HCMC UNIVERSITY OF TECHNOLOGY AND EDUCATION

FACULTY OF CHEMICAL AND FOOD TECHNOLOGY

*******************

REPORT TEMPLATE
PRACTICE OF FOOD ANALYSIS

Lecturer: Phạm thị Hoàn

Group: 3
Student: Ngô Thị Phương An _ 22116001
Huỳnh Lưu Tuyết Minh _ 22116015
Nguyễn Minh Ngọc _ 22116018
Lê Minh Nhật _ 22116019
Class:
Time: 22/2/2024 – 14/3/2024
 LESSON: DETERMINE OF MOISTURE CONTENT

I. Principle
 The gravimetric method determines moisture content based on the decrease in
mass of a sample when heated in an oven for a sufficiently long period of time.
This method offers several advantages, including: low cost, simplicity, high
accuracy.
II. Apparatus and Chemicals
1) Tools and equipment
 Petri dishes and lids
 Desiccators (with dried desiccant)
 Plastic gloves (or tongs)
 Electric stove
 Oven dryer
 Analytical balance

2) Samples and chemicals

 Jinju rice cake

III. Experiments procedure

 Clean the laboratory equipment thoroughly.

 Place the Petri dishes and lids in the oven and dry at 105°C for 30 minutes.
 Remove the Petri dishes and lids from the oven and let them cool in a desiccator to
room temperature.
 Weigh the Petri dishes and lids and record the values.
Petri dish + lid (1): 63,2546g
Petri dish + lid (2): 65,2480g
Petri dish + lid (3): 68,3808g
 Grind the Jinju rice cake sample quickly in a mortar and pestle to prevent moisture
absorption from the environment, which can affect the accuracy of the results.
 Weight Jinju rice cake sample
Sample 1: 4,0057g
Sample 2: 4,0021g
Sample 3: 4,0058g
  Place the Petri dish containing the sample in the oven and close the lid.
 Dry the sample at 105°C for 3 hours.
 After 3 hours, remove the sample from the oven and place it in a desiccator to cool
to room temperature.
 Weigh the Petri dish and sample and record the value.
 Caculate the moisture content.

IV. Result and comment:

1) Result

Mass of Mass of Wet Moisture

Sample Remaining Average
petri dish sample mass Conten
number mass (g) NC (%)
& lid (g) (g) (g) %
1 63,2546 4,0057 67,1341 0,1262 3,15
2 65,2480 4,0021 69,1355 0,1146 2,86 3,02
3 68,3808 4,0058 72,2638 0,1228 3,06

% Moisture content = 100 * ¿ ¿ ¿

[(¿❑ 4,0057+63,2546)−(67,1341)]
1st %% Moisture content = 100 * ❑ ¿ = 3,15 %
[(¿ 4,0057+63,2546)−63,2546 ]¿
[(¿❑ 4,0021+ 65,2480)−(69,1355)]
2nd %% Moisture content = 100 * ❑ ¿ = 2,86 %
[(¿ 4,0021+ 65,2480)−65,2480]¿
[(¿❑ 4,0058+68,3808)−(72,2638)]
3th %% Moisture content = 100 * ¿ = 3,06 %
[(¿❑ 4,0058+68,3808)−68,3808]¿

√
2 2 2
SD = (3 , 15−3 , 02) +(2 , 86−3 , 02) +(3 , 06−3 , 02) = 0,1485
2
0,1485
CV = .100 = 4,71 %
3 , 15
3 ,02−6
% E rel = .100 = -49,66 %
6

2) Comment
 The results are consistent with the initial target.
 The moisture content measurement results are in line with the labeled moisture
content of the Jinju rice cake (≤6%)
 LESSON: DETERMINE OF ASH CONTENT
I. Principle
 Ash content: Ash is the remaining component of food after burning all organic
substances. Ash is the only mineral salt found in food (hence the term ash total)
 The gravimetric method for ash determination is based on the mass of the
remaining white ash after the sample is heated at a high temperature for a long
time to ensure that all organic matter has been completely decomposed.

