UNIVERSITI KUALA LUMPUR

MALAYSIAN INSTITUTE OF CHEMICAL & BIOENGINEERING

TECHNOLOGY
LABORATORY TECHNICAL REPORT
SUBMISSION FORM

To: Dr. Khairul Faizal Bin Pa’ee Code Subject: CFB 31203
From: Student ID. No.:
1. NUR SYAFIQAH BINTI AMINUDIN 1. 55218220043
2. MOHD ADAM BIN MOHD SOFIAN 2. 55218220016
3. HELMY ISKANDAR BIN AZHAR 3. 55218220046
No. of Group: 1 Date of Experiment: 29th November 2021
Title of Experiment: Crude Protein Determination- Kjedahl Method

Received by: Dr. Khairul Faizal Bin Pa’ee Date of Submission: 26th December 2021

Note: Late submission will not be accepted.

*To be filled by the marker*

VERY VERY
POOR GOOD EXCELLEN
CRITERIA POOR GOOD
2 3 T5
1 4
1.0 ABSTRACT & OBJECTIVES (HALF 2 4 6 8 10
PAGE ONLY) (TOTAL: 10%)
1. State the summary to the experiment
conducted.
2. State the objectives of the experiment
(point form)
2.0 PROCEDURES (TOTAL: 5%) 1 2 3 4 5
1.Methodology is presented in suitable and
understandable flowchart.
3.0 RESULTS (TOTAL: 10%) 2 4 6 8 10
1.Data are presented as deemed suitable with
complete label and units in tables and/or graphs.

4.0 DISCUSSIONS (MAXIMUM 1 PAGE) 3 6 9 12 15

(TOTAL: 15%)
1. Explanations of the referred tables and/or
graphs are presented after it.
2. Discuss on the findings and relations to the
theory and objective of experiment.
5.0 CONCLUSIONS (TOTAL: 5%) 1 2 3 4 5
1. Summary of the results to relate the findings or
results with the theory applicable to
the experiment.
6.0 REFERENCES (TOTAL: 5%) 1 2 3 4 5
1. Minimum of 4 references.

TOTAL MARKS
 Lab Technical Report
Course code/name

1.0 Abstract & Objective(s):

1.1 Abstract

The purpose of this experiment was to determine the amount of crude protein in a cream cracker
using the Kjeldahl method and compare the acquired result to previously published data on the protein
content of cream crackers. In this experiment, 2g of crushed cream crackers were used as a sample to
analyse the protein content, and the sample was prepared in triplicate. Based on the result of the
experiment, the %protein in the Hup Seng cream cracker was 2.7802
± 0.0608 or 2.7194% to 2.841%, and the coefficient of variation was 2.1869%. However, the result
acquired was less than those obtained in the previous study, with the percentage of protein in a
cream cracker were around 7-8%. This is most likely due to a systemic error in the usage of contaminated
equipment, and the Kjeldahl method has its drawbacks because it is an indirect protein measurement
method that measures the amount of nitrogen rather than the amount of protein.

1.2 Objectives

The objectives of this experiment were:

 To determine the amount of crude protein in a cream cracker using the Kjeldahl method.
 To compare the acquired result to previously published data on the protein content of cream
crackers.
2.0 Methodology:

A. Digestion

2 g of sample is weighed
with padded on the filter The sample is inserted into
The digestion block is
Three kjeltabs (1000 kjeltabs The digestion is started after
paper. a digestion tube along with
preheated to 420 The tubefilter
rack is removed and
Cu/ 3.5) and 12 ml of H2SO4 are placing the digestion tube with paper.
°C.
B. Distillation (The sample was prepared left the has been digested
carefully added into the the racks into the digestion
digestion tube. in triplicate with one sample cools for 20 minutes,
block. The process takes 75
blank.) and it turns from clear green
(Shake to wet the sample with minutes and is completed
25ml of 4% boric acid (containing bromocresol greeninto
andclear
methyl
bluered indicator) is added to a 250ml o
solution.
acid.) when the sample turns into a
The samples are diluted with 80 ml deionized water.
clear green solution. The digestion tube is attached to the rubber holder, and th

