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Clinical Chemistry 30: 707-711, 1984;
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Clinical Chemistry, Vol 30, 707-711, Copyright © 1984 by American Association for Clinical Chemistry

Quantification of pepsin A activity in canine and rat gastric juice with the chromogenic substrate azocoll

PC Will, WE Allbee, CG Witt, RJ Bertko and TS Gaginella

The activity concentration of pepsin may be quantified by using azocoll as a chromogenic substrate. The measured enzyme activity is constant between pH 1.2 and 3.4 and is proportional (r = 0.61) to the activity measured with hemoglobin as substrate. The activity of purified porcine pepsin is inhibited by pepstatin A with an apparent Ki of 115 nmol/L. The azocoll method is useful for measuring changes in pepsin secretion in response to pharmacological agents. For example, pepsin activity of canine gastric juice is decreased by 80% after in vivo administration of 0.5 mg of the synthetic trimethyl prostanoid Ro 22-6923 per kilogram of body weight. The method is sufficiently sensitive to measure the pepsin activity in 0.2 microL of canine gastric juice with a CV of approximately 10%, is simpler than the hemoglobin-substrate methods, and the substrate is commercially available.




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