UNIVERSITI KUALA LUMPUR
MALAYSIAN INSTITUTE OF CHEMICAL & BIOENGINEERING
TECHNOLOGY
LABORATORY TECHNICAL REPORT
SUBMISSION FORM
To: Dr Khairul Faizal Bin Pa’ee Code Subject: CFB 20703
From: Student ID. No.:
1. Emelya Natra bt Mohd Khayry 55218118193
2. Hanim Amira bt Hairol Annuar 55218118195
3. Mastura bt Abdullah 55218118130
No. of Group: LO1-P1 (2) Date of Experiment: 22/8/2019
Title of Experiment: CALORIMETRIC ANALYSIS OF REDUCING SUGAR USING
DINITROSALICYLIC (DNS) ASSAY
Received by: Date of Submission: 29/8/2019
Note: Late submission will not be accepted.
*To be filled by the marker*
VERY VERY
POOR GOOD EXCELLENT
CRITERIA POOR GOOD
2 3 5
1 4
1.0 ABSTRACT & OBJECTIVES (HALF PAGE 2 4 6 8 10
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) (TOTAL: 3 6 9 12 15
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
Abstract & Objective(s):
The dinitrosalicylic acid (DNS) method gives a rapid and simple estimation of the extent of
saccharification by measuring the total amount of reducing sugars in the hydrolysate and allows the
analysis of a relatively high number of samples. However, 3,5-dinitrosalicylic acid (DNS) reduced to
3-amino,5-nitrosalicylic acid under alkaline conditions because oxygen can be dissolved in
interfering with glucose oxidation, sulphite, which itself is not necessary for the colour reaction that
added in the reagent to absorb the dissolved oxygen. In addition, the type of side reaction depends
on the exact nature of the reducing sugars. When different reducing sugars, generally the different
colour intensities occur, thus, it is necessary to calibrate for each sugar. Based on the result obtained
in this experiment, when the amount of concentration is higher, the value of absorbance also
increases. The data was interpreted in the graph that shows the absorbance was directly
proportional to the concentration where more molecules passes through the light.
Objectives
1. To test this method for the presence of free carbonyl group (C=O).
Methodology:
Using a suitable flowchart, state the steps involve in this lab work.
Serial dilution with concentration of 20, 40, 60, 80 and 100 ppm were prepared
while the blank solution was prepared by pipette 1.5 ml of DNS reagent and
1.5ml of distilled water (0.05 g of DNS in 500 ml distilled water for all group).
1.5ml of blank solution were added in 10 test tube, sample 1 (Mirinda), sample
(Mountain dew), 5 serial dilution with different concentration and 2 unknown
solutions. Then, 1.5 ml of DNS reagent were added into each test tube.
All the test tube was boiled for 5 minutes and was cooled in the water. After that,
10ml of distilled water were pipetted into all the test tube.
The reaction mixture was analyzed after the preparation is done by using UV-Vis
Spectrophotometer to determine the value of R with absorbance at 540nm.
Result:
Wavelength λ: 540nm
Table 1: Result of photometric scan
No Concentration (ppm) Absorbance
1 20.000 0.093
2 40.000 0.086
3 60.000 0.102
4 80.000 0.106
5 100.000 0.118
Unknown 1 37.475 0.093
Unknown 2 59.319 0.101
Sample 1 (Mirinda) 2.528 2.5282
Sample 2 (Mountain Dew) 2.671 2.6710
Absorbance vs Concentration (ppm)
0.12
0.115 R² = 0.8113
0.11
Absorbance
0.105
0.1
0.095
0.09
0.085
0.08
0 20 40 60 80 100
Concentration, ppm
Graph 1: Graph of Absorbance against Concentration (ppm)
Discussion
The objective of this experiment was to test the presence of free carbonyl group called
reducing sugars. This experiment also involving the oxidation of the aldehyde functional group such
as glucose and ketone functional groups in fructose. Then, under the alkaline condition, the
aldehyde group will be reduced to carboxyl group such as 3,5-dinitrosalicylic acid reduced to 3-
amino,5-nitrosalicylic acid. Different reducing sugars generally yield different colour intensities. By
enhancing the intensity of developed colour, carboxymethyl cellulose can affect the calibration
curve. The amount of sugar in unknown sample given were determined by spectrophotometric
method which also called as colorimetric method. This method is used to measure the absorbance
value but not the concentration of substance. Besides, to determine the concentration of the
substance in a solution is by using standard curve which shows the absorbance values into
concentration.