The analyzer button is pressed. The distillation

50 ml
process
of 40%will
NaOHtakeis5pumped
minutes.into the tubeThe
andprocess
steam is
is completed
created in the
when
steam
the receiver
generator.
solution turns

C. Titration

The distillate is titrated with

The volume of 0.1N HCl is
0.1N HCl until the color of the recorded as Vsample when
distillate change from light
the color of distillate
green to slight pinkish. changes.
3.0
Table 3.1 Sample Information

Sample Name Hup Seng Ping Pong Cream Crackers

Company Hup Seng Perusahaan Makanan (M) Sdn. Bhd.
Sample Description The sample has a creamy, buttery taste and a crunchy texture.

Ingredients include Wheat Flour, Vegetable Oil (Palm Oil), Corn Starch,
Glucose Syrup, Leavening Agent (Ammonium Bicarbonate), Sugar, Full
Cream Milk Powder, Salt, Yeast.

Table 3.2 Test Data

Sample Data Replicate 1 Replicate 2 Replicate 3

Sample Weight, A (g) 2.0003 2.0084 2.0054
Amount HCl used, Vsample (ml) 6.3 6.6 6.5
Amount HCl used, Vblank (ml) 0.1 0.1 0.1
Normality of HCl, Nacid (N) 0.1 0.1

Table 3.3 Standard Deviation calculation data

Replicates Protein (%) Deviation from the Square (Xi-X)2

mean, (Xi-X)
1 2.7138 -0.0664 0.0044
2 2.8331 0.0529 0.0028
3 2.7938 0.0136 0.0002
Total 8.3407 ∑Xi-X= 0.0001 (∑Xi-X)2= 0.0074

Table 3.4 Test Results

Test Data Replicate 1 Replicate 2 Replicate 3

𝑉𝑠𝑎𝑚𝑝𝑙𝑒 − 𝑉𝑏𝑙𝑎𝑛𝑘 × 1.4007 × 𝑁𝑎𝑐𝑖𝑑 0.4342 0.4533 0.4470
%𝑁
𝐴
% 𝑃𝑟𝑜𝑡𝑒𝑖𝑛 = (𝑓𝑥 %𝑁) 2.7138 2.8331 2.7938
Mean of protein 2.7802
Standard deviation 2.7802 ± 0.0608
Coefficient of Variation, CV (%) 2.1869
4.0
There are various methods to determine the protein content in food. The type of sample determines
the method because different methods are based on different analytical principles. Mæhre et al. (2018)
stated that those methods could be classified into direct and indirect protein determination. The direct
approach calculates the protein using amino acid residues, whereas indirect protein determination uses
nitrogen content or chemical reactions with functional groups in the protein to figure out the protein
content.

The Kjeldahl method is one of the examples of indirect protein determination. It is usually used in
industries because it is recognised by the Association of Official Agricultural Chemists (AOAC)
International as the official method for food protein determination Mæhre et al. (2018). The Kjeldahl
procedure involves three steps: digestion, where the organic nitrogen is converted into ammonium, NH +;
distillation or neutralisation,
4
where the ammonia, NH is distilled and retained in a receiver vessel; and
titration, where the nitrogen is determined (Goulding et al., 2020). This method releases the nitrogen at a
high temperature, 420°C, into a strong acid, ammonium sulphate (NH4)2SO4, and the content was
measured after the neutralisation and titration. The volume of HCl used to change the colour of the
distillate from light green to slightly pinkish is used to calculate the amount of nitrogen by the formula
below:

𝑉𝑠𝑎𝑚𝑝𝑙𝑒 − 𝑉𝑏𝑙𝑎𝑛𝑘 × 1.4007 ×

%𝑁
𝑁𝑎𝑐𝑖𝑑
𝐴

Once the nitrogen concentration has been calculated, the crude protein content can be calculated by
multiplying the total nitrogen by the conversion factor, commonly used for all types of food, 6.25 as
formula followed:

% 𝑃𝑟𝑜𝑡𝑒𝑖𝑛 = (𝑓𝑥 %𝑁)

Conversion factor 6.25 is based on the assumption that the general nitrogen content in food proteins is
16% and that all nitrogen in foods is protein-bound.

