CH 502 Experimental & Analytical

Methods

Supplement - 1
Glass Membrane Electrodes
• pH electrode • Ion Selective Field Effect
Transistor (ISFET) electrode

csrg.ch.pw.edu.pl/tutorials/isfet/
www.ph-meter.info
Spectroscopic Methods

• Based on the interaction between electromagnetic radiation

(EMR) and matter
• EM spectrum ranges from Gamma rays to Radio waves
• The energy associated with the rays is related to the
wavelength by the Planck’s law equation
– E = hc / λ
• h – Plancks Constant c – velocity of light λ - wavelength
• Gamma rays have the lowest wavelength and hence the
highest energy
Electromagnetic Spectrum
• Gamma Rays: < 0.01 nm
• Hard X-rays: 0.01 – 0.1 nm
• Soft X-rays: 0.1 – 10 nm
• Vac. UV: 10 – 180 nm
• Visible: 380 – 700 nm
• Near UV: 180 – 380 nm
• Near IR: 700 – 1400 nm
• Mid IR: 1400 – 15000 nm
• Far IR: 15 – 1000 μm
• Microwave: 1 – 10 mm
• Radio waves: > 10 mm

http://nationalatlas.gov/articles/mapping/IMAGES/
Types of Interaction between EMR and Matter
• NMR (Radio frequency):
– Change of Spin – absorption on RF energy
• Microwave
– Change of Orientation
• Infrared
– Change of Configuration – rotational/vibrational
• UV/Vis
– Change in electronic structure
• X-rays
– Change in electronic structure
• Gamma Rays
– Change in nuclear configuration/mutation/isotopes
 Instrumentation for Spectroscopy
Absorption Spectroscopy

Source Sample
Wavelength Detector Data
Selector Readout

Emission Spectroscopy
Sample

EMISSION Wavelength
Selector

EXCITATION

Wavelength Detector Data

Selector Readout

Source
 Optical Materials

• Transmittance is the main property for selecting material for

optical elements
– LiF – transmittance between 150 – 7000 nm
– Borosilicate (normal glass) absorbs below ~380 nm
– Quartz or Fused Silica is ideal for UV applications and have very high
transmittance
– IR optical elements – halide salts (AgCl, KBr, NaCl), polymers have high
transmittance
 Sources

• Continuous Sources
– Xenon Arc Lamp – 250 – 600 nm
• Molecular Fluorescence
– Hydrogen or Deuterium Lamp – 180 – 380 nm
• UV Molecular Absorbance
– Tungsten/Halogen – 240 – 2500 nm
• UV/Vis/NIR – Molecular Absorbance
– Tungsten – 350 – 2200 nm
• Vis/NIR
– Nernst Glower – 400-20,000 nm
• IR – molecular absorbance
– Nichrome – 750 – 20,000 nm
– Globar – 1200 – 40,000 nm
– Tunable Dye Lasers
 Sources

• Continuous Source Lamp Spectra & Irradiance

From “Oriel Corporation”: (www.lot-oriel.com)

 Sources
• Line Sources
– Narrow Discrete Spectra

From “Oriel Corporation”: (www.lot-oriel.com)

 Sources

• Pulsed Sources
– Lasers (Light Amplification by Stimulated Emission of Radiation)
• Permits light of low divergence
• Well defined wavelength
• Low Energy and High Energy
• Nd: YAG Lasers
 Wavelength Selectors

• Radiation is measured in ‘bands’

– Selectivity – narrow bands
– Sensitivity – broader bands – allow more energy
• Devices to select wavelength
• Absorption/Emission Wavelengths
– Monochromator
– Band Filters
– Interference Filters
Wavelength Selectors

• Monochromator
– Czerny-Turner
– A – incident beam
– B – Entrance Slit
– C – Mirror 1
– D – Diffraction
Grating (Rotating)
• Blazed Grating
• Prism
• Concave
• Holographic
– E – Mirror 2
– F- Exit Slit

Image:
http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
 Wavelength Selectors

• Blazed Diffraction Grating

From www.edmundoptics.com
 UV Absorption Spectroscopy
• Transmittance, T = I / I0
• Absorbance, A = -log T
• Beer’s Law
– A = ε.L.C
– ε = absorption coefficient
– L = path length
– C = concentration of analyte
• Example Absorption
Spectra shown here

Image:
http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
 UV Absorption Spectroscopy

• Double Beam Spectrophotometer (figure above)

– Analysis of Reference and Sample
Simultaneously
• Single Beam
– Only one detector – reference and sample
cuvettes move to come in line to the path of
light.
Infra-red Spectroscopy

Structural Features:
• Stretching or Bending Vibrations
http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
• Organic or Inorganic Bonds
IR Vibrational Spectroscopy
Formaldehyde – Gas Phase IR spectra

Some Common Structural Features

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
 IR Vibrational Spectroscopy
Typical Example of Infrared Absorption Frequencies
Stretching Vibrations Bending Vibrations
Intensit
Functional Class Range (cm-1) Intensity Assignment Range (cm-1) Assignment
y
Alkanes 2850-3000 str CH3, CH2 & CH 1350-1470 med CH2 & CH3 deformation
2 or 3 bands 1370-1390 med CH3 deformation
720-725 wk CH2 rocking

