ABSTRACT

The objective for this experiment is to define absorbance of solutions at different

concentrations.After some experiments that we have been done,we can conclude that who
have the higher and the lower absorbance of substance.After that,from this experiment we
need to determine the concentration of the given sample by using spectrophotometer to
measure the absorbance at800nm of the stock solution and the diluted solution.

From the procedure,we need to prepare CuSO4 solution of 14g in 500 ml distilled water
and we need to prepare four sample of solution in 100ml volumetric flasks by diluting the
stock solution using sample concentrations of 14,7,3.5 and 1.75 g/l.After that,we labelled the
volumetric it with C1,C2,C3,C4,C5 and measure the absorbance value at 800nm of the stock
solution(C1) and of each the diluted solutions from C1 until C5.We also need to prepare
CA,CB,CC to use small amount of NaCl between 0.1> x >0.5 mg a 100 ml volumetric flask and
fill the flask with the 14 g/l solution.Then, measure the absorbance for sample CA,CB,CC.After
all the procedure have been done,we get the final result for all sample.

The conclusion for this experiment,the absorbance increased if the concentration of

solution increased.UV-vis Spectrophotometer need to use to determine the different
concentration from all the sample because we need to get the best final result because it can’t
determine it with human eye.
INTRODUCTION

This experiment introduce the function of UV-vis Spectrophotometer, it is to measure the

absorbance value in different concentration.An UV-vis Spectrophotometer is an instrument
for measuring the transmittance or absorbance of a sample as a function of the wavelength of
electromagnetic radiation.A certain amount of the light get absorb by the chemical which
cause electrons to be increase from one energy level to another.The amount of light that reach
the detector known as spectrum.Spectrum is a band of colours as seen in a rainbow produced
by separation of the components of light by their different degrees of refraction according to
wavelength.
 1.CuSO4 stock solution of 14g was prepared in 500ml distilled water.

2.Four sample solutions was prepared in the 100ml volumetric flask by diluting the stock
solution using sample concentrations of 14, 7, 3.5 and 1.75 g/l.

3.The absorbance of distilled water was measure and use it as a reference.

4.The absorbance values was measured at 800 nm of the stock solution (C1) and of each of the
diluted solutions. Label as C1, C2, C3, C4 and C5.

5.Small amount of NaCl between 0.1 > x > 0.5mg a 100ml volumetric flask and fill the flask with the
14g/l solution. Label this sample as CA, CB and CC.

6.The absorbance was measured as mentioned is step 4.

 Sample Solution Concentration (g/l) Absorbance (AU)
C1 28.00 1.183
C2 14.00 0.713
C3 7.00 0.404
C4 3.50 0.134
C5 1.75 0.118
Table 1
Sample Solution Mass of NaCl (mg) Absorbance (AU)
CA 0.29 0.657
CB 0.50 1.401
CC 0.28 0.393
Table 2
Calculate concentrations of CuSO4 stock of 14g in 500ml distilled water.
14g0.5l = 28.00g/l
Calculate concentrations of CuSO4 in 100ml volumetric flask
M1V1 = M2V2
M1 = Stock concentration
V1 = Volume of CuSO4 needed
M2 = Concentration of CuSO4 used
V2 = Volume of volumetric flask 100ml

Calculate volume of C1
28(V1) = 28 (100)
V1 = 100ml of CuSO4
Calculate volume of C2
28(V1) = 14(100)
V1 = 50ml of CuSO4
Calculate volume of C3
28(V1) = 7 (100)
V1 = 25ml of CuSO4
Calculate volume of C4
28(V1) = 3.5(100)
V1 = 12.5ml of CuSO4
Calculate volume of C5
28(V1) = 1.75(100)
V1 = 6.25ml of CuSO4

DISSCUSION
1.Plot absorbance of C1 until C5 as a function concentration.Explain the relationship between
the absorbance and the concentration?

Concentration vs Absorbance
1.4

1.2

1
Absorbance (AU)

0.8

0.6

0.4

0.2

0
1.75 3.5 7 14 28
Concentration (g/l)

When the concentration in solution increase,the absorbance also will be increased.So, the
absorbance is propotional with the concentration of the CuSO4 solution.

(1.183−0.713)
m= y = mx+c
(28−14)
=0.0336 1.183 = 0.0336(28)+c
C=0.243
2.Determine the concentrations of sample CA,CB and CC.

Sample Solution Concentration (g/l)

CA 12.32

CB 34.46

CC 4.46

Calculate concentration of CA
Y = mx+c
0.657 = 0.0336(CA)+0.243
CA = 12.32

Calculate concentration of CB
Y = mx+c
1.401 = 0.0336(CB)+0.243
CB = 34.46

Calculate concentration of CC
Y = mx+c
0.393 = 0.0336(CC)+0.243
CC = 4.46
3.Discuss the cause of differences in the absorbance values for stock solution and when
adding small amount of NaCl.
From the experiment result,the factor to affect the absorbane values are according the
concentration of the solution.When a we adding a small amount of NaCl into the solution,the
concentration of thesolution will be increased.So,from the experiment,we could know that the
absorbance value change when NaCl was mixed with the solution and that make the
concentration of the solution increased.
CONCLUSION

From this experiment,we can conclude the relationship between the absorbance values and
concentration of a solution. The objective for this experiment was to determine the
absorbance values of solutions using different concentration of given samples. First of all, we
prepared 14g of CuSO4 in 500ml of distilled water. Four solutions has been prepared in the
100ml volumetric flasks by diluting the solution using sample concentrations of 14, 7, 3.5,
and 1.75 g/l and labeled it C1, C2, C3, C4 and C5. The result received using an UV-vis
Spectrophotometer. From the result, the absorbance values is directly proportional to the
concentration of the CuSO4 solution. Other than that, we prepared NaCl that range between
0.1 > x < 0.5 mg a 100ml volumetric flask and fill the flask with the 14 g/l solution. This
sample is label as CA, CB, CC. We also need to use UV-vis Spectrophotometer to determine the
absorbance values. From the result, the absorbance values will be increase when we increase
the amount of NaCl into 14 g/l solution. We can use the y = mx + c to calculate the
concentration of CA, CB and CC .