The purpose of this experiment was to determine the amount of crude protein in a cream cracker
using the Kjeldahl method and compare the acquired result to previously published data on the protein
content of cream crackers. In order to achieve the objectives, 2g of crushed cream crackers were used as a
sample to analyse the protein content. The sample was made in triplicate to acquire the standard deviation
and coefficient of variation values, which were used to determine the precision of the data obtained. Based
on table 3.4, the results obtained for the protein percentage of Hup Seng cream cracker was 2.7802 ±
0.0608 or 2.7194% to 2.841%. Meanwhile, the coefficient of variation, CV, for the data obtained from
those three replicates of the samples was 2.1869% which is considered as acceptable because 2.1869% is
less than 5%. Therefore, it shows a high level of precision or repeatability for all three replicates.
However,
based on calculations from Hup Seng - Cream Crackers Calories, Carbs & Nutrition Facts (n.d.), the
percentage of protein content in 1 sachet of Hup Seng cream cracker is 7%. This calculation is reliable
because a study from Sykes & Davidson (2020) claimed that the protein content for cream crackers is in
the range of 7-8%.

This implies that the results acquired from this experiment are lower than those obtained from the
previous study. This is most likely due to a systemic error in the usage of contaminated equipment. In
addition, the Kjeldahl method has its drawbacks because it is an indirect method of protein measurement
that measures the amount of nitrogen rather than the amount of protein (Mæhre et al., 2018). As a
recommendation, the Kjeldahl method can be replaced with the amino acid analysis method as it is a direct
protein determination method. It is also the only protein analysis method whose results will not be
disturbed by contamination of other substances (Mæhre et al., 2018).

5.0 Conclusion:

The experiment was conducted to determine the amount of crude protein in a cream cracker using
the Kjeldahl method and compare the acquired result to previously published data on the protein content of
cream crackers. In this experiment, the protein determination was conducted by using the Kjeldahl method
and Hup Seng cream cracker as the sample. The sample was prepared in three replicates in order to
increase the accuracy and precision level. The result obtained for the %protein in the Hup Seng cream
cracker was 2.7802 ± 0.0608 or 2.7194% to 2.841%, and the coefficient of variation was 2.1869%.
However, the result acquired was less than those obtained in the previous study, with the percentage of
protein in cream cracker in between 7-8%. In conclusion, the results of this experiment indicate that the
Kjeldahl method can be used to estimate the %protein in cream cracker precisely since its CV value is less
than 5%. Nevertheless, it is not accurate in terms of accuracy compared to past research from journals
because the %protein value is found to be lower than in prior studies. This is due to the fact that the
Kjeldahl method has its drawbacks because it is an indirect method of protein measurement that measures
the amount of nitrogen rather than the amount of protein (Mæhre et al., 2018). As a recommendation, the
Kjeldahl method can be replaced with the amino acid analysis method as it is a direct protein
determination method. It is also the only protein analysis method whose results will not be disturbed by
contamination of other substances (Mæhre et al., 2018).
6.0 References:

Goulding, D. A., Fox, P. F., & O’Mahony, J. A. (2020). Milk proteins: An overview (pp. 21–98).

Elsevier. http://dx.doi.org/10.1016/b978-0-12-815251-5.00002-5

Hup Seng - Cream Crackers calories, carbs & nutrition facts. (n.d.). MyFitnessPal. Retrieved

December 26, 2021, from https://www.myfitnesspal.com/food/calories/cream-crackers-

1651777581

Mæhre, H., Dalheim, L., Edvinsen, G., Elvevoll, E., & Jensen, I.-J. (2018). Protein determination—

method matters. Foods, 7(1), 1–3. MDPI.

https://doi.org/10.3390/foods7010005

Sykes, G. B., & Davidson, I. (2020). Crackers. In Biscuit, Cookie and Cracker Process and Recipes

(pp. 1–31). Elsevier. http://dx.doi.org/10.1016/b978-0-12-820598-3.00001-9

 UniKL MICET
Psychomotor performance

Course code/name : CHEMICAL FOOD ANALYSIS (CFD 31203)

