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How Does a Spectrophotometer Work?

Brian Lew
Winter 2007
February 2, 2007

I. Introduction What is a Spectrophotometer?
II. Components of the Spectrophotometer
III. How a Spectrophotometer Works
A. The Light Path
B. The Charge-Coupled Device (CCD)
C. The Interpreter
IV. Different Types of Spectrophotometers
A. Single Beam vs. Double Beam
B. Visible Light
C. Ultraviolet Light
D. Infrared Light
V. Uses of a Spectrophotometer


A spectrophotometer is a device that measures light intensity as a function of
wavelength. It does this by diffracting the light beam into a spectrum of wavelengths,
detecting the intensities with a charge-coupled device, and displaying the results as a
graph. There are different types of spectrophotometers for different purposes.

I. Introduction What is a Spectrophotometer?
A spectrophotometer is a device to measure light intensity at different
wavelengths. It produces light with a light source, and after the light passes through a
subject, the light is diffracted into a spectrum which is detected by a sensor and
interpreted into results we can use.
The output of a spectrophotometer is usually a graph of light intensity versus
wavelength. The data collected to generate this graph can typically be saved as a table of
wavelengths and intensities. The y values of the graph can be represented as either
transmittance or absorbance.

II. Components of the Spectrophotometer
There are four main parts of a spectrophotometer: the light source, subject,
detector, and interpreter. Some examples of light sources are visible, infra red, and
ultraviolet light. The light created by the light source passes through the subject where
some light is usually absorbed, is received by the sensor, and is interpreted into an output
such as a graph.

III. How a Spectrophotometer Works
A. The Light path
In the Ocean Optics USB2000 Spectrophotometer, light is produced by a violet
LED-boosted tungsten lamp. The light passes through the sample (usually a solution in a
cuvet) and enters the spectrophotometer through a slit. The narrow slit disperses the
light, spreading it out. The light reflects off of a concave collimating mirror and is
reflected to a dispersion grating. The dispersion grating reflects the light and also
disperses it towards a second concave mirror. This focusing mirror focuses the light onto
a detector.

Figure 1. Schematic Optical Path of the Ocean Optics USB2000 Spectrophotometer

Figure 2. Drawing of Ocean Optics Spectrophotometer

B. The Charge-Coupled Device (CCD)
At the end of the light path is the detector. In most spectrophotometers, it is a
linear charge-coupled device (CCD). A CCD is a type of image sensor that detects light.
It is an integrated circuit made up of an array (or in this case, a linear arrangement) of
linked/coupled light-sensitive capacitors. The light-sensitive capacitors detect the
intensity of light received and convert it into an electrical signal.

The linear CCD detector corresponds to the range of wavelengths on a hand held
spectrophotometer. Each pixel on the CCD represents a specific wavelength of light, and
the more photons absorbed, the more electrical signal generated. Therefore, the electrical
signal output by the CCD at each pixel is proportional to the light intensity at each
corresponding wavelength.

C. The Interpreter
Spectrophotometers can have their own display for output, but it is more common
for them to be connected to a computer where software manipulates the data and displays
it in a usable for, like a graph of transmittance or absorbance versus wavelength.

Figure 3. Sample Output Graph of a Spectrophotometer
IV. Different Types of Spectrophotometers
A. Single Beam vs. Double Beam
There are two classes of spectrophotometers: single and double beam. The single
beam spectrophotometer was the first invented, and all the light passes through the
sample. In this case, to measure the intensity of the incident light, the sample must be
removed so all the light can pass through. This type is cheaper because there are less
parts and the system is less complicated. Later, the double beam spectrophotometer was
invented. In this type, the light source is split into two separate beams before it reaches
the sample. One beam is used for reference and the other passes through the sample.
This is advantageous because the reference reading and sample reading can be taken at
the same time. In some double beam spectrophotometers, there are two detectors and the
sample and reference beams can be measured simultaneously. Other double beam
spectrophotometers that have only one detector use a beam chopper. This device inside
blocks one beam at a time and the detector alternates between measuring the sample and
reference beams.
B. Visible Light
The visible region of light is about 400-700nm. Visible region
spectrophotometers vary in accuracy. Some have CCD detectors with enough pixels to
take reading every 10nm, while others can take several reading per nanometer. These
spectrophotometers can use incandescent, halogen, LED, or a combination of these
sources. For example, the Ocean Optics USB2000 Spectrophotometer uses an LED
boosted tungsten bulb.

Figure 5. Ocean Optics USB4000 Spectrophotometer

Figure 6. Spectrum of a Halogen lamp, using Ocean Optics USB400

C. Ultraviolet Light
UV spectroscopy is most commonly used for liquids, but can also be used for
gases and even solids. Samples are placed in a cuvette, a small rectangular container,
usually 1cm in width. These are can be made of plastic, glass, or quartz (listed in
increasing expense). Plastic and glass absorb UV, so they can only be used for visible
light spectroscopy.

Figure 4. Spectrum of Ethyl Alcohol

D. Infrared Light
Infrared spectroscopy is used to study molecules and the vibrations associated
with their structures. Different chemical structures vibrate in different ways in response
to different wavelengths, due to the varying energies associated with each wavelength.
For example, mid-range infrared tends to cause rotational vibrations, while the near
infrared (higher energy) tends to cause whole molecule harmonic vibrations like
stretching, and rocking.

V. Uses of a Spectrophotometer
Spectrophotometers are directly used to measure light intensity at different
wavelengths, and this can be represented as percent of incident light transmitted or
absorbed. Using this information and comparing it to other data obtained or known,
spectroscopy can be used as a tool. One example is comparing spectra to determine
concentrations of a solute in solution. This can be done by recording
transmittance/absorbance at a specific wavelength (a wavelength that the solute absorbs)
and known concentration. Then analysis of a solution of unknown concentration can be
compared to the known data, and be interpolation the concentration can be calculated.
This can even be done with solutions containing multiple solutes, however is it most
accurate when the different solutes absorb different wavelengths.
Spectrometers that do not have a light source, but generate spectra based on the
incoming light can be used in a similar way to identify light sources. The spectra graph
obtained from an unknown light source (or mixture of sources) can be compared to a
database of graphs for different known light sources to identify the unknown light source.
Another application of the spectrophotometer is to determine the equilibrium
constant of a reaction involving ions, which takes place in aqueous solution. Starting a
solution containing only one reactant, the spectrum is measured. Then small, measured
amount of the other reactant is added and after each addition, the spectrum is measured
again. This method works best if there is a known wavelength that the product absorbs.
Then, as more products are formed from adding more reactant, more light will be
absorbed. When the solution becomes saturated and the reaction reaches net equilibrium,
the increase in light absorption will level out, indication equilibrium.


Balch, Alan. Chemistry 2BH Laboratory Manual. Davis, CA: University of California,
Davis, 1999.

Charge-coupled device. Wikipedia. 5 Feb. 2007. 10 Feb. 2007

Infrared spectroscopy. 9 Feb. 2007. 10 Feb. 2007

Spectrophotometry. Wikipedia. 8 Feb. 2007. 10 Feb. 2007

Oxtoby, D. W.; Gillis, H.P.; and Nachtrieb, Norman H. Principles of Modern Chemistry.
Fifth edition. South Melbourne, Australia: Thomson Learning, 2002.

Ultraviolet-visible spectroscopy. Wikipedia. 2 Feb. 2007. 10 Feb. 2007