RESULTS AND DISCUSSIONS

The table below shows the corresponding colors of each wavelength. At 350 nm, no color
is visible because this is still at ultraviolet radiation. 400 nm until 650 nm corresponds to the colors
listed below.

Wavelength Color
350 Not visible/UV
400 Violet
450 Blue
500 Blue green
550 Green
600 Orange
650 Red
Table 1. Visible colors in each wavelength

The wavelength corresponding to the maximum absorbance as determined from the plot,
was noted and was then used for measuring the absorbance of residual concentration.

Wavelength Absorbance Wavelength Absorbance Wavelength Absorbance

375 0.0112 540 0.391 690 0.0206
400 0.0293 550 0.3026 700 0.0176
410 0.0367 560 0.2216 710 0.0147
420 0.0477 570 0.1497 720 0.0122
430 0.0682 580 0.0957 730 0.0102
440 0.1026 590 0.0624 740 0.0084
450 0.1532 600 0.0462 750 0.0072
460 0.219 610 0.0391 760 0.0063
470 0.2802 620 0.0352 770 0.0062
480 0.3208 630 0.0331 780 0.0062
490 0.3548 640 0.0319 790 0.0066
500 0.3865 650 0.0307 800 0.006
510 0.4308 660 0.0291
520 0.4571 670 0.0269
530 0.4431 680 0.0239
Table 2. Data on the reported absorbance measurement in wavelengths 375 nm-800 nm

SHERALD G. AGUSTIN
 The wavelength noted is 520 nm corresponding to a maximum absorbance reading
of 0.4571. The data for absorbance readings were plotted against the wavelengths as shown in
the graph below.

Wavelength at Maximum Absorbance

0.5
0.45 0.4571
0.4
0.35
Absorbance

0.3
0.25
0.2
0.15
0.1
0.05
0
375 410 430 450 470 490 510 530 550 570 590 610 630 650 670 690 710 730 750 770 790
Wavelength

Absorbance

Figure 1. Plot of absorbance readings in each wavelength

The figure below shows the plot of the standard curve based on Beer-Lambert’s Law.

Beer's Law Plot Standard Curve

0.25
y = 0.0358x - 0.0279
0.2 R² = 0.9882
Absorbance

0.15

0.1

0.05

0
0.005 0.01 0.02 0.03 0.04 0.05
Concentration

Figure 2. Plot of absorbance readings in each initial concentration of the sample tested

SHERALD G. AGUSTIN
 The Beer-Lambert law is based on a linear relationship between the absorbance and the
concentration, molar absorption coefficient, and the optical path length. By plotting the reported
values in a standard curve, our goal is to determine the linearity of the data. From the standard
curve obtained, the generated working equation of the line is y = 0.0358x – 0.0279 with the value
of regression coefficient, R2 =0.9882.

Concentration Absorbance
0.005 0.019
0.01 0.036
0.02 0.073
0.03 0.113
0.04 0.15
0.05 0.193
Table 3. Absorbance readings in each concentration for Beer’s law plot

In order to determine the unknown concentration, we can easily use a formula from the
MS Excel which is the TREND option. This could be used to calculate a linear trend line through
a given set of dependent y-values and, optionally, a set of independent x-values and return values
along the trend line. Using this, the corresponding concentration for the absorbance of 0.095 is
0.0252 M. This proved to be true as value of this concentration is expected to lie between 0.02
and 0.03 as could be inferred from the given table of data above.
Quantitatively, the unknown concentration could also be determined by using the Beer’
law general equation:

A = 𝟄Cl Equation 1
Where:
A = absorbance
𝟄 = Molar absorption coefficient (/M cm)
C = Molar concentration (M)
l = optical path length (cm)

This could also be related to the linear equation of y = mx where y is the value of
absorbance, m corresponds to 𝟄l which is the slope, and x is the value of the concentration. Based

SHERALD G. AGUSTIN
on the graph, the obtained slope of the line is 3.858630137. To determine the unknown
concentration with the absorbance reading of 0.095:

A = (𝟄l) C
0.095 = (3.858630137) C
C = 0.0246 M

| 0.0246−0.0252 |
% Relative error = | 0.0246 |
(100%)

% Relative error = 2.44%

By following the formula given, the obtained value of the unknown concentration is 0.0246
M. This may have a little deviation from the value obtained directly from the MS Excel sheet with
a relative error of 2.44%.

SHERALD G. AGUSTIN