In cellular studies of patients with lipid related disorders such as mammary cancers, leukemia, and artheroscierosis, separation of molecular species of oxidized phosphatidylcholine (PC) can be an important assistance in research or diagnosis. Goals of this project were to optimize separation of oxidized and unoxidized PC molecular species in a single HPLC chromatogram followed by in line identification of hydroperoxides. Oxidized egg PC's were produced using UV light exposure in air. Separations were performed on an Ultrasphere ODS column and an Asahipak ODPVA column using a Waters 2695 system with photodiode array. The ODPVA column routinely gave 10 times larger plate numbers. Various mobile phase mixtures (methanol, acetonitrile, water) and gradients were tested. The optimum gradient on our system is (1) 5 minutes, 47 % methanol/40 % acetonitrile/13 water in a linear gradient to (2) 17 minutes, 49 % methanol/40 % acetonitrile/11 % water to (3) 18 minutes, 29 % methanol/60 % acetonitrile/11 % water linearly to (4) 50 minutes, 31 % methanol/60 % acetonitrile/9 % water continued isocratically to 110 minutes. Oxidized hydroperoxides were detected by fluorescence using a post column reaction with diphenyl-1 pyrenylphosphine (DPPP). Both iron (III) and pyridine were tested as catalysts for this reaction. / Department of Chemistry
| Identifer | oai:union.ndltd.org:BSU/oai:cardinalscholar.bsu.edu:handle/187935 |
| Date | January 2005 |
| Creators | Weddle, Carolyn A. |
| Contributors | Pattison, Scott E. |
| Source Sets | Ball State University |
| Detected Language | English |
| Format | viii, 77 leaves : ill. ; 28 cm. |
| Source | Virtual Press |
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