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L/2/10

VIOLETTA DZIEDZIEJKO

SEPARATION, IDENTIFICATION AND CHARACTERIZATION OF ALKALINE
PHOSPHATASE ISOENZYMES IN RATS USING HIGH-PERFORMANCE LIQUID
CHROMATOGRAPHY (HPLC) AND OTHER ANALYTICAL METHODS


Summary
The aim of this work was to develop a separation method, determine organ specificity of alkaline phosphatase (ALP) isoenzymes in rat serum, compare rat ALP isoenzymes with their human counterparts, and assess the usefulness of some analytical methods studied for the specific measurement of ALP isoenzyme and isoform activities in rats. The study material included sera and samples of the intestine, bones and liver of rats, as well as human sera and tissue-specific ALP standards. Chromatography with post-column reaction detection enabled identification in rat sera of three isoforms of intestinal ALP and one bone isoform of non-specific ALP isoenzyme. Electrophoresis disclosed some macromolecular forms of the enzyme. Separation was complemented with non-separation methods to determine organ source of serum ALP isoforms. The properties of rat and human bone isoforms differ little whether studied in serum or bone. Detailed understanding of properties and function of the highly heterogenic ALP enzyme in man and animals should be based on a combination of many contemporary analytical methods.

K e y w o r d s : alkaline phosphatase – isoenzymes - high-performance liquid chromatography (HPLC) – agarose electrophoresis – rats.

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