Professional Documents
Culture Documents
ANALYTICAL CHEMISTRY
(PART 2)
THE
ANALYTICAL PROCESS
THE STEPS IN
A CHEMICAL ANALYSIS
1
Steps of the Analytical Process
6 basic main steps in chemical analysis
1. Problem definition - Identify the problem and
formulate questions
2. Method selection - Select method of analysis
3. Sampling - Obtain bulk sample and extract
smaller laboratory sample from bulk
4. Sample Preparation - Prepare sample for
analysis
5. Analysis Analytical measurement of the
analyte in question
6. Calculation and reporting - Calculate results,
interpret/present data and draw conclusion
2
(1) Problem Definition
Need to translate general questions into specific
questions - to be answered by chemical measurement
Example: Why did Singapore Gov. reject our crop
shipments last year?
Formulate the Question: How much pesticide residue is
left on the crop
What information is needed?
What type of sample to be analysed?
How sensitive must the method be?
What degree of accuracy and
precision required?
How are interferences eliminated?
The answers will help determine the specific
techniques to be adopted
3
(2) Method Selection
Selection of method depends on a
number of factors:
Sample type, sample size & preparation
required
Skill and training of analyst
Tools/instruments available
Selectivity, precision, sensitivity required
Cost (budget) and speed
Time required/target deadlines
Availability of methods or standard
methods (chemical literature: Books,
journals, manuals, etc)
4
Standard methods of analysis are available in
published materials such as the following:
Journals
Analytical Chimica Acta Analytical Abstracts
Analytical Chemistry Analyst
Analytical Communications Talanta
J. of Association of Official Analytical Chemists (AOAC)
Journal of Chromatography
Journal of Chromatographic Science
Trends in Analytical Chemistry
Books
ASTM Book of Standards
Official Methods of Analysis of the AOAC
5
(3) Sampling
Sampling is the process to obtain a small
representative and homogeneous sample
- Representative - content of analytical sample
reflects content of bulk sample
- Homogeneous content is the same throughout
the whole sample
Sampling is the most critical step because it can
limit the accuracy of measurements
Requires storage and preservation steps
Sampling methods depends on the samples
- type, size & homogeneity
- physical state (solid, liquid, gas)
- chemical state
6
Sampling Step/Designing A
Sampling Plan
Sampling plan is a plan that ensures that
a representative sample is collected.
7
Gross sample representative
portions of the material to be
tested.
Laboratory sample- a small
portions of gross sample and
made homogeneous
Analysis sample- small portion
of lab sample that actually
analysed
8
Sampling Steps
General guideline:
- Homogeneous parent samples Simple grab
sample approach taken at random and assumed
representative
- Heterogeneous parent samples - Several
samples have to be taken
10
SAMPLING SOLIDS
11
Homogenization of Solid Samples
Crushing
Pulverizing
Grinding
Rendering the sample into a thoroughly
mixed powder
The smaller the particle size of the sample, the lower
the error in analysis
Sample mixtures
Homogeneous samples may become
inhomogeneous upon standing
It should be thoroughly mixed before an
aliquot is taken for analysis
12
Example sampling techniques to obtain
representative gross sample
Stockpile of cereals: take increment from
surface and work into the interior
Compact solids (metals and alloys): random
drilling or sawing across the metal at random
intervals and collect the `sawdust
Bulky material (ore, grain, coal): obtain a
random sample while the material is in motion
(eg conveyor belt), and periodically transfer
portion into a sample container
Shipment/cargo of bagged material (bags of
grain, cement, etc): obtain a small sample from
each bag or every 10th bag, and combine
samples
13
Cone and Quarter Method
18
Continued.
Examples of liquid sampling techniques
Large stationary liquids (eg lakes, rivers) are
sampled at different depths using a thief
sampler (a special device for obtaining
aliquots at different level)
The separate aliquots of liquids can be
analyzed individually or can be combined
into one gross sample (composite sample)
For biological fluids, the timing of sampling
is very important (eg, Blood sample is
collected after the patient has fasted for a
number of hours to analyze for sugar)
19
20
SAMPLING GASES Devices for gas
sampling
Gases tend to be homogeneous
and a large volume of sample is
required (bcause of low density)
Examples
- Air analysis: Use `Hi-Vol
sampler that contain filters to Air Sampling Filters
collect particulates
- Liquid displacement method:
Sample must be slightly
soluble in the liquid & does not
react with it Dust Sampler
- Breath sample: Subject blows Air/Hi Vol
into an evacuated bag Samplers
21
Hi Volume sampler
SAMPLE STORAGE AND PRESERVATION
Gross sample must be transported from the
sampling point to the analytical laboratory
without a physical or chemical change in its
characteristics
Preservation is can be carried out right at the
sampling point or in the laboratory
Sample preservation help minimize:
- physical changes such as adsorption, diffusion,
volatilization
- chemical changes such as oxidation and
microbiological degradation
23
Methods of Preservation & Storage
Method Description
Keep in sealed - Prevent decomposition of
containers or biological samples from
under vacuum or bacterial action or
nitrogen sensitive samples from
Refrigerate or oxidation
freeze or protect - Protect from light
from light - Avoid thermal degredation
of thermal labile samples
- Prevent loss of water from
hygroscopic sample
- Prevent loss of volatile
analytes from sample
24
Method Description
Add chemical - Prevent sample
stabilizers: decomposition by
antibacterials or bacteria
antioxidants - Prevent sample oxidation
Adsorption on a - To immobilize or
solid phase stabilize the analyte
Store in - Teflon (PTFE) for ionic
appropriate analyte
containers - Glass for organic analyte
Acidify (Add 10% - Prevent precipitation of
HNO3 upon metals from water
collection, pH<2) samples
25
26
(4) Sample Preparation
Sample preparation (sample pretreatment) is a step
in chemical analysis where the sample is brought
into the correct size or form for analysis
General principles:
Determine amount (Weight or Volume)
Prepare replicate samples for statistics
- Replicates samples analyzed in the same way, same
size, at the same time
Bring the analyte into the best chemical form for
assay method used Dissolve in solution
Bring analyte into the best concentration range
for the chosen method
Pretreat, separate, pre-concentrate, eliminate
interferences
27
Maximize RECOVERY During Sample Preparation
Recovery:
A measure of the amount of analyte in the assay
relative to the amount of analyte in the sample
Percentage Recovery or % recovery
= (analyte concentration in assay) x 100
(analyte concentration in sample)
How analytes can be lost from the sample during
sample preparation?