The procedure of this experiment was prepared the stock solution of glucose for standard
solution 20ppm, 40ppm, 60ppm, 80ppm and 100ppm, unknown solution 1, unknown solution 2,
sample 1 (Mirinda) and sample 2 (mountain Dew). Next, the solutions were mixed with DNS in
different test tubes. Then, the solution was heated for 5 minutes and distilled water was added after
the solution cool. The stock solution of glucose for standard solution used was 500g/ml. Based on
the Graph 1, the absorbance was directly proportional to concentration. This graph show that the
higher the concentration, the higher the absorbance value. The coefficient of determination in the
linearity was 0.8113. The result shows the absorbance value at 20 ppm, 40 ppm, 60 ppm, 80 ppm
and 100ppm which was 0.093, 0.086, 0.102, 0.106 and 0.118 respectively. Based on result above,
the absorbance value at the concentration 40 ppm decrease because the solution was not rinse
properly before measure the absorbance.
Based on the result obtained, the absorbance for the unknown solution 1 and unknown
solution 2 was 0.093 and 0.101 respectively. Next, the absorbance for sample 1 which is Mirinda
and sample 2 which is Mountain Dew was 2.528 and 2.671 respectively. This shows that unknown
solution 2 and sample 2 has higher sugar content than unknown 1 and sample 1.
Conclusion
By the end of this experiment, can be concluded that higher amount of concentration, the
value of absorbance also increases. This law states that the absorbance of a light material is
proportional to its concentration in solution. This relationship is expressed by the Lambert-Beer Law,
which is commonly known as Beer’s Law. The absorbance is based primarily on those three factors
which are molar absorptivity constant, path length and concentration. Based on the graph obtain,
the absorbance was directly proportional to the concentration. If the concentration of solution is
increased, then there are more molecules for the light to hit when it passes through. As the
concentration increase, there are more molecules in the solution, and more light is blocked. This
causes the solution to get darker because less light can be through.
References:
1. Miller, G.L., Use of dinitrosalicylic acid reagent for determination of reducing sugar, Anal.
Chem., 31, 426, 1959.
2. Wang.N.S. Glucose Assay By Dinitrosalicylic Calorimeteic Method. Accessed on 25/8/2019.
Retrieved at: https://user.eng.umd.edu/~nsw/ench485/lab4a.htm
3. Real Tech Talk (April 17, 2018), What is The Relationship Between Absorbance and
Concentration. Retrieved from http://realtechwater.com/blog-post/what-is-the-relationship-
between-absorbance-and-concentration/
4. S. Suzanne Nielsen, Food Analysis Third Edition. Retrieved from:
https://books.google.com.my/books?id=8S1QLUsDOwgC&lpg=PA150&dq=what%20does
%20dinitrosalicylic%20acid%20do&pg=PA150#v=onepage&q=what%20does%20dinitrosal
icylic%20acid%20do&f=false
APPENDICES
Figure 1: Standard solution and Samples before undergo boiling process
Figure 2: UV-Vis Spectrophotometer
UniKL MICET
Psychomotor performance rubric
Course code/name : Food Chemistry (CFB 20703)
Experiment title: CALORIMETRIC ANALYSIS OF REDUCING SUGAR USING DINITROSALICYLIC (DNS) ASSAY
Student 's name: Emelya Natra bt Mohd Khayry
Date : 29/8/2018
Advanced Proficient Developing
No Criteria (5 points) (3 points) (1 points) Points
Successfully completes experiment Successfully completes experiment Cannot completes tasks and
procedures independently. procedures with minimal supervision. standard procedures
Successfully performs
1
experiments without supervision. Shows excellent understanding Shows good understanding of Show no understanding of
of procedure and theory taught. experiments procedure and theory experiments procedure and
taught theory taught.
