efinitions: Analyte- the component of interest in the sample Matrix- the remainder of the sample Examples: Lead in drinking water Benzene in soil Salt in blood LEVELS OF ANALYTICAL METHODOLOGIST

1. UNIVERSITY OF NORTHERN PHILIPPINES
COLLEGE OF HEALTH SCIENCES
Bachelor of Science in Medical Laboratory
Science
ANALYTICAL
CHEMISTRY
2ND
SEMESTER
INTRODUCTION TO
ANALYTICAL TECHNIQUES
Definitions:
Analyte- the component of interest in
the sample
Matrix- the remainder of the sample
Examples:
Lead in drinking water
Benzene in soil
Salt in blood
LEVELS OF ANALYTICAL
METHODOLOGIST
Graphic Furnace Atomic
Techniques:
Absorption Spectroscopy (GFAAS)
TECHNIQUE- it’s any chemical or
physical principle that can be used to
study an analyte
METHOD- the application of a
technique for a specific analyte in a
specific matrix
* We use different methods because
they have different matrices
* If you have different matrices means
you have different indifferences
PROCEDURE- a set of written
directions telling us how to apply a
method to a particular sample
Protocol- a set of shins get guidelines
specifying a procedure that must be
followed if an agency is to accept the
results
Common Analytical techniques
Methods:
Procedures:
Protocols:
Pb in soil Pb in water Pb in
blood
APHA ASIM
EPA
TOTAL ANALYTICAL
TECHNIQUE (CLASSICAL)
-measure mass or the volume
- Gravimetry
- Titrimetry
SPECTROSCOPY-the combination of
both principles collectively
APHA- American Public Health
Association
ASTM- American Society for Testing
and Materials
EPA- Environmental Protection Agency
CONCENTRATIONTECHNIQUES
(INSTRUMENTAL)-
-measure concentrations and normally
it’s based from the form of an electrical
or optical signal. It requires specialized
instrument
- Spectroscopy
- Chromatography

2. UNIVERSITY OF NORTHERN PHILIPPINES
COLLEGE OF HEALTH SCIENCES
Bachelor of Science in Medical Laboratory
Science
ANALYTICAL
CHEMISTRY
2ND
SEMESTER
Gravimetry
- a measurement of mass or a change I
mass provides quantitative information
about the analyte
Generalprocedure
Preparation of solution containing the
analyte
Separation of analyte from the sample
Drying and weighing of the isolated
analyte
Calculations
Common Equipment (Materials
Needed)
- Analytical Balance
- Top-loading balance
- Crucibles
- Evaporating dish
- Aluminum gas
- Gravity filtration setup
- Suction filtration setup
- Lab oven
- Muffle furnace
- Desiccators
Types of Gravimetry
1. Particulate Gravimetry- the analyte
is already present as in a particulate
form that is easy to separate from its
matrix
Application 1: determination of total
suspended solids in treated wastewater.
The method uses a glass-fiber filter to
retain the suspended soils.
- After filtering the
sample, the filter is dried
to constant weight at
103-105 C
- Method 25400 in
Standard Method for the
Examination of Waters
and Wastewaters, 20th
edition (APHA, 1998)
Application 2: Determination of crude
total fat in chocolate
- Samples are extracted
with ether for 16 hours
then the lipid extract is
evaporated to dryness at
95- 100 C and weighed
Application 3- measurement of total
suspended particulates in atmosphere
- Total airborne
particulates are
determined using a high-
volume air sampler
equipped with either
cellulose fiber or glass
fiber filters
2. Volatilization Gravimetry- thermal
or chemical energy decomposes the
sample containing the analyte
Measurements: the mass of residue
remaining after decomposition
- Mass of volatile product
collected using a
suitable trap

3. UNIVERSITY OF NORTHERN PHILIPPINES
COLLEGE OF HEALTH SCIENCES
Bachelor of Science in Medical Laboratory
Science
ANALYTICAL
CHEMISTRY
2ND
SEMESTER
- Change in mass due to
the loss of volatile
materials
Application 1 - determination of
organic ash content of pullulan
- Residue in ignition test
Application 2- loss on drying test
for drug substances (USP< 731>)
3. Precipitation Gravimetry-
insoluble compound forms when we
add a precipitating reagent, or
precipitant to a solution containing
our analyte
- In most methods, the
precipitant is the product
of a simple double-
displacement reaction
between the analyte and
the precipitant
Application 1- assessment of
the accuracy of other methods of
analysis
- Verification of the
composition of standard
reference materials
- Identification of organic
and organic analytes
Titrimetry
- Analytical technique in which
volume serves as the analytical
signal
Titration- deliver a measured
volume of a solution of known
concentration (the titrant) into
another solution (titrand)
Note:titrand normally contains the
analyte
Equivalent Point- the point at
which enough titrant has been added
to react exactly with the analyte
(stoichiometric point)
Indicator- a substancethat changes
with or in responseto a chemical
change
Endpoint- the point at which the
indicator changes color so you tell
the equivalence point has been
reached
Stoichiometric Ratio-mole ratio
between the titrant and the analyte
based on a blance chemical reaction
Equipment/ Materials Needed:
- Burette
- Erlenmeyer flask or beaker
- Iron stand and clamp
- Automation titration set- up
GeneralProcedure
- Preparation of analyte and indicator
(if applicable)
- Preparation of titrant
- Gradual addition of titrant to analyte
until endpoint is reached
- Calculations
Moles of titrant= Mt x Veq
Moles of analyte= Kx moles of titrant

