Session 3

Optical Spectroscopy: Introduction/Fundamentals Atomic and molecular spectroscopies Instrumentation

Overview

Physical basis of absorption and emission Atomic spectra Molecular spectra Instrumentation: components of optical systems for spectrometers Common techniques in atomic spectroscopy: AAS and ICP-OES Calibration

Useful websites for spectroscopy

http://www.shsu.edu/~chm_tgc/sounds/fl ashfiles/ICPwCCD.swf http://www.thespectroscopynet.com/Inde x.html?/ http://teaching.shu.ac.uk/hwb/chemistry/t utorials/molspec/ http://www.chemguide.co.uk/analysis/uvvi siblemenu.html

See also individual citations on slides
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Electromagnetic radiation
Spectroscopy = interactions between light & matter
E = hn = hcl n = frequency; l = wavelength

http://www.spectroscopynow.com/coi/cda/detail.cda?id=18411&type= EducationFeature&chId=7&page=1 This primer also contains a wavelength-energy converter

Fundamentals

Absorption and emission of light by compounds is generally associated with transitions of electrons between different energy levels
DE2 excited states DE = hn = hc/l DE1 ground state
E2

E2

DE2
E1

E1

E0

DE1
E0

Emission: Sample (in an excited state) produces light/looses energy

Absorption: sample takes up energy Consumes light of appropriate wavelength

Atomic spectra: line spectra provide specificity: each element has its own pattern, as each element has its own electronic configuration http://physics.nist.gov/PhysRefData/ASD/lines_form.html
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Fundamentals

The population of different states is given by the Boltzmann equation:

N1 g1 e N0 g0
N0: number of atoms in ground state N1: number of atoms in excited state g1/g0 : weighting factors

E kT

Note: Equation contains temperature: Excitation can be achieved by providing thermal energy

Atomic emission: Flame spectroscopy

Observation
Persistent golden-yellow flame Violet (lilac) flame carmine-red flame Brick-red flame Crimson flame Yellowish-green flame Green flame Blue flame (wire slowly corroded)

Caused by...
Sodium Potassium, cesium Lithium Calcium Strontium barium, molybdenum Borates, copper, thallium Lead, arsenic, antimony, bismuth, copper

Lithium

Cesium

Qualitative method

Sodium

A simple spectroscope

Spectroscope: Device for qualitative assessment of a sample E.g. used in flame analysis E.g. used in gemmology

Atomic Spectroscopies - Synopsis

Techniques for determining the elemental composition of an analyte by its electromagnetic or mass spectrum

Optical spectroscopies

Mass spectrometries

AES

Others See table

Fluorescence Spectroscopy ICP-OES

AAS ICP-MS SIMS

Others (L. 6)

Flame AAS

GFAAS

Atomic spectroscopies
Technique
Atomisation/Excitation

Sample etc

Arc/spark
Laser microprobe Glow discharge ICP-OES Flame photometry (atomic emission) AAS Atomic fluorescence X-ray fluorescence ICP-MS

e
e e e e a fe fe -

Electric arc/spark
Laser Glow discharge lamp Electromagnetic induction Flame UV/Vis light UV/Vis light X-radiation n/a

Solid sample on carbon electrode

Solid sample on support Solid sample disc Liquid sample, sprayed into gas plasma Liquid sample, sprayed into flame Liquid sample, sprayed into flame or furnace Liquid sample, sprayed into flame or furnace Solid or liquid Liquid sample, sprayed into gas plasma
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Atomic spectroscopies

Common principles:

Sample introduction: Nebulisation, Evaporation Atomisation (and excitation or ionisation) by flame, furnace, or plasma Light source (can be sample itself - Only AA requires external light source Optical system (or mass spectrometer) Detector
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Spectrometer components:

Atomic spectra vs molecular spectra:

Lines

Bands

(nm)

Typical atomic spectrum e.g. acquired by AAS

Two typical molecular spectra Acquired by UV-Vis spectroscopy

Y axes: intensity of absorbed light. Under ideal conditions proportional to analyte concentration (I c; Beers law).

