Date of Award:
Master of Science (MS)
Nutrition, Dietetics, and Food Sciences
Nutrition and Food Sciences
A great deal of variability has been evident in reported pantothenic acid blood values. The purpose of .the study was to establish optimum enzyme treatment for liberation and assay of pantothenic acid in blood and sera.
There were eight major findings . First, it was found that there was an active pantetheinase in sera, negating the need for hog kidney peptidase or pigeon liver enzyme in whole blood. The second finding (related to the first) indicated that there was significant autolysis of the peptidase bond between the pantothenic acid and cysteamine moie- ties of coenzyme A in whole blood. The third finding was that Clarase (a monoesterase) liberated equivalent amounts of pantothenic acid in fresh whole blood when compared to alkaline phosphatase, but less in frozen blood. The optimum time for enzymatic liberation of pantothenic acid from whole blood was determined in the fourth. When using alkaline phosphatase alone, or alkaline phosphatase plus a pantetheinase, maximum liberation of pantothenic acid occurred at 4 to 5 hours. The fifth find-ing was that in sera, acid or base hydrolysis and boiling increased pantothenic acid liberation by approximately lQO nanograms over and above that liberated by enzyme treatment with alkaline phosphatase and hog kidney peptidase. In whole blood, both boiling and acid hydrolysis increased pantothenic acid liberation by approximately 100 nanograms; however, base hydrolysis did not yield a similar increase in pantothenic acid. In the sixth, it was found that substantial autolysis of bound forms of pantothenic acid (to liberate free pantothenic acid) occurred in fresh and frozen whole blood at 37°C and 23°C . The autolysis did not occur in sera or in boiled blood. The results from the seventh finding indicated that blood frozen for 15 months showed less bound (released by alkaline phosphatase plus hog kidney peptidase) pantothenic acid and a greater amount of free pantothenic acid than did fresh blood. The eighth was actually a series of findings in which it was established that a protease digestion of whole blood with trypsin, chymotrypsin , papain , or pepsin increased pantothenic acid liberation from whole blood. Of the four proteases used, trypsin was found to be optimal for pantothenic acid liberation. The pantothenic acid liberated by a pro-tease digestion was released as free pantothenic acid, and was from a different source than that liberated by alkaline phosphatase plus hog kidney peptidase . When assaying trypsin digested whole blood with both the radioimmuno and microbiological assay, the r2 between assays was 0.901 . The source of the pantothenic acid which was liberated by the trypsin digestions is unknown .
Pearson, Jan, "Optimum Enzyme Treatment for Liberation and Assay of Pantothenic Acid in Blood" (1980). All Graduate Theses and Dissertations. 5243.
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