Bai Giang Chung PDF

Chemistry 208
Lecture Notes

2006

David E. Henderson
1
8/15/2006 Chem208Spring2006 1
Chem 208
Introduction to Analytical Chemistry
8/15/2006 Chem208Spring2006 2
Chem 208
Most Useful Course
Everyone in science does Analytical work
Summer J obs
Research preparation
Most Important area of Chemistry?
8/15/2006 Chem208Spring2006 3
How AC Saved the Human
Genome Project
CapilaryElectrophoresis J orgenson UNC
4 Color Fluorescence Det. Smith U. Wis.
Reusable Capillaries Karger Northesastern
Look at all Capillaries at once Yeunge
IowaState
HGP Finished 3-5 years early
Cost $300 million instead of $3 billion
Anal Chem74 (1) p. 23A (2002)
2
8/15/2006 Chem208Spring2006 4
Blackboard - Handouts and
prelabs
All Handouts on Blackboard
Prelabs- Read handout first
Watch Videos
Answer questions
MUST COMPLETE FOR ADMISSION TO
LAB
8/15/2006 Chem208Spring2006 5
Materials Required
Textbook
Lab Notebook - duplicate page type
Calculator (TI-86, 83 or 82)
Least Squares
Quadratic Equation solver
8/15/2006 Chem208Spring2006 6
Review Blackboard Handouts
Plagiarism Handout
Syllabus
Course Schedule
Isis Draw
Project Lab
TI Interactive
3
8/15/2006 Chem208Spring2006 7
Project Lab
Culmination of Course
Real Analytical Problemto solve
Literature Research
Experiment Design
Mini-Research Project
Public Presentation of Results
You will be guided through this
Community Service Opportunity Alta Lash
8/15/2006 Chem208Spring2006 8
HISTORY OF ANALYTICAL
CHEMISTRY - 1950's
pH meters and single wavelength
spectrophotometers, and
electrochemical techniques.
Lots of titrations, gravimetric analysis.
Data was primarily one dimensional.
Experiment ====>Number
8/15/2006 Chem208Spring2006 9
CHEMISTRY -1960s
Gas Chromatography
Atomic Absorption spectrophotometry
Analysis of ppm and ppb levels of
metals and organics in the environment
begins.
Thus data representations were now two
dimensional.
Experiment ===>Graph
4
8/15/2006 Chem208Spring2006 10
CHEMISTRY -1970s
Liquid chromatography
Mass spectrometry
Sensitivity allowing part per trillion trace
analysis.
We begin to find virtually everything almost
everywhere. Surface analysis of thin layers
becomes common.
problems of the environment, energy, and
biological and physiological analysis.
8/15/2006 Chem208Spring2006 11
CHEMISTRY -1980s
Hyphenated methods
GC-MS, LC-MS, GC-IR, etc.
Computers control instruments, manipulate
data, and run experiments
Robots conduct complete analytical schemes
Multidimensional data representations
Single Atom detection
Three dimensional data presentations
become important (MS-MS and 2D NMR).
Experiment ===>3D Graphic
8/15/2006 Chem208Spring2006 12
CHEMISTRY -1990s
Sample preparation is a major focus
old, time consuming methods of solvent
extraction are replaced with fast, automated
procedures.
Multi-channel analysis becomes the
major thrust (396 well plates, parallel
analysis).
LC/MS brings MS to biochemistry
5
8/15/2006 Chem208Spring2006 13
PRESENT OF ANALYTICAL
CHEMISTRY -2000s
Single molecule detection in living cells
Multi-channel hypenated methods
combined with combinatorial processes.
96 channel CE-ESI/MS.
High speed analysis (1000 samples per
day per operator for HPLC/MS).
LC/NMR/MS Instruments
Commercialized
8/15/2006 Chem208Spring2006 14
THE PROCESS OF
ANALYTICAL CHEMISTRY
I. Define The Question -
What information is needed?
How accurately must the answer be
known?
What legal constraints apply to the results?
8/15/2006 Chem208Spring2006 15
THE PROCESS OF
II. Select the best available method for
the analysis
This is the most difficult step and requires
the most background.
What Differentiating Characteristicdoes
the analyte have? - Get a Handle on the
Molecule
What tools are available to exploit that
characteristic?
Analytical chemists tend to be
generalists.
6
8/15/2006 Chem208Spring2006 16
THE PROCESS OF
III. How will the method be validated
Are Standard Reference Materials
Available
QA/QC of your method
8/15/2006 Chem208Spring2006 17
THE PROCESS OF
IV. Sampling -The sample to be
analyzed must be representative of the
bulk of material of interest.
It must not be contaminated or altered prior
to analysis.
How much sample, what particle size, how
many replicates.
You must measure the sample
accurately.
8/15/2006 Chem208Spring2006 18
THE PROCESS OF
V. Sample Preparation.
Most samples do not come ready for
analysis.
The process of getting them ready for a
measurement is, in many respects, more
important than the measurement itself.
Without accurate preparation, the
measurement is useless.
7
8/15/2006 Chem208Spring2006 19
THE PROCESS OF
VI. Separation - remove interferences
not always be required
Most separations involve an equilibration between
phases.
Solid-Liquid (precipitation, SPE), Gas-Liquid (distillation,
GC), Liquid-Liquid (extraction, HPLC).
Modern chromatographic methods often allow
the separation of as many as 1000
components in a mixture.
8/15/2006 Chem208Spring2006 20
THE PROCESS OF
VII. Measurement-Analysis.
step most often associated with Analytical
Chemistry.
often done by specialists when the specific
measurement involves complex
instrumentation.
Robotic systems are also becoming
important here.
8/15/2006 Chem208Spring2006 21
THE PROCESS OF
Chemical Measurements
cheap and accurate
require no calibration.
However
often slow, insensitive to trace amounts,
and difficult to automate.
Titrations.
Gravimetric Analysis.
8
8/15/2006 Chem208Spring2006 22
THE PROCESS OF
Physical Measurements.
high sensitivity, continuous or automated
operation, speed (low cost).
However- require careful calibration and
offer lower precision than the chemical
methods.
Spectroscopy.
Electrochemistry.
Physical property measurement.
8/15/2006 Chem208Spring2006 23
THE PROCESS OF
Biological Measurements.
extremely specific
often slow and of low precision and
accuracy.
Growth studies.
Inhibition studies.
Enzymatic assays.
Immunoassay.
8/15/2006 Chem208Spring2006 24
THE PROCESS OF
IX. Evaluation Of Data.
calculating the result
evaluating the significance of the result.
QA/QC - how well the experiment was done and
how much confidence can be placed in the
answer. (decisions effecting lives and money are
at stake)
Chemometrics is a growing field within Chemistry
which deals entirely with data and experimental
optimization.
9
8/15/2006 Chem208Spring2006 25
HOW TO SURVIVE CHEM
208 (OR BETTER YET DO
WELL)!!
LEARN TO DISCRIMINATE WHAT
IS IMPORTANT FROM WHAT IS NOT.
This is at the heart of your success in
learning Analytical Chemistry.
You cannot get by with just memorizing
everything.
Appropriate problem solving is the
name of the game here.
8/15/2006 Chem208Spring2006 26
Appropriate Problem Solving
In lab
when to use a Class A volumetric pipet and
when to just add a few drops.
balance being too careful and never
getting finished with being too sloppy and
getting terrible results. (do you need an
exact weight or just to know the weight
exactly?)
In the class
when to approximate an answer and when
you need a result which is exact.
8/15/2006 Chem208Spring2006 27
Solving Equilibrium Problems
What chemicals react with what. Surprise!
You need to learn some chemistry. Only after
you identify the reactions can you begin to
look for equilibria.
Learn to look for all reactions that may be
present.
Which reactions go to completion and which are
equilibria?
write out a complete scheme of chemical
equations to describe what is happening when
dealing with these problems.
10
8/15/2006 Chem208Spring2006 28
Solving Equilibrium Problems
Work through all material with a pencil
and paper. It is usually not sufficient to
just read this material. You will need to
work it through yourself to see each
step.
8/15/2006 Chem208Spring2006 29
WORKING MEMORY
OVERLOAD!!
A.H. J ohnstone."NewStars for the Teacher
to Steer By?"J .Chem.Ed.61: 847-49 (1984).
WHAT IS IT?
each of us has a limited and essentially
constant amount of working memory
entire problem must fit into this memory.
8/15/2006 Chem208Spring2006 30
Working Memory Overload
Larger problems must be broken down
into smaller pieces which will fit in the
available space.
A problem which must be broken down
into smaller problems will necessarily
take longer to solve than one which will
fit in memory as a single problem.
11
8/15/2006 Chem208Spring2006 31
"Every learner has to analyze the
information coming in and organize it for
him- or herself, if the learning is to
become a part of him or her. If her or she
tries to take the teacher's organization
and structure, he or she has to resort to
memorization which certainly does not
guarantee understanding."
1
Thus only by working through problems
in a way you understand and developing
your own mental organization can you
expect to deal with the problems you will
encounter in this course.
1
8/15/2006 Chem208Spring2006 1
Chem 208
Chapter 1 Basic Tools
8/15/2006 Chem208Spring2006 2
Using the Text
Read with pencil and paper
Do assigned problems
Detailed answers will be posted to Blackboard
before exams.
Review any problems you got wrong
Quizzes, classroom exercises, problem
seminars
8/15/2006 Chem208Spring2006 3
.Calculations and Units -
Use Factor Label Method!!
Unit Conversions
How many moles of CH
2
Cl
2
are in 20.2 ml?
20.2 ml * 1.37 gm/ml * 1 mol/85.1 gm=0.324 mol
2
8/15/2006 Chem208Spring2006 4
Units of measurement for
concentration
Molarity - Moles per liter of solution
C
A
Formal or Analytical concentration
eg. C
HOAc
=1.00 M or 1.00 F means 1.00 moles
added to make 1.00 l of solution.
[ ] Equilibrium Concentration. Quantity of a
species present at equilibrium.
eg. [HOAc] [H+] [OAc
-
] We will spend a lot of
time and effort getting from analytical
concentration to equilibrium concentration.
8/15/2006 Chem208Spring2006 5
concentration
C
w/w
or % w/w Percent weight/weight
grams analyte/grams solution * 100%
20% w/wNaCl soln =20 gm NaCl +80 gm H
2
O
C
w/v
or % w/v percent weight/volume
gm analyte/ml solution * 100%
20% w/v NaCl soln =20 gm NaCl 100 ml soln
C
v/v or
% v/v percent volume/volume
ml analyte / ml solution * 100%
20% v/v ethanol =20 ml 100% ethanol 100 ml
solution
8/15/2006 Chem208Spring2006 6
concentration
CAUTION: in water where 1 gm =1 ml
treat w/w and w/v calculations as interchangeable.
Rarely strictly true.
How accurate is the result?
How close is the solution to pure water?
river water =pure water
sea water, blood pure water
organic solvents pure water
Density correction ml sample * gm/ml =gm
sample
Do solution volumes add ideally?
3
8/15/2006 Chem208Spring2006 7
concentration
ppm useful for trace analysis ppm=
gm analyte/10
6
gm solution (~10
6
ml)
gm analyte/1000 liters solution (water)
mg analyte/liter solution
g analyte/ml solution
8/15/2006 Chem208Spring2006 8
concentration
ppb ultratrace analysis
ppb=gm analyte/(10
9
gm solution)
g analyte/liter solution (water)
ng/ml
pg/l
8/15/2006 Chem208Spring2006 9
Converting ppm Molarity
How much Na
2
SO
4
would you use to
prepare a 100 ppm sodium sulfate?
100 ppm=100 mg/l
How much to prepare 100 ppm sulfate ion?
100 mg/l sulfateX (142 g Na
2
SO
4
/mol)/(96 g SO
4
/mol) =148 mg
Molarityof 100 ppmsulfate?
100 mg/l x 1 mol/96 g x 1 g/1000 mg =1.04x10
-3
M
4
8/15/2006 Chem208Spring2006 10
Weighing Samples
Weigh by Difference
Samples at Room Temperature
Fingerprints weight
Weighing Bottles for dried samples
Exact weight vsweigh exactly
When to use a low precision balance
8/15/2006 Chem208Spring2006 11
Making Solutions
Volumetric Flasks TC
Add sample quantitative transfer
Dissolve before you fill
Mix 10 inversions
Dont heat sample in Vol. Flask
8/15/2006 Chem208Spring2006 12
Dilution
C
o
V
o
= C
d
V
d
Quantity in original portion =quantity in
dilute portion
Works for any concentration unit
5
8/15/2006 Chem208Spring2006 13
Solution Stoichiometry
1-34 How many g of 0.491% w/wAqueous HF
are required to provide a 50% excess to react with
25 ml of 0.0236 M Th
4+
Th
4+
+4 F- ThF
4
Moles Thx 4 mol F-/Mol Thx 1/.50 =mol F-
5.9x10
-4
x 4 x 2 =0.00472
Mol F- x 19 g/1 mol x 1 g solution/0.00491 g F- =g solution
0.00472 x 19 / 0.00491 =18.3 g solution
1
Chapter 2 & 3
How do we measure things
Chem 208
8/15/2006 Chem208Spring2006 2
RECORDING DATA - USE OF
SIGNIFICANT FIGURES
INTEGERS (Things you count)
ALWAYS EXACT VALUES eg.you did the
experiment three times.
Precision of these numbers is infinite.
REALS (Things you measure)
ALWAYS CONTAIN AN UNCERTAINTY.
Assume error is +/- 1 in the last place unless specified.
For the number 1.0034 the actual value lies between
1.0033 and 1.0035.
8/15/2006 Chem208Spring2006 3
METHODS FOR
EXPRESSING
UNCERTAINTY
ABSOLUTE UNCERTAINTY.
Buret reading 40.34 +/- .01 ml
RELATIVE UNCERTAINTY.
0.01 ml/40.34 ml =.00025 relative
.00025 * 100% =.025% rel.unc.
.00025 * 1000 ppt=.25 ppt rel.unc. =0.2 ppt
2
8/15/2006 Chem208Spring2006 4
SIGNIFICANT FIGURES
KEEP ONLY THE FIRST UNCERTAIN
DIGIT IN A RESULT.
Assume that data is presented this way
by other responsible scientists.
8/15/2006 Chem208Spring2006 5
SIGNIFICANT FIGURES
ROUND OFF SUPERFLUOUS
DIGITS.
5 ROUNDS UP IF ODD DOWN IF EVEN.
Thus the result of rounding the digit 5 is always
a number ending in an even digit. eg..435 =>
.44 .425=>.42
Round off only once
8/15/2006 Chem208Spring2006 6
SIGNIFICANT FIGURES
ADDING OR SUBTRACTING
KEEP ONLY THE NUMBER OF DECIMAL
PLACES IN THE LEAST PRECISE
RESULT.(Remember these numbers will
always have the same units.)
Note- the number of significant digits in a
number can change dramatically in
subtraction.
3
8/15/2006 Chem208Spring2006 7
SIGNIFICANT FIGURES
Absolute uncertainty
42598 21.1
-42595 2.035
3 +6.12
29.255
29.3 0.1
8/15/2006 Chem208Spring2006 8
SIGNIFICANT FIGURES
MULTIPLYING OR DIVIDING
Rule of Thumb Keep the same