II. Apparatus and Chemicals

1) Tools and equipment

 Analytical balance
 Dryer
 Crucibles
 Desiccator
 Temperature adjustable furnace

2) Samples and chemicals

 Black bean powder

 HNO3/H2O2

III. Experiments procedure

 Wash crucibles and experimental equipment with distilled water.

 Place the crucible in a furnace. Heat it at 105 oC. Maintain this temperature until
the weight of the crucible stabilizes.
 Remove the crucible from the furnace and let it cool in a desiccator.
 Weigh the cooled crucible on an analytical balance. Record its weight.
 Add approximately 5g of the ground black bean sample to the crucible. Weigh the
crucible and sample. Record their weight.
  Place the crucible and sample in the muffle furnace. Gradually heat them at 500 -
600 °C. Maintain this temperature until the ash turns white (about 6-7 hours).
 Wait for the muffle furnace temperature to drop to around 250 °C.
 Remove the crucible from the muffle furnace and place it in a desiccator to cool to
room temperature.
 Weigh the crucible and ash. Record their weight.

IV. Result and comment:

1) Result
- Formula: X = (G2 – G) x 100 / (G1-G)
G: Crucible weight (g)
G1: Weight of crucible and sample before burning (g)
G2: Weight of crucible and sample after burning (g)
Before burning (G1) After burning (G2) % Ash
Sample number crucible(G) Sample Sum
1 44,2347 3,0278 47,2625 44,3745 4,61%
2 43,4992 3,0422 46,5414 43,6355 4,48%
3 37,7285 3,0161 40,7446 37,8693 4,66%

4 ,61+ 4 , 48+ 4 , 66
Mean = 3
= 4,58

√
2 2 2
SD = (4 , 61−4 ,58) +(4 , 48−4 , 58) +(4 ,66−4 , 58) = 0,0930
2
0,0930
CV = 4 , 58 *100 = 2,0305%
4 ,58−4 ,6
%Erel = 4 ,6
∗100 = -0,4347%
 2) Comment

The experiment results deviate from the true value due to the following reasons:
- Positive error:
+ The sampling procedure involving removal from the desiccator for weighing
causes the sample to absorb moisture in reverse.
- Negative error
+ Sample loss due to handling
Error reducing method:
- Practice improving accuracy in handling.
 LESSON: PROTEIN NITROGEN DETERMINATION
BY KJELDALH METHOD
I. Principle
 Determining nitrogen content in food samples through three stages of digestion,
distillation, and titration
II. Apparatus and Chemicals
1) Tools and equipment

 Kjeldahl system
 Kjeldahl tube
 Micropipette
 Pipette 2 mL, 10 mL
 Erlen 250 mL
 100 mL volumetric flask
 Burette 25 mL
 Becher 100 mL, 250mL

2) Samples and chemicals

 Hung Thinh Fish Sauce

 Concentrated H2SO4
 K2SO4
 CuSO4
 Distilled water
 NaOH 40%,
 H3BO3
 Bromocresol
 Methyl red
 HCl 0,1N

III. Experiments procedure

Digestion:
 Preparing the sample in a digestion vessel by adding 2ml distilled water, 10 ml of
concentrated H2SO4 and catalysts (1g CuSO4, 3g K2SO4) to a test tube
  Pipetting exactly 2 ml of Hưng Thịnh fish sauce using a micropipette into two
tubes used for digestion in the Kjeldahl system.
 Add 10 ml of concentrated H2SO4 and catalysts ( aproximatelly 1g CuSO4, 3g
K2SO4)
 Placing the samples in the digestion block and running the Kjeldahl system in 3
hours
 After the digestion is complete, the sample will become transparent, indicating that
the digestion process is complete

Food + H2SO4 -> (NH4)2SO4 + CO2 + SO2 + H2O ( catalyst: K2SO4, CuSO4 )
Distillation:
 After digestion, take the tube containing the digestion solution and place it in the
distillation system.
 Dissolve 8.0019g of H3BO3 acid and make up to 200 ml. Then add 1.4 ml of
methyl red and 2 ml of bromocresol (the resulting mixture has a deep pink-red
color). Use a pipette to add 30 ml of the mixture to a 500 ml receiving flask and
place it at the outlet of the distillation system.
 Distill for about 30 to 45 minutes with NaOH 30%- 40%.
 Check the outlet of NH3 with litmus paper. If the litmus paper at the outlet of the
tube is still blue, continue distilling. If there is no blue color, the distillation
process is complete (the solution obtained after distillation is neon green).

