SPECTROMETRY

(process) used to assess the concentration or amount of a given chemical substance

SPECTROSCOPY
Energy = Frequency (directly proportional); Conc. = Absorbance
Wavelength = Wave number
(study) interaction between radiation and matter as a function of wavelength

1) ABSORPTION SPECTROCOPY – measures the absorption of radiation

2) EMISSION SPECTROSCOPY – examine wavelengths of photons emitted by atoms during transition from an excited energy state – lower energy state
3) SCATTERING SPECTOSCOPY – measures amnt of light subs. scatters at certain wavelengths, incident angles & polarization
TYPES PRINCIPLE APPLICATION INSTRUMENTATION
A. Despersive B. Fourier Transform
Quantitative - Grating/Scanning Infrared (FTIR)
measures the bond techniques Interoferometer:
- Detects molecular
vibration frequencies in a 2 TYPES OF VIBRATION: 1. Light source - Source
impurities/ additives
molecule and is used to
 Stretching – - Analysis of formulations - Nernst Glower - Beamsplitter
determine the functional - Global Source - Two mirrors
vibration/oscillation along (eg. insecticides)
INFRARED group 2. Monochromator - Laser
line of bond Qualitative
SPECTROSCOPY - Prism of alkali halides - Detector
 Bending – change of - Identification of
Atomic wt. = Frequency 3. IR Transducers Advantages:
angle; vibration not along compounds by matching
Bond energy = Frequency - Thermal - nondestructive, increase
the line spectrum of unknown
subs.; functional groups; - Pyroelectric sensitivity
polymers, plastics & - Phytoconducting - precise measurement
resins
3. Sample cells
1. Light source
containers for the sample
BEER LAMBERT LAW - Deuterium (200-400nm)
and reference solution
Absorbance = Conc. - Xenon Arc lamps (200-1000nm)
Cuvettes:
(directly proportional) - Tungsten lamp (350-2500nm)
- Visible (plastic/glass)
measures the absorption of 2. Monochromator - UV (quartz/fused silica
Aλ = ελbc - Determination of
radiation in ultraviolet- wavelength selector cuvettes)
UV-VIS SPECTROSCOPY ελ = molar absorptivity solutions of transition
visible spectral region
UV (190-380nm) (L∙mole-1∙cm-1) metal ions
Path length= Absorbance
VIS (380-780nm) b = sample path length in - Identification of inorganic
molecules undergo
(cm). and organic species
electronic transitions 4. Detector
c = concentration of the
compound in the sample, in
molarity (mol L-1)

ION SOURCE
- Electronspray ionization
HIGH VACUUM SYSTEM - Matrix Assisted Laser
- Bioavailability studies Desorption Ionization
measures mass-to-charge IONIZER - Pharmacokinetics MASS ANALYZER
ratio (m/z) of charged Ion source - Characterization of - Time of flight (TOF)
MASS SPECTROSCOPY particles by which drugs ions are formed in pulses;
compounds can be MASS ANALYZER - Drug degradation smaller ions = faster travel
determined by their separates ions product analysis - Quadrople Mass Analyzer
masses - Screening of drug RF + DC voltages to operate
DETECTOR candidates mass filters
presents information DETECTOR
- Photomultiplier
DATA SYSTEM
 - Sun SPARK/DEC Station
3. Shim coils
measures the number and Shimming: avoids
1. Magnets
types of atoms in a fluctuation
- Magnetic resonance alignment of orientation
molecule by studying the 4. Sample probe
imaging for medical - Permanent magnets
NUCLEAR MAGNETIC spin changes at the nuclear 4MHz – 750 MHz (0.4-75m holds sample in fixed
diagnosis - Conventional magnets
RESONANCE (NMR) level when radio wavelength) position
- MR Microscopy in - Superconducting magnets
frequency is absorbed in 5. Transmitter coils
research 2. Field lock
the presence of magnetic generates pulse to provide
constant magnetic field
field RF
6. Detector

