The Analytical Process

1) Selecting a method
2) Sampling
3) Preparing the laboratory sample
4) Defining replicate samples
5) Preparing of solutions of samples
6) Eliminating interferences
7) Measurement of analyte concentration
8) Calculations
9) Determining reliability of data
Selecting the method
Method used must be fit for the purpose
> Ex. databank of trends, quality assurance,
goods assessment, forensics
Source of methods:
1. In-house method
2. Methods published in open scientific lit
3. Methods supplied by trade organizations
4. Methods in books published by professional
organizations (Ex. Royal Academy of
Chemistry)
5. Methods from standards organizations (Ex.
US-EPA)
6. Methods from statutory organizations
Factors to consider in choosing a method:
1. Limit of detection
Important for trace analysis
Should ideally be at least 1/10th of the
concentration to be measured

 Ex. legal limit of Pb in tap water is 50

ppb, therefore analytical limit of detection
should be 5 ppb
2. Accuracy
Very often NOT important in trace
analysis
3. Precision
4. Speed to minimize effort and cost
5. Equipment required
6. Sample size
Especially in clinical chemistry and
forensics
Ex. blood samples
Linked to limit of detection
7. Cost
8. Safety
9. Specificity degree of discrimination
between analyte and other substances
present or extracted from the matrix
Making your choice: Will the method selected
be adequate for the decision you have to take
when the result is available?
Sampling
Process of selecting a portion of material, in some
manner, to represent or provide information about
a larger body of material
Types of samples
1. Representative sample sample that is typical
of parent material for the characteristic under
inspection

 must take into account the state of the

parent material:
a) homogeneous Ex. vegetable oil at 40oC,
filtered aqueous solution
b) heterogeneous Ex. palm oil at 15oC,
muesli breakfast cereal
c) static system composition of material is
permanent wrt position in space and
stable in time
Ex. sample of oil in a drum, warehouse
stock of food
d) dynamic conditions parent material is
changing with time; removal of portion at
any instant represents only a snapshot of
that moment in time and in that particular
location
Ex. estuarine or riverine water
2. Selective sample sample deliberately chosen
by using a sampling plan that screens out
materials with certain characteristics and/or
selects only material with other relevant
characteristics; directed or focused sampling
Ex. suspected contamination of sample due to
rat feces
3. Random sample sample selected by a random
process to eliminate questions of bias in
selection and/or to provide a basis for statistical
interpretation of measurement data
Types:
a) simple random sampling any sample
has an equal chance of selection
Ex. raffle draw

b) stratified random sampling lot is

subdivided/stratified and a simple
random sample is selected from each
stratum Ex. regional raffle draws
c) systematic sampling first sample is
selected at random then the subsequent
samples are taken according to a
previously arranged interval (every 5th,
10th, etc.)
Ex. take every 50th bottle in a production
line for quality control analysis
4. Composite sample consists of 2 or more
portions of material (collected at the same time)
selected so as to represent the material being
investigated
ratio of components taken to make up
composite can be in terms of bulk, time or
flow
components of the composite sample are
taken in proportion to the amount of the
material they represent Ex. food samples
may be bulked together in proportion to the
amount normally consumed
Preparing the laboratory sample
Crushing and grinding of lab samples
required to decrease the particle size of solid
samples
tend to alter composition of sample, hence
particle size should be reduced no more than
is required for homogeneity and ready attack
by reagents

 factors that can cause appreciable changes in

samples composition as a result of grinding:

heat can generate loss of volatile
components
increase in surface area increases
susceptibility to reaction with atmosphere
Ex. Fe content of rx can decrease by as
much as 40% during grinding
water content altered substantially during
grinding
differences in hardness of the component
softer components ground to fine particles
more easily and be lost as dust
intermittent screening often increases
efficiency of grinding
tools employed:
large samples with large lumps: jaw
crushers, disk puverizers
medium-size samples and particles: ball
mills useful device for grinding of solids
that are not too hard
small materials: mortars
> Plattner diamond mortar used for
crushing hard, brittle materials
Mixing Solid Lab Samples
essential to ensure a random distribution of
the components in the analytical samples
common method: rolling sample on a sheet of
glazed paper

 others: rotating sample for some time in a ball

mill or a twin-shell V-blender

Note: long standing, finely ground
homogeneous materials may segregate on
the basis of particle size and density

Types of Moisture in Samples

Essential water part of the chemical identity
of the sample; present in stoichiometric
amounts
o Water of crystallization/hydration
Ex. BaCl2 2H2O
o Water of constitution
present as H and OH groups which
combine to give H2O
takes higher T to expel from samples
Ex. C6H12O6 6C + 6H2O
Non-essential water superficially held water
or water bound to the sample physically and
removed at a much lower T range (100-120oC)
o Adsorbed water retained on surface of
solid
o Sorbed water condensed water in
interstices of capillaries of colloidal solids
o Occluded water liquid water entrapped in
microscopic pockets spaced irregularly
throughout solid crystals