number of significant digits in the result
as in the value with the lowest number
of sig. Figs.
This often doesnt work out
101/99 =1.02
8/15/2006 Chem208Spring2006 9
SIGNIFICANT FIGURES
EXAMPLE
21.1 * 0.029 * 83.2 =50.91008
Rel.Unc 0.1/21.1 0.001/0.029
0.1/83.2
0.0047 0.034 0
.0012
.03 * 100% =3% rel. unc.

4
8/15/2006 Chem208Spring2006 10
SIGNIFICANT FIGURES
COMMON ERROR 0.7834 gm KHP =
0.0034 moles KHP
This error appears in far too many lab
notebooks.The student has taken a
number with a relative uncertainty of
1/7800 and reduced it to a number with
an apparent uncertainty of 1/34.
8/15/2006 Chem208Spring2006 11
Common error
1.234*(40.35-40.31)=0.04936
How many significant figures should we
have?
1.234*0.04=0.04936
0.05
8/15/2006 Chem208Spring2006 12
SIGNIFICANT FIGURES
DURING CALCULATIONS
ONE ADDITIONAL INSIGNIFICANT DIGIT
SHOULD BE CARRIED IN
INTERMEDIATE RESULTS.
EXCEPTION.The calculation of sums
and sums of squares requires you to
keep all digits
5
8/15/2006 Chem208Spring2006 13
LOGS
log consists of two parts,
characteristic
eg.pH=1.03 characteristic =1 mantissa =.03
The characteristic is a power of ten and is an
integer (counting decimal places).
mantissa.
Measured value - real
The significant figures which denote the
uncertainty of the value are those in the
mantissa.
8/15/2006 Chem208Spring2006 14
LOGS - General rule
Number of Sig. Figs is number of digits to
right of decimal
A two digit mantissa corresponds to a relative
error of approximately 2%,
A three digit mantissa to 2 ppt, etc.
EXAMPLE
pH 1.03 [H
+
]=9.3x10
-2
pH 13.03 [H
+
] =9.3x10
-14
1.02 <pH <1.04 13.02 <pH <13.04
9.1x10
-2
<[H
+
] <9.5x10
-2
9.1x10
-14
<[H
+
] <
9.5x10
-14
Note that the relative uncertainties are the
same.+/- 2/93 =2% rel.unc.
8/15/2006 Chem208Spring2006 15
SIGNIFICANT FIGURES
Uncertainty of sum or difference
40.34 ml +/- .01 ml
-1.03 ml +/- .01 ml
39.31 ml +/-0.014 ml
2 2 2
c b a y
s s s s
c b a y
+ + =
+ + =
014 . 0 0002 . 0 =
6
8/15/2006 Chem208Spring2006 16
SIGNIFICANT FIGURES
MULTIPLYING OR DIVIDING
UNCERTAINTY OF THE RESULT IS
THE SUM OF THE RELATIVE
UNCERTAINTIES.
2 2
2
*
=
=
c
s
b
s
a
s
s
c
b a
y
c b a
y
8/15/2006 Chem208Spring2006 17
SIGNIFICANT FIGURES
EXAMPLE
21.1 * 0.029 * 83.2 =50.91008
Rel.Unc 0.1/21.1 0.001/0.029
0.1/83.2
0.0047 0.034 0
.0012
.03 * 100% =3% rel. unc.

