April 19, 2020

Experiment No. 5

Determination of Phosphate in Detergent Samples: A Molecular Spectroscopy Application

Members:
Butac, Jazzlei Nyah

Comia, Josiah Jarenee

Gojar, Xyrill

Objectives:

1. To familiarize and learn the operation of UV-VIS spectrometer through online videos.
2. To apply data analysis on the raw data given and also apply spectroscopic calculations to
determine the amount of phosphate in the sample given by the raw data.

Flow Diagram

Sample Preparation: Hydrolysis of detergent.

Weigh 0.500g sample Add two or more Add 15 mL of 2M Add boiling chips
and place it to 100 ml drops of antifoaming and attach reflux
erlenmeyer flask. agent H2SO4 into the flask. condenser into it.

Add one more drop

of antifoaming agent Cool the mixture and Add distilled water in
Reflux the mixture
and dilute to the filter it into 125 mL order to keep the
for 30 mins
mixture with distilled volumetrc flask. volume constant.
water
Sample Preparation: Sample color development via Molybdenum blue method.

Transfer 5 mL of the Add 1 mL of

prepared solution into a Transfer 5 mL of this molybedate solution
100 mL volumetric solution into a 10mL and 0.4 mL reducing
flask and dilute with volumetric flask solution. Use
distilled water graduated pipet

Remove the cover and Cover the volumetric Dilute the

allow the solution to flask and mix the
stand for 20 mins in the solution by repeated mixture to the
dark. inversions mark.

Determination of Phosphate Content in sample using Standard Calibration Method

Dissolve the KH2PO4 in Prepare six standards in

Prepare 0.0250 g of 50mL solution, 0.25
500 mL of 0.1% H2SO4. ppm, 0.50 ppm, 1.00
dried KH2PO4 This will serve as your ppm, 2.00ppm, 4.00ppm
stock solution (50ppm) and 8.00ppm.

Pipet 5 mL of the
Mix and let it stand for Add 0.4 mL of reducing solution to a beaker and
20 minutes. solution add 1 mL of molybdate
solution

Using the UV-VIS Using the UV-VIS Repeat the process for
instrument, measure the instrument, measure the the other concentration
absorbance of the absorbance of the and do three readings
standards. solution prepared in A2. per concentration.
Determination of Phosphate Content in sample using Constant Volume Standard Addition
Method

Pepare the following solutios Allow the solution to

in a 25 mL volumetric flask. stand for 20 mins in the
(See table 1) dark

Measure the absorbance

using the UV-VIS
instrument

Table 1. Preparation of Solutions for Constant Volume Standard Addition Method

Volume (mL)
Solution
Molybdate
Sample Standard Reducing Agent
Solution
1 10 0 2 1
2 10 2 2 1
3 10 4 2 1
4 10 6 2 1
5 10 8 2 1
6 10 10 2 1
Data Entries

PART 1: Standard Calibration Method

Table 1.1. Standard Calibration Method    
Reading
Standard Concentration, ppm
R1 R2 R3
1 0.25 0.019 0.019 0.019
2 0.50 0.139 0.139 0.139
3 1.00 0.200 0.200 0.200
4 2.00 0.360 0.360 0.360
5 4.00 0.898 0.898 0.898
6 8.00 1.706 1.706 1.706

Table 1.2. Concentration and Absorbance (SCM)

Standard Concentration, ppm Average Reading

1 0.25 0.019
2 0.50 0.139
3 1.00 0.200
4 2.00 0.360
5 4.00 0.898
6 8.00 1.706

Concentration V Absorbance
1.800
1.600 f(x) = 0.22 x − 0.01
R² = 1
1.400
1.200
Absrobance

1.000
0.800
0.600
0.400
0.200
0.000
0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00
Concentration, ppm

Figure 1. Standard Calibration Method

PART 2: UV - VIS
 Table 2.1. Sample Readings using UV-VIS  
Sample Readings Trial 1 Trial 2 Trial 3
1 0.033 0.035 0.035
Bar 2 0.033 0.035 0.035
3 0.034 0.036 0.036
1 0.013 0.017 0.014
Liquid 2 0.013 0.017 0.014
3 0.014 0.016 0.015
1 0.020 0.021 0.021
Powder 2 0.021 0.022 0.022
3 0.020 0.021 0.021
XTable 2.2. Average Absorbance Readings per Trial
Sample Trial 1 Trial 2 Trial 3
Bar 0.0333 0.0353 0.0353
Liquid 0.0133 0.0167 0.0143
Powder 0.0203 0.0213 0.0213

