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Clinical Chemistry 31: 1478-1480, 1985;
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Clinical Chemistry, Vol 31, 1478-1480, Copyright © 1985 by American Association for Clinical Chemistry

Fluorescence depolarization assay for quantifying alpha-amylase in serum and urine

M Hofman and M Shaffar

We have developed a new method for quantifying alpha-amylase (EC 3.2.1.1) in serum and urine by fluorescence depolarization. Amylase in the sample catalyzes the hydrolysis of the substrate, a fluorescein- labeled amylose. This results in decreased fluorescence polarization, owing to the increased rate of rotation of the amylose fragment relative to the intact substrate. The TDx amylase assay is calibrated with six human-serum-based pancreatic amylase calibrators. Amylase activities are determined by interpolation from the calibration curve, which is stored in the TDx analyzer's memory. Results correlate well with those by the Du Pont aca assay and the Beckman "DRI-STAT" assay. Endogenous glucose does not interfere. CVs are less than 6%, and the reagents are stable in liquid form.




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Copyright © 1985 by the American Association for Clinical Chemistry.