Role of Separation

in Quantitative Analysis
 What is Analytical Chemistry?
 “What analytical chemists do”
 Science of chemical measurement
— Design of new experimental methods
— Optimization of existing methods
 Many People Use Analytical Chemistry
 Environmental Monitoring
 Diabetics for glucose monitoring
 Smoke detectors
 Wear sunglasses
 Radar detectors
 Two Major Types of Analysis
 Qualitative Analysis
— to find WHAT constituents are present
in a sample
 Quantitative analysis
— HOW MUCH of a given substance is
in a sample
 Topics of Interest to Analytical Chemists
 Average composition
— overall chemical composition
— foods (e.g. fat, protein, vitamins, minerals, contaminants)
 Surface composition
— ball bearing failure (surface analysis for mode of failure)
 Bulk composition
— how does it differ from the surface composition
— may give insight into processes such as corrosion
 Spatially-resolved composition
— certain areas in a river with high levels of contamination (point source)
 Time-resolved composition
— variation of composition on some timescale (years, days, minutes s, ms, s ns and ps)
 3 Major Classes of Analytical Methods
 Gravimetric Analysis
— based on a mass measurement
 Volumetric Analysis
— based on a volume measurement
 Physicochemical Analysis
— based on a physical/chemical property
— Modern or Instrumental
 Modern Analytical Chemistry
 Spectroscopy
 Electrochemistry
 Spectroscopy and
Electrochemistry
• Solid sample
• Liquid Sample
Excitation Chemical • Gas sample
Source Sample • In vivo
• In vitro
• In lab
• Out in Field
 Spectroscopy and
Electrochemistry

Excitation Chemical Chemical

Source Sample Transducer

 Chemical response -->

Electrical Signal
 Spectroscopy and
Electrochemistry

Excitation Chemical Chemical

Source Sample Transducer

Data Recording
and Processing
 Modern Analytical Chemistry
 Spectroscopy
 Electrochemistry

Separation Science

Remove impurities

Isolate analyte

Identification
Fish are dying at a fish farm
north of here. We think
there is something wrong in
the water. Do you think you
can tell us what it is?
 Fish are dying at a fish farm north of here. We think there is
something wrong in the water. Do you think you can tell us what it is?

? ASK QUESTIONS
? ?
?
 Look at the fish, are they mauled
? ?
 Is the diet ok ?
?  Change in food supplier
Are only the young dying ?

?
? Change in weather -- sudden cold
? ?
? ?
?
 Analytical Process
 Definition - What is the analytical problem?
 Method - Sensitivity, detection limit, cost, ease of use
 Sampling - Obtain a REPRESENTATIVE sample
 Separation - Isolate analyte of interest from the bulk
 Measurement - Quantitative or Qualitative
 Analysis (Measurement)
 Evaluate and Report - internally
 Conclusions and report - publish
 Future

= technician = supervisor
 Sampling
 Obtain a representative sample.
— Type of sample and Type of analysis
— What is the question?

 How much caffeine is in a chocolate bar?

— Sample different brands?
— More than one bar (of each)?

 We can not analyze the entire Population!

 How do we get a representative sample?
Most samples are Heterogeneous
Need Bulk sample from Lot
 Obtaining the Sample
Step 1. Obtain a representative bulk sample.
Step 2. Extract from the bulk sample a smaller,
homogeneous laboratory sample.
Step 3. Convert the laboratory sample into a
form suitable for analysis, a process that
usually involves dissolving the sample.
Step 4. Remove or mask species that will
interfere with the chemical analysis
(Separation!).
 Size of the Bulk Sample
 As small as possible
 Determined by
— Uncertainty that can be tolerated
— Degree of heterogeneity

— Particle size
 Homogeneous vs.
Heterogeneous
 Gas
 Liquid - Suspension or Dissolved
 Solid
— Mixture of solid particles
— Particles have finite size

Homogeneous Random Segregated

Heterogeneous Heterogeneous
 Random Heterogeneous

 Divide into 20000 segments

 Choose 100 at random
 Combine to form Bulk sample
Segregated Heterogeneous material

 Need composite sample

 Divide into 20000 segments
 Draw randomly until correct % from each region
 Combine to form Bulk sample
 Homogeneous Lab Sample
Solid
 Grind to powder
 Mix
 Keep smaller bottle in lab

Liquid
 Homogeneous solution
 Centrifuge out sediment or filter
 Homogeneous Lab Sample

Chocolate
 Freeze
 Grind to powder
 Mix
 Prepare for analysis
 Homogeneous
Lot Heterogeneous

Sampling

Bulk
Storage
Preparation

Aliquot
Lab Aliquot

Aliquot Aliquot Aliquot

 Storage
 Temperature – decomposition
 Humidity – chemical reaction
 Air – oxidation
 Container
— Glass – ion exchange
— Plastics – absorb reagents
 Sample Handling
 Manual Sampling or Automatic
 Type of material
— Inorganics, Trace Metals, Extractable Organics, Volatile
Organics, Nutrients, Microbiological,
 Cleaning of the storage container
 Preservation and Lifetime
 Collect from least to most contaminated site.
 Wear Latex Gloves. New each time.
 Proper Labeling and documentation
“Unless the complete history of
any sample is known with
certainty, don’t bother with it”

“it did not happen if it is

not documented”
 Pre-processing

Technique specific
Pre-concentrate — Filtering
Labeling (radioactive, fluorescent)
Remove impurities
—

— Load onto support

— NMR – IR prep
 Quantitative Transfer
 Transfer analyte in its matrix
— by mass
— by volume
 Analyzing the Sample
Step 5. Measure the concentration of the analyte
in several aliquots.
Step 6. Interpret your results and draw
conclusions.
Chemical Separation
Chromatography - Spectroscopy
 Calibration Curve

20ul
50ug each

 Known standard concentrations

 Best fit line
 Different response for each analyte
Chemical Analysis

 Qualitative
— rt
 Quantitative
— area
 Calibration
 Initial Calibration
— Typically 3 standards in range specified
— Correlation coefficient > 0.995

— Initial calibration verification standard (ICVS) 10%

 Continuing Calibration
— during the analysis after every XX samples
— CCVS - continuing calibration standard 15%

 Reference standard -
— once every YY samples
— certified standard! 10%
— Verifies your in house standards
When does quantification begin?
 Analytical result = 78 ppm caffeine in chocolate
 g analyte moles analyte
vs
g of so ln L of so ln

vol or mass vol or mass vol or mass vol or mass

injected filtered centrifuged of solvent

mass mass mass mass

ground extracted “defatted” dissolved
When does quantification begin?

here
fro m
Caffeine here

e
Cam
Same Concentration

Same Concentration
 Evaluation
78 g caffeine
x g H 2 O dissolved in
g solution
g extracted

%RSD %RSD
0.51% 70%
6.0% 150%
 Compare to Soda?

g per 100g

0.0952
0.0025
0.0250
0.0029
0.1235
0.0705
0.0212
0.0137

Use of literature values!

 Analytical Process
 Definition - What is the analytical problem?
 Sampling - Obtain a REPRESENTATIVE sample
 Method - Sensitivity, detection limit, cost, ease of use
 Separation - Isolate analyte of interest from the bulk
 Measurement - Quantitative or Qualitative
 Evaluation - Are the results meaningful / Reporting

Future - Draw Conclusions / Chain of command