4. ANALYTICAL
CHEMISTRY
2ND
SEMESTER
UNIVERSITY OF NORTHERN PHILIPPINES
COLLEGE OF HEALTH SCIENCES
Bachelor of Science in Medical Laboratory
Science
Ma= moles of analyte
Va
Where:
Mt= molarity of the titrant (normally
determined via standardization)
Veq= volume of titrant used to reach the
equivalent point
K= mole ratio between the titrant and the
analyte based on a balanced chemical
equation
Ma= molarity of analyte (mol/ L)
Va= volume of analyte (L)
Acid base titrimetry
- Acid (or base) f known
concentration neutralizes a base or
an acid of unknown concentration
- The titration progress can be
monitored by visual indicators pH
electrodes, or both
HCl + NaOH ----at equivalence point---
NaCl= H2O
Acid base at equivalent point salt
1 mole of HCl = 1 mole of NaOH
Application – kjeldahl analysis for
nitrogen
- Caffeine + saccharine in
pharmaceutical products
- Proteins in foods
- Analysis of nitrogen in fertilizers,
sludges and sediments
Complexometric Titrimetry- those
reactions of simple ion is transformed
into a complex ion and the equivalence
point is determined byusing complex ion
and the equivalence point is determined
by using metal indicators or
electrometrically
Redox Titrimetry- involves oxidation-
reduction reactions between the titrant
and the analyte
Electron
Reducing oxidizing oxidized reduced
Agent agent
Where: reducing agent & oxidize =
oxidation (electron loss)
Oxidizing agent & reduced = reduction
(electron gain)
- The titrant can serve as its own
indicator if tis oxidized and reduced
forms differ significantly in color
Iodometry- an oxidizing agent is added
to excess iodide to produce iodine, and
the iodine produced is determined to/ by
titration with sodium thiosulfate
Iodimetry- is a reducing agent is titrated
directly with a standard iodine solution

5. ANALYTICAL
CHEMISTRY
2ND
SEMESTER
UNIVERSITY OF NORTHERN PHILIPPINES
COLLEGE OF HEALTH SCIENCES
Bachelor of Science in Medical Laboratory
Science
Spectroscopy
- is a technique due to the interaction of
matter which electromagnetic radiation
Basic components of a Spectroscopic
Instrument
Energy Source
EMR sources
Thermal sources
Chemical sources
WavelengthSelector
Filters
Monochromators
Interferometers
Detectors
Photon transducers
Thermal transducers
SignalProcessors
Analog/ digital meters
Recorders
Computers
Infrared Spectroscopy
- IR Range: 780nm- 1mm
- Exploits the fact that molecules
absorb frequencies that are
characteristic of their structure
- This technique is commonly used for
analyzing samples with covalent
bonds
Application – identification of
functional groups and structural
elucidation
- Identification of substances
- Studying the progress reaction
- Detection of impurities
UV- Vis Spectroscopy
Range: 200 nm (near UV) – 800 nm
(very near IR)
- Ultraviolet & visible radiation
interacts with matter which causes
electronic transitions (promotion of
electrons from the ground state to a
high energy state)
Beer’s Law
At= 𝐸 �
�
1 1
- Absorbance= - logo I/Io
Io= incident beam intensity
I = transmitted beam intensity
- Optical path length
- Solution concentration (M/L)
- Molar Absorptivity (L/Mcm)
Atomic Absorption Spectroscopy
- analyte is atomized prior to
detection. Two common types of
atomizers and electrothermal
atomizer
- spectroscopyofatoms or ions
involve promoting an electron from
a ground state to a higher empty
atomic state orbital
Application: analysis of trace metals
Inductively Coupled Plasmas (ICP)

6. ANALYTICAL
CHEMISTRY
2ND
SEMESTER
UNIVERSITY OF NORTHERN PHILIPPINES
COLLEGE OF HEALTH SCIENCES
Bachelor of Science in Medical Laboratory
Science
- is an ionization sourcethat fully
decomposesa sample into its
constituent elements and transforms
those elements into ions.
- composed of argon gas, energy is
“coupled” to it using an induction
coil to form the plasma
ICP-MS:Application
- determination of trace metals in
drinking water, wine, food and those
bound to proteins
- analysis of soil samples for crime
investigations
- determination of nutrients level in
agricultural soils
Chromatography
- refers to a number of application
techniques where molecules are
distributed between 2 phases:
stationary phase, often a
chromatography resin, and a mobile
phase or element
Gas Chromatography
- for separating and analyzing
compounds that can be vaporized
without decomposition
High Performance Liquid
Chromatography
Common detectors:
- UV-vis detector
- PhotodiodeArrays (PDA)
- Mass Spectrometer
Application: HPLC can be used for:
- Checking productpurity during
pharmaceutical development
- Detention of illegal drugs in urine
- Confirming results of synthesis
reactions for research
- Analysis of Vitamin D in plasma for
clinical studies
- Determination of fluoxetine (Prozac)
in serum
- Determination of polyaromatic
hydrocarbons in waste water
Size Exclusion Chromatography
(SEC)
- which also is known by the terms
molecular sieve or gelpermeation
chromatography(GPC)- the
separation of solutes depends upon
their ability to enter into the pores of
the stationary phase
Application:
- measuring both size and
polydispersity of a synthesized
polymer
- absolute molecular weight
measurement when used in tandem
with a viscometer
- analysis of protein structure (tertiary
and quaternary)
Common Detectors:
- UV-vis detector
- Refractive index
- Multi-angle light scattering