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22-90-01 Measurement of Urease Activity

This document outlines a method for measuring urease activity in soybean products. It describes preparing samples, reagents, and procedures for conducting tests and blanks in test tubes in a water bath. The pH difference between the test and blank after 30 minutes is a measure of the urease activity in the sample. Intralaboratory precision is 0.03-0.10 pH units depending on if the sample is cooked or uncooked. Care must be taken to prevent contamination and ensure pH instruments give prompt, stable readings.

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Ng Mun Yee
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288 views2 pages

22-90-01 Measurement of Urease Activity

This document outlines a method for measuring urease activity in soybean products. It describes preparing samples, reagents, and procedures for conducting tests and blanks in test tubes in a water bath. The pH difference between the test and blank after 30 minutes is a measure of the urease activity in the sample. Intralaboratory precision is 0.03-0.10 pH units depending on if the sample is cooked or uncooked. Care must be taken to prevent contamination and ensure pH instruments give prompt, stable readings.

Uploaded by

Ng Mun Yee
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 2

Enzymes AACC International Method 22-90.

01
Page 1 of 2

Measurement of Urease Activity


Final approval May 1, 1969; Reapproval November 3, 1999

Objective
The method is a modified Caskey-Knapp method that yields residual urease
present in soybean products under conditions of the test. It is applicable to soy-
bean meals, soy flour, and millfeeds.

Apparatus
1. Water bath capable of being maintained at 30 ± 0.5°.
2. pH meter equipped with glass and calomel electrodes and with provision
for testing 5 ml of solutions. It should be a precision instrument with sensitivity
of ±0.02 pH units or better, and with temperature compensator.
3. Test tubes, 20 × 150 mm, fitted with rubber stoppers.

Reagents
1. Phosphate buffer solution, 0.05M. Dissolve 3.403 g KH2PO4 in approxi-
mately 100 ml freshly distilled water. Dissolve 4.355 g K2HPO4 in approxi-
mately 100 ml water. Combine the two solutions and make to 1000 ml. Adjust
pH of solution to 7.0 with solution of strong acid or base before using. Useful
life of buffer solution, prepared as described, is less than 90 days.
2. Buffered urea solution. Dissolve 15 g urea in 500 ml phosphate buffer solu-
tion. Add 5 ml toluene to serve as preservative. Adjust pH of urea solution to
7.0 as for reagent 1.

Procedure
1. Grind sample as fine as possible without raising temperature. It is
preferable that at least 60% of sample pass a no. 40 U.S. Standard sieve. Soy
flour requires no further grinding.
2. Weigh 0.200 g of prepared sample into test tube. Add 10 ml buffered urea
solution, stopper, mix, and place in water bath at 30°. Note time. (Test sample.)
3. Prepare blank by weighing 0.200 g of prepared sample into test tube. Add
10 ml phosphate buffer solution, stopper, mix, and place in water bath at 30°.
Note time. (Blank.)
4. Allow 5-min interval between preparing test and blank.
5. In both test and blank runs, mix contents of tubes every 5 min during diges-
tion period, remove tubes from water bath at end of 30 min, and determine pH
of supernatant liquids.
6. Take pH reading at exactly 5 min after removal of tubes from bath. See
Note 1.

Calculation
Record difference in terms of pH between test and blank runs as pH increase
or urease activity.

doi: 10.1094/AACCIntMethod-22-90.01
Enzymes AACC International Method 22-90.01
Page 2 of 2

Measurement of Urease Activity (continued)

Precision
Intralaboratory precision is 0.03 and 0.10 pH units on cooked and uncooked
material, respectively. Interlaboratory precision should be 0.05 and 0.15 pH
units, respectively.

Note
Care must be exercised to prevent contamination of all glassware or elec-
trodes. Should the pH instrument fail to deliver a prompt stable reading, check
into the difficulty. Occasionally trouble is caused by a coating from the soluble
fraction of the soybeans on the fiber in the calomel electrode.

Reference
Caskey, C. D., Jr., and Knapp, F. C. 1944. Method for detecting inadequately heated soybean oil
meal. Ind. Eng. Chem., Anal. Ed. 16:640.

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