(NH4)2SO4 + 2NaOH = 2NH3 + Na2SO4 + H2O

NH3 + H3BO4 -> NH4+ + H2BO3-

Titration:
 Add 25 ml of 0.1N HCl solution to a 25 ml buret and titrate with the neon green
solution obtained from the distillation process. To know when the titration is
complete, a drop will cause the solution to change from neon green to light pink.
 Record the volume of HCl used in the buret and calculate the % Nitrogen and %
Protein.

H2BO3- + H+ -> H3BO3

IV. Result and comment:
1) Result

Blank Tube 1 Tube 2

Corrected acid volume

0,2 8,3 8,5
(ml)

Sample (ml) 2 2 2

Formula:
Content (%) of total nitrogen in the sample:
Corrected acid volume 14
%N = NHCL x x x 100
sample 1000

Content (%) of protein in the sample:

%P = %N x 6,25
( 8 ,3−0 , 2 ) 14
%N in tube 1 = 0,1 x x x 100 = 0,567
2 1000

%P in tube 1 = %N x 6,25 = 3,54

( 8 ,5−0 , 2 ) 14
%N in tube 2 = 0,1 x x x 100 = 0,581
2 1000

%P in tube 2 = %N x 6,25 = 3,63

3,5875+3,7187
Mean = = 3,6531
2

√
2 2
SD = (3,5875−3,6531) + ( 3,7187−3,6531 ) = 0,0928
1
0,0928
CV = .100 = 2,5403 %
3,6531
 3,6531−3 , 56
% E rel = .100 = 2,6152 %
3 , 56

2) Comment

 When determining the protein content in a 2ml sample (Hung thinh fish sauce), it
was found that the sample contained approximately 3.6% protein (equivalent to
0.036g of protein).
 After analysis, it was found that the 200ml sample contained approximately 3.6g
of protein, which is consistent with the nutritional information labeled on the
200ml bottle of fish sauce.
 LESSON: DETERMINATION OF SOLUTION PROTEIN
BY BIURET METHOD

I. Principle
 Biuret is an analytical method used to determine soluble protein.
 The Biuret method is based on the formation of a characteristic color complex of
Cu2+ and peptide chains in the control medium. The resulting complex will be
measure by absorption spectroscopy at 540 nm.
 The more protein, the more difference in the color of the complex. The amount of
soluble protein will be evaluated by the standard curve.
II. Apparatus and Chemicals
1) Tools and equipment
 Curvet
 Test tube
 Beaker 500ml
 Volumetric flask 250ml
 Micropipette 01mL
 Vortex
 UV-vis spectrophotometer

2) Samples and chemicals

 Black soybeans solution ( 111,5g)
 CuSO4. 5H2O
 NaKC4H4O6.4 H2O (NatriKali tartrate)
 NaOH 10%
 BSA

III. Experiments procedure

 a. Prepare Biuret reagent
 Add 0.375g CuSO4. 5H2O and 1.51g NaKC4H4O6 dissolved in 100mL of
distilled water and stirred to ensure that the mixture were completely dissolved.
 Then, add 100 mL of 10% NaOH solution to the mixture.
 Finally, transfer to a volumetric flask 250ml then top up to the mark.

b. Prepare the standard curve

 Dilute 5 solution of BSA with 5 different concentration: 2,4,6,8,10 ( mg protein/

mL )

 Put the solutions into the test tube one by one according to the ratio as shown in
the table below.

 Take 1ml of protein solution with 5ml of biuret reagent into a test tube

 Shake well for 15 minutes at room temperature

 Then take the test tubes to measure by absorption spectroscopy at 540 nm.