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Origin of bands in molecular spectra

Molecules have chemical bonds Electrons are in molecular orbitals Absorption of light causes electron transitions between HOMO and LUMO Molecules undergo bond rotations and vibrations: different energy sub-states occupied at RT and accessible through absorption: many transitions possible: A band is the sum of many lines

LUMO Vibrational substates HOMO

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rotational substates

Quantitative analysis by molecular absorption: Colorimetry

Because absorption spectroscopy is widely applicable, sensitive (10-5-10-7 M), selective, accurate (0.1-3% typically), and easy:

95% of quantitative analyses in field of health performed with UV/Vis tests Hemoglobin in blood

First step in analysis: establish working conditions

Select l Selection, cleaning and handling of cells Calibration: determine relationship between absorbance and 14 concentration

Instrument components
AAS Spectrometer Light Source Sample Monochromator Detector Readout/Data system

ICP-OES Spectrometer

Sample = light source

Monochromator

Detector

Readout/Data system

UV-Vis Spectrometer: Light Source Monochromator Readout/Data system

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Sample

Detector

UV-Vis spectrophotometer (dual beam)

Monochromator Diffraction grating

Slit
Mirror Light sources

Slit Filter

HalfMirror Mirror

Reference

Detector
Sample

http://www.spectroscopynow.com/coi/cda/detail.cda?i d=18412&type=EducationFeature&chId=7&page=1

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Example for a dual beam spectrometer

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Single beam

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UV-Vis spectroscopy practicalities: Referencing

Matrix (solvent, buffer etc) might also have absorbance: Must be taken care of In dual beam:

Simultaneous measurement of reference cell eliminates absorbance of background Recording of baseline recommended Requires measurement of reference spectrum, can be subtracted from sample spectrum Preferentially in same cuvette
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Single beam:

Light sources
Wavelength(nm) 100 200 UV 400 Visible 700 2000 4000 7000 IR 10,000 20,000 40,000

VAC Spectral region

Ar lamp

Near IR

Far IR

Continuum

Xe lamp D2 lamp Tungsten lamp Nernst glower (ZrO2 + Y2O3) Nichrome wire

Line

Globar (SiC) Hollow cathode lamps Lasers 20

Example of a continuum source: Output from Tungsten lamp

Widely applied in UV-Vis spectrometers

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Hollow cathode lamp

Used in AAS Filled with Ne or Ar at a pressure of 130-700 Pa (1-5 Torr). When high voltage is applied between anode and cathode, filler gas becomes ionised Positive ions accelerated toward cathode Strike cathode with enough energy to "sputter" metal atoms from the cathode to yield cloud with excited atoms Atoms emit line spectra

22

Example: Output from iron hollow cathode lamp

Small portion of spectrum from Fe hollow cathode lamp Shows sharp lines characteristic of gaseous atoms Linewidths are artificially broadened by monochromator (bandwidth = 0.08 nm)

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Wavelength selectors: dispersive elements and filters

Wavelength(nm) 100 200 UV 400 Visible 700 2000 4000 7000 IR 10,000 20,000 40,000 VAC Spectral region Near IR Far IR

Fluorite prism
Fused silica or quartz prism Glass prism NaCl prism KBr Prism 3000 lines/nm

Continuous

Gratings
Interference filters Interference wedge Glass filters

50 lines/nm

Discontinuous

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Monochromators

Consist of

Entrance slit Collimating lens or mirror Dispersion element (prism or grating) Focusing lens or mirror Exit slit
Mirrors Common in UV-Vis spectrometers

Czerny-Turner grating monochromator:

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Dispersers

Separate polychromatic light into its components

Prism Diffraction grating: patterned surface which diffracts light

Prisms

Blazed diffraction grating

Holographic grating
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Echellette grating:

Extra pathlength travelled by wave 2 must be multiple of l for positive interference: nl = d(sin i + sin r) for UV 1000-2000 lines/mm: d = 0.5-1 mm

echelle: French for ladder

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Bandwidth of a monochromator

Spectral bandwidth: range of wavelengths exiting the monochromator Related to dispersion and slit widths Defines resolution of spectra: 2 features can only be distinguished if effective bandwidth is less than half the difference between the l of features

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Effect of slit width on peak heights

29

Components of optical system in an ICP-OES spectrometer

spherical and cylindrical lenses flat and spherical mirrors parallel planes optical path under vacuum or controlled nitrogen atmosphere (necessary for wavelengths <200 nm; air absorbs far UV light) Disperser(s)
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Old models: Sequential type

Can only measure one wavelength at a given time: Slow

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Newer: Simultaneous type

CCD detector: 2D detector

Echelle cross disperser (polychromator): Consists of Echelle grating and prisms/ echellette: separates lights in 2 dimensions

This combination allows high-speed measurement, providing information on all 72 measurable elements within 1 to 2 minutes
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Detectors
Wavelength(nm) 100 200 UV 400 Visible 700 2000 4000 7000 IR 10,000 20,000 40,000

VAC Spectral region

Near IR

Far IR

Photographic plate Photomultiplier Phototube Photocell Silicon diode Charge-coupled device (170-1000)