XTable 2.3. Mass of Sample

Table 2.4. Concentration of each sample per Trial,
Sample Mass, g
ppm
Bar 0.987
Liquid 1.034 Sample Trial 1 Trial 2 Trial 3
Powder 0.914 Bar 0.2201 0.2294 0.2294
Liquid 0.1277 0.1431 0.1323
Powder 0.1600 0.1647 0.1647

Table 2.5. Concentration of PO4 per sample, per

Trial, ppm Table 2.6. Weight of PO4 per sample, per Trial, g
Sample Trial 1 Trial 2 Trial 3 Sample Trial 1 Trial 2 Trial 3
Bar 5.5029 5.7340 5.7340 Bar 1.38E-03 1.43E-03 1.43E-03
Liquid 3.1924 3.5775 3.3079 Liquid 7.98E-04 8.94E-04 8.27E-04
4.0011 4.1166 4.1166 Powder 1.00E-03 1.03E-03 1.03E-03
Powder
Table 2.7. %Weight of Phosphate per trial
Sample Trial 1 Trial 2 Trial 3
Bar 0.139% 0.145% 0.145%
Liquid 0.077% 0.086% 0.080%
Powder 0.109% 0.113% 0.113%
Table 2.8. Descriptive Statistics for %Weight of Phosphate per sample    
Sample Mean SD n df C.L. tstat C.I.
Bar 0.1433% 3.38E-05 3 2 0.95 4.303 8.39E-05
Liquid 0.0812% 4.78E-05 3 2 0.95 4.303 1.19E-04
Powder 0.1115% 1.82E-05 3 2 0.95 4.303 4.53E-05

PART 3: SAM Liquid

Table 3.1. Constant Volume (25mL) Readings for Liquid Detergent Sample using 4ppm standard

Volume of Standard, Trial 1 Trial 2 Trial 3

mL
R1 R2 R3 R1 R2 R3 R1 R2 R3
0.00 0.0052 0.0052 0.0052 0.0053
0 0.005 5 0.005 5 5 5 0.00533 3 0.00533
0.01 0.0136 0.0136 0.0136 0.0138
2 0.013 3 0.013 5 5 5 0.01385 5 0.01385
0.02 0.0210 0.0210 0.0210 0.0213
4 0.020 0 0.020 0 0 0 0.02132 2 0.02132
0.03 0.0315 0.0315 0.0315 0.0319
6 0.030 0 0.030 0 0 0 0.03197 7 0.03197
0.04 0.0462 0.0462 0.0451 0.0468
8 0.044 4 0.043 0 0 5 0.04689 9 0.04583
0.05 0.0546 0.0546 0.0546 0.0554
10 0.052 2 0.052 0 0 0 0.05542 2 0.05542

Table 3.2. Average reading per volume of standard, mL

Volume of Standard, mL Trial 1 Trial 2 Trial 3
0   0.005 0.00525 0.00533
2 0.013 0.01365 0.01385
4 0.020 0.02100 0.02132
6 0.030 0.03150 0.03197
8 0.044 0.04585 0.04654
10   0.052 0.05460 0.05542

Trial 1
0.06
0.05
f(x) = 0 x + 0
Absorbance

0.04 R² = 0.99
0.03
0.02
0.01
0
0 2 4 6 8 10 12
VStd
 Figure 2.1. Constant Volume Reading for Liquid Detergents (Trial 1)

Trial 2
0.06
0.05 f(x) = 0.01 x + 0
R² = 0.99
0.04
Absorbance

0.03
0.02
0.01
0
0 2 4 6 8 10 12
VStd

Figure 2.2. Constant Volume Reading for Liquid Detergents (Trial 2)

Trial 3
0.06
0.05 f(x) = 0.01 x + 0
0.04 R² = 0.99
Absrobance

0.03
0.02
0.01
0
0 2 4 6 8 10 12
VStd

Figure 2.3. Constant Volume Reading for Liquid Detergents (Trial 3)

