7.

3 Chromic Oxide Content (Acid-digestion Method)

Introduction
Chromic oxide is frequently used as an inert reference marker to measure the
digestibility in animal feeds. The method is quite accurate and precise when used in
pelleted feeds.
Principle
The method involves oxidation of organic material in the sample using concentrated
nitric acid followed by the oxidation of the insoluble (green) chromium III in chromic oxide
to soluble (yellow) chromium VI which is then determined spectrophotometrically.
Materials:
Dried (homogeneous) sample
Equipments:
1. Kjeldahl digestion tubes
2. Digester block
3. Pipette (5 mL)
4. Volumetric flask (100 mL)
5. Spectrophotometer (350 nm) and cuvette
Reagents:
1. Concentrated nitric acid
2. Concentrated perchloric acid
Procedure:
1. Weigh accurately (5 d.p.) 50 - 100 mg of sample into a foil.
2. Transfer the sample carefully to a numbered Kjeldahl tube and re-weigh the foil.
Aquaculture Nutrition and Feed Technology Laboratory Manual 22
3. Add 5- 6 mL conc. nitric acid washing the sample down from the side of the tube.
4. Switch on the fume cupboard and switch on the digester to 100 C.
5. Place stand and tubes in the preheated digester.
6. Digest samples for 20 minutes ensuring that it does not boil dry and allow tubes to
cool.
7. Add 3 mL of perchloric acid to the tube with great care **(Perchloric acid can be
explosive).
8. Place stand and tubes back in the preheated digester.
9. Boil the sample until the colour changes from green to yellow, orange or brown and
continue to boil for a further 10 minutes.
10. Allow flask to cool and if the colour reverts to green, go back to "h".
11. Carefully wash the contents into a 100 mL volumetric flask with distilled water and
make up to 100 mL.
12. Transfer an aliquot to a spectrophotometer cuvette and read the absorbance at 350
nm with distilled water as the blank.
Calculation:
Weight of sample = W0 (mg)
Absorbance = Y
Weight of chromic oxide in the sample = = X (mg) 2089 .00032 .0 y
% Chromic oxide in the sample = X/W0 x 100
Reference:
Furukawa, A. and H. Tsukahara. 1966. Bulletin of the Japanese Society of Scientific Fisheries. Vol.
32, No.6, 1966.