ĐC 1 2 3 4 5 M1 M2
Dd chuẩn 0 0.2 0.4 0.6 0.8 1 0 0
(BSA )
(mL)
distilled 1 0.8 0.6 0.4 0.2 0 0 0
water
(mL)
Sample 0 0 0 0 0 0 1 1
(mL)
Biuret 5 5 5 5 5 5 5 5
(mL)
Emission 0.112 0.195 0.263 0.319 0.428 0.224 0.235
at 540nm

 Protein Standard Curve
0.45
0.428
0.4 f(x) = 0.378 x + 0.0366
R² = 0.989069865640337
0.35
0.319
0.3
0.263
0.25

0.2 0.195

0.15
0.112
0.1

0.05

0
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1

IV. Result and comment:

1) Result
Equation: y = 0.378x + 0.0366

Sample 1:
Average protein concentration in peptone sample: 0.224 = 0.378x + 0.0366
=> x = 0.4842

We dilute black soybean solution 5 times => 0.4842 x 5 = 2.4212 ( mg/mL )

Dissolve 4g black soybean powder into 111.5mL solution => 2.4212 x 111.5

= 269.9625 (mg protein )

269.9625 x 100
% Protein in sample = = 6.749 %
4000

Sample 2:
Average protein concentration in peptone sample: 0.235 = 0.378x + 0.0366
=> x = 0.5249

We dilute black soybean solution 5 times => 0.5249 x 5 = 2.6243 ( mg/mL )

Dissolve 4g black soybean powder into 111.5mL solution => 2.6243 x 111.5

= 292.614 (mg protein )

292.614 x 100
% Protein in sample = = 7.314 %
4000

6.749+7.314
Mean = = 7.0315
2

( 6.749−7.0315 )2 +(7.314−7.0315)2
SD = √ = 0.2861
1

0.2861
CV = X 100 = 4.07%
7.0315

2) Comment

a/ The cause of the error

_ The test weighing may not be completely exact due to outside natural components or
insulant clean test planning gear.
_ The cuvette may not have been altogether cleaned, or there may be buildups amid
dealing with and transportation.
_ The capacity of the Biuret reagent isn't entirely controlled.

b/ The solution

_ Limiting the fondness of encompassing environment (Clean the test contamination,

weigh test within the encased space)
_ Carefully not to touch the clean confront of the cuvette (avoid unique finger impression
lead to blunder in spectrophotometry.
_ Limiting the contact of the Biuret reagent with discuss and light.
 LESSON: DETERMINATION OF TOTAL LIPID IN SOLID
SAMPLES BY SOXHLET METHOD
I. Principle
 Fat is extracted with organic solvent which is heated and volatilised then
condensed above the sample. Solvent drips onto the sample, soaks it to extract the
fat. Final fat content is measured by weight loss of the sample.

II. Apparatus and Chemicals

1) Tools and equipment
 Bleck beans powder
 Filter paper
 Analytical balance
 Soxhlet extractor

2) Samples and chemicals

 Diethyl ether
 Petroleum ether
III. Experiments procedure

 Weigh 3g of blackbean powder. Then, pack it in filler paper and folded it small

 Place it into the extraction cylinder. Then pour solvent (diethyl ether and
petroleum ether) from the top of the condenser through the extraction cylinder and
finally down into the solvent flask. Turn on the water to cool the condenser and
turn on the heat source to evaporate the solvent. The volatile solvent will be cooled
by a condenser and condenser tube system, flowing down the extraction cylinder
and then the solvent tank. This circulation process will help to extract all the fat in
the sample after about 8 hours. Alternatively, we can determine the stopping point
of the experiment by applying a few drops of solvent from the extraction cylinder
onto the filter paper. When the solvent evaporates without leaving an oil slick, the
experiment is terminated.

 After extraction is complete, remove the sample from the system and dry it in the
Hood cabinet. Then, we proceed to dry the solvent mixed in the thimbles and the
sample during extraction at 102–104 degree Celsius to constant weight. Cool in a
desiccator and weigh (total weight of sample).

