Masters Theses

Date of Award

5-2013

Degree Type

Thesis

Degree Name

Master of Science

Major

Chemistry

Major Professor

Shawn R. Campagna

Committee Members

Michael J. Sepaniak, Robert N. Compton

Abstract

Two biological systems were studied using LC/ESI/MS/MS on a triple quadrupole operated in SRM (selected reaction monitoring) scan mode. The first bacterium system is aquatic and microscopic in size known as Roseobacter. The second mammalian system is terrestrial and large in size relative to humans known as Holstein cows.

Roseobacter is a clade of marine bacteria abundant in the ocean. Roseophages are viruses that infect Roseobacter and cause viral lysis. Sulfitobacter sp. 2047 was isolated and infected with Roseophages, and the fold change in the metabolic pool relative to a control was studied at discrete time points. The absolute concentration of glutamate and glutamine in the infected and control was determined at each time point using an external calibration curve. Flux analysis through the addition of 13C-acetate at early and late post infection was compared to the control.

Holstein cows are a breed of cattle known to be the world’s highest producers of milk. Twelve Holstein dairy cows were selected, and samples of blood and milk were taken at different weeks of lactation. The fold change in the phospholipid pool relative to the first week of lactation was studied from early, mid, and late lactation. The absolute concentration of lipids at each week of lactation was determined using isotope dilution mass spectrometry with the exception of GPC (glycerophosphocholine) where an external calibration curve was used due commercial unavailability of an isotope-labeled standard.

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