Photon detectors

Thermal detectors

Photoconductor Thermocouple Golay pneumatic cell Pyroelectric cell

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Photomultiplier: detects one wavelength at a time

Based on photoelectric effect Photocathode and series of dynodes in an evacuated glass enclosure

Photons strike cathode and electrons are emitted Electrons are accelerated towards a series of dynodes by increasing voltages Additional electrons are generated at each dynode Amplified signal is finally collected and measured at anode
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Photodiode arrays: measure several wavelengths at once

linear array of discrete photodiodes on an integrated circuit (IC) chip Photodiode: Consists of 2 semiconductors (n-type and p-type)

Light promotes electrons into conducting band: generates electron-hole pair Concentration of these electron-hole pairs directly proportional to incident light a voltage bias is present and the concentration of lightinduced electron-hole pairs determines the current through 35 semiconductor

Detection in simultaneous ICP-OES:

CCD: Charge-coupled device

Also integrated-circuit chip Contains an array of capacitors that store charge when light creates electron-hole pairs Accumulated charge is read out at given time interval Each wavelength is detected at a different spot Much more sensitive than photodiode array detectors
http://www.chemistry.adelaide.edu.au/external/soc-rel/content/ccd.htm
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Lecture 4
AAS and ICP-OES
Sample preparation Interferences Calibration

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Crucial steps in atomic spectroscopies and other methods

Laser ablation etc. Nebulisation
Solution

Solid/liquid sample

Sample preparation

M+

X-

Desolvation Vaporisation

Molecules in gas phase

MX(g)

M(g) + X(g) Sputtering, etc.

M+
Ions
Atoms in gas phase

Atomisation= Dissociation Excitation

Excited Atoms

Ionisation

ICP-MS and other MS methods

AAS and AES, X-ray methods

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Adapted from www.spectroscopynow.com (Gary Hieftje)

Sample Introduction: liquid samples

Often the largest source of noise Sample is carried into flame or plasma as aerosol, vapour or fine powder Liquid samples introduced using nebuliser

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Sample preparation for analysis in solution: Digestion

Digestion in conc. HNO3 and mixtures thereof (e.g. aqua regia) Br2 or H2O2 can be added to conc. acids to give a more oxidising medium and increase solubility Certain materials require digestion in conc. HF Common to use microwave digestion
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Microwave digestion

Rotor

Supplied with dedicated vessels (e.g. PTFE) Closed vessel digestion minimises sample contamination Faster, more reproducible, and safer than conventional 41 methods

Sample preparation and sample handling for trace analysis

As always sample preparation is key Ultra-trace: Contaminations introduced during sample processing can seriously limit performance characteristics Points to consider:

Purity of reagents Chemical inertness of reaction vessels and any other material samples come into contact with Working environment

Preparation of standards and blanks crucial Also measure a process blank:

Important for determination of LOD and LOQ

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Common Units in trace analysis

ppm, ppb, ppt, ppq..: parts per million etc. ppm: mg/kg; often also used as mg/L ppb: mg/kg ppt: ng/kg ppq: pg/kg

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Atomic absorption spectroscopy

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Atomic Absorption Spectroscopy

Flame AAS has been the most widely used of all atomic methods due to its simplicity, effectiveness and low cost First introduced in 1955, commercially available since 1959 Qualitative and quantitative analysis of >70 elements Quantitative: Can detect ppm, ppb or even less Rapid, convenient, selective, inexpensive
H Li Be B Al Sc Y La Ti Zr Hf V Cr Mn Fe Co Ni Pd Pt Cu Zn Ag Cd Au Hg C Si N P O S Se Te Po F Cl Br I At He Ne Ar Kr Xe Rn

Na Mg K Rb Cs Fr Ca Sr Ba

Ga Ge As In Tl Sn Pb Sb Bi

Nb Mb Tc Ta W Re

Ru Rh Os Ir

Ra Ac

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Flame AA Spectrometer
Hollow cathode lamps with characteristic emissions

Burner

Flame fuelled by (e.g.) acetylene and air

Nebuliser and Spray chamber

Hollow cathode lamps available for over 70 elements Can get lamps containing > 1 element for determination of multiple species
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Schematic

I0
Light Source

It
Monochromator Detector Amplifier

E.g. Hollow cathode lamp

Atomiser

Fuel (e.g. acetylene) Air

Analyte solution

Nebuliser, spray chamber, and burner

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Flame atomisation: Laminar flow burner - components

Nebuliser: converts sample solution into aerosol Spray chamber: Aerosol mixed with fuel, oxidant and burned in 5-10 cm flame Fuel: Acetylene or nitrous oxide Oxidant: Air or oxygen Burner head: Laminar flow: quiet flame and long pathlength But: poor sensitivity (not very efficient method, most of sample lost)

from: Skoog

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Structure of a flame

Relative size of regions varies with fuel, oxidant and their ratio

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Electrothermal atomisation: GFAAS