Table 3.3. Concentration of Sample per trial, ppm

Table 3.4. Mass of
Volume of Sample
Trial 1 Trial 2 Trial 3
Standard, mL
Sample Mass, g
0   0.2664029 0.2664029 0.2664029 Bar 0.987
2 Liquid 1.034
4 Powder 0.914
6
 8
10  

Table 3.5.Concentration of PO4 per trial, ppm

Volume of Standard, mL Trial 1 Trial 2 Trial 3

0  
2
4
16.6501813 16.6501813 16.6501813
6
8
10  

XTable 3.6.Weight of PO4 per trial, g

Volume of Standard, mL Trial 1 Trial 2 Trial 3

0  
2
4
0.00416255 0.00416255 0.00416255
6
8
10  

PART 4: SAM Powder

Table 4.1. Constant Volume (25 mL) Readings for Powder Detergent Sample using
4 ppm standard      
Trial 1 Trial 2 Trial 3
Volume of Standard, mL
R1 R2 R3 R1 R2 R3 R1 R2 R3
0 0.008 0.007 0.008 0.00840 0.00735 0.00840 0.00853 0.00746 0.00853
2 0.017 0.018 0.017 0.01785 0.01890 0.01785 0.01812 0.01918 0.01812
 4 0.033 0.033 0.034 0.03465 0.03465 0.03570 0.03517 0.03517 0.03624
6 0.045 0.046 0.045 0.04725 0.04830 0.04725 0.04796 0.04902 0.04796
8 0.053 0.053 0.053 0.05565 0.05565 0.05565 0.05648 0.05648 0.05648
10 0.066 0.066 0.065 0.06930 0.06930 0.06825 0.07034 0.07034 0.06927

Table 4.2. Average reading per volume of standard, mL

Volume of Standard, mL Trial 1 Trial 2 Trial 3

0   0.008 0.00805 0.00817

2 0.017 0.01820 0.01847
4 0.033 0.03500 0.03553
6 0.045 0.04760 0.04831
8 0.053 0.05565 0.05648
10   0.066 0.06895 0.06998

Trial 1
0.07
0.06 f(x) = 0.01 x + 0.01
R² = 0.99
0.05
Absrobance

0.04
0.03
0.02
0.01
0
0 2 4 6 8 10 12
VStd

Figure 3.1. Constant Volume Readings for Powdered Detergent (Trial 1)

 Trial 2
0.08
0.07
f(x) = 0.01 x + 0.01
0.06 R² = 0.99
Absorbance

0.05
0.04
0.03
0.02
0.01
0
0 2 4 6 8 10 12
VStd

Figure 3.2. Constant Volume Readings for Powdered Detergent (Trial 2)

Trial 3
0.08
0.07
f(x) = 0.01 x + 0.01
0.06 R² = 0.99
Absorbance

0.05
0.04
0.03
0.02
0.01
0
0 2 4 6 8 10 12
VStd

Figure 3.3. Constant Volume Readings for Powdered Detergent (Trial 3)

Table 4.3. Concentration of Sample per trial, ppm

Table 4.4. Mass of
Volume of Sample
Trial 1 Trial 2 Trial 3
Standard, mL
Sample Mass, g
0   Bar 0.987
2 Liquid 1.034
4 Powder 0.914
0.53681065 0.53681065 0.53681065
6
8
10  
 Table 4.5.Concentration of PO4 per trial, ppm

Volume of Standard, mL Trial 1 Trial 2 Trial 3

0  
2
4
33.5506656 33.5506656 33.5506656
6
8
10  

Table 4.6.Weight of PO4 per trial, g

Volume of Standard, mL Trial 1 Trial 2 Trial 3
0  
2
4
0.00838767 0.00838767 0.00838767
6
8
10  

Table 4.7. %Weight of Phosphate per trial, and Descriptive Statistics

  Trial 1 Trial 2 Trial 3
%Wt 0.9177% 0.9177% 0.9177%
Mean 0.9176878%
SD 2.45327E-18
n 3
df 2
C.L. 0.95
tstat 4.303
C.I. 1.34558E-18

