1) The document analyzes the chemical composition of rice bran and rice straw, including their moisture content, carbohydrate, lignin, and cellulose content. 2) Tests were performed to determine the enzyme activity of samples, including CMC-ase activity and activity on filter paper and glucose. 3) The results found that rice bran is high in nutrients but rice straw alone is not sufficient to maintain animal health without supplementation. The analysis provided experience in processing and testing methods.

1. MANIPAL INTERNATIONAL UNIVERSITY
ANALYZING THE COMPONENT OF RICE BRAN & RICE
STRAW TO PRODUCE THE ANIMAL FEEDS
Present by : AMIN RAHMAN BIN SABEER MOHAMED
Matrix no : 1000113
Industrial Supervisor : EN. MOHD FIRKHRY FADHLY BIN ABDUL
GHANI
Academic Supervisor : PROF. DR. AROKIARAJ PAPPUSAMY
Department : INDUSTRIAL BIOTECHNOLOGY RESEARCH CENTRE
(IBRC)

2. COMPANY PROFILE
Sirim Berhad
 Standards and Industrial Research Institute of Malaysia
(SIRIM).
 A corporate organization wholly owned by the Malaysian
Government.
 Main headquarters – Shah Alam, Selangor

3. OBJECTIVES
The objectives of SIRIM are:
1) To innovate and develop processes, product
and technologies for industry.
2) To promote standardization and quality.
3) To provide technical services for industry and
the public.

4. INTRODUCTION
Rice Bran
 A by product of rice milling industry.
 Research by Mohammed et al. (2014) revealed that
rice bran is an incredible source of the:
 Vitamins
 Minerals
 Amino acids
 Essential fatty acids
 Dietary fibre and
 more than 100 antioxidant nutrients
that helps to fight against disease and promote good
health

5. CONT’
Rice Straw
 The vegetative part of the rice plant.
 Cut at grain harvest or after.
 It may be burned and left on the field before the next
ploughing.
 Good source of energy, but is poor in protein (Heuze & Tran,
2013).
 It has low digestibility.
 Low in energy with a crude protein content of only 3–
5 percent.

6. OBJECTIVE
-To analyze the content of rice bran
and rice straw.
-To gain the working experience.

7. TASKS
Moisture Content test
Carbohydrate test
Lignin test
Cellulose test
Enzyme Activity
Enzyme Kinetic

8. METHODOLOGY
Moisture Content test- Oven- Dry method.
Carbohydrate test- Phenol Sulphuric Acid method.
Lignin test- Tappi 222 test method.
Cellulose test- Tappi 203 test method.
Enzyme activity- FP Assay, CMC Assay, Glucose
Enzyme Kinetic- Method of CMC assay (different
concentration of CMC solution)

9. Moisture Content test
1. The Petri dish was weighed before place the
sample.
2. 1 gram of sample was added and incubated at
90°C for 16 hours.
3. The sample was weighed and minus the weight
of original Petri dish.

10. PHENOL SULPHURIC ACID METHOD (TOTAL
CARBOHYDRATE)
 100 mg of sample weighed into boiling tube.
 The boiling tube was kept in boiling water bath for 3
hours with 5 mL of 2.5 NHCL and cooled to room
temperature.
 Solid sodium carbonate was added to neutralized the
mixture.
 The volume was made up to 100 mL and
centrifuged.
 0.2, 0.4, 0.6, 0.8 and 1 mL of working standard were
pipette out into a series of test tube.
 0.1 and 0.2 mL of the sample solution were pipette
out in two separate test tubes. Made up the volume
in each tube to 1 mL with distilled water.

11. CONT’
 The blank was set with 1 mL of distilled water.
 1 mL of phenol solution was added to each tube.
 5 mL of 96% sulphuric acid was added to each tube
and shake well.
 After 10 minutes the content in the test tubes were
shake and placed in a water bath at 25- 30 ̊C for 20
min.
 The color was read at 490 nm.