.0047
2
+.034
2
+.0012
2
= 0 . 034
8/15/2006 Chem208Spring2006 18
CALCULATION OF
MAXIMUM EXPERIMENTAL
ERROR.
The sum of all errors is best approximated by adding
the squares of all errors and taking the square root of
the resulting sum.
Follow each step in the procedure
Add the square of the absolute uncertainties
during addition and subtraction.
Add the square of the relative uncertainties
during multiplication and division.
You should obtain an absolute or relative
uncertainty for the final result.
7
8/15/2006 Chem208Spring2006 19
CALCULATION OF
MAXIMUM EXPERIMENTAL
ERROR -Examples.
Addition and subtraction
Volume of titrant =Final buret reading -
initial reading
V=43.24 0.01 ml - 0.230.01 ml =
43.01 ml
error= 0.01
2
+ 0.01
2
= 0.014ml
8/15/2006 Chem208Spring2006 20
CALCULATION OF MAXIMUM
EXPERIMENTAL ERROR -
Examples.
Multiplication and division
Molarity of titrant =grams acid*(mol /
gm)/vol. titrant
M=0.5675.0001 gm* 1 mol/204.23.02 gm /
43.010.014 ml * 1 l./1000 ml
M=.0646066 Mol/l
error = (
0.0001gm
.5675gm
)
2
+ (
.02gm/ mol
204.23 gm/ mol
)
2
+ (
0.014 ml
43.01 ml
)
2
8/15/2006 Chem208Spring2006 21
CALCULATION OF MAXIMUM
EXPERIMENTAL ERROR -
Examples.
error =0.00038 =0.0004 relative error
0.0004 * 1000 =0.4 ppt
0.0004 * 0.0646066 Mol/l =0.0000258
Mol/l
M =0.064610.00002 mol/l
error = (
0.0001gm
.5675gm
)
2
+ (
.02gm/ mol
204.23 gm/ mol
)
2
+ (
0.014 ml
43.01 ml
)
2
8
8/15/2006 Chem208Spring2006 22
SIGNIFICANT FIGURES
a
s
x
y
s
a y
a
y
x
=
=
Partial Derivative Calculus- how does y change
with small changes in the value
of the measured quantity a
a
s
s
a y
a
y
303 . 2
) log(
=
=
a
y
a
s
y
s
y
303 . 2
10
=
=
8/15/2006 Chem208Spring2006 23
Stoichiometry
Reactants and Products related by
Coefficients
Many Analytical Procedures relate analyte
to a reaction product or another reactant
Titrate HCl with NaOH
H
+
+OH
-
H
2
O
Measure OH with buret
Moles OH =moles H
8/15/2006 Chem208Spring2006 24
Stoichiometry
Conservation of Mass
Mass of each atomis conserved in all Reactions
Types of Reactions
Acid Base = Protons Conserved
Redox =electrons conserved
Complexation=electron pairs conserved
Precipitation =Charge conserved
9
8/15/2006 Chem208Spring2006 25
Stoichiometry
Conservation of Mass
C
4
H
10
+ O
2
4 CO
2
+5 H
2
O
Moles CO
2
=4 x moles C
4
H
10
Moles H
2
O =5 x moles C
4
H
10
2 x Moles H
2
O =10 x moles C
4
H
10
8/15/2006 Chem208Spring2006 26
Stoichiometry Example
C
10
H
20
N
2
S
4
+oxidizing agent 4 SO
2
SO
2
+H
2
O
2
H
2
SO
4
H
2
SO
4
+2 NaOH 2 H
2
O
Measure moles of NaOH (buret)
Overall Ratio 1:8
Anabuse mol
4
1
1
1
2
1
NaOH moles
2 4 2
2 4 2
=
molSO
molAnabuse
x
SO molH
molSO
x
molNaOH
SO molH
x
8/15/2006 Chem208Spring2006 27
A Look Inside What We Do
Usually dont talk about these things
explicitly.
Need to convert quantities into NUMBERS
Counting - M&M colors
Numbered Scales - Buret
Digital Displays - Balance
10
8/15/2006 Chem208Spring2006 28
Resolution
How finely can the quantity be measured
4 place balance - Resolution 0.0001 gm
Sartoriusbalance - Resolution 0.00001 gm
50 ml Buret - resolution 0.01 ml
Relative resolution depends on quantity
measured
8/15/2006 Chem208Spring2006 29
Relative resolution -
uncertainty of measurement
1.0000 gm weighed on the balance
0.0001/1.0000 * 1000 =0.1 ppt
150.0000 gm weighed on same balance
0.0001/150.0000 * 1,000,000 =0.7 ppm
0.0100 gm
0.0001/0.0100 * 1000 =10 ppt
30.00 ml titrant
0.01/30.00 * 1000 =0.3 ppt
8/15/2006 Chem208Spring2006 30
Conversion devices
Readout Conversion
Intermediate conversion device
Transducer
Sensor
Data Logger on PC
Calibrated T readout
Serial Interface Box
12 Bit A/D
Thermistor
Temperature probe
Quantitybeing measured
11
8/15/2006 Chem208Spring2006 31
Conversion devices
Linear Operating Range
Transfer Function
Relationship between input and output
Sensitivity - Slope of function
8/15/2006 Chem208Spring2006 32
Linear Scales
Interpolation
Estimate position between smallest division
Bias in Interpolation reading
Look at Buret data from lab
8/15/2006 Chem208Spring2006 33
Verniers
Devices to make interpolation more
accurate
Measure an object with Caliper
12
8/15/2006 Chem208Spring2006 34
Volume to Length Converters
Buret
Pipet
Graduate Cylinder
Sensitivity depends on inner diameter
Transfer function =length=V/pi r
2
Sensitivity =1/pi r
2
smaller r=greater sensitivity =more length
change per unit volume - Examples
8/15/2006 Chem208Spring2006 35
Pipet Two 10 ml pipets one has 6 mm ID
and the other has 3 mm ID
Sensitivity =1/pi r
2
1/3.1415x3
2
=0.0354
1/3.1415x 1.5
2
=0.1415 4 times more change
in length for a given volume
smaller r=greater sensitivity =more length
change per unit volume
8/15/2006 Chem208Spring2006 36
Class A Pipets 1 ppt
Class B Pipets 2 ppt
Electronic Pipetters 2-5 ppt
Graduated Pipets 5-10 ppt
Graduate Cylinders 5-10 ppt
Graduate Beakers and Flasks 20-100 ppt
13
8/15/2006 Chem208Spring2006 37
Challenge of Digital Data
A/D converters electrical signal to number
Computers work in Binary
Thermistor Temperature Measurement
12 bit A/D 111 111 111 111
Decimal equivalent 2
12
=4096
Serial Interface 0-5 volts ==> 0 to 4096
Resolution 5 volts/4096 =0.0012
volts/digit
8/15/2006 Chem208Spring2006 38
Thermistor Temperature probe
0.073 deg/digit resolution
0.073 * 4096 =299 degrees possible range
thermistor only good for about 150 degree
range
output varies from 0 to 2.5 volts
Digital Music - Display colors - etc.
16 bit vs24 bit
8/15/2006 Chem208Spring2006 39
A/D - Discrete Data
Convert continuous function to discrete
function
Stairstep
Potentiometric titration
Sequence of discrete points
Sampling signal at specific times
Discrete Calculus
14
8/15/2006 Chem208Spring2006 40
Null Measurements
Classic Balance
Put weights on reference pan =sample weight
Electronic Equivalent
Use solenoid to apply restoring force
amount of current required converted to Digital
Value
Resolution of A/D
Sensitivity of null detector
balances to 0.000001 gm
8/15/2006 Chem208Spring2006 41
Using EXCEL
Excel powerful tool
Calculations
Graphing
Curve fitting
Learn to use it well
Learn to document your spreadsheet
Do your own!!
1
8/15/2006 Chem208Spring2001 1
Chem 208
Chapter 3 - Lecture 1
What to Look for in an
Analytical Method
8/15/2006 Chem208Spring2001 2
Accuracy
ACCURACY - This is how close the
experimental value is to the "true value"
if this is known
Only measurable if you know true
valueof measured quantity
Estimate using statistics - more later
Measure using Standard Reference
Materials
8/15/2006 Chem208Spring2001 3
Precision
PRECISION - This tells how close
together the various experimental
results are.
Assessed using statistical methods
(standard deviation)
compared with the maximum
experimental error calculated by
analysis of experiment.
2
8/15/2006 Chem208Spring2001 4
Accuracy and Precision
Accurate and Precise Not Accurate but
Precise
Not Accurate and Not
Precise
Accurate but Not
Precise
8/15/2006 Chem208Spring2001 5
Sensitivity
Method Signal =K Quantity Measured
K=slope
Sensitivity =slope of Analytical Function
8/15/2006 Chem208Spring2001 6
Selectivity
Discrimination between Analyte and
Interference
Signal =K
A
C
A
+K
i
C
i
Only works if K
A
C
A
>>K
i
C
i
Either K
A
>>K
i
or C
A
>>C
i
Selectivity Coefficient
K
A,I
=K
i
/K
A
Signal =K
A
(C
A
+K
A,I
