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Date of Award


Document Type

Thesis - Pacific Access Restricted

Degree Name

Master of Science (M.S.)



First Advisor

Patrick R. Jones

First Committee Member

O. David Sparkman

Second Committee Member

James W. Blankenship


A rapid method was developed for the quantitative analysis of N, N'hexamethylenebisacetamide (HMBA) and its deacetylated metabolites, N-acetyl-1, 6- diaminohexane (NADAH) and 1, 6-diaminohexane (DAH). The method used flowinjection liquid chromatography (LC)-atmospheric pressure chemical ionization (APCI)selected ion monitoring at low/high resolving power (LR/HR-SIM)-mass spectrometry (MS). Samples were extracted from murine erythroleukemia (MEL) cell culture and were analyzed without chemical derivatization. The limits of detection (LOD) for HMBA, NADAH and DAH measured by HR-SIM were 0.05, 0.1 and 0.2 picomole, respectively. Calibration curves were constructed by using tetradeuterated DAH as an internal standard for both SIM methods. The ratio of the analyte concentration in the extract of a MEL cell culture treated with HMBA for 120 hours was found to be 1. 7, 27 and 17 times that of the control for DAH, NADAH and HMBA, respectively. The HRSIM that used a lock-mass technique helped eliminate chemical interferences that had mlz values similar to the one of interest. Although this technique was developed using HMBA, NADAH and DAH, it is expected to work for natural polyamines and their acetylated metabolites.

Qualitative analysis was performed by both positive electrospray (ES)- and APCIMS under full mass range scanning. The mass spectra of polyamine standards were characterized. HMBA exhibited an ability to form mono- and bi-molecular adducts with metal ions, itself and with other polyamine molecules in ES-MS. The LODs estimated by ES-MS were consistently higher than the APCI-SIM methods that is described here.



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