- Absorption (eg. metal ions can absorb on the glass
surface, organic compounds can absorbed on plastic
containers)
- Decomposition (eg. organic compounds can
decompose by oxidation)
28
Sample Preparation for Solids
(1) Grind and sieve to a suitable size to obtain
required homogenous size
Homogenize
- Solid sample is placed in a blender (solvent
may be added) and sample is homogenized to
finely divided particles
- Useful for plant and animal tissue, food, and
environmental samples
- Aqueous or organic solvent can be used
- Finely divided particles promote more
efficient extraction
29
(2) Dry samples to remove moisture @ 105-
110oC for 1-2 hr
- Side reactions or decomposition of samples
must be considered
- Follow by storage in a desiccator
34
Types of flux
Base flux: Na carbonates, hydroxides,
borate for alkaline : pyrosulfates, boric
oxide, fluoride acids
Oxidizing Flux: Sodium peroxide or
nitrate/alkaline metal + Sodium Carbonate
Disadvantages: Contamination by flux
material, high salt content may complicate
analysis, high temperature result in loss of
analyte (evaporation), sample container
may react with flux material
35
Errors In Sample Dissolution
36
To Measure Analyte containing
Interfering species
a) Use a selective analytical technique (eg
ion selective eletrode)
b) Use selective derivatization that converts
the analyte into another chemical species
that can be measured more easily
c) Use Standard Solution of high-purity
standard materials
d) Remove the analyte from the sample
matrix by a separation or extraction
process
Separation and Preconcentration
Separation of analyte from matrix -
Eliminate interferences from samples*
Provide suitable selectivity
Required if many analytes are present
Cause preconcentration of analyte - good for
more sensitive or accurate measurement
Preconcentration -
Required if amounts of analyte(s) too small in
bulk material (eg trace element analysis)
Extraction + preconcentration (10-100 times)
- Improves sensitivity
- Eliminate interferences from the sample matrix*
38
Summary of Separation Techniques
Soxhlet extraction
Liquid-liquid extraction
Chromatography : Gas chromatography,
liquid chromatography, gel permeation
chromatography, TLC, Ion exchange
chromatography, Paper
chromatography
Electrophoresis : liquid/solid
Dialysis
Precipitation : liquid/solid
Addition of Masking agent
Soxhlet Extraction
Classical extraction method
(Apparatus named after developer)
40
40
Dilution and Matrix Matching
After dissolution and separation, the sample
may not be suitable for direct measurement
(1) The instrument or method requires a particular
type of sample form, concentration, solvent,
oxidation state, etc
(1) Some samples require modifications:
- pH adjustment with acid/base/buffer
- Adjustment of oxidation state using an
appropriate oxidising agent
- Dissolving in acid (trace metal analysis by
spectroscopic techniques often requres
nitric acid to avoid mass-spectral
interferences with chloride solution)
(5) Analytical Measurement
Measurement is often the simplest
stage of the analytical process
Use reagents of high purity (eg. analytical
grade, reagent grade, etc)
A blank measurement must be performed
for trace analysis (The results of the blank will
be subsequently subtracted from the raw
analytical measurement)
Choice of Analytical Measurements:
- Classical methods
- Instrumental methods
42
Classical methods (gravimetric, volumetric)
- Accurate and precise to approx. 0.1% but
require large (mmol, mg) amount of analyte
43
(6) Calculating Results and Reporting
(1) Determine the concentration of the
analyte in the analytical sample solution
(2) Use results to calculate the amount of
analyte in the original (bulk) sample
Evaluate the results
- Requires appropriate use of statistics
- Must be reasonable, reliable and related to the
problem as originally stated
Data presentation must be understood,
conclusions clearly shown
Report results with accuracy & precision
(include standard deviation, mean value, etc)
Verify reports (Professional/charted chemist)
44
ILLUSTRATIONS AND
PHOTOGRAPHS
OF SAMPLING
TECHNIQUES
AND APPARATUS
45
Consider the different
flowrates, mixing and
turbulance (eg rainy and
dry season)
46
GOOGLE EARTH
47
In-situ testing
(Parameters such as pH, DO, turbidity)
48
Composite Water Grab Sampler
Sampler (with (Van Dorn Sampler)
rechargable battery, able
to adjust to desired
sample: 25 mL to 600
mL, time 5 min to 4 hrs)
49
Water Sampling
(How much water do you need?)
Depends on parameters
to be analyzed
Chlorophyll and TSS often
require the greatest
volume (> 1L)
Better be safe to have
too much water rather
than too little
Depends on the system
and how practical it is
to carry out large
volumes of water
50
Water sampling
(Microbes)
Grab samples taken
with sterile containers
51
Examples of Gas Samplers
Hi-vol sampler
[Particulate (PM10) and Underground gas
microbes on filters] sampler
(sampling gas leaks,
methane decomposition
in landfills, etc)
Gas
canister
52