Alkenes 3020-3100 med =C-H & =CH2 (usually sharp) 880-995 str =C-H & =CH2
1630-1680 var C=C (symmetry reduces 780-850 med (out-of-plane bending)
intensity) 675-730 med cis-RCH=CHR
1900-2000 str
C=C asymmetric stretch

• Examples of Functional classes that exhibit IR absorbance

– S compounds
• S-H, S-OR, S-S, C=S, S=O
– P compounds
• P-H, (O=)PO-H, P-OR, P=O
– Si Compounds
• Si-H, Si-OR, Si-CH3
– Oxygen-Nitrogen
• NOH, N-O, N=O

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
Michelson Interferometer

http://www.biophysik.uni-freiburg.de/Spectroscopy/Time-Resolved/spectroscopy.html
IR Spectrophotometer

www.thermonicolet.com
Fourier Transform

FTIR

Processor

Absorption Spectra
Interferogram

• The interferogram is the signal that is received by the detector (which is a

thermal detector).
• This interferogram is processed using a fourier transform to obtain spectral
information (or an absorption spectrum) shown on the right side.
• The transmission is the usual representation of IT spectra – absorption can
be converted to transmission easily.

http://www.biophysik.uni-freiburg.de/Spectroscopy/Time-Resolved/spectroscopy.html
NMR Spectroscopy
• Nuclei of elements have a characteristic spin – I
– Integral: e.g. 1,2,3… (# protons and # nuetron are odd)
– Half integral: e.g. ½, 3/2, 5/2…(#protons + # nuetrons are odd)
– Zero (#protons and #nuetrons are even)
• Spinning charge generates a small magnetic field resulting in a
magnetic moment
• In the presence of an external magnetic field, B0 , two spin
states exist, +1/2 and -1/2.
• The magnetic moment of the lower (+1/2) energy spin state is
aligned with the external magnetic field and that of the higher
(-1/2) is aligned in the opposite direction to B0.
 Spin states (two energy) in external magnetic
field
No Magnetic
Field

B0 – Magnetic Field Strength

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/nmr/nmr1.htm
Magnetic Fields
• Earth’s Magnetic field – 10-4 T (Tesla)
• Modern NMR instruments use magnetic fields 1 – 20 T.
• Even at high B0, the energy difference observed between the two spin
states is less than 0.1 cal/mole
– IR (1-10 cal/mole), Electronic transitions are 100 times higher
• Energy difference is expressed in terms of frequency in MHz
– Ranging from 20 – 900 MHz depending on the field and the molecule
• Irradiation of the sample with Radio Frequency energy corresponding to
this energy difference between the two spin states causes absorption and
excitation from the +1/2 to the -1/2 state.
• For ½ spin nuclei, the energy difference is proportional to their magnetic
moments at a given B
• Common nuclei used in NMR spectroscopy are
– 1H, 13C, 19F, 31P,
– Most common is the 1H NMR also referred to as Proton NMR.
Proton NMR Spectroscopy

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
Proton NMR Spectroscopy
• Consider a sample of water
– External magnetic field B = 2.3487 T
– 100 MHz - radiation
• If B is increased slowly to 2.3488 T
– H nuclei will absorb RF energy and a resonance signal will
appear

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/spectro.htm#contnt
Chemical Shift
• Different H nuclei have different signals at different frequencies
– Electron density around a nucleus produce a magnetic field that opposes the
much stronger external field.
– This secondary field shields the nucleus from the external field
– To compensate for this shielding B must be increased to achieve resonance
– Scanning of B at a RF frequency produces a SHIFT
– Compounds that have higher electron density have the resonance at higher
energy frequencies
Chemical Shift
• The characteristic frequencies are dependent on the magnitude of B and the radio
frequency
• No two magnets have the same field and resonance frequencies vary accordingly
– Reference proton: TMS (Trimethylsilane)
– Shift is measured as shown below in terms of ppm
• Solvent – CDCl3
Chemical Shift
NMR Spectroscopy – examples

