ATOMIC SPECTROMETRY

I N S T R U M E N T A T I O N M E T H O D S
C H A P T E R 7 C

A R T S A N D S C I E N C E S D E PA R T M E N T
LEARNING OBJECTIVES
After studying this chapter, you should be able to:

❑ Define and explain the principles behind Atomic Absorption Spectroscopy

(AAS), Atomic Emission Spectroscopy (AES), and Flame Emission
Spectroscopy (FES) in the context of clinical laboratory analysis.
❑ Identify and describe the key components of AAS, AES, and FES
instrumentation.
❑ Calculate the concentration of analytes (e.g., calcium, sodium, potassium)
in biological fluids.
LEARNING OBJECTIVES
After studying this chapter, you should be able to:

❑ Compare and contrast AAS, AES, and Flame Emission Spectroscopy in

terms of sensitivity, specificity, clinical applications, and limitations in the
analysis of elements.
REVIEW
REVIEW
REVIEW
Instrument Components

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
REVIEW

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Introduction
➢ Atomic spectroscopic methods are used for qualitative and
quantitative determination of more than 70 elements.
➢ The two major types are
▪ optical atomic spectroscopy
▪ atomic mass spectroscopy.
➢ Atomization, in which a sample is volatilized and decomposed to
produce gas-phase atoms and ions, is required as the first step in
all atomic spectroscopic procedures.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Introduction
➢ The most common atomization methods are
• inductively coupled plasmas (ICPs)
• flames
• electrothermal atomizers
➢ Atomic absorption (AA) spectroscopy uses flames and
electrothermal atomizers.
➢ Optical emission and atomic mass spectrometry use
inductively coupled plasma.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Optical Atomic Spectroscopy
➢ Optical atomic spectroscopy encompasses several techniques used
to analyze the elemental composition of a sample by studying the
light emitted or absorbed by atoms.
➢ The main types include:
• Atomic Emission Spectroscopy (AES),
• Atomic Absorption Spectroscopy (AAS), and
• Atomic Fluorescence Spectroscopy (AFS).

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Emission Spectra
• Atomic Emission Spectroscopy (AES), is used to analyze
the elemental composition of a sample by examining the light
emitted when atoms are excited to higher energy levels.
• Analyte atoms are excited by heat or electrical energy.
• Energy is typically supplied by a
▪ plasma
▪ flame
▪ low-powered discharge
▪ high-powered laser

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Emission Spectra
• Before the external energy source is
applied, the atoms are usually in their
ground state (lowest-energy).
• Applied energy causes atoms to
momentarily enter an excited state
(higher-energy).
• These excited electrons are unstable
and eventually return to lower energy
levels, releasing the absorbed energy in
the form of photons (light particles).

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Emission Spectra
• A resonance transition is a
transition to or from the ground state.
• A resonance line is the spectral line
produced as a result.
• Figure shows emission spectra of
several elements along with the
spectrum of white light.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Absorption Spectra
• Atomic Absorption Spectroscopy (AAS), is used to
measure the concentration of elements by detecting
the absorption of light by free atoms in a gaseous
state
• an external source of radiation interact on the analyte
vapor.
• if the source radiation is of the appropriate frequency
(wavelength), it can be absorbed by the analyte atoms
and promote them to excited states.
• Figure a shows three of several absorption lines for
sodium vapor.
• Figure b shows the source of these spectral lines.
Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Sample Introduction Systems for Atomic
Spectroscopy
• There are two classes of atomization devices:
▪ Continuous atomizers such as plasmas and flames that introduce
samples as a continuous stream.
▪ Discrete atomizers such as the electrothermal atomizer that are
used to inject individual samples using a syringe or autosampler.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Sample Introduction Systems for Atomic
Spectroscopy
• Figure shows the processes that must occur to produce free
atoms or elementary ions.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Flame Atomizers
• A flame atomizer, the most common used atomization technique,
consists of a pneumatic nebulizer that converts the sample solution
into a mist or aerosol that is introduced into a burner.
• The high-pressure gas is usually the oxidant with the aerosol-
containing oxidant being mixed subsequently with the fuel.
• The burners are generally premixed, laminar flow burners.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Flame Atomizers
• Figure shows a typical commercial
laminar flow burner for atomic
absorption spectroscopy.
• The aerosol flows into a spray
chamber where it encounters a series
of baffles that remove all but the finest
droplets.
• The sample spray is mixed with fuel
and oxidant gas in the spray chamber
before these components are burned
in a slotted burner.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Flame Atomizers
• Regions of a Flame When a nebulized sample is
carried into a flame, the droplets are desolvated in
the primary combustion zone.
• The resulting finely divided solid particles are carried
to a central inner core. This is the hottest part of the
flame. Particles are vaporized and converted to
gaseous atoms, elementary ions, and molecular
species and excitation of atomic spectra takes place.
• The atoms, molecules, and ions are carried to the
outer edge (outer cone) where oxidation may occur.
This is called the secondary combustion zone.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 Flame Atomizers
• The temperature of the flames produced is relatively low, so the technique is
only suitable for the elements that are easily excited such as alkali and alkali
earth elements.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Flame Emission Spectroscopy (FES)
▪ also known as flame photometry, it is a specific type of AES where a flame
is used to excite the atoms in the sample.
▪ It is used to determine the concentration of certain elements by
measuring the light emitted when they are introduced into a flame.
▪ simple and inexpensive method for analyzing certain elements,
particularly alkali and alkaline earth metals
▪ often used in clinical and environmental laboratories for measuring the
concentration of specific ions, such as sodium and potassium.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Flame Emission Spectroscopy (FES)
▪ A sample containing the analyte of interest
is introduced into a flame.
▪ The heat of the flame excites the atoms of
the analyte, causing their electrons to move
to higher energy levels.
▪ When these excited electrons return to
their ground state, they emit light at specific
wavelengths.
▪ The intensity of the emitted light is directly
proportional to the concentration of the
analyte in the sample.
Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Flame Emission Spectroscopy (FES)