C
i
)
3
8/15/2006 Chem208Spring2001 7
Detection and Quantitation
Conc vs Abs usi ng 1/16 as st d
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
0 0.00005 0.0001 0.00015 0.0002
M/l
A
b
s
CorrrectedAbs calc Abs
Sample characteristic
Absorbs light at 235
nm
Detector Response
Less Light received
Encoded as electrical
current
Converted to
Absorbance
8/15/2006 Chem208Spring2001 8
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
0 0.00005 0.0001 0.00015 0.0002
M/l
A
b
s
Detection Limit
Minimumsample
quantity which
produces a signal 2X
the noise
Noise can be standard
deviation of signal or
the blank
YES or NO answer to
question Is it there?
8/15/2006 Chem208Spring2001 9
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
0 0.00005 0.0001 0.00015 0.0002
M/l
A
b
s
Quantitation Limit
Minimumsample
quantity which
produces a signal that
produces a quantitative
result
Usually 10X noise
How much is there?
4
8/15/2006 Chem208Spring2001 10
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
0 0.00005 0.0001 0.00015 0.0002
M/l
A
b
s
MaximumMeasurable
amount. - Detector at
maximumoutput
Range -
Max mount/Detection
limit
Analytical Methods
10 to 10
14
range
8/15/2006 Chem208Spring2001 11
Interferences
False Positive - Positive response in
absence of true Analyte
False Negative - no response in presence of
analyte
8/15/2006 Chem208Spring2001 12
Scale of Operation
How much sample is available?
How much analyte is in sample?
How much analyte is needed to get a signal?
Major component analysis 1-100%
Minor component analysis 0.01-1%
Trace analysis 0.1 to 100,000 ppm(definitions vary)
Ultratraceanalysis below 0.1 ppm
5
8/15/2006 Chem208Spring2001 13
Equipment, Time, Cost
Do you have the needed Equipment?
Can you pay someone else to do it on theirs?
How Long will Analysis Require
Most time spent in sample preperation
What does it cost per analysis
8/15/2006 Chem208Spring2001 14
Interferences
Are there components present that produce
unwanted signals?
Interferences in Sample
Can they be removed by separation step?
Are they constant?
Can we measure themindependently?
Interference in reagents
Method Blank
8/15/2006 Chem208Spring2001 15
Calibration - Standardization
Calibration - Equipment operating correctly
Calibrate Balances
Spectrometers
Electrodes
Standardization Compare output to known
concentrations
Calibration Curves Working Curves
6
8/15/2006 Chem208Spring2001 16
QA/QC
How Good is the Result?
Standard Reference Materials
Known Values
How close is the result to True Value
Control Charts
Monitor day to day quality
8/15/2006 Chem208Spring2001 17
Control Chart Example
Control Chart
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0 5 10
Experi ment Day
R
e
s
u
l
tData
+2 stdev
-2 stdev
8/15/2006 Chem208Spring2001 18
Identification and Separation
Identification - Not part of 208
Chem211 Organic
Chem312 instrumental
Separation - We spend lots of time on this
Remove Interferents
Quantitation of Mixtures
Identification of Mixtures
7
8/15/2006 Chem208Spring2001 19
Sample Prepartion
Crucial first step in all analysis
Ask Series of Questions
1. Is sample solid or liquid
2. Do you want to analyze all or only a few
components
3. Is sample size appropriate
4. Is concentration appropriate for detector
8/15/2006 Chem208Spring2001 20
Solids
Is the particle size appropriate for
dissolution or extraction?
If not, mill, grind, chop, blend etc.
Is the sample homogeneous?
Homogenize
Take a representative sample - sample
size required is a function of particle
size.
Lab exercise
8/15/2006 Chem208Spring2001 21
Solids - A Few Components
Are they the volatile components.
Headspace analysis, Solid Phase
microextraction, or purge and trap.
Non-volatile components
separation is necessary by boiling, soxhlet
extraction, sonication, microwave
digestion, supercritical fluid extraction.
8
8/15/2006 Chem208Spring2001 22
Solids - All Components
Sample usually must be dissolved.
What is the sample soluble in?
Water
organics, etc.
What if the sample is insoluble?
intact high polymers.
Inorganic analysis -Ashing techniques - Wet vs
Dry Ashing
Fusions with Carbonate and Borate
8/15/2006 Chem208Spring2001 23
Strange Techniques
Soxhlet Extraction -
Solid Phase Extraction-
Solid Phase Microextraction-
Headspace Sampling-
Purge and Trap analysis-
Microwave methods-
Sonication Methods-
8/15/2006 Chem208Spring2001 24
Liquids- Few components
Separation is necessary by extraction or
chromatography.
Is the concentration of the analytes
appropriate for the measurement
technique?
If not, dilute or concentrate with extraction,
evaporation, lyophilization (freeze drying).
9
8/15/2006 Chem208Spring2001 25
Liquids - Few Components
Is sample unstable
If yes, derivatize, cool, freeze, store in dark
Is the liquid or solvent compatible with
the analytical method?
If not, do solvent exchange with extraction,
distillation, lyophilization.
1
8/15/2006 Chem208Spring2002 1
Chem 208
Chapter 4 Lecture 1
Meaurement and Calculation
Accuracy and Precision
8/15/2006 Chem208Spring2002 2
Experimental Uncertainty
KHP Titration
0.8 0.0001 g KHP
40 0.014 ml (two reading of buret)
204.23 0.02 g/mol KHP
M
NaOH
=0.8 g x 1 mol/204.23g /0.040 l =0.09793 mol / liter
Uncertainty on value
0.09793 0.0004 or 0.0979 0.0004
About 4 ppt undertainty
0004 . 0
40
014 . 0
23 . 204
02 . 0
8 . 0
0001 . 0
y undertaint
2 2 2
=
=
8/15/2006 Chem208Spring2002 3
Experimental Uncertainty
Calculated uncertainty is worst case
scenario
Assumes all errors are in the same direction
Multiple determinations produce average
closer to correct value
Average =Mean
2
8/15/2006 Chem208Spring2002 4
Error vs Uncertainty
Uncertainty is inherent in all measurements
Related to significant figures available from
measurement devices
Possible to calculate the Uncertainty of a
measurement
8/15/2006 Chem208Spring2002 5
Precision
PRECISION - This tells how close together
the various experimental results are.
Assessed using statistical methods (standard
deviation)
compared with the maximum experimental
error calculated by analysis of experiment.
8/15/2006 Chem208Spring2002 6
MEANSOF MULTIPLE
EXPERIMENTAL
DETERMINATIONS
THE STANDARD DEVIATION MAY BE
USED TO ASSIGN UNCERTAINTY
INSTEAD OF EXPERIMENTAL
UNCERTAINTY.
The mean of many values should be more
accurate than any of the individual values.
CAUTION - never more than one extra digit
beyond the individual values.This digit is in
fact the extra one carried in the intermediate
calculations.
3
8/15/2006 Chem208Spring2002 7
EVALUATION OF ERROR
PRECISION - This tells how close together
the various experimental results are.
Assessed using statistical methods
Standard deviation
Compared with the maximumexperimental
error predicted fromerror analysis.
8/15/2006 Chem208Spring2002 8
Standard Deviation
Assume Gaussian Distribution of
Experimental Values
Mean
+1 std.
dev.
2 std dev
-1 std
dev
8/15/2006 Chem208Spring2002 9
Standard Deviation
Standard Terms associated with statistical
treatment of data.
Mean - arithmetic average.
Median - middle value of the set
Mode - most common value of the set

x =

n
( x
i
)
n
4
8/15/2006 Chem208Spring2002 10
Standard Deviation
Standard Deviation - s
x
(sample)
or
x
(population >24)
Variance =square of standard deviation
V =s
x
2
or V =
x
2
1
2