Experiment title: CRUDE PROTEIN DETERMINATION- KJEDAHL METHOD
Student 's name: NUR SYAFIQAH BINTI AMINUDIN
Date : 29th November 2021

Advanced Proficient Developing

No Criteria (5 points) (3 points) (1 points) Points
Successfully completes experiment Successfully completes Cannot completes tasks and
procedures independently. experiment procedures with standard procedures
Successfully performs minimal supervision.
1 experiments without
Shows excellent understanding Shows good understanding of Show no understanding of
supervision.
of procedure and theory taught. experiments procedure and experiments procedure and
theory taught theory taught.
Ability to organise, performs Practices safely, can work independently Practices most procedures Fails to notice important
experiments and take initiative as safely conforms to the lab information and safety
safely and aware of priorities in well as cooperating effectively in a regulations with factors in the workplace.
2
the laboratory. team. minimal cooperating effectively
in a
team.
Ability to show engagement in Shows excellent performance Shows good performance with Performs with little energy,
conducting with unusual energy, is very focused, confident, energy and focus and no commitment
3
experiment. shows confidence and full commitment. commitment. needs support.

Ability to demonstrates Always demonstrates respect and Good demonstration of respect No demonstration of respect
4 care and respect in equipment care for equipment. and and care for equipment.
set-up. care for equipment.

Total points
 UniKL MICET
Psychomotor performance

Course code/name : CHEMICAL FOOD ANALYSIS (CFD 31203)

Experiment title: CRUDE PROTEIN DETERMINATION- KJEDAHL METHOD
Student 's name: HELMY ISKANDAR BIN AZHAR
Date : 29th November 2021

Advanced Proficient Developing

No Criteria (5 points) (3 points) (1 points) Points
Successfully completes experiment Successfully completes Cannot completes tasks and
procedures independently. experiment procedures with standard procedures
Successfully performs minimal supervision.
1 experiments without
Shows excellent understanding Shows good understanding of Show no understanding of
supervision.
of procedure and theory taught. experiments procedure and experiments procedure and
theory taught theory taught.
Ability to organise, performs Practices safely, can work independently Practices most procedures Fails to notice important
experiments and take initiative as safely conforms to the lab information and safety
safely and aware of priorities in well as cooperating effectively in a regulations with factors in the workplace.
2
the laboratory. team. minimal cooperating effectively
in a
team.
Ability to show engagement in Shows excellent performance Shows good performance with Performs with little energy,
conducting with unusual energy, is very focused, confident, energy and focus and no commitment
3
experiment. shows confidence and full commitment. commitment. needs support.

Ability to demonstrates Always demonstrates respect and Good demonstration of respect No demonstration of respect
4 care and respect in equipment care for equipment. and and care for equipment.
set-up. care for equipment.

Total points
 UniKL MICET
Psychomotor performance

Course code/name : CHEMICAL FOOD ANALYSIS (CFD 31203)

Experiment title: CRUDE PROTEIN DETERMINATION- KJEDAHL METHOD
Student 's name: MOHD ADAM BIN MOHD SOFIAN
Date : 29th November 2021

Advanced Proficient Developing

No Criteria (5 points) (3 points) (1 points) Points
Successfully completes experiment Successfully completes Cannot completes tasks and
procedures independently. experiment procedures with standard procedures
Successfully performs minimal supervision.
1 experiments without
Shows excellent understanding Shows good understanding of Show no understanding of
supervision.
of procedure and theory taught. experiments procedure and experiments procedure and
theory taught theory taught.
Ability to organise, performs Practices safely, can work independently Practices most procedures Fails to notice important
experiments and take initiative as safely conforms to the lab information and safety
safely and aware of priorities in well as cooperating effectively in a regulations with factors in the workplace.
2
the laboratory. team. minimal cooperating effectively
in a
team.
Ability to show engagement in Shows excellent performance Shows good performance with Performs with little energy,
conducting with unusual energy, is very focused, confident, energy and focus and no commitment
3
experiment. shows confidence and full commitment. commitment. needs support.

Ability to demonstrates Always demonstrates respect and Good demonstration of respect No demonstration of respect
4 care and respect in equipment care for equipment. and and care for equipment.
set-up. care for equipment.

Total points