IV. Result and comment:

1) Result
a. Calculation
 Paper bag Weight of paper bag
Sample weight and and sample after lipid Initial sample mass
initial sample extraction and drying

1 5.4256 5.3685 3.0136g

2 5.4302 5.3742 3.0135g

3 5.0093 4.9543 3.0209g

Fat content (%)

( MI −M 2)
X= x 100
m
MI: mass of paper bag and initial sample (g)
M2: weight of paper bag and sample after lipid extraction and drying (g)
m: initial sample weight (g)
m: initial sample mass (g)

Test results:

Sample Fat content (%)

1 1.89

2 1.86

3 1.82

1.89+1.86+1.82
Mean = = 1.856
3

( 1.89−1.856 )2 + ( 1.86−1.856 )2 + ( 1.82−1.856 )2

SD = √ = 0.0351
2
 0.0351
CV = X 100 = 1.89 %
1.856
1.856−1.7
% E rel = .100= 0.0917 %
1.7

3) Comment

A . The factors affect the result

Weighing samples is not very accurate because it can be influenced by experimental
Weighing tests isn't exceptionally exact since it can be affected by exploratory
variables or since the sample preparation gear isn't exceptionally clean all through the
weighing prepare.
The sample's dissolvable or dampness cannot be totally dispensed with by drying.

B . Solutions
Reduce the affect of the environment (can weigh tests in a totally contained area),
completely clean test arrangement hardware.
Extricate the sample's fat after protracting the drying handle.

C . Ways to speed up the extraction process.

Dry the sample before extraction to diminish dampness, which is able offer assistance
the dissolvable enter the test more quickly and minimizes blunders.
To get tall extraction proficiency, utilize the suitable dissolvable for the test.
For uniform dissolvable infiltration and fabulous extraction viability, finely smash the
strong fabric.
 LESSON: DETERMINATION OF THE TOTAL LIPID OF LIQUID BY ADAM
ROSE-GOTTLIEB METHOD
I. Principle
 Lipids in coconut milk were extracted using four different solvents: NH3,
C2H5OH, diethyl ether, and petroleum ether.
 NH3 + C2H5OH: This solvent mixture precipitates and removes proteins from the
fat globules.
 Diethyl ether + petroleum ether: This solvent mixture extracts lipids and other fat-
soluble components.

II. Apparatus and Chemicals

1) Tools and equipment
  Analytical balance
 Desiccator
 Separatony funnel
 Temperature adjustable furnace

2) Samples and chemicals

 Yolo coconut milk

 NH3
 C2H5OH
 Diethyl ether
 Petroleum ether
III. Experiments procedure

 Weigh approximately 10g of Yolo coconut milk into a 250ml Erlenmeyer flask.
 Add 1.5ml of NH3 and 10ml of C2H5OH to the Erlenmeyer flask, and shake the
mixture well for 1 minute.
 Add 20ml of diethyl ether to the Erlenmeyer flask, and shake well for 5 minutes.
 Add 20ml of petroleum ether to the Erlenmeyer flask, and shake well for 5
minutes.
 Transfer the mixture to a separatory funnel and let it stand for 30 minutes. The
mixture will separate into two phases:
+ Upper phase: Diethyl ether, petroleum ether, fat.
+ Lower phase: Precipitated protein, C2H5OH, NH3, and other components
in coconut milk.
 Weigh a Petri dish and record the data.
 Use the Petri dish to collect the upper phase after extraction.
 Gently heat the Petri dish containing the upper phase to remove the solvent.
 Place in an oven and dry until constant weight is obtained (about 1 hour).
 Remove the Petri dish from the oven and place it in a desiccator.
 Weigh the Petri dish containing the sample and record the result.

IV. Result and comment:

1) Result
 Coconut milk density: 1,03g/mL
 Formula: TF=(M-m) *100/v
 TF (total fat): The fat content (g/100mL)
M: Weight of petri dish and sample after drying
m: Weight of petri dish
v: The volume of milk for analysis (mL)
Sample number m M sample v TF
1 63,3067 65,3118 10,3014 10,0013 20,0483
2 64,5552 66,5902 10,3173 10,0167 20,3160

20,0483+20,3160
Mean = 2
= 20,1821

√
2 2
SD = (20,0483−20,1821) +(20,3160−20,1821) = 0,1892
1
0,1892
CV = 20,1821 ∗100 = 0,9374%

20,1821−21
%Erel = 21
∗100 = -3,8947%
 2) Comment
The experiment results deviate from the true value due to the following reasons:
- Negative error
+ Not able to transfer the entire sample between containers.
+ The sample weight is insufficient to achieve a volume of 10 ml
Error reducing method:
- Practice improving accuracy in handling.
LAB REPORT : DETERMINATION OF REDUCING SUGAR BY DNS METHOD