Provides enhanced sensitivity

entire sample atomised in very short time atoms in optical path for a second or more (flame 10-4 s)

Device: Graphite furnace

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Sensitivity and detection limits in AAS

Sensitivity: number of ppm of an element to give 1% absorption. Limit of detection: dependent upon signal:noise ratio: S/N Light intensity reaching detector

S/N=3.2

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Interferences in AAS

Broadening of a spectral line, which can occur due to a number of factors (Physical) Spectral: emission line of another element or compound, or general background radiation from the flame, solvent, or analytical sample

Background correction can be applied

Chemical: Formation of compounds that do not dissociate in the flame Ionisation of the analyte can reduce the signal Matrix interferences due to differences between surface tension and viscosity of test solutions and standards

Another caveat: Non-linear response common in AAS

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Physical interferences: Atomic line widths/ line shapes

Very important in atomic spectroscopy Narrow lines increase precision, decrease spectral interferences Lines are broadened by several mechanisms:

Natural broadening Doppler effect Pressure broadening

Figure taken from http://www.cem.msu.edu/~cem333/ Week03.pdf

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Natural linewidths

Width of an atomic spectral line is determined by the lifetime of the excited state Consequence of the Heisenberg uncertainty principle

For example, lifetime of 10-8 seconds (10 ns) yields peak widths of 10-5 nm
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Doppler Effect
Photon detector

Due to rapid motion of atoms in gas phase Atom moving toward the detector absorbs / emits radiation of shorter l than atom moving perpendicular to detector. Atom moving away from the detector absorbs / emits radiation of longer l: detector perceives fewer oscillations

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Pressure broadening

Results from collisions of absorbing/emitting species

With analyte atoms or combustion products of fuel Deactivates the excited state shorter lifetime - broader spectral lines Increases with concentration and temperature E.g. in flame, Na absorbance lines broadened up to 10-3 nm.

Doppler and pressure effects broaden atomic lines by 1-2 orders of magnitude as compared with their natural linewidths

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Background correction in AAS

particularly important in GFAAS Use beam chopper to distinguish the signal due to flame from desired atomic line at the same wavelength (old method)
Lamp and flame emission reach detector

Only flame emission reaches detector

Resulting signal
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Background correction in AAS

High energy Deuterium background corrector

Detector Hollow cathode lamp

Beam combiner

Lamps are pulsed out of phase with each other

Sample Deuterium lamp

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Minimising the effect of Matrix Interferences

The term "matrix" refers to the sum of all compositional characteristics of a solution, including its acid composition Calibration standards and samples must be matrix-matched in terms of composition, total dissolved solids, and acid concentration of the solution Also advisable for ICP-OES and -MS
Effect on K concentration on measured Sr
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Specialised applications in AAS: Flameless cold vapour methods

Mercury: has sufficient vapour pressure at RT Hydride generation technique for determination of As, Sb, Bi, Se, Te, Ge, Pb, and Sn

Generation of volatile metal hydrides (As, Sb, Bi, Se, Te, Ge, Pb, and Sn) Reduction by NaBH4 to form volatile hydride (e.g. SnH4) Hydrides carried into light path by argon gas Decomposed into elemental vapour by injection into (electrothermally) heated silica cell

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Calibration some practical aspects

61

Principles

Recap: Measured quantity must change with analyte concentration in systematic and defined way Can be determined by calibration, using defined standards Stock solutions of standards can either be prepared or purchased Working solutions are best prepared by weighing the amounts of stock solution and matrix (rather than using volumetric ware) NEVER extrapolate: concentration of sample must be in same range as standards
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Calibration in AAS
In theory, Beers law applies for dilute solutions In practice, deviation from linearity is usual Small dynamic range Possible to use non-linear curve fitting for calibration Reasons: Self-absorption:

excited atoms emit light that can also be absorbed instead of that of source: on average, less light per number of atoms is absorbed

Linear range

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Alternative to matrix-matching: Method of standard additions

Extensively used in absorption spectroscopy, accounts for matrix effects Several aliquots of sample
Sample (1): diluted to volume directly Samples (2,3,4,5): known amounts of analyte added before dilution to volume

BUT: Only makes sense if the added standard closely matches the analyte present in the samples chemically and physically

if simple, dissolved ions are analysed

64

Method of standard additions

If linear relationship exists between measured quantity and concentration (must be verified experimentally) then:

kVx c x kVs cs AT VT VT

Vx, Cx: volume and concentration of analyte Vs: variable volume of added standard Cs: concentration of added standard VT: total volume of volumetric flask k: proportionality constant (= l) Ax, AT: absorbances of standard alone and sample + standard addition,
respectively.
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Method of standard additions

