Characterization of a spectrophotometric assay for angiotensin converting enzyme.

1981 ◽  
Vol 27 (4) ◽  
pp. 530-534 ◽  
Author(s):  
K W Ryder ◽  
S J Jay ◽  
S A Jackson ◽  
S R Hoke

Abstract Although serum angiotensin converting enzyme (EC 3.4.15.1) activity has generally been shown to be increased in patients with sarcoidosis, considerable variation in the diagnostic usefulness of this test has been reported. We investigated the possibility that this variation may be the result of inhibition of the widely used spectrophotometric assay by various substances. We also prospectively examined the predictive value of measurements of this enzyme in serum from 100 patients being evaluated for sarcoidosis. The following did not significantly affect the results: storage at 4 or 25 degrees C for one week, hemoglobin, lipoproteins, or corticosteroid medications. Bilirubin, at concentrations of 20 mg/L of serum or move, significantly inhibited the assay. Sera with increased activity showed nonlinear reaction rates during the usual 60-min reaction interval, This was corrected by shortening the reaction time to 30 min. We found a predictive value of 88% when the serum angiotensin converting enzyme test was applied to the diagnosis of active sarcoidosis.

1979 ◽  
Vol 25 (7) ◽  
pp. 1259-1262 ◽  
Author(s):  
S G Chiknas

Abstract I describe modification of the spectrophotometric assay described by Cushman and Cheung [biochem. Pharmacol. 20, 1637 (1971)] for serum angiotensin-converting enzyme, with use of "high-pressure" liquid chromatography to measure the hippuric acid end product. After reaction of 10 microL of untreated serum with the angiotensin-converting enzyme substrate analog hippuryl-L-histidyl-L-leucine, the hippuric acid produced is extracted into ethyl acetate and quantitated, relative to an added internal standard, by liquid chromatography. Total chromatographic running time is 3 min per sample, with a within-run CV of 4.3% and a day-to-day CV of 6.6%, for aliquots of plasma supplemented with 1 mmol of hippuric acid per liter. The measurements are linear for hippuric acid in amounts up to 20 nmol per assay. The presence of large quantities of lipid in the serum did not affect the accuracy of the determination.


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