A. Filter Fluorometer B. Spectrofluorometer

- measures the ability of a - uses fluorescent properties
sample to absorb light at one of some compounds in order
wavelength and emit light at a to provide information
longer wavelength. regarding their
- Biomedical, medical, and
detecting and measuring - sensitive quantitative concentration and chemical
chemical research fields
fluorescence in measurements are desired for environment in a sample
for analyzing organic
FLUORESCENCE compounds that uses specific compounds.
compounds
SPECTROSCOPY ultraviolet light stimulating 1. Light source
- Differentiation on - Emission
the compounds, causing - lasers (high energy)
malignant and bashful filters a wavelength specific
them to emit visible light - photoiodides
skin tumors from benign to emission
- lamps
3. Sample cuvette/cell
2. Monochromator
- Borosilicate
- Excitation
- Quartz glass
selects wavelength to react
4. Detector
with the sample
FLAME ATOMIZATION
Desolvation – liquid solvent
determining the is evaporated, dry sample
concentration of a remains
particular metal element in Vaporization – solid sample
a sample by using a flame vaporizes to a gas
to atomize the sample (eg. Volatilization – compounds AAS
ATOMIC ABSORPTION
graphite furnace) are broken down into atoms 2. Monochromator
SPECTROSCOPY 1. Light Source – emits
(ground state atoms)
ATOMIZATION: ELECTROTHERMAL wavelength required for
analysis 3. Detector
1. Flame atomization ATOMIZATION - Determine the metal - Photomultiplier tube
2. Electrothermal Drying (above 110˚C) present in the sample Hollow cathode lamps
atomization Ashing (up to 1000 ˚C) (toxicities) Electrodeless Discharge
Atomization (2000-3000 ˚C) lamps
Cleanout (up to 3500 ˚C)
1. Flame atomizer:
- Absence of the light
Nebulizer
quantitative measurement source
Flame/Burner – excites
of optical emission from - The temperature in the
ATOMIC EMISSION atoms to higher energy
excited atoms to atomizer is big enough to
SPECTROSCOPY levels
determine analyte atomize the analyte atoms
2. Monochromator
concentration and excite them to a higher
3. Detector
energy level
4. Readout meter
 measures the scattered
suspended particles should be
light by the particle at an
*small with respect to the
angle (usually 90o) as a - Determination of
NEPHELOMETRY wavelength used
function of the suspended matter in
concentration of the water.
*light scattered symmetrically
dispersed phased - Determination of the
clarity of beverages and
measures concentration of pharmaceutical
a substance in a solution preparations.
suspended particles should
by measuring the loss in - Measurement of
be *larger than the
intensity of a light beam bacterial concentrations.
wavelength of light
TURBIDIMETRY through a solution that - Immunoassays
contains suspended
*light mostly/preferentially
particulate matter or
scattered forward
measures transmitted
light at 180 o

1. Gas inlets – filter the

gases to ensure purity
Carrier (H2, He, N2)
Make-up gas (H2, He, N2)
Detector Fuel Gas
2. Pneumatic controls –
- Analyzes residual correct pressure & then
FACTORS AFFECTING GC
solvents in raw materials supply to various parts
SEPARATION
separating and analyzing and finished products 3. Injector – sample is
- Boiling point
GAS compounds that can be - Urine drug screens for volatilized then carried into
- Polarity
CHROMATOGRAPHY vaporized without barbiturates and the stream
- Column temp.
decomposition underivatized drugs 4. Column coated w/ immobilized liquid
- Flow rate of gas
- Flavors and fragrance Packed columns SP
- Length of column
(Food industry) packed w/ a solid support 5. Column oven – regulates
coated with immobilized liquid temperature
stationary phase material of 6. Detector – amplifies
various film thickness response and generates
Capillary column electronic signal
long hollow silica tubes w/ the 7. Data system – receives
inside wall of the column the signal