Date of Award

Fall 1990

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Program/Concentration

Biology

Committee Director

Lloyd Wolfinbarger, Jr.

Committee Member

Mark S. Elliot

Committee Member

Deborah A. Waller

Call Number for Print

Special Collections LD4331.B46 L4

Abstract

The purpose of this study was to assess the effectiveness of various solvents m removal of lipids from human bone, and to determine the effects of lipid extraction on subsequent acid solubilization of the mineral components of bone. Specifically, the following processing steps were studied; 1) ground human bone was de fatted in various ethanol / water mixtures, and / or chloroform / methanol ( 2: 1, v:v ), 2) ground human bone which had lipids extracted ( in absolute ethanol ) and bone which had not had lipids extracted were demineralized using vanous concentrations of hydrochloric acid to assess the value of lipid extraction on subsequent demineralization. For most of the studies, ground bone was placed into columns and the various combinations of solvent systems allowed to flow through the bone matrix while lipid and calcium concentrations were monitored in the eluent solutions. The data demonstrate that chloroform / methanol ( 2:1, v:v ) removes bulk lipid more effectively than various combinations of ethanol / water mixtures. Lipid removal prior to demineralization with HCl improves subsequent demineralization of bone and the rate and extent of demineralization were significantly affected by the concentration of HCl used in the demineralization process. The process of demineralization was monitored using a calcium-specific electrode or by measuring the pH of the eluent solutions. All data were statistically evaluated.

The results indicate that the capacities of solvents for lipid extraction from human bone matrix did not differ significantly among the various solvents tested. However, lipid extraction using column extraction methods and chloroform / methanol mixtures required less volume than absolute ethanol. Generally, lipid extraction of bone prior to demineralization made the calcium more extractable than from non lipid extracted bone. Higher concentrations of HCl solution usually required less time for calcium extraction from bone matrix, although 0.3 N HCl extracted more calcium than 0.5 N HCL The present data indicate that pH values of eluent acidic solutions remain between pH 3.0 and 4.0 while the demineralization process occurs. This value declines to below or near pH 1.0 when demineralization is completed.

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DOI

10.25777/4qvh-kj26

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