Graphical evaluation
slope = m = (lcs) / VT

intercept = b = (lVxcx) / VT

Limitations The calibration graph must be substantially linear since accurate regression cannot be obtained from non-linear calibration points. Caution: The fact that the measured part of the graph is linear does not always mean that linear extrapolation will produce the correct results It is also essential to obtain an accurate baseline from a suitable reagent blank

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Most simple version of standard addition: spiking

Spiking means deliberately adding analyte to an unknown sample Involves:

preparation of sample and measurement of absorbance Addition of standard with known concentration, measurement of absorbance From difference in absorbance, calculate e From reading of sample alone, calculate amount of analyte (use Beers law for calculations)
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Other uses for spiking

Add spike at beginning of sample preparation Process sample with and without spike Difference should correspond to amount spiked Deviation allows to calculate recovery factor

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Atomic emission spectroscopy

69

Atomic emission spectroscopy

Historically, many techniques based on emission have been used (See Table on p. 4) Flame and electrothermal methods now widely superseded by Inductively-Coupled Plasma (ICP) method

Developed in the 1970s Higher energy sources than flame or electrothermal methods

70

ICP-AES/OES
Inductively coupled plasma-atomic emission spectroscopy (or optical emission spectroscopy)

Offer several advantages over flame/electrothermal:

Lower inter-element interference (higher temperatures) With a single set of conditions signals for dozens of elements can be recorded simultaneously Lower LOD for elements resistant to decomposition Permit determination of non-metals (Cl, Br, I, S) Can analyse concentration ranges over several decades (vs 1 or 2 decades for other methods) More complicated and expensive to run Require higher degree of operator skill

Disadvantages:

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Modern ICP-OES spectrometer

Over 70 elements (in principle simultaneously) Including non-metals such as sulfur, phosphorus, and halogens (not possible with AAS) ppm to ppb range Principle: Argon plasma generates excited atoms and ions; these emit characteristic radiation
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ICP-AES Instrumentation

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Components for sample injection and the ICP torch

Up to 7000C

www.cleanwatertesting.com/news_ NR149.htm

www.midwestrefineries.com /refiningandassaying.htm
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Meinhard nebuliser

Caution: The capillary is easy to block and difficult to unblock

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ICP torch

water cooled induction coil powered by RF generator (2 kW power at 27 MHz)

concentric quartz tubes

11-17 L/min d=2.5 cm 76

Torch Ignition Sequence

Ionisation of Argon initiated by spark from Tesla coil Start gas flow Switch on RF power

After leaving injector, sample moves at high velocity Punches hole in centre of plasma
Plasma generated
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Atomisation / Ionisation

In plasma, sample moves through several zones

Preheating zone (PHZ): temp = 8000 K: Desolvation/evaporation Initial radiation zone (IRZ): 6500-7500 K: Vaporisation, Atomisation Normal analytical zone (NAZ): 6000-6500 K: Ionisation

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Advantages of plasma

Prior to observation, atoms spend ~ 2 sec at 4000-8000 K (about 2-3 times that of hottest combustion flame)

Atomisation and ionisation is more complete Fewer chemical interferences Prevents side-product (e.g. oxide) formation

Chemically inert environment for atomisation

Temperature cross-section is uniform (no cool spots)

Prevents self-absorption Get linear calibration curves over several orders of magnitude

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Radial and axial observation

Axial

Radial. Can achieve higher sensitivity

Combined viewing expands dynamic range

http://las.perkinelmer.com/content/relatedmaterials/brochures/bro 80 _atomicspectroscopytechniqueguide.pdf

Applications

ICP-OES used for quantitative analysis of:

Soil, sediment, rocks, minerals, air

Geochemistry Mineralogy Agriculture Forestry Fornensics Environmental sciences Food industry

Elements not accessible using AAS

Sulfur, Boron, Phosphorus, Titanium, and Zirconium

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Homework for revision

Read http://las.perkinelmer.com/content/ relatedmaterials/brochures/bro_ato micspectroscopytechniqueguide.pdf

82

Lab Experiment 3

Analyse a Chromium complex for [Cr] in three ways:

UV (absorbance & extinction coefficient) Titration (moles Cr and charge) AAS (Cr standard curve and unknown concentration)

AAS data analysis

Fit standards to quadratic equation

A=a[Cr]2 + b[Cr] + c

Use a, b, and c to calculate unknown concentration

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