=

n
x x
s
n
i
x
n
x x
n
i
x

8/15/2006 Chem208Spring2002 11
Standard Deviation
Relative standard deviation.
% relative st. dev =Coefficient of
Variation (CV)
% s
x
=
s
x
x
100 %
ppt s
x
=
s
x
x
1000 ppt
8/15/2006 Chem208Spring2002 12
Significance of Standard
Deviation
One standard deviation is the distance from
the mean in which 69.3% of all values are
expected to lie.
95.5% of all values are expected to lie
within 2 std. dev. from the mean
99.8% within 3 std. dev.
As a general rule, virtually all experimental
data should lie in the range 4 s.
5
8/15/2006 Chem208Spring2002 13
Relative Standard Deviation
Standard deviation is fine for
comparing things that are the same. It is like
absolute error and has the same units as the
quantity being measured.
Relative standard deviation useful
comparing the deviations of data of differing
magnitude to see which has the smallest
deviation.
8/15/2006 Chem208Spring2002 14
Relative Standard Deviation
eg. One student reports 25.34% KHP in an
unknown with a standard deviation of 0.05
while another reports 64.34% KHP with a
standard deviation of 0.09.Which is a better
result? Calculate the relative standard
deviation for each. The smallest relative
standard deviation gets the best grade.
(2.0 ppt and 1.4 ppt)
8/15/2006 Chem208Spring2002 15
Confidence Limits
-for a small number of values where the true
value is not known, the true value may be
estimated from the mean and the standard
deviation.
=truevalue
t
n
=Students t for theparticular C.L. and number of replicates
n =number of replicates
s
x
=std. dev.
x bar =mean value
n
s t
x
x n
=
6
8/15/2006 Chem208Spring2002 16
Confidence Limits
Assumes there is no determinate error
present.
Different confidence levels include different
areas of the gaussiandistribution.
90% confidence. This means the confidence
limit calculated will include 90% of the total
area under the distribution.10 % is not included,
5% on each side
8/15/2006 Chem208Spring2002 17
Confidence Limits
The 99% confidence interval includes a
larger area than the 90% and hence the
confidence limits will be greater.
Note that the larger the value of n, the
smaller the confidence limit. This is due to
the fact that larger n not only increases the
denominator but also decreases Student's t.
8/15/2006 Chem208Spring2002 18
Confidence Limits
Example mean =144 ppm s
x
= 10.2 ppm
n =4 t
90%
=2.353 t
95%
=3.182
t
99%
=5.841

90%
= 144 12 ppm
confidence that true value lies in the interval
132 ppm< <156 ppm

