SPECTROPHOTOMETER

UV/VIS

Wavelength

Wavelength and Color of the Light

Colors and Complementary Colors

Ultra Violet /VIS Spectrum

Lambert Beers Law

 At this time, I/Io is called transmittance (T) , I/Io x 100 is

percent transmittance (T%) and 1/T = log (Io/I) is called
absorbance (Abs).

T = I/Io = 10.k l c
Abs = log (1/T) = log (Io/I) = k l c
log T = - l c or log T = l c
log T is also defined as absorbance (A) or optical density
(OD), thus log T = A = l c.
is defined as the molar absorbtivity and the concentration
is given in mol/liter (also called the molar extinction
coefficient).
If the concentration is given in grams/liter, is replaced by a,
the specific absorbtivity.

Qualitative and Quantitative

Analysis
To analyze what this substance is and
what substances are mixed are called
qualitative analysis, while to analyze the
amount of these substances is called
quantitative analysis.

Qualitative Analysis
Spectrum and Chemical Structure

Colorimetric Analysis
(Quantitative Analysis)
The methods for performing quantitative
analysis by comparing color darkness of
substance is called colorimetric analysis.
When the substance is transparent, if
absorbance exists in the visible and
ultraviolet, it is measured.

The quantitative method to measure

concentration of the sample with
unknown concentration from absorption
of the sample with known concentration
is provided in two methods :
1. calibration curve method
2. standard additive method

Calibration Curve

The main steps to analyze with

spectrophotometer uv/vis
1. Formation of molekuls which can
absorp uv/vis light
2. Selection of wave length, max and Absmax
3. Preparation of calibration curve
4. Measurement of samples absorbance (A)

Spectrophotometer types
Spectrophotometer can be
categorized by optical system as
follows :
Measurement
optical System

Single beam
configuration
Double beam
configuration

Single Beam Configuration

Double Beam Configuration

ATOMIC ABSORPTION
SPECTROPHOTOMETER
Atomic Emission
EXCITATION
Energy

+
Ground state
atom

Excited state
atom

DECAY

+
Excited state
atom

Ground state
atom

Light Energy

 Atomic emission can be used when

no lamp is available
Flame emission has better detection
limits than atomic absorption for
several elements including Al, Ba, Li,
K, Na, and V

Atomic Absorption

Light Energy

+
Ground state
atom

Excited state
atom

Atomic Emission vs Atomic

Absorption
There are some basic differences between
atomic emission and atomic absorption.
In atomic emission, the processes which
involved are excitation and decay.
In atomic absorption, the ground state atom
absorbs light energy of a specific wavelength
as it enters the excited state.

Atomic Absorption Instrumentation

There are five basic components of an
atomic absorption instrument :
1. The light source which emits the spectrum of
the element
2. An absorption cell in which atoms of the
sample are produced (flame, graphite
furnace)
3. A monochromator for light dispersion
4. A detector which measures the light intensity
and amplifies the signal
5. A display that shows the reading after it has
been processed by the instrument electronics

THE USE OF FLAME IN AAS

In atomic emission :
It converts the sample aerosol into an
atomic vapor and then thermally
elevates the atoms to an excited state.
When these atoms return to ground
state, they emit light which is detected
by the instrument. The intensity of light
emitted is related to the concentration of
the element of interest in solution.

In atomic absorption :
The only function of the flame is to convert
the sample aerosol into atomic vapor
which can then absorb light from the
primary light source (hollow cathode lamp
or discharge lamp).

FLAMES
There are oxidant and fuel gas
combination used in atomic absorption
is air acetylene.
Other flames that can be used are air
hydrogen, nitrous oxide acetylene
and argon hydrogen entrained air.

AIR - ACETYLENE
Can be used for determination of
approximately 35 elements
The temperature is about 2300 oC
The acetylene flow is about 4 liters/minute
The air flow is about 24 liters/minute
For many elements, the fuel/oxidant gas
ratio must be adjusted for maximum
sensitivity.