PART 5: SAM Bar

Table 5.1. Constant Volume (25 mL) Readings for Bar Detergent Sample using 4
ppm standard      
Volume of Standard, Trial 1 Trial 2 Trial 3
mL
R1 R2 R3 R1 R2 R3 R1 R2 R3

0 0.01100 0.01100 0.01200 0.01155 0.01155 0.01260 0.01172 0.01172 0.01279

2 0.01700 0.01800 0.01800 0.01785 0.01890 0.01890 0.01812 0.01918 0.01918

4 0.02400 0.02400 0.02400 0.02520 0.02520 0.02520 0.02558 0.02558 0.02558

6 0.03200 0.03100 0.03200 0.03360 0.03255 0.03360 0.03410 0.03304 0.03410
8 0.03900 0.03900 0.03900 0.04095 0.04095 0.04095 0.04156 0.04156 0.04156

10 0.05000 0.04800 0.04900 0.05250 0.05040 0.05145 0.05329 0.05116 0.05222

Table 5.2. Average reading per volume of standard, mL

Volume of Standard, mL Trial 1 Trial 2 Trial 3
0   0.011 0.01190 0.01208
2 0.018 0.01855 0.01883
4 0.024 0.02520 0.02558
6 0.032 0.03325 0.03375
8 0.039 0.04095 0.04156
10   0.049 0.05145 0.05222

Trial 1
0.06

0.05
f(x) = 0 x + 0.01
0.04 R² = 0.99
Absorbance

0.03

0.02

0.01

0
0 2 4 6 8 10 12
VStd

Figure 4.1. Constant Volume Readings for Bar Detergent (Trial 1)

 Trial 2
0.06

0.05
f(x) = 0 x + 0.01
0.04 R² = 0.99
Absorbance

0.03

0.02

0.01

0
0 2 4 6 8 10 12
VStd

Figure 4.2. Constant Volume Readings for Bar Detergent (Trial 2)

Trial 3
0.06

0.05
f(x) = 0 x + 0.01
0.04 R² = 0.99
Absorbance

0.03

0.02

0.01

0
0 2 4 6 8 10 12
VStd

Figure 4.3. Constant Volume Readings for Bar Detergent (Trial 3)

Table 5.3. Concentration of Sample per trial, ppm

Table 5.4. Mass of
Volume of
Trial 1 Trial 2 Trial 3 Sample
Standard, mL
Sample Mass, g
0   Bar 0.987
2 Liquid 1.034
4 Powder 0.914
1.0991453 1.0991453 1.0991453
6
8
10  
Table 5.5.Concentration of PO4 per trial, ppm

Volume of Standard, mL Trial 1 Trial 2 Trial 3

0  
2
4
68.6965812 68.6965812 68.6965812
6
8
10  

Table 5.6.Weight of PO4 per trial, g

Volume of Standard, mL Trial 1 Trial 2 Trial 3

0  
2
4
0.01717415 0.01717415 0.01717415
6
8
10  

Table 5.7. %Weight of Phosphate per trial, and Descriptive Statistics

Trial 1 Trial 2 Trial 3
%Weight 1.7400% 1.7400% 1.7400%
Mean 1.7400350%
SD 0
n 3
df 2
C.L. 0.95
tstat 4.303
C.I. 0
PART 6.1: TRIAL COMPARISON (Standard Calibration Method)
Table 6.1.1. t-Test between Bar Trials
  Trial 1 vs Trial 2 Trial 1 vs Trial 3 Trial 2 vs Trial 3
0.0353333
Mean 0.03333333 0.03533333 0.03333333 0.03533333 3 0.03533333
Variance 3.3333E-07 3.3333E-07 3.3333E-07 3.3333E-07 3.3333E-07 3.3333E-07
Pearson Correlation 1 1 1
df 2 2 2
t Stat 65535 65535 #DIV/0!
P(T<=t) one-tail #NUM! #NUM! #DIV/0!
t Critical one-tail 2.91998558 2.91998558 #DIV/0!
P(T<=t) two-tail #NUM! #NUM! #DIV/0!
t Critical two-tail 4.30265273   4.30265273   #DIV/0!  