12. LIGNIN TEST
1 gram of
sample was
weighed
Placed in a
beaker and 15
ml of 72% of
sulfuric acid
was added
The sample
was stirred in
acid until
fully
dissolved.
Transfered into
a schott bottle
and distilled
water was
added until the
total volume
reached 575 ml.
The solution was
autoclaved and
allowed it to
cool
The solution
was filtered
The filtrate
washed using
hot water (3
times)
And dried in an
oven for 2 days at
90°C
The sample placed
in vesicator for 1 day
The dried sample
weighed.

13. ALPHA- CELLULOSE
 25 mL of the filtrate and 10.0 mL of 0.5N
potassium dichromate solution were pipette into
a 250-mL flask.
 Add cautiously, while swirling the flask, 50 mL of
concentrated H₂SO₄.
 Allowed the solution to remain hot for 15 min,
then add 50 mL of water and cool to room
temperature.
 2 to 4 drops of Ferroin indicator were added and
titrated with 0.1N ferrous ammonium sulfate
solution to a purple color.
 Make a blank titration substituting the pulp
filtrate with 12.5 mL of 17.5% NaOH and 12.5
mL of water.

14. BETA- AND GAMMA- CELLULOSE
 50 mL of the pulp filtrate was pipette into a 100-mL graduated
cylinder having a ground glass stopper.
 50 mL of 3N H₂SO₄ was added and mix thoroughly by inverting.
 The cylinder was heat submerged in a hot water bath at about
70-90C for a few minutes to coagulate the beta-cellulose.
 Allowed the precipitate to settle for several hours, preferably
overnight, then decant or filter, if necessary, to obtain a clear
solution.
 50 mL of the clear solution and 10.0 mL of 0.5N K₂Cr₂O₇ was
pipette into a 300-mL flask.
 90 mL of concentrated H2SO4 was cautiously added.
 The solution was allowed to remain hot for 15 min, then proceed
with titration as outlined in alpha cellulose method.
 Made a blank titration substituting the solution with 12.5 mL of
17.5% NaOH, 12.5 mL of water and 25 mL of 3N H₂SO₄.

15. ENZYME ACTIVITY TEST FOR CMC-ASE
Stock dilute
solution
1 ml dilute
sample was
added in test
tube
Add 1 ml of
citrate buffer pH
4.8 and add 1 ml
of CMC
Incubate at 50°C
for 1 hour
Add 3 ml of
DNS
Incubate at
boiling
temperature for
15 minutes
Add 1 ml of
40% potassium
tartarate
Add 20 ml of
Distilled Water
Cool down the
sample in room
temperature
Absorbance at 540 nm using
spectrophotometer.

16. Enzyme activity test for FP- ase method
1. 0.5 mL of diluted enzyme solution was taken into the
test tube.
2. 1 mL 0.1 mM citrate buffer pH 4.8 and 50 mg (1x6
cm strip) of Whatman No. 1 filter paper that has
been curled round a glass rod was added.
3. Test tube was incubated for 1 hour at 50 ̊C.
4. After incubation, 3 mL DNS reagent was added.
5. Then boiled for 15 minutes and 1 mL of 40% sodium
potassium tartarate was added.
6. 20 mL of distilled water was added.
7. After cooling to room temperature, absorbance was
measured at 550 nm.

17. ENZYME ACTIVITY TEST FOR GLUCOSE
METHOD
 10 mM glucose solution were taken and diluted
to final volume of 1 mL.
 In 1 mL standard solution, 2 mL 0.l M citrate
buffer, pH 4.8 was added.
 3 mL DNS reagent was added and boiled for 15
minutes.
 After boiled, 1 mL 40% sodium tartarate was
added in hot tubes.
 20 mL of distilled water was added.
 The tubes were cooled at room temperature and
the absorbance was measured at 550 nm.

