Use of purified amyloglucosidase for the specific determination of total carbohydrate content of rat liver homogenate in a single step
Johnson, J.A.; Fusaro, R.M.
Analytical Biochemistry 98(1): 47-52
1979
ISSN/ISBN: 0003-2697 PMID: 120688 Document Number: 149636
In prior studies, a crude amyloglucosidase preparation, in conjunction with glucose oxidase reagent, was used to determine total carbohydrate in liver and muscle homogenates by a 2-step procedure. Glycogen content was determined by subtracting tissue glucose (determined separately). Using a purified amyloglucosidase, the accuracy of two-step assay of rat liver homogenate with crude amyloglucosidase was verified. The reliability of glucose oxidase detection of glucose in amyloglucosidase hydrolysates was established by comparing results with those obtained with hexokinase/glucose-6-phosphate dehydrogenase reagent. The availability of purified amyloglucosidase made it feasible to assay total carbohydrate content of homogenate in a single step, by incubating aliquots in the presence of both amyloglucosidase and glucose oxidase reagents. Results obtained with this 1-step assay agreed with those of 2-step analysis. This is the 1st report of direct enzymic assay of total carbohydrate in rat liver homogenate, in a single step.