Evaluation of a glucose oxidase peroxidase method ...



Title Evaluation of a glucose oxidase peroxidase method for indirect measurement of glycogen content in marine mussels (Mytilus edulis)
Author(s) Shelley A. Burton, A. L. MacKenzie, T. Jeffery Davidson, N. MacNair
Journal Journal of Shellfish Research
Date 1997
Volume 16
Issue 2
Start page 435
End page 439
Abstract A colorimetric method (glucose oxidase/peroxidase) for indirect measurement of glycogen concentrations in tissue homogenates of marine mussels (Mytilus edulis) was evaluated. This method uses a conversion of glycogen to glucose by amyloglucosidase. Varying the buffer pH (4.5, 5.0; 5.5) and the amyloglucosidase concentration (160, 80, 40, 20, 10, 5, 1, and 0.5 mg/mL) did not appreciably optimize glycogen concentration. Coefficients of variation (n = 10) for mussel homopenates with mean glycogen concentrations of 94 and 334 mg/dL had within-run values of 0.75 and 0.96%, respectively. The between-run coefficients of variation (n = 10) for the same homogenates were 2.10 and 1.10%, respectively. When mean glycogen concentrations of thawed mussel homogenates were compared with those of initial fresh homogenates, a significantly (p less than or equal to 0.05) lower glycogen concentration was seen in samples thawed after 1 day, but not in samples thawed after 1 h, 1 wk, or 1 me. Glycogen recovery percentages of 99.3, 99.0, and 95.6% were obtained with mixed solutions containing 103.8, 95.2, and 10.8 mg/dL glycogen, respectively. The lower limit of sensitivity for the procedure was approximately 10 mg/dL. Because dilutions of a mussel homogenate with a high glycogen concentration (413.1 mg/dL) gave observed results within 5% of expected results, the assay was considered to be linear to at least 413.1 mg/dL. Glycogen concentrations based on analysis of wet tissue and lyophilized samples from 20 mature mussels were compared, resulting in a significant (p less than or equal to 0.05) correlation coefficient of 0.52. An initial laboratory range (43-91 mg/g) for tissue glycogen based on wet weights (3.9-12.4 g) was determined with 20 mature mussels during July from the Morell region, Prince Edward Island, Canada. It was concluded that the colorimetric assay offered a reliable indication of tissue concentrations of glycogen in marine mussels (M. edulis).
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