Date of Award
Doctor of Philosophy (PhD)
Chemistry and Biochemistry
John Van Norman
The most significant problem in maintaining platelet viability during storage is the decrease in pH resulting from accumulation of lactic acid formed by platelet anaerobic glycolysis. This study investigates nutrient alternatives to glucose for maintenance of morphology and functional integrity during platelet storage in a well-defined synthetic medium and correlates this data with purine nucleotide levels determined by high-performance liquid chromatography (HPLC) and other in vitro measures of platelet quality. Platelets prepared from pooled ABO- and Rh- identical platelet-rich plasma were stored as concentrates in the synthetic medium without added nutrients (control) and with added substrates.
The HPLC procedure developed for this study permits simultaneous analysis of purine nucleotides, nucleosides, and bases. The separation is performed on a C18 column (end-capped) using 0.1 M phosphate buffer, pH 5.4, containing dibutylamine phosphate (D-4) ion pair reagent and a non-linear, concave gradient to 0.1 M phosphate buffer, pH 5.4, with D-4 reagent in water:acetonitrile (75:25, v/v). The results for metabolic and storage nucleotide pools on day 1 of storage were consistent with previously reported levels in platelets
Keegan, Thais E..
"Platelet Function and Energy Metabolism During Storage in a Well-Defined Experimental Medium"
(1989). Doctor of Philosophy (PhD), dissertation, Chemistry and Biochemistry, Old Dominion University, DOI: 10.25777/0hff-jk73