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Optimization of the urinary acid alpha-glucosidase determination
Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Fur Klinische Chemie Und Klinische Biochemie
|December 1, 1985
Summary
This study optimized acid alpha-glucosidase determination in human urine using a synthetic substrate. The improved method shows high precision and excellent correlation with existing techniques.
Area of Science:
- Biochemistry
- Clinical Chemistry
- Enzyme Assays
Background:
- Acid alpha-glucosidase is a key enzyme.
- Accurate measurement in human urine is important for diagnostics.
- Existing assay methods may require optimization for precision and reliability.
Purpose of the Study:
- To optimize the assay method for determining acid alpha-glucosidase activity in human urine.
- To investigate the impact of specimen storage and pretreatment on enzyme activity.
- To validate the optimized method using a synthetic substrate.
Main Methods:
- Utilized 4-nitrophenyl-alpha-D-glucopyranoside as a synthetic substrate.
- Investigated optimal conditions for human urine specimen storage and pretreatment.
- Employed gel filtration as part of the analytical procedure.
- Assessed assay precision using within-run coefficients of variation (CV).
- Correlated results with a method using maltose as a substrate.
Main Results:
- The optimized method demonstrated good precision (within-run CV of 7.4% for normal and 3.7% for elevated samples).
- Storage and pretreatment conditions were optimized for reliable enzyme determination.
- An excellent correlation (r = 0.9893) was observed between the optimized method and the maltose-based method.
Conclusions:
- The optimized method for acid alpha-glucosidase determination in human urine is precise and reliable.
- The use of 4-nitrophenyl-alpha-D-glucopyranoside offers a viable alternative for enzyme activity measurement.
- This optimized assay provides a valuable tool for clinical chemistry applications.