Ability to organise, performs Practices safely, can work Practices most procedures safely Fails to notice important
experiments independently and take initiative as conforms to the lab regulations with information and safety
2
safely and aware of priorities in well as cooperating effectively in a minimal cooperating effectively in a factors in the workplace.
the laboratory. team. team.
Ability to show engagement in Shows excellent performance Shows good performance with confident, Performs with little energy,
conducting with unusual energy, is very focused, energy and commitment. focus and no commitment
3
experiment. shows confidence and full needs support.
commitment.
Ability to demonstrates Always demonstrates respect and Good demonstration of respect and No demonstration of respect
4 care and respect in equipment care for equipment. care for equipment. and care for equipment.
set-up.
Total points
UniKL MICET
Psychomotor performance rubric
Course code/name : Food Chemistry (CFB 20703)
Experiment title: CALORIMETRIC ANALYSIS OF REDUCING SUGAR USING DINITROSALICYLIC (DNS) ASSAY
Student 's name: Hanim Amira bt Hairol Annuar
Date : 29/8/2018
Advanced Proficient Developing
No Criteria (5 points) (3 points) (1 points) Points
Successfully completes experiment Successfully completes experiment Cannot completes tasks and
procedures independently. procedures with minimal supervision. standard procedures
Successfully performs
1
experiments without supervision. Shows excellent understanding Shows good understanding of Show no understanding of
of procedure and theory taught. experiments procedure and theory experiments procedure and
taught theory taught.
Ability to organise, performs Practices safely, can work Practices most procedures safely Fails to notice important
experiments independently and take initiative as conforms to the lab regulations with information and safety
2
safely and aware of priorities in well as cooperating effectively in a minimal cooperating effectively in a factors in the workplace.
the laboratory. team. team.
Ability to show engagement in Shows excellent performance Shows good performance with confident, Performs with little energy,
conducting with unusual energy, is very focused, energy and commitment. focus and no commitment
3
experiment. shows confidence and full needs support.
commitment.
Ability to demonstrates Always demonstrates respect and Good demonstration of respect and No demonstration of respect
4 care and respect in equipment care for equipment. care for equipment. and care for equipment.
set-up.
Total points
UniKL MICET
Psychomotor performance rubric
Course code/name : Food Chemistry (CFB 20703)
Experiment title: CALORIMETRIC ANALYSIS OF REDUCING SUGAR USING DINITROSALICYLIC (DNS) ASSAY
Student 's name: Mastura bt Abdullah
Date : 29/8/2018
Advanced Proficient Developing
No Criteria (5 points) (3 points) (1 points) Points
Successfully completes experiment Successfully completes experiment Cannot completes tasks and
procedures independently. procedures with minimal supervision. standard procedures
Successfully performs
1
experiments without supervision. Shows excellent understanding Shows good understanding of Show no understanding of
of procedure and theory taught. experiments procedure and theory experiments procedure and
taught theory taught.
Ability to organise, performs Practices safely, can work Practices most procedures safely Fails to notice important
experiments independently and take initiative as conforms to the lab regulations with information and safety
2
safely and aware of priorities in well as cooperating effectively in a minimal cooperating effectively in a factors in the workplace.
the laboratory. team. team.
Ability to show engagement in Shows excellent performance Shows good performance with confident, Performs with little energy,
conducting with unusual energy, is very focused, energy and commitment. focus and no commitment
3
experiment. shows confidence and full needs support.
commitment.
Ability to demonstrates Always demonstrates respect and Good demonstration of respect and No demonstration of respect
4 care and respect in equipment care for equipment. care for equipment. and care for equipment.
set-up.
Total points