Table 6.1.2. t-Test between Liquid Trials

  Trial 1 vs Trial 2 Trial 1 vs Trial 3 Trial 2 vs Trial 3
0.0133333 0.0166666 0.0133333 0.0143333 0.0166666 0.0143333
Mean 3 7 3 3 7 3
3.3333E- 3.3333E- 3.3333E- 3.3333E- 3.3333E- 3.3333E-
Variance 07 07 07 07 07 07
Pearson Correlation -1 1 -1
df 2 2 2
t Stat -5 #DIV/0! 3.5
0.0188747 0.0364136
P(T<=t) one-tail 8 #DIV/0! 8
2.9199855 2.9199855
t Critical one-tail 8 #DIV/0! 8
0.0377495 0.0728273
P(T<=t) two-tail 5 #DIV/0! 5
4.3026527 4.3026527
t Critical two-tail 3   #DIV/0!   3  

Table 6.1.3. t-Test between Powder Trials

  Trial 1 vs Trial 2 Trial 1 vs Trial 3 Trial 2 vs Trial 3
Mean 0.02033333 0.02133333 0.02033333 0.02133333 0.02133333 0.02133333
Variance 3.3333E-07 3.3333E-07 3.3333E-07 3.3333E-07 3.3333E-07 3.3333E-07
Pearson
Correlation 1 1 1
df 2 2 2
t Stat #DIV/0! #DIV/0! #DIV/0!
 P(T<=t) one-tail #DIV/0! #DIV/0! #DIV/0!
t Critical one-tail #DIV/0! #DIV/0! #DIV/0!
P(T<=t) two-tail #DIV/0! #DIV/0! #DIV/0!
t Critical two-tail #DIV/0!   #DIV/0!   #DIV/0!  
PART 6.2: TRIAL COMPARISON (Standard Addition Method)
Table 6.4. t-Test between Bar Trials
  Trial 1 vs Trial 2 Trial 1 vs Trial 3 Trial 2 vs Trial 3
Mean 0.02877778 0.03021667 0.02877778 0.03066992 0.03021667 0.03066992
Variance 0.00019447 0.00021441 0.00019447 0.00022089 0.00021441 0.00022089
Pearson Correlation 1 1 1
df 5 5 5
t Stat -5.0547773 -5.0547773 -5.0547773
P(T<=t) one-tail 0.00195867 0.00195867 0.00195867
t Critical one-tail 2.01504837 2.01504837 2.01504837
P(T<=t) two-tail 0.00391735 0.00391735 0.00391735
t Critical two-tail 2.57058184   2.57058184   2.57058184  

Table 6.5. t-Test between Liquid Trials

  Trial 1 vs Trial 2 Trial 1 vs Trial 3 Trial 2 vs Trial 3
Mean 0.02727778 0.02864167 0.02727778 0.02907129 0.02864167 0.02907129
Variance 0.00032806 0.00036169 0.00032806 0.00037262 0.00036169 0.00037262
Pearson Correlation 1 1 1
df 5 5 5
t Stat -3.6889775 -3.6889775 -3.6889775
P(T<=t) one-tail 0.00708034 0.00708034 0.00708034
t Critical one-tail 2.01504837 2.01504837 2.01504837
P(T<=t) two-tail 0.01416068 0.01416068 0.01416068
t Critical two-tail 2.57058184   2.57058184   2.57058184  

Table 6.6. t-Test between Powder Trials

  Trial 1 vs Trial 2 Trial 1 vs Trial 3 Trial 2 vs Trial 3
Mean 0.02727778 0.02864167 0.02727778 0.02907129 0.02864167 0.02907129
Variance 0.00032806 0.00036169 0.00032806 0.00037262 0.00036169 0.00037262
Pearson Correlation 1 1 1
df 5 5 5
t Stat -3.6889775 -3.6889775 -3.6889775
P(T<=t) one-tail 0.00708034 0.00708034 0.00708034
 t Critical one-tail 2.01504837 2.01504837 2.01504837
P(T<=t) two-tail 0.01416068 0.01416068 0.01416068
t Critical two-tail 2.57058184   2.57058184   2.57058184  

PART 7: Method Comparison

Table 7.1. t-Test between method on Bar
  SCM vs SAM
Mean 0.00143287 0.01740035
Variance 1.1417E-09 0
Observations 3 3
Pearson Correlation #DIV/0!
Hypothesized Mean Difference 0
df 2
t Stat -818.50641
P(T<=t) one-tail 7.4632E-07
t Critical one-tail 2.91998558
P(T<=t) two-tail 1.4926E-06
t Critical two-tail 4.30265273  

