620 is the λmax at which maximal absorbance occurs in the above graph
620 is the λmax at which maximal absorbance occurs in the above graph
620 is the λmax at which maximal absorbance occurs in the above graph
Visible spectroscopy is also called as colorimetry. This means it measures colored light
(color+metry= color+ measurement).
Every colored substance absorbs light radiation of different wavelengths. When we plot
a graph of the wavelength of radiation vs. absorbance, a curve is derived which shows
the wavelength at which maximum absorption takes place. This is termed as “λmax”
and is unique for every compound. This λmax is qualitative aspect and helps to identify
the substances.
620 is the λmax at which maximal absorbance occurs in the above graph
When the sample is exposed to the visible radiation, it absorbs light of a particular
wavelength. If the concentration of the sample is high, the absorbance if high and vice
versa.
Beer’s law: The intensity of the beam of monochromatic (single colored) light
decreases exponentially with an increase in the concentration of the absorbing
substance arithmetically.
Lambert’s law states that the rate of decrease in intensity of monochromatic light with
the thickness of the medium is directly proportional to the intensity of incident light.
Further details can be seen in the principle of spectroscopy.
Instrumentation of visible spectroscopy:
There are two types of instruments like simple colorimeters and sophisticated
spectrophotometers. Their sensitivity and range differ, but the basic components for
both remain the same.
1. Source of light: Any lamp which can produce a visible spectrum in the range of 400
to 800nm can be used. But it should have an adequate intensity and be free from
fluctuations.
The common sources of light are tungsten lamp and carbon arc lamps.
The carbon arc lamp can also produce light over the entire range of the visible
spectrum. But it is difficult to maintain and handle compared to the tungsten lamp.
2. Filters and monochromators: The visible white light has many wavelengths and
colors. So filters and monochromators are used to filter unwanted wavelengths. The
filters are of two types as
a) Absorption filters:
b) interference filters:
Absorption filters: These filters are less expensive and find a place in most of the
colorimeters. They are made of either glass or dyed gelatin. They absorb unwanted
radiation and transmit the remaining radiation.
These filters are easy to use and construct and also inexpensive.
The problem with them is less accuracy due to high bandpass of ± 30nm. For example,
if we want a wavelength of 530nm, these filters pass radiation ranging between 500nm
or 560nm.
Interference filters:
Here in between two parallel reflecting silver films, a dielectric spacer film is present.
This spacer film is made of CaF2, MgF2 or SiO.
When light radiation is allowed to pass through the filters, the incoming radiation gets
interfered by reflected radiation from the second film.
The order number mentioned in the equation indicates the thickness of the dielectric
spacer films made of either CaF2, MgF2 or SiO.
These interference filters are efficient than the absorption filter as they have a lesser
bandpass of ± 10nm only.
These filters are also inexpensive, and use of additional filters can cut of undesired
wavelengths. But, the transmission and intensity of radiation could become low.
Monochromators
These are more sophisticated and accurate than the filters. They are of two types as
They are expensive but very accurate. They have a bandpass of ± 0.1nm and have a
higher range like 400 to 1000nm wavelength.
Sample cells: These are cuvettes which hold the sample under test. They are made of
color corrected fused glass or polystyrene so that the light is not absorbed. Sample
cells made of glass can be used for both aqueous and nonaqueous samples while
those made of polystyrene cannot be used for organic solvents.
The sample cell should be completely transparent such that light falls and passes
through it. These cells are mostly either cylindrical or rectangular. The internal path
length is 0.1 cm or 1 cm or rarely even 10cm. They can hold 0.5ml to 5ml of liquid
samples.
Detectors: Once the monochromatic lights pass through the sample cell, part of it is
absorbed by the sample. The unabsorbed light is allowed to fall onto a detector for
measurement. The detectors are of different types like
1. Photovoltaic cells
2. Phototubes
3. Photomultiplier tubes.
Of the above three, photomultiplier tubes are highly sensitive and accurate. So they are
used in spectrophotometers while others are used in colorimeters.
When the photon from the unabsorbed radiation falls on to the detector, it is converted
into an electrical signal and measured.
Types of instruments
The visible spectroscopy instruments are of two types as single beam and double beam
types.
The single beam types are again as colorimeters and spectro-colorimeters. These
single beam spectrophotometers have a single cell for sample alone.
While the double beam photometers have two sample cells, one cell is meant for the
test sample and other is meant for reference solutions. This includes
spectrophotometers with advanced filters, detectors and recorders.