95%
=144 16 ppm 128 < <160 ppm

99%
=144 +30 ppm 114 < <174 ppm
95% confidence is most commonly used.
7
8/15/2006 Chem208Spring2002 19
Types of Errors
Random Errors - Indeterminate errors
Have no definite cause
randomerrors in reading a buret
Result in decreased precision and wider
confidence limits
8/15/2006 Chem208Spring2002 20
Types of Errors
SystemmaticErrors - Determinate errors
Have a definite cause
Balance not calibrated
Solution at wrong temperature for volumetric work
Result in measured values which fall outside
the confidence interval of the measurement
May be constant relative or absolute error
Vary Sample size to evaluate
8/15/2006 Chem208Spring2002 21
Types of Errors
Sampling Errors
Sample not representative of whole
M&M experiment
Method error
Interferences
Mesurement Errors
Temperature not 20 deg for Volumetric flask
8
8/15/2006 Chem208Spring2002 22
Types of Errors
Personal Errors
Analyst makes a mistake
8/15/2006 Chem208Spring2002 23
Finding Errors
Constant errors
Balance reads 0.002 gmhigh
Proportional errors
Balance reads 0.1% high
Try different sample quantities and observe
the effect of the error
Use Standard Reference Material to test
method
8/15/2006 Chem208Spring2002 24
ELIMINATION OF
QUESTIONABLE RESULTS
These are the only methods you are to use
with your experimental data for the course.
Errors occurred in the experiment.
Definable errors. eg. you spilled some of the
solution out of the flask during a titration or
obviously overran an endpoint. Note the error
in the notebook beside the data and draw a
diagonal line through the data. Do not erase or
obscure the data.
9
8/15/2006 Chem208Spring2002 25
ELIMINATION OF
Undefinable error. These are of two types.
Errors due to learning to do the experiment
correctly or due to problems in start up. These
are common in student experiments. You may
discard all data prior to any point and call that
data practice. Note this in the notebook with the
data.
8/15/2006 Chem208Spring2002 26
ELIMINATION OF
Undefinable error. These are of two types.
Errors due to deterioration of reagents, operator
patience, or equipment. These occur at the end
of a series of experiments. You may discard all
data after any given data set and assume some
unknown problemoccurred.
You may NOT discard data in the middle of
a data set unless there is a definable error!!!
8/15/2006 Chem208Spring2002 27
ELIMINATION OF
Q-test.
measure of the range between an outlying
value and its nearest neighbor compared to the
total range of the data.
EXAMPLE
0.1013, 0.1014, 0.1016 , 0.1024,
95% confidence Q
4
=0.83 >0.73
Cannot discard value
low high
nearest outlier
x x
x x
Q
=
exp
727 . 0
1013 . 0 1024 . 0
1016 . 0 1024 . 0
=
= Q
10
8/15/2006 Chem208Spring2002 28
Significance Testing
Assume Gaussian Distribution
Assign Probability to two possible
outcomes
NULL HYPOTHESIS
Differences observed are due to random,
indeterminate errors
ALTERNATIVE HYPOTHESIS
Differences observed are real
8/15/2006 Chem208Spring2002 29
NEVER PROVE ANYTHING IS TRUE
Prove NOT FALSE
Use alpha =the area of the distribution not
included usually 0.05
95% of area is included and 5% excluded
8/15/2006 Chem208Spring2002 30
T-test
Compare mean to true value
Compare Replicate measurements
Comparing two methods of analysis
Useful for comparing new method to existing method
Do both methods givethesameresult?
F-Test
Compare sample variance to population variance
Compare variance of two data sets to see if the same
11
8/15/2006 Chem208Spring2002 31
T-test example
Measured pH of natural and machine made
snow. Are they the same or different?
Various t-tests
Variance on each data set is the same
Variance on each data set is different
Data in two sets are paired or not
Excel Tools Data Analysis
8/15/2006 Chem208Spring2002 32
2.364624 t Critical two-tail
0.010426 P(T<=t) two-tail
1.894579 t Critical one-tail
0.005213 P(T<=t) one-tail
3.468549 t Stat
7 df
0 Hypothesized Mean Difference
7 8 Observations
0.065229 2.211829 Variance
4.885714 6.74 Mean
Variable 2 Variable 1
Natural Machine
t-Test: Two-Sample Assuming Unequal Variances
8/15/2006 Chem208Spring2002 33
2.100922 t Critical two-tail
0.158074 P(T<=t) two-tail
1.734064 t Critical one-tail
0.079037 P(T<=t) one-tail
-1.47281 t Stat
18 df
0 Hypothesized Mean Difference
14 7 Observations
0.450411 0.065229 Variance
5.185714 4.885714 Mean
Variable 2 Variable 1
Mixed Natural
t-Test: Two-Sample Assuming Unequal Variances
12
8/15/2006 Chem208Spring2002 34
What does this mean
P<0.05 95% confidence that values are
different.
One Tail all 5% area under curve on one side
Use when reason to expect one value to be smaller
and the other larger
Two tail 2.5 % of area at each end of curve
More rigorous test
Use when no reason to predict the order of values
8/15/2006 Chem208Spring2002 35
Snow data
Acid Rain Natural snow should be acidic
Machine Made snow made from surface
water therefore less acidic
Use one Tail t-test P=0.005
Significant difference
Mixed Snow one tail test also appropriate
P=0.07 Not significantly different
1
8/15/2006 Chem208Spring2002 1
Chapter 5
Calibration, Standardization
Blank Correction
8/15/2006 Chem208Spring2002 2
S
meas
= kC
Analyte
+ S
reagent
Calibration necessary for all measurements
What to use as STANDARDS
Primary Standards
Known stoichiometry
Known purity
Stable for long periods
List in Appendix 2 text
Ex. KHP for Titration
8/15/2006 Chem208Spring2002 3
S
meas
= kC
Analyte
+ S
reagent
Secondary Standards
Determined relative to a Primary Standard
Ex. EDTA
.
2H
2
O in Zn determination
2
8/15/2006 Chem208Spring2002 4
Standard Solutions
Weigh Primary Std. Into Volumetric Flask
Dilute to mark only after all dissolved
Do not heat
Mix at least 10 times
8/15/2006 Chem208Spring2002 5
Standard Solutions
Serial Dilution for Calibration or dilute
Pipet portion of stock solution into Vol.
Flask and dilute
C
o
V
o
=C
d
V
d
Many ways to get to specific concentration
Errors not the same for each
problem4-11
8/15/2006 Chem208Spring2002 6
Calibration Methods
Single Point
External Standard Method Calibration
Curve
Internal Standard Method
Standard Addition Method
3
8/15/2006 Chem208Spring2002 7
Single Point Calibration
k=S
stand
/C
s
One Standard
Assumes response factor k is constant over
entire range
Assumes linear relationship
Ignores Blank due to reagents
Generally not a good idea
8/15/2006 Chem208Spring2002 8
External Standard Method
Using a Standard Curve
Known concentrations vsSignal
Intercept need not be Zero
Identify non-linearity if present
8/15/2006 Chem208Spring2002 9
Vanilla Extraction Lab (S=Absorbance)
Fluoride Lab (S=Potential)
In 111-112 Fe[SCN] Lab (S=Absorbance)
Can also be applied to most other types of
signals.
4
8/15/2006 Chem208Spring2002 10
Chromatography
make calibration curve
run sample and compare
Limited by injection accuracy
2-4% manual
1-2% auto
0.5% valve injections
8/15/2006 Chem208Spring2002 11
LINEAR REGRESSION
ANALYSIS
(LEAST SQUARES).
Best linear relationship for a set of data.
Can be extended to find higher order
polynomial relationships
Can be extended to data where the assumptions
below are known not to hold
8/15/2006 Chem208Spring2002 12
How to Draw the Calibration
Line?
The best linear relationship is that which
minimizes the sum of the squares of the
deviations on the y axis of all points from
the line
X values are assumed to be Exact
DO NOT USE LEAST SQUARES
WHERE BOTH X AND Y DATA
CONTAIN LARGE ERRORS
5
8/15/2006 Chem208Spring2002 13
ASSUMPTIONS
Values for the x axis are relatively free fromerror.
Concentration =independent variable
Values for the the y axis and may contain some
error.
Signal =what you measured
The standard least squares method assumes all
have the same error
more sophisticated approaches allow weighing factors
for cases where errors differ greatly.
8/15/2006 Chem208Spring2002 14
Calculating Linear Regression
on a TI Calculator
LIST EDIT
Enter values into XSTAT (Independent variable)
Enter values into YSTAT (Dependent variable)
Enter 1 into Fstat column for each Xstat value
EXIT
STAT
CALC
LinR (Enter) Gives slope, intercept and Correlation
Coefficient
TwoVar (Enter) Gives sums of squares needed for error
calculations
8/15/2006 Chem208Spring2002 15
Computing Least Squares
Excel -
Make XY scatter Graph
Click on data in graph
Right click Select Add Trendline
Options -
display equation
display R squared
6
8/15/2006 Chem208Spring2002 16
Excel Getting values for Calculations
=Slope(yvalues, xvalues) b
o
=Intercept(yvalues, xvalues) b
1
=RSQ(yvalues,xvalues) R
2
8/15/2006 Chem208Spring2002 17
Excel Alternate Approach
Tools ==>AddIns==>Analysis Pack
Tools==>Data Analysis==>Regression
Select Range for x and Y data
Select where results go
Choose options (some dont work quite right for
graphics)
Get lots of stuff - std. Dev of slope and intercept
8/15/2006 Chem208Spring2002 18
Excel Least Squares
Least Squares
y=0.9624x+0.0763
R
2
=0.9974
0
2
4
6
0 2 4 6
Independent Variable
D
e
p
e
n
d
e
n
t

V
a
r
i
a
b
l
e
Y values
Linear (Y
values)
X values Y values
0 0.1
1.1 1
2 2.1
3.98 4
5.2 5
7
8/15/2006 Chem208Spring2002 19
Residuals
X values Y values Y calc Residuals
0 0.1 0.0763 -0.0237
1.1 1 1.13494 0.13494
2 2.1 2.0011 -0.0989
3.98 4 3.906652 -0.09335
5.2 5 5.08078 0.08078
Residuals
-0.2
-0.1
0
0.1
0.2
0 2 4 6
X values
R
e
s
i
d
u
a
l
s
Residuals
8/15/2006 Chem208Spring2002 20
Residuals
Random distribution ==>Data is inherently
linear
Non-Random ==>Data not linear
Use a different model
Errors Greater at one end of Residuals
Errors in measurement are not constant.
8/15/2006 Chem208Spring2002 21
Non-linear data
Xvalues Y values Y calculateResiduals
0 -0.05 0.0669 0.1169
1.1 1.15 1.12257 -0.02743
2 2.14 1.9863 -0.1537
3.98 3.89 3.886506 -0.00349
5.2 4.99 5.05734 0.06734
Non-linear Data
y=0.9597x+0.0669
R
2
=0.9974
-2
0
2
4
6
0 2 4 6
X values
Y

v
a
lu
e
s
Y values
Linear (Y
values)
Same R squared
as previous data
8
8/15/2006 Chem208Spring2002 22
Residuals
Note pattern in residuals
Residuals
-0.2
-0.1
0
0.1
0.2
0 2 4 6
X value
R
e
s
i
d
u
a
l
s
Residuals
8/15/2006 Chem208Spring2002 23
Calculation of Least Squares
Line
y =b
0
+b
1
x equation of line
S
xx
= SUM ( mean x - x
i
)
2
S
yy
= SUM ( mean y - y
i
)
2
Sxy=SUM ( mean x - x
i
)( mean y - y
i
)
Compute sum of squares of deviations from
average x, average y and xyproduct
8/15/2006 Chem208Spring2002 24
Least Squares Line
xx
xy
S
S
b slope = =
1
x b y b
o
* intercept
1
= =
9
8/15/2006 Chem208Spring2002 25
Special Precautions
When computing sums of squares, it is
essential that ALL DIGITS be kept in
calculations.
Large differences in results occur based on
small round off or truncation errors.
Different results depending on how carefully
you use calculators and which one you use.
8/15/2006 Chem208Spring2002 26
Calculating an Unknown
The value for an unknown result x
c
is calculated
frommeasured value y
c
b
1
=slope b
o
=intercept
1
b
b y
X
o s
s
=
8/15/2006 Chem208Spring2002 27
Precision of Unknown
m =number of observations of unknown
n =number of calibration points
point for residual ) (
2
) (
) (
) ( 1 1
2
2 2
1
2
1
=
+ + =
i i
i i
r
i
x r
x
y y
n
y y
s
x x b
y y
n m b
s
s
10
8/15/2006 Chem208Spring2002 28
Equation hard to use. Errors occur if any
sum value is not taken with all digits
point for residual ) (
2
) (
) (
) ( 1 1
2
2 2
1
2
1
=
+ + =
i i
i i
r
i
x r
x
y y
n
y y
s
x x b
y y
n m b
s
s
8/15/2006 Chem208Spring2002 29
Precision of Regression
s
x
function has a minimumvalue when the mean
value for the unknown is close to the mean value
of the calibration curve.
(y
x
bar - y bar) =0
The standard deviation of the unknown can then
be used to calculate a confidence limit for the
value of the unknown in the usual manner using
Students t and the equation given previously.
8/15/2006 Chem208Spring2002 30
Confidence Limit for unknown
x true
ts x x =
11
8/15/2006 Chem208Spring2002 31
Error in b
1
and b
o
Std. Dev. of slope and intercept define an
envelop
This envelop is narrow at the mean of the
calibration curve and expands on both sides.
8/15/2006 Chem208Spring2002 32
95% Convidence Envelope
-1
0
1
2
3
4
5
6
7
0 2 4 6
X Val ues
Y