18. ENZYME KINETIC
Stock dilute
solution
1 ml dilute
sample was
added in test
tube
1 ml of citrate
buffer pH 4.8
and 1 ml of
different
concentration of
CMC was added.
Incubated at
50°C for 1 hour
3 ml of DNS
was added.
Incubated at
boiling
temperature for
15 minutes
1 ml of 40%
potassium
tartarate was
added.
20 ml of distilled
water was added.
The sample was
cooled at room
temperature
Absorbance at 540 nm using
spectrophotometer.

19. RESULTS

20. MOISTURE CONTENT TEST
Rice Bran Rice Straw
Weight of Petri Dish 94.39 gram 93.62 gram
Sample 1 gram 1 gram
Weight after incubation 95.25 gram 94.52 gram
Difference of weight 0.84 gram 0.90 gram
Moisture % 14% 10%
Moisture% = Weight of original sample – weight of dried sample x 100
weight of original sample

21. RESULT OF LIGNIN TEST
Sample Weigh of
sample
Weight of
Petri Dish
before
incubation
Weight of
Petri Dish
after
incubation
Differences
in Weight
(%)
Rice Bran 1 gram 2.69 3.78 0.09 gram
(9%)
Rice Straw 1 gram 2.74 3.70 0.08 gram
(8%)

22. ALPHA, BETA AND GAMMA CELLULOSE
α- cellulose β- cellulose γ- cellulose
Rice Bran 62.34877666 26.26777657 11.38344693
Rice Straw 59.28866667 25.21842677 9.49290666

23. GLUCOSE IN ENZYME ACTIVITY TEST
0.0909
0.1828
0.9263
1.2463
1.9699
2.4862
y = 0.5045x - 0.6154
R² = 0.9738
-0.5
0
0.5
1
1.5
2
2.5
3
0 1 2 3 4 5 6 7
Absorbance
[Glucose], mg/ml
Abs (550nm)
Abs (550nm)
Linear (Abs (550nm))

24. [CMC HTec],
mg/ml
Abs (540nm)
Standard Factor CMC-ase Activity
Blank 0.4262 - -
1 0.8128 1.23 1.641856
2 0.9263 2.02 1.871126
3 1.2463 2.41 2.517526
4 1.9699 2.02 3.979198
5 2.48862 2.02 5.0270124
[CMC CTec],
mg/ml
Abs (540nm)
Standard Factor CMC-ase Activity
Blank 0.3574 - -
1 1.5441 0.65 2.177181
2 1.8277 1.04 2.577057
3 2.1311 1.41 3.004851
4 2.7854 1.41 3.927414
5 3.7484 1.33 5.285244

25. [Filter Paper Assay HTec], mg/ml
Abs (550nm)
Blank 0.07
1 0.9722
2 1.7059
3 2.0465
4 2.438
5 3.5793
[Filter Paper Assay CTec], mg/ml
Abs (550nm)
Blank 0.0747
1 0.8794
2 1.4484
3 1.5541
4 1.9328
5 2.673

26. [Enzyme Kinetics HTec], mg/ml Abs (540nm)
Blank 0.0875
0.8 0.4899
1 0.6062
1.2 0.8879
1.4 2.1804
1.6 2.6088
1.8 3.1877
2 4.461
[Enzyme Kinetics CTec], mg/ml Abs (540nm)
Blank 0.0904
0.8 0.3191
1 0.9931
1.2 1.5585
1.4 1.7958
1.6 2.3257
1.8 2.9981
2 4.3732

27. CONCLUSION
 Diet of rice straw alone is not sufficient even to
maintain the animals weight unless supplementary
protein is provided. (Food and Agriculture
Organization)
 All of the nutrients in rice bran were significantly
effective and can be useful in food application as
edible oil extraction and food supplement
(Mohammed et al. 2014).
 Learn about the process and method of analysis the
chemical content in sample.
 Know how to use equipment in proper way.