I. Principles

The method based on the reducing sugar reduces 3,5-dinitrosalicylic (DNS) acid to 3
amino 5 nitro salicylic acid which is orange-red in alkaline solution

The color intensity is measured by UV-Vis spectrophotometer and converted to a numerical value

Based on the standard curve equation to calculate the concentration of reducing sugar in soft drink 7Up
sample

II. Apparatus and Chemicals

1. Chemicals
- DNS reagent includes:

Sodium hydroxide: 1.6255 gram

Sodium potassium tartrate: 30.0265 gram

3,5-dinitrosalicylic: 1.0072 gram

Distilled water

Stir the mixture completely with a vortex mixer and makeup to the 100mL mark of
volumetric flask.

- Standard glucose solution 0.1%

Stir completely 1g glucose and makeup to the 100mL mark of the volumetric
flask.
 - Distilled water

2. Tools and equipment

- Test tubes

- Beaker 100ml, 250ml

- Pipette 5 mL, 10 mL

- Micropipette

- Glass Funnel

- Volumetric Flask 100ml

- Cuvet

- Vortex mixer

- Spectrophotometer

III. Experiment procedure

1. Decarbonate: place the sample into Becher and use the glass rod to stir until no
observable carbon dioxide bubbles appear
2. Dilute 1000 times the decarbonated sample: use a micropipette to take 1000 μL
sample into volumetric flask and make up to 100mL mark
3. Prepare test tubes: Add sample solution and reagents into 7 test tubes (labeled)
following the table below.
 Test tube Blank 1 2 3 4 5 6 7
Sample Sample
1 2
Glucose solution 0 0.2 0.4 0.6 0.8 1 0 0
0.1% (mL)
Sample solution 0 0 0 0 0 0 1 1
(mL)
Distilled water 3.0 2.8 2.6 2.4 2.2 2 2 2
(mL)
DNS reagent 1 1 1 1 1 1 1 1
(mL)

4. Put the test tubes into the water bath and boil for 5 minutes, then cool down to
room temperature
5. Take samples into cuvettes and observe the absorbance spectra 540nm. Zero the
spectrophotometer with the blank
6. Record the result
7. Draw the standard curve according to data collected from the experiment and
determine the concentration of reducing sugar.

IV. Calculation and discussion

1. Result

Sample Sample
Test tube Blank 1 2 3 4 5
1 2
Glucose 0,1% 0 0,2 0,4 0,6 0,8 1,0 0 0
 (mg/mL)
OD540nm 0 0,134 0,192 0,231 0,281 0,323 0,219 0,223

Glucose standard curve (OD540nm)

2. Calculation

Amount of reducing sugar in the sample by the equation

y=2,335 x +0,0921

0,219+0,223
Average absorbance: =0,221
2

Average glucose concentration in sample: 0,221=2,335 x +0,0921 → x=0,0552(mg/mL)

Glucose content in the initial sample: 0,0552 ×1000=55,2034(mg/mL) = 5,5204

(g/100mL)

√
2 2
Standard deviation (SD): ( 0 ,219−0 , 221 ) +(0,223−0,221) =0,0028284
2−1
 0,0028284
Coefficient of variation (CV) = ×100 %=1,2798 %
0,1286

E|¿| 5 ,5204−6 , 8936

Erel = ¿= =−0 ,1992
T 6 ,8936
E|¿|
%Erel = × 1 00 %=−19 , 92% ¿
T
Comments
Conclusion on result and comparison with information on packaging

- Total carbohydrate in 7Up sample: 5,5204g/100mL ~5,5204%

- Total carbohydrate on nutrition facts on packaging: 16,2g/235ml ~6,8936%
 There is a little error between the experiment result and information of total
carbohydrate on nutrition facts

Cause of error:

- Error in measuring equipment

- Making mistakes while weighing

Suggestion to minimize errors

- Regularly check the accuracy of experimental equipment (precision scale,

micropipette)
- Take the chemicals carefully
 LESSON: DETERMINATION OF TOTAL CARBOHYDRATE BY
PHENOL-SULFURIC ACID METHOD