V
a
l
u
e
s
Exp. Data
+95% CL
-95% CL
8/15/2006 Chem208Spring2002 33
95% CL best case
-1
0
1
2
3
4
5
6
0 2 4 6
X val ues
Y

c
a
l
u
e
s
Exp. Values
-b+m
+b-m
12
8/15/2006 Chem208Spring2002 34
Non-Linear LS
Data obtained in a real experiment may not
be linear even though it should be in theory.
Use a second order fit:
y=a +bx+cx
2
Calculation of Unknown gives 2 values
Quadratic Solution Required
8/15/2006 Chem208Spring2002 35
Figures of merit for LS
Correlation coefficient
=RSQ(known_y's,known_x's)
Known_y's is an array or range of data points.
Known_x's is an array or range of data points.
R
2
correlation coefficient 0.0 to +1.0
Variance or standard deviation of the fit
8/15/2006 Chem208Spring2002 36
Figures of merit for LS
A poor fit may be due to one of two causes.
If the data is fundamentally nonlinear, there
will be definite trends in the residuals,
either bowing up or down.
If the data is just bad but linear, the
distribution of the deviations will be
random.
13
8/15/2006 Chem208Spring2002 37
Detection Limits
S
dl
=S +z sigma
Z=3 gives 99.86% confidence that
substance is present
Z=2 97.7% confidence
8/15/2006 Chem208Spring2002 38
Quantitation Limit
S
dl
=S +10 sigma
Gives quantitation with 10% rel error
8/15/2006 Chem208Spring2002 39
Limit of Linearity
Data deviates by 10% from Least SQ line
Usually Negative
14
8/15/2006 Chem208Spring2002 40
Non-Linear Data
If an obvious trend is seen, then data may
be non-linear. -
Use other statistical tools (higher order fit)
Drop data from non-linear part and repeat
regression analysis.
Common if you exceed the Range of the
Method
8/15/2006 Chem208Spring2002 41
Calculating Standard Deviation
on a TI Calculator
LIST EDIT
Enter values into XSTAT
Enter 1 into Fstat column for each Xstat
value
EXIT
STAT
CALC
OneVa(Enter)
8/15/2006 Chem208Spring2002 42
Standard Addition
Useful if matrix of sample has background
absorbance which cannot be accounted for
in a blank or calibration curve
Quick if only one or two samples are to be
run
15
8/15/2006 Chem208Spring2002 43
Standard Addition
Three approaches to Std. Add.
Dilute unknown and standard additions to
constant volume.
50 ml sample +10 ml std 100 ml
Add micro amounts of standard and ignore
dilution.
100 ml sample +0.1 ml std.
Add standard and correct for dilution.
8/15/2006 Chem208Spring2002 44
Analysis Methods
Standard Addition Methods
S
Samp
=kC
A
V
o
/V
f
S
Spike
=k(C
A
V
o
/V
f
+C
s
V
s
/V
f
)
Assume k is equal in both equations
and solve
f
S
S
f
o
A
Spike
f
o
A
samp
V
V
C
V
V
C
S
V
V
C
S
8/15/2006 Chem208Spring2002 45
Methods of Quantitation
Prepare solutions by adding known amounts of
analyte directly to the unknown
one or more different additions
For one addition
+
+
+
=
s o
S
s o
o
A
Spike
A
samp
V V
Vs
C
V V
V
C
S
C
S
16
8/15/2006 Chem208Spring2002 46
Multi-point Standard Addition
Check for Linearity
Reduces Error
Graph S
spike
vsC
s
C
s
is calculated ignoring C
A
C
s
=C
std
V
s
/V
f
8/15/2006 Chem208Spring2002 47
Graphical treatment of std.
addition
X-Intercept=-C
A
V
o
/V
f
-Cx Unk. Unk+S1
U k S2
Absorbance
8/15/2006 Chem208Spring2002 48
Std. Addition as Check on
External Calibration
Compare value of k (slope) for external
standard and standard addition calibrations
IF EQUAL Method is good and there are
not matrix effects
IF NOT EQUAL Matrix error present in
external standard calibration
17
8/15/2006 Chem208Spring2002 49
Methods of Application of
ISE's
Standard Addition NOTE S=k Log C
A
Measure sample
E =Q - .0591log C
x
n
Measure sample +Std.
E =Q - .0591log (C
x
+C
s
)
n
8/15/2006 Chem208Spring2002 50
ISE's
Std. allows compensation for unknown
matrix
eg. ionic strength and E
j
effects minimized
E
x
=Q +.0591log C
x
n
E
s
=Q +.0591log
V
s
C
s+
V
x
C
x
n V
s
+V
x
8/15/2006 Chem208Spring2002 51
ISE's
Take inverse log
rearrange
x s
x x s s
x
s x
V V
C V C V
C
E E n
+
+
=
log log
05916 . 0
) (
10
n(E
x
E
s
)
0.0592
=
C
x
( V
s
+V
s
)
V
s
C
s
+V
x
C
x
C
x
=
C
s
V
s
(V
x
+ V
s
) 10
n(E
x
E
s
)/ 0.0592
V
x
18
8/15/2006 Chem208Spring2002 52
Analysis Methods
eliminates injection error in Chromatorgraphy
- only integration error is left
Eliminates need for careful sample workup in
many methods
add standard to each sample use it to correct for
variation in injection and in sample workup etc.
Single point - determine peak ratio with one standard
mixture
Calibration curve - plot area ratio vs mass ratio
8/15/2006 Chem208Spring2002 53
Data from GC experiment
C22:1
y =0.6548x - 0.0804
R
2
=0.9989
0
1
2
3
4
5
0 2 4 6 8
Series1
Linear
(Series1)
C16:0 y =1.0405x -
0.3135
R
2
=0.9991
0
5
10
15
20
0 10 20
Series1
Linear
(Series1)
8/15/2006 Chem208Spring2002 54
Selecting an Internal Standard
Chemically Similar to Analyte
HPLC Lab Theophyllinefor Caffeine
Isomers or close homologs
GC Lab C
17
fatty acid
Once internal Std and Sample are made
homogeneous all further steps need not be
quantitative (within limits of linearity and
detection)
19
8/15/2006 Chem208Spring2002 55
Selecting an Internal Standard
Mass Spectrometry Isotopic labels
D,
13
C,
15
N, etc
All chemical properties same as Analyte
Mass Spec gives separate signals
Cocaine methods for Chem312
1
8/15/2006 Chem208Spring2002 1
Chem 208
Chapter 6
Chemical Equilibrium
8/15/2006 Chem208Spring2002 2
Chemical Equilibrium
Bottom of energy well
Change in concentration of any species
requires input of energy
Any external change away from equilibrium
is adjusted back to new equilibrium
Marble in a bowl analogy
8/15/2006 Chem208Spring2002 3
Equilibrium constant
Free Energy change is related to chemical
potential
Note difference from Eq6.4
Ratio of activities to Std. St.
Dimensionless
Q RT G G
o
ln + =
b
o
b
b
a
o
a
a
d
o
d
d
c
o
c
c
Q
2
8/15/2006 Chem208Spring2002 4
At Equilibrium
eq
o
K RT G ln =
values m equilibriu are s a' where
b
o
b
b
a
o
a
a
d
o
d
d
c
o
c
c
eq
K
8/15/2006 Chem208Spring2002 5
Normally write using Concentrations
All values are dimensionless (ratio to std state)
Ignore Activity effects in most cases
(Chapter 8)
b
eq
a
eq
d
eq
a
eq
eq
B A
D C
K =
8/15/2006 Chem208Spring2002 6
Properties of K
eq
Reverse Reaction Invert K
Cu +4 NH
3
Cu(NH
3
)
4
Cu(NH
3
)
4
Cu +4 NH
3
4
3
2
4 3
] ][ [
] ) ( [
NH Cu
NH Cu
K
f +
=
] ) ( [
] ][ [ 1
4 3
4
3
2
NH Cu
NH Cu
K
K
f