Table 7.2. t-Test between method on Liquid  

  SCM vs SAM
Mean 0.0008122 0.00402567
Variance 2.2828E-09 1.5422E-35
Observations 3 3
Pearson Correlation -0.7245031
Hypothesized Mean Difference 0
df 2
t Stat -116.49337
P(T<=t) one-tail 3.684E-05
t Critical one-tail 2.91998558
P(T<=t) two-tail 7.368E-05
t Critical two-tail 4.30265273  

Table 7.3. t-Test between method on Powder

  SCM vs SAM
Mean 0.00111545 0.00917688
Variance 3.3284E-10 6.0185E-36
Observations 3 3
Pearson Correlation -0.4082483
Hypothesized Mean Difference 0
df 2
t Stat -765.34368
P(T<=t) one-tail 8.536E-07
 t Critical one-tail 2.91998558
P(T<=t) two-tail 1.7072E-06
t Critical two-tail 4.30265273  
Sample Calculations:
Table 1

Mean=
∑ x = 0.019+0.019+ 0.019 =0.01 9
N 3

Diagram for Post Run Calculations

0.900 - 1.100 g of sample + 15

mL 2M H2SO4

250 mL

5.00 mL

100 mL

5 mL + 1 mL Molybdate Solution
+ 0.4 mL Reducing Solution

10 mL Sample

Diagram for Standard Calibration Method

 250 mL

10 mL + Standard
and Molybdate
Solution

25 mL

Diagram for Standard Addition Method

Table 2 (SCM)

Trial 1 of Bar Sample

μg 5 mL 100 mL

%Wt=
(ppm , ) (
mL 10 mL 5 mL )( )( 2505 mLmL )( 1005 mLmL )( 250 mL ) ( 1 x 101 g μg ) x 100 %
6

g , actual sample

%Wt=
(0.2201) ( 105 mLmL )( 1005 mLmL )( 2505 mLmL )( 1005 mLmL ) ( 250 mL) ( 1 x 110g μg ) x 100 %
6

0.987 g
%Wt=0.139 %

Table 3, 4, & 5 (SAM)

Trial 1 of Liquid Detergent Sample

μg 25 mL

%Wt=
(ppm , ) (
mL 10 mL )( 25100mLmL ) ( 250 mL )( 1 x 101 g μg ) x 100 %
6

g , actual sample
 %Wt=
(0.266) ( 2510 mL 250 mL
mL )( 10 mL )
( 250 mL )
( 1 x 101 g μg ) x 100 %
6

1.034 g
%Wt=0.4026 %
Summary of Results

Table 8. Percent Weight and Amound of PO43- in the sample.    

  Standard Calibration Method Standard Addition Method
  Liquid Powder Bar Liquid Powder Bar
Mass PO43-, g 0.000840 0.001020 0.001414 0.004163 0.008388 0.017174
%Wt of PO43- 0.08122% 0.11155% 0.14329% 0.40257% 0.91769% 1.74003%

Table 8.1. Descriptive statistics for Percent By Weight of PO43-, Standard

Calibration Method    
Sample Mean SD RSD n df tstat C.I.
Bar 0.001433 3.3789E-05 2.3581% 3 2 4.303 8.39E-05
Liquid 0.000812 4.7779E-05 5.8826% 3 2 4.303 1.19E-04
Powder 0.001115 1.8244E-05 1.6356% 3 2 4.303 4.53E-05

Table 8.2. Descriptive statistics for Percent By Weight of PO43-, Standard

Addition Method    
Sample Mean SD RSD, % n df tstat C.I.
Bar 0.01740035 0.000E+00 0.00E+00 3 2 4.303 0
Liquid 0.00402567 3.9271E-18 9.76E-14 3 2 4.303 2.154E-18
1.3456E-
Powder 0.00917688 2.4533E-18 2.67E-14 3 2 4.303 18
Grading Table

Objectives/s  5   
Diagram  15   
Data Entries  40   
Performance  30   
Summary of
results  10   
Merits/Demerits     

Total Rating  100   

Checked By:

Dr. Kathlia De Castro-Cruz