https://apollo.nvu.vsc.edu/classes/met130/notes/chapter2/aurora_atom.html
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Flame Emission Spectroscopy (FES) Components
➢ Sample Introduction System - Nebulizer: Converts the liquid sample into a fine
mist (aerosol). Atomizer: Carries the aerosol into the flame.
➢ Flame (Excitation Source) - Commonly a mixture of air-acetylene or nitrous
oxide-acetylene. Provides the thermal energy to excite the electrons of metal
atoms
➢ Monochromator (Wavelength Selector) - Disperses the emitted light into its
component wavelengths. Allows selection of a specific wavelength for
detection.
➢ Detector - Photomultiplier tube (PMT) or photodiode detects light intensity at
the selected wavelength. Converts light into an electrical signal.
➢ Readout Device/Data System - Displays or records the intensity data, often in
terms of emission intensity vs. concentration.
Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Flame Emission Spectroscopy (FES)

https://namrataheda.blogspot.com/2016/05/spectrophotometry-flame-photometry.html
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Flame Emission Spectroscopy (FES)

Gandhi, K., Sharma, N., Gautam, P.B., Sharma, R., Mann, B., Pandey, V. (2022). Atomic Absorption Spectroscopy and Flame Photometry. In:
Advanced Analytical Techniques in Dairy Chemistry. Springer Protocols Handbooks. Springer, New York, NY. https://doi.org/10.1007/978-1-
0716-1940-7_11
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Atomic Absorption Spectroscopy (AAS)
▪ Used to measure the concentration of elements by detecting the absorption of
light by free atoms in a gaseous state.
▪ When light of a particular wavelength is passed through a sample containing
these atoms, the atoms will absorb the light, and the amount of light
absorbed is proportional to the concentration of the element in the sample.
▪ Relatively low equipment and operating costs. Only a small amount of sample
is needed.
▪ Can only analyze one element at a time. Not suitable for ultra-trace levels
(typically detects ppm to low ppb).

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Atomic Absorption Spectroscopy (AAS)
▪ The sample is first converted into free atoms in the gaseous phase. This is
typically done by introducing the sample into a flame or a graphite furnace.
▪ A light beam from a hollow cathode lamp (HCL) specific to the measured
element is passed through the atomized sample.
▪ The atoms of the element in the sample absorb light at their specific
wavelength from the HCL beam.
▪ A detector measures the amount of light that passes through the sample, and
the amount of light absorbed is calculated.
▪ The amount of light absorbed is directly proportional to the concentration of
the analyte.
Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Atomic Absorption Spectroscopy (AAS) - Components
▪ Light Source: hollow cathode lamp (HCL), which emits light at a specific
wavelength that can be absorbed by the measured element.
▪ Atomizer: This is where the sample is converted into free atoms. The sample
is introduced into either a flame or a graphite furnace.
• A flame uses a mixture of fuel and oxidant gases to create heat,
atomizing the sample.
• A graphite furnace uses an electrically heated graphite cylinder to
vaporize the sample.
▪ Monochromator
▪ Detector
▪ Readout Device
Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY
Atomic Absorption Spectroscopy (AAS)

http://biomedicalinstrumentationsystems.com/flame-atomic-absorption-spectrometry/
INSTRUMENT USED IN ATOMIC SPECTROSCOPY

https://chem.libretexts.org/Bookshelves/Analytical_Chemistry/Instrumental_Analysis
 INSTRUMENT USED IN ATOMIC SPECTROSCOPY

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
Assignment: Short Bond paper
You are a Medical Laboratory Scientist working at the Biochemistry Section of a tertiary hospital.
The attending physician requests serum calcium analysis for five patients with varying clinical
symptoms, including muscle cramps, cardiac arrhythmias, and confusion. You proceed to analyze
the samples using Atomic Absorption Spectroscopy (AAS) and obtain the following data:
Calibration Standards:

Standard Ca²⁺ Concentration (mg/L) Absorbance

Std 1 0.0 0.000
Std 2 2.5 0.135
Std 3 5.0 0.265
Std 4 7.5 0.375
Std 5 10.0 0.525
Std 6 12.5 0.755

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
Assignment: Short Bond paper
Patient Sample Results:
Patient Absorbance Questions: Show calculation and report answer in
A 0.140 four significant figures.
B 0.330 1. Construct the calibration curve equation from
C 0.480 the standard data.
D 0.095 2. Find the value of the slope and intercept.
E 0.210 3. Find the value of r-squared.
4. Calculate the calcium concentration (mg/L) for
each patient.
5. Convert the calcium concentrations to mg/dL.

Skoog, West, Holler, and Crouch, Fundamental of Analytical Chemistry, 10e. © 2022 Cengage
REFERENCES :
▪ Fundamentals of
Analytical Chemistry 10th
Edition: Skoog, et al.,
Cengage Learning (2021)
▪ Quantitative Chemical
Analysis: Harris, D.C.
(2020)