NUTRITION

BIOCHEMISTRY
LABORATORY

EXPERIMENT 4:

DETERMINATION OF
URINARY IODINE
CONCENTRATION

Instructor Bui Ngoc Yen Tram

WEDNESDAY, 23 TH OCTOBER, 2019

GROUP 3:
1. Nguyễn Hữu Anh Minh – BTBCIU16081
2. Lê Thiện Thanh Đan– BTBCIU16040
3. Võ Minh Hạnh– BTBCIU16083
4. Hoang Nguyen Minh Anh– BTBCIU16008

1
I) Principle of the test
In this experiment, iodine concentration in urine was determined by Sandell–Kolthoff
reaction. The principle of this method is using iodide as a catalyst in the reduction of
ceric ammonium sulfate (yellow) to the colorless cereus form in the presence of
arsenious acid. Before performing the test, the urine must be rid of any interfering
substances by digestion with ammonium persulfate. The absorbance is then measure
at 420nm.

II) Method:
Firstly , preparing the urine and types of reagents. After that, iodine standard curve will
be made first. Diluting the iodine solution with seven different concentration 0 ,
50,100,150,300,500 and 1000µg/l. Putting 250 µl from seven concentration for each
test tube . Each concentration will be used two test tube Next , the urine was added in
new test tube about 250 µl for each test tube. Liking the iodine solution each urine was
used two test tube. 1 ml 1.0 M ammonium persulfate will be added for each test tube (
note using the silver paper to cover test tube). Taking all twenty- two test tube to the
water path and put it into this about 60 minutes at 100oC .After 60 minutes , those test
tube was taken out and cooled to room temperature. Arsenious acid solution was
added in each test tube with 2.5 ml per test tube. Waiting for 15 minutes and ceric
ammonium sulfate solution was added in each test tube about 300 µl ( note each test
tube will be added after 30 seconds compared with the previous ones). After ceric
ammonium was added completely , all test tube were taken nearly the UV-vis
spectrophotometer. At exactly 30 minutes from stop watch all solution in test tube will
be tested from 1 to 22 test tube. Each test tube will be test after 30 seconds compared
to the previous. Recorded the absorbance for making the standard curve as well as
calculating the amount of iodine in each urine in the report.