I. Principle

 Carbohydrates (simple sugars, oligosaccharides, polysaccharides, and their

derivatives) react in the presence of strong acid and heat to generate furan
derivatives that condense with phenol to form stable yellow-gold compounds
 The color intensity is measured by UV-Vis spectrophotometer and converted to a
numerical value

II. Apparatus and Chemicals

1) Tools and equipment

 Cuvettes

 Erlenmeyer flask, 100 ml, for dd water

 Becher

 Micropipette with plastic tips

 Test tubes

 Test tube rack

 Volumetric flasks, 100 ml

 Pipette, 5 ml, 10ml

 Spectrophotometer
3) Samples and chemicals

 7Up sample
 Standard glucose solution
 Phenol 8%
 Concentrated sulfuric acid

III. Experiments procedure

1. Decarbonate: place the sample into Becher and use the glass rod to stir until no
observable carbon dioxide bubbles appear
2. Dilute 1000 times the decarbonated sample: use a micropipette to take 1000 μL
sample into volumetric flask and make up to 100mL mark
3. Prepare test tubes: Add sample solution and reagents into 9 test tubes (labelled)
following the table below. Test tube 1-6 are used to collect data for drawing the
standard curve, and test tubes 7-9 contain diluted sample 3 times repeat

Test tube 1 2 3 4 5 6 7-9

Standard glucose solution
0 0,1 0,2 0,3 0,4 0,5 0
(mL)
Distilled water (mL) 0,5 0,4 0,3 0,2 0,1 0 0
Diluted sample (mL) 0 0 0 0 0 0 0,5
Phenol 5% (mL) 0,5 0,5 0,5 0,5 0,5 0,5 0,5
Concentrated H2SO4 (mL) 2,5 2,5 2,5 2,5 2,5 2,5 2,5

4. Shake well all the test tubes after the addition of samples and reagents
5. Put all the test tubes on the test tube rack and chill for 10 minutes
6. Take samples into 9 cuvettes and observe the absorbance spectra 490nm. Zero the
spectrophotometer with the blank
7. Record the result
 8. Draw the standard curve according to data collected from the experiment and
determine the concentration of total carbohydrates.

IV. Result and comment:

1) Result
Test tube 1 2 3 4 5 6 7 8 9
Glucose 0,1%
0 0,1 0,2 0,3 0,4 0,5 0 0 0
(mg/mL)
OD540nm 0 0,061 0,073 0,106 0,141 0,190 0,151 0,157 0,165

2) Calculation

Amount of reducing sugar in the sample by the equation: y=3,4914 x+ 0,0077

0.151+0.1 57+ 0,165

Average absorbance = = 0.1576
3

Average glucose concentration in sample: 0,1576=3,4914 x +0,0077 → x=0,04293(mg/mL)

Glucose content in the initial sample: 0,04293 ×1000=42 , 93(mg/mL)=¿4,293(g/100mL)

 √
2 2 2
Standard deviation (SD): ( 0,151−0,1576 ) +(0,157−0,1576) +(0,165−0,1576) =0,007024
3−1

0,007024
Coefficient of variation (CV): ×100=4,457 %
0,1576

E|¿| 3 ,359−6 , 8936

Erel = ¿= =−0,3772
T 6 ,8936
E|¿|
%Erel = ¿ x 100% = -37,72%
T

2) Comment
a. Comparison to data labeled on packaging

Total carbohydrate in 7Up sample: 3,359g/100mL ~3,359%

Total carbohydrate on nutrition facts on packaging: 16,2g/235ml ~6,8936%

 There is a large error in the experiment result compared to the information on the
label.

b. The cause of the error

 Improper use of experimental equipment (use of micropipette) causing

inconsistency

 The cuvette may not be clear, affecting the absorbance result

 Making mistakes when titration phenol, glucose solution

 Error in measuring equipment

c. The solution
 Clean the cuvette before using and use gloves while holding
 Read the experimental equipment usage instructions carefully
 Regularly check the accuracy of experimental equipment (precision scale,
micropipette)