Date of Award

Fall 1991

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

Program/Concentration

Chemistry

Committee Director

Patricia A. Pleban

Committee Member

Mark S. Elliot

Committee Member

Myuang H. Kim

Committee Member

Roy L. Williams

Call Number for Print

Special Collections LD4331.C45C48

Abstract

The objective of this research was to develop a method for the separation of different types of collagen using a large pore size (100 nm) reversed-phase C-8 column. Three different types of collagen (type I, III, and V) were sufficiently separated on the column using a mobile phase containing water-acetonitrile-trifluoroacetic acid. Collagens found in each HPLC peak were identified using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to assay the column effluents.

Trifluoroacetic acid (TFA) was used as an ion pair reagent. This compound improved the chromatographic profile for these proteins on a reversed-phase column. It was observed that with higher TFA concentration, the collagens eluted more quickly.

Type I and type III collagen were also isolated from placental tissue both by DEAE-anion exchange cellulose column and by salt precipitation. The isolated proteins were applied to the reversed phase HPLC column and were detected using SDSPAGE.

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DOI

10.25777/vn89-jq66

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