Red peaks indicate different characteristic groups of the chemical,

which exhibit a certain chemical shift with respect to a reference such
as TMS.
Chemical Shift
• Signal Strength
– Number of Hydrogen atoms
– Concentration
• OH proton signal
– Aliphatic or aromatic
• Π-electrons
• Solvent effects
• Spin-Spin Interactions
– J Coupling
J-Coupling
• Signal Splitting
– Doublet
– Triplet
– Quartet
– Quintet
– Concentration
13
C NMR Spectroscopy
• For Molecules that lack significant C-H bonds – one
cannot use proton (or 1H NMR spectroscopy)
– Polychlorinated compounds
– Triple bond compounds
– Polycarbonyl ass
• 13 C isotope is a viable alternative
– Less abundant  less sensitive signal splitting due to
interaction with H
• Solid State NMR
– anisotropy
Raman Spectroscopy
• Used in chemistry,
– Vibrational information is very specific
for the chemical bonds in molecules.
– Provides a fingerprint by which the
molecule can be identified. The
fingerprint region of organic molecules
is in the range 500-2000 cm-1.
– Study changes in chemical bonding, e.g.
when a substrate is added to an enzyme.
– Raman gas analyzers have many
practical applications, for instance they
are used in medicine for real-time
monitoring of anaesthetic and
respiratory gas mixtures during surgery.
– Characterize materials – measure T
– Crystallographic Orientation of a sample
• Inelastic Scattering or Raman scattering
of monochromatic light usually from a
laser in the visible, NIR or near UV
range.
• Holographic diffraction gratings and
multiple dispersion stages to separate
the weak inelastic Raman scattering and
the strong Rayleigh scattering
• Spontaneous and stimulated RS
Atomic Spectroscopy
• Qualitative/Quantitative determination of elements
– Analysis of metals in water, soils, solids, tissue
• Ionization/Atomization of sample
– Flame Atomization
– Plasma
• Analysis
– Atomic Optical Spectrometry
– Atomic Mass Spectrometry
• Generic Instruments
– Atomic Absorption (AA) - Flame
– ICP-AES/OES - Plasma
– ICP-MS - Plasma

SAMPLE Sample Spray Flame or Analyzer

Processin /Vapor Plasma
g (Nebulizer
or
Vaporizer
Atomic Absorption Spectroscopy - AA
• Source – Hollow Cathode Lamp
– Tungsten anode/Cylindrical Cathode in quartz with inert gas
– Cathode is made of the analyte metal or has a coating of the metal
– Line spectra (narrow)
– Individual lamps for different metals – about 70 elements
• Sample is aspirated into the atomizing zone as a spray –
– Nebulizer
• Ionization/Atomization of sample
– Flame – Burner – 1700 – 3500 C
• Fuel + air (natural gas, Hydrogen, acetylene)
• Flow rate is high – efficiency of atomization is not good
• Higher T increases sensitivity
– Graphite Furnace
• Graphite tube with electric current
• Thermal energy causes electronic transition
– Light Absorption
• Monochromator
• Detector
– Photometers
– Spectrophotometer – UV/Vis monochromator
Atomic Absorption Spectroscopy - AA
• Source of Error
– Flame Emission
– Modulation – Chopper/Reflected/Sample
– Alternating signal from the lamp and continuous signal from the flame
– Correction is made at the data analysis.
• Background Correction
– Hollow cathode lines are narrow so very little chance of interferences
– Molecular species in the range of the wavelength absorbed can cause interference
– AT = AA + AB
– Continuous Source Background Correction
• Part of the flame used for quantitative measurement
– Primary combustion zone (very close to nozzle) - hottest
– Interzonal region
– Secondary combustion zone (outer ring) – oxidation may occur
– Flame position must be adjusted since the response of each metal varies with it.
• Beer’s Law
• Internal Standards
• Hg – Cold Vapor AAS
• Detection limits
– 0.2 – 30 ppb (g/L) (for different elements)
Plasma For Atomization
• Plasma tube
– 3 concentric quartz tubes
– Argon Gas – as ionizing gas
and for cooling
– RF Coils – 2KW energy at
27/40 MHz
– Temperature – 6000 – 8000 K
– Ionization efficiency is high
– Tangential Gas flow allows for
thermal separation of plasma
zone from the region before it
ICP-AES
ICP-AES
• Interference
– Blank interference
• MilliQ water – relative
• Blank – MDL and IDL
– Spectral interference
– Analyte interference
• Physical/chemical effects
• Operating conditions
• Detection limits
ICP-AES
• Wavelength selection
• Monochromator
– Sequential analysis/scan of wavelengths and
a PMT
• Spectrograph
– Large aperture for a band and detection
using a CCD or array detector
• Polychromator
– Grating + Simultaneous detection using a
CCD or array detector
ICP-MS

icpms.ucdavis.edu/InstrumentationQuad.html
ICP-MS
• Operates under Vacuum – 10-6 torr
• After ionization sample is introduced to the
analyzer zone through two orifices
– Skimmer cone
– Sampler cone
• Ion Optics
• Mass Analyzer
• Selection on the basis of Isotope (or
Mass/Charge ratio)
– Capable of isolating “isotopes”
• Very low detection limits.
ICP-MS – Mass Analyzer
• Ion Optics
– Focus the ions in a
barrow beam towards
the Mass Analyzer
• Mass Analyzer
– Quadrupole
– Allows only a certain
M/Z set to pass through
to the detector • Other Types of analysers
• Detector – Ion Trap
– PMT/Dynode – Time of Flight(TOF) MS
– Interferences
• Ar+, ArO+ , ArH+, H2O+,
Ar2+ or combination
• Same isotope
• Matrix effects
ICP-MS
• During analysis, the use of blank is important
since it accounts for contamination
– in the instrument (previous sample memory effects)
– the surroundings (cleanliness of the lab).
– For very sensitive (or low concentration) work, a clean
room (with very low particulate contamination) may be
required.
• Internal standard is recommended in all samples
(same amount in all samples, standards and
blanks)
– The internal standard should be an element that is not
present in the sample
– Selection of internal standard depends on the nature
of the analysis matrix