification matrix: Validation of the DART-MS method was performed to determine the limits of detection/quantitation, investigate matrix effects with respect to instrumental signal suppression when matrix-diluted standards were used, and perform recovery studies for accuracy and precision. Statistical analysis was used to compare calibration curves and recovery results generated from matrix-free and matrix-diluted sugars standards.
The resulting DART-MS method for glucose analysis was found to be precise, fast, and robust for the quantitation with saccharification samples. Since DART-MS requires little to no sample preparation, this technique becomes an attractive option and could be the choice in the quantitation of sugars for biofuel advancement.
Copyright 2011 Daudi Sayialel Saang'onyo
Saang'onyo, Daudi Sayialel, "Optimization And Validation Of Direct Analysis In Real Time Mass Spectrometry (dart-Ms) For Quantitation Of Sugars To Advance Biofuel Production" (2011). Online Theses and Dissertations. 44.