1) What is Spectrophotometry?

(2pts)

 Spectrophotometry is a method to measure how much a chemical substance absorbs light by

measuring the intensity of light as a beam of light passes through sample solution.
 Spectrophotometry is a branch of electromagnetic spectroscopy concerned with the
quantitative measurement of the reflection or transmission properties of a material as a
function of wavelength.

2) Why is Spectrophotometry important? (3pts)

 Spectrophotometry is widely used for quantitative analysis in various areas (e.g., chemistry,
physics, biology, biochemistry, material and chemical engineering, clinical applications,
industrial applications, etc). Any application that deals with chemical substances or materials
can use this technique. In biochemistry, for example, it is used to determine enzyme-catalyzed
reactions. In clinical applications, it is used to examine blood or tissues for clinical diagnosis.
There are also several variations of the spectrophotometry such as atomic absorption
spectrophotometry and atomic emission spectrophotometry.

3) What is a spectrophotometer? Illustrate its basic parts and describe each function. (10pts)

 A spectrophotometer is an instrument that measures the amount of photons (the intensity of

light) absorbed after it passes through sample solution. With the spectrophotometer, the
amount of a known chemical substance (concentrations) can also be determined by measuring
the intensity of light detected.
4) Explain the principle of spectrophotometry. (5pts)

 The basic principle is that each compound absorbs or transmits light over a certain range of
wavelength. This measurement can also be used to measure the amount of a known chemical
substance.

5) Differentiate transmittance from absorbance. (2pts)

 Absorbance is the amount of light that is taken in as it passes through an object. Absorbance
is extremely difficult to measure because its value is almost always less than 1. There are
many instances where absorbance is applicable in real life.
 Transmittance is defined as the light passing through an object that is not reflected or
absorbed. It’s relatively easy to determine an object’s transmittance levels. You can either
figure this out using an equation or a transmittance measurement device. Transmittance is
very applicable to many real-life situations.

6) Explain Beer-Lambert Law. Cite an example how it is calculated. (3pts)

 The Beer-Lambert law relates the attenuation of light to the properties of the material
through which the light is traveling. This page takes a brief look at the Beer-Lambert Law and
explains the use of the terms absorbance and molar absorptivity relating to UV-visible
absorption spectrometry.
 Beer-Lambert Law (also known as Beer's Law) states that there is a linear relationship
between the absorbance and the concentration of a sample. For this reason, Beer's Law can
only be applied when there is a linear relationship. Beer's Law is written as:
 A=ϵlc

There is a substance in a solution (4 g/liter). The length of cuvette is 2 cm and only

50% of the certain light beam is transmitted. What is the extinction coefficient?

Solution

Using Beer-Lambert Law, we can compute the absorption coefficient. Thus,

−log(ItIo)=−log(0.51.0)=A=8ϵ−log⁡(ItIo)=−log⁡(0.51.0)=A=8ϵ

Then we obtain that

ϵϵ = 0.0376