Common Practices
1) remove moisture from samples prior to weighing
or to bring water content to some reproducible
level
2) determine water content at the time samples are
weighed for analysis so that results can be
corrected to a dry basis
Example: A food sample analyzed as received, is
found to contain 14.57% protein and 10.4% water.
Calculate % protein on a dry basis.
Example: 1.000 gram of soil as received, gave a
moisture content of 14.0%. The oven-dried sample,
completely moisture-free, showed 18.00% K. Find
the % K in the sample as received.
Methods of Moisture Determination
1) Application of Heat
a. Drying done to make sample anhydrous or
to remove adsorbed moisture but retain
chemically combined water
i. Air-drying
ii. Oven-drying
1. conventional oven
2. vacuum oven 50-100 mm Hg
3. microwave oven
iii. Dessicator
iv. IR lamps

Disadvantages of drying:
Procedure not specific for water
Large (+) systematic errors in samples
that
yield
volatile
decomposition
products upon heating
Time-consuming
b. Freeze-drying or lyophilization
For biological materials and unstable
natural products
Water from frozen sample is evaporated in
a high vacuum system, T kept below 0oC
Nearly dry sample further dried at high T in
order to attain a minimum weight in the
shortest possible time
c. Distillation with a high boiling liquid
immiscible with water
Total vapor pressure above a binary
system of immiscible liquids = sum of
vapor pressure of the 2 liquids
Two liquids boil and distill over a lower T
than they would if they were not in a
mixture
Common carrier liquids: toluene, o-xylene,
m-xylene, p-xylene

2) Chemical Methods: Karl Fischer Titration

KF Reagent:
o Solution A: pyridine, methanol, SO2 +
sample
o Solution B: I2 in methanol (titrant)
Reactions:

C5H5NI2+C5H5NSO2+C3H5N+H2O2C5H5NHI+C5H5N
SO3

C5H5NSO3 + CH3OH C5H5N(H)SO4CH3

important because C5H5NSO3 is also capable of
consuming water
C5H5NSO3+ H2O C5H5NHSO4H
undesirable because not specific for water; prevented
by excess CH3OH
Endpoint:
o yellow light reddish brown color of
excess I2
also be determined using
o may
electrometric methods
Interferences: compounds that react with
one of the components of the reagent to
produce water
o carbonyl compounds (RCHO)
o metal oxides react with HI
o oxidizing or reducing substances that
reoxidizes iodide produced or reduces
I2 in the reagent

Preparing of solutions of samples: Decomposing

and Dissolving the Sample
Sources of error:
Incomplete dissolution of analytes ideal
sample treatment should dissolve sample
completely
Losses of analyte by volatilization
Introduction of analyte as a solvent
contaminant
Introduction of contaminants from reaction
of the solvent with the vessel walls
Decomposing Samples by Inorganic Acids in Open
Vessels
Most common reageant: mineral acids
Hydrochloric acid excellent for inorganic
samples but finds limited application for
decomposing organic materials; widespread
use for dissolving many metal oxides and
metals more easily oxidized than hydrogen
Nitric acid strong oxidant that dissolves all
common metals except Al and Cr; alone or
in combination with other acids and
oxidizing agents, widely used for
decomposing organic samples (wet ashing)
Sulfuric acid many materials are
decomposed and dissolved by hot,
concentrated sulfuric acid; effectiveness
due to its high boiling point
Perchloric acid attacks a number of iron
alloys and stainless steel that are

intractable to other mineral acids; problem:

explosive when hot, concentrated perchloric
acid comes in contact with organic
materials or easily oxidized inorganic
substances
Oxidizing mixtures
Aqua regia 3:1 mixture of HCl:HNO 3
Br2 or H2O2 hasten oxidation of
organic materials
HNO3 + HClO4 useful also for
organics
Hydrofluoric acid primary use is for
decomposition of silicate rocks and
minerals in the determination of species
other than silica
Microwave Decompositions
May be carried out in open or closed
vessels, but the latter is preferred since
higher temperatures are realized because
of the higher pressures.
Evaporative losses are also avoided. Less
reagents required, less risk of interferences
due to contamination.
Loss of volatile components of sample
avoided.
Easy to automate. Main advantage: speed
Vessels for moderate-pressure
digestions
High-pressure microwave vessels
designed to operate at 80 atm or 10

times the pressure that can be

tolerated by moderate-pressure
vessels
Microwave ovens designed to heat
simultaneously 12 of the moderatepressure vessels
Microwave furnaces for performing
fusions and for dry ashing samples
containing large amounts of organic
materials; temperatures of 1000oC
reached in 2 minutes!

Elimination of Interferences
Interference arises whenever a species in the
sample matrix either produces a signal that is
indistinguishable from that of the analyte or,
alternatively, attenuates the analyte signal
2 Methods:
1) use of a masking agent immobilizes or
chemically binds the interfering species in a
form in which it no longer contributes to or
attenuates the signal from the analyte
>
Ex. use of F- to prevent Fe3+ from interfering
in the iodometric determination of Cu2+
2) Converting either the analyte or the
interferences into a separate phase that can be
separated mechanically
> Most common method: chromatography
> Others: precipitation, extraction, ionexchange, distillation