I.

Introduction
In spectrophotometry the amount of light absorbed is
measured. The instrument in its simplest form consists of:
radiant energy source, means of obtaining monochromatic light,
cell to hold a sample or blank detection and readout device.
Transmittance, T, is the ratio of the light transmitted through
the sample and the light entering the sample. Absorbance, A, is
the amount of light absorbed from the initial incoming light.
Absorbance is the base 10 logarithm of the reciprocal of
transmittance. The intensity of the transmitted light decreases
exponentially as the path length of the sample increases.
If the path length of the sample cell is constant, the
absorbance ill increase linearly ith an increase in
concentration and the transmittance ill decrease exponentially
ith an increase in concentration. Absorptivity is a fundamental
constant for a specific chemical species at a specific
avelength. Absorptivity cannot be changed.
If the concentration is in moles!liter, absorptivity is
called molar absorptivity. It is designated by the symbol:
A = bc
= molar absorptivity = liters/mole cm
b = cell path length = cm
c = concentration = moles/liter
"eer#s la assumes that each absorbing particle is
absorbing independently of every other absorbing particle. The
la also assumes that the species being measured remains the
same and the concentrations are constant.
There are real and apparent limitations to "eer#s $a. %eal
limitations are the necessity of using lo analyte
concentrations. At higher concentrations &'0.01(), the distance
beteen molecules diminish thus affecting the charge
distribution to its neighbor. Thus possibly diminishing the
ability of the neighbor to absorb a given avelength of
radiation. The higher concentration may also change the
refractive index of the medium &rare at concentrations loer
than 0.01().
II. Objectives
To determine the relationship beteen the concentration
of a light absorbing species.
To investigate the intensity of the transmitted test.
III. Materials:
*olorimeter
10+m$ hack test+tubes ! scre cap and test tube rack
0., ( *u-.
/
0,1
2
.
10+m$ pipet and aspirator
IV. Methodology
The purpose of this experiment is to determine the
concentration of a light absorbing species and to
investigate the intensity of the transmitted test. The
instrument that as used to determine the absorbance of the
solution as colorimeter. It as setup ith a 300+nm
absorption filter. There ere , test tubes for each 2.0,
/.0, 3.0, 4.0 and 10.0 m$ 0.,( *u-.
/.
It as then diluted to
10+m$ ith distilled ater and as mixed thoroughly. A
blank 10+m$ distilled ater as also prepared and served as
a blank. 5ith the aid of colorimeter the absorbance as
determined for each solution including the blank.
An unknon concentration as given and ith the help
of colorimeter and by using least+s6uares method, its
concentration and absorbance as determined.
V. Results and Discussion
Table 1 showing the volume and concentration o !u"O
#

Volume$m%& ' # ( ) 1*
!oncentration $M& *.1 *.' *.+ *.# *.,
Table ' showing the values o concentration and absorbance
o !u"O
#
!oncentration $M& -bsorbance $-&
*.1 *.1(1
*.' *.'),
*.+ *.+./
*.# *.#.)
*., *.(*'
The absorbance of the unknon concentration is 0.7/3.
The concentration can be calculated using least+s6uares
method.
8rom the table above, the slope m 9 1.0:, and the y+
intercept 9 0.0301. 5ith &x) as the concentration and &y) as
absorbance, e can find the value of &x) 9 concentration.
Table + showing the concentration and absorbance o blan0
and 1 solution.
!oncentration $M& -bsorbance $-&
2lan0 $Distilled 3
'
O& *.**
4 5 6n0nown 5 *.+ *.+#(
7ra8h showing the relationshi8 between concentration and
absorbance
;sing (
1
<
1
9 (
2
<
2
e can calculate each concentration as
seen in table 1. And ith the aid of colorimeter e can
determine the absorbance of each solution because the
solution absorbs at the fre6uency the colorimeter is set at.
An unknon concentration as determined by using least+
s6uares method and it is determined to be 0.23 (,
approximately 0.7 (. The reading of the blank is 0 absorbance
so there is no need to subtract or counter it since it ould
give the same result. 5e can see in the graph the
relationship beteen concentration and absorbance. As the
concentration increases the absorbance also increases. There
is a positive correlation beteen energy absorbance and the
concentration of the solution doing the absorption.
8urthermore, the relationship is linear &first order) ithin
a certain range. Also, the "eer+$ambert la tells us the
linear relationship beteen absorbance and concentration of
an absorbing species.
VI. !onclusion
The concentration of a light+absorbing species can be
determined using least+s6uares method ith absorbance given
already by the aid of a colorimeter. The concentration of the
unknon species as found out to be 0.23 ( = 0.7 ( ith
absorbance e6ual to 0.7/3. 5e can conclude that the
relationship beteen absorbance and concentration is directly
proportional, as concentration increases, absorbance also
increases.
VII. Reerences
http:!!teaching.shu.ac.uk!hb!chemistry!tutorials!molspec!beer
s1.htm
http:!!.chm.davidson.edu!vce!spectrophotometry!beersla.htm
l
http:!!.ncsu.edu!labrite!res!gt!gt+reg+home.html
Mindanao University of Science and Technology
C.M. Recto, Lapasan, Cagayan de oro City
Analytical Chemistry 1 Laboratory
!periment "o. #
$etermination of Concentration of Light%Absorbing Species
Allen Mar& T. Librado
'S%Chemistry (
)ro*p 1
S.+. (,1-%(,1.