i
+
= =
3
8/15/2006 Chem208Spring2002 7
Properties of K
eq
When Reactions are added, the K
eq
of the
sum is the product of the Ksof the separate
reactions
CH
3
COOH +H
2
O H
3
O
+
+CH
3
COO
-
K
a
CH
3
COO
-
+H
2
O CH
3
COOH +OH
-
K
b
2H
2
O H
3
O
+
+OH
-
K
w
=K
a
x K
b
8/15/2006 Chem208Spring2002 8
Precipitation Reactions
AgCl
(s)
Ag
+
+Cl
-
K
sp
=[Ag
+]
[ Cl
-
]
Note solid does not appear in different phase
Only works for a few species
Silver halides
BaSO
4
All others have complex chemistry which make
the simple treatment useless.
8/15/2006 Chem208Spring2002 9
PbCl
2(s)
Pb
+2
+2Cl
-
K
sp
=[Pb
+2
][ 2Cl
-
]
Also need to consider
PbCl
+
, PbCl
3
-
, and PbCl
4
2-
Simple equilibriumgives wrong answers
4
8/15/2006 Chem208Spring2002 10
BRONSTED-LOWRY ACID
BASE CONCEPT
We use this concept because it applies to any
solvent.
ACID - donates proton to solvent
BASE- accepts proton from solvent.
HCl + H
2
O <===> H
3
O
+
+ Cl
-
acid + base conj.acid conj.base
8/15/2006 Chem208Spring2002 11
Look at half reactions
Acid
1
==>H
+
+Base
1
Base
2
+H
+
==>Acid
2
Sum these to get
Acid
1
+Base
2
==> Base
1
+Acid
2
conjugate Acid-Base pairs (subscripts)
8/15/2006 Chem208Spring2002 12
Acid-Base Strength
Relative
Strong acids have greatest energy release
when losing proton
Strongest bases have greatest energy release
when accepting a proton
Products must be weaker than reactants
HCl + H
2
O ==>Cl
-
+ H
3
O
+
Stronger acid+stronger base==>weaker base +weaker acid
5
8/15/2006 Chem208Spring2002 13
Acid-Base Strength
Relative
In water - H
3
O
+
is strongest acid
Anything above it reacts with
water to produce H
3
O
+
HCl, H
w
SO
4
, HBr, HI, HClO
4
Anything below water is weak acid
Farther below water the weaker it gets
None react completely
Equilibrium
8/15/2006 Chem208Spring2002 14
Strong Acids
Above H
3
O
+
HCl +H
2
O <===> Cl
-
+H
3
O
+
Cl
-
very weak proton acceptor
HCl verystrong proton donor.
Thus there is a complete proton transfer.
K
a
>>>100
8/15/2006 Chem208Spring2002 15
Weak acids
Acetic acid for example:
CH
3
COOH + H
2
O <===>CH
3
COO
-
+
H
3
O
+
K
a
=1.75x10
-5
acetate ion stronger proton acceptor
than water so proton tends to remain on
the acetate (making it acetic acid).
6
8/15/2006 Chem208Spring2002 16
Strong bases
O
2-
+ H
2
O <===> 2 OH
-
K
b
>>>100 strong base
Anything which yields OH
-
ions in
solutions
NaOH, KOH, Ba
2
OH - must be soluble
Cu(OH)
2
insoluble not strong base
8/15/2006 Chem208Spring2002 17
Weak bases
NH
3
+H
2
O <===> NH
4
+
+ OH
-
K
b
=1.7x10
-5
weak base
Amines, conjugate bases of weak acids
phosphates, carbonates, acetate, etc.
8/15/2006 Chem208Spring2002 18
Acid and Conjugate Base
Strengths are related to the solvent.
CH
3
COOH +H
2
O <===> CH
3
COO
-
+ H
3
O
+
CH
3
COO
-
+H
2
O <===> CH
3
COOH + OH
-
K
a
=
[H
+
][OAc
]
[HOAc]
K
b
=
[OH
][HOAc]
[OAc
]
7
8/15/2006 Chem208Spring2002 19
Add reactions
CH
3
COOH +H
2
O +<===> CH
3
COO
-
+
H
3
O
+
CH
3
COO
-
+ H
2
O <===> CH
3
COOH + OH
-
+ add two reactions
2 H
2
O <===>H
3
O
+
+ OH
-
Solvent Autoprotolysis
Equilibrium constant K
w
8/15/2006 Chem208Spring2002 20
Acid- Base Pair Relationship
K
a
x K
b
= K
w
This relationship is true for any conjugate acid
base pair in water.
K
a
K
b
=K
s
For any solvent
If you know the K
a
you can always get the K
b
.
Also note that for a solvent of different K
s
the
relationship between the K
a
and K
b
will be
different.
8/15/2006 Chem208Spring2002 21
Temperature Effects
K
a
and K
s
vary with temperature
Temperature(C ) Kw
0 0.114x 10
-14
25 1.01x10
-14
50 5.47x10
-14
100 49x10
-14
37 3.2x10
-14
8
8/15/2006 Chem208Spring2002 22
What is a neutral solution?
25 deg C K
w
=1.0x10
-14
=[H
3
O
+
] [OH
-
]
[H
3
O
+
]=1.0x10
-7
pH =7.00
At 50 deg C neutral water pH=6.63
Ice water neutral pH =7.42
Very important for biochemist who work at
37 C ( normal human body temp) pH=6.74
8/15/2006 Chem208Spring2002 23
Solubility Equilibria
AgCl <==> Ag
+
+Cl
-
K
sp
=[Ag
+
] [ Cl
-
]
Solubility =how many moles dissolve in 1 liter of
water
S=[Ag
+
] =[Cl
-
] =K
sp
1/2
Al(OH)
3
<Al
3+
+3 OH
-
S=[Al
3+
] =1/3 [OH
-
] so [OH
-
] =3S
K
sp
=[Al
3+
] [OH
-
]
3
=S x (3S)
3
=9S
4
8/15/2006 Chem208Spring2002 24
Its not so simple
Most slightly soluble salts have complicated
chemistry
Anions act as weak bases- must consider
fraction in simple form
anions act as complexingagents- must consider
fraction of metal in uncomplexedform
9
8/15/2006 Chem208Spring2002 25
Bottom Line
AgCl, AgBr, AgI and BaSO
4
are the only
cases which work even tolerably well
Silver halides form higher complex ions
AgCl
2
-
, AgCl
3
-2
, AgCl
4
-3
, AgCl
(aq)
8/15/2006 Chem208Spring2002 26
Coordination Complexes
Lewis Acid Base Theory
Acids =electron pair acceptor
H
+
, Metal ions Fe
3+
, Cu
2+
, etc.
Base =Ligand=electron pair donor
OH
-
, Cl
-
, :NH
3
, CN
-
, H
2
O, etc.
Acid +Base ==>Complex ion
8/15/2006 Chem208Spring2002 27
Complex ions Co complex lab
All ions in water are hydrated
Cations =Lewis acids ==>accept electron pair
from:O on H
2
O
Fe
3+
+6 H
2
O <==> [Fe(H
2
O)
6
]
3+
hexaaquoiron (III) ion
Formation of other complex ions by
Displacement
10
8/15/2006 Chem208Spring2002 28
Displacement
Fe
3+
+6 H
2
O <==> [Fe(H
2
O)
6
]
3+
[Fe(H
2
O)
6
]
3+
+6Cl
-
<==>[FeCl
6
]
3-
+6H
2
O
Cu(H
2
O)
5
2+
+4 NH
3
<==>Cu(NH
3
)
2+
+5H
2
O
DEMO
8/15/2006 Chem208Spring2002 29
Ligands
LigandCharacteristics
How many binding sites
1=Monodentate
2=Bidentate
3=Tridentate
4=tetradentate
6=hexadentate
8/15/2006 Chem208Spring2002 30
Ligands
Ligand Type Examples
Monodentate Cl
-
, NH
3
, Br
-
, thiocynate SCN
-
, cyanide CN
-
Bidentate ethylenediamine (en), Oxalate
C
2
O
4
2-
, acac
Tridentate NTA nitrilotriacetic acid
Hexadentate EDTA
ethylenediaminetetracetic acid
11
8/15/2006 Chem208Spring2002 31
Ligands
LigandCharacteristics
Strength of ligandfield
Electron density of ligand
Strength of bonding
Ligandsare bases (Lewis and Bronsted)
Acid Base Chemistry important
8/15/2006 Chem208Spring2002 32
Metals
Characteristics of Metal ions
Coordination number (how many ligands)
Maximumcoord. No.
Rate of ligandexchange
Labile =Fast up to 10
-9
sec Na
+
, Mg
2+
Inert =Slow up to 10
6
sec Cr
3+
, Pt
2+
Hydrolysis of metal aquocomplex
Fe(H
2
O)
6
3+
<==>Fe(OH)(H
2
O)
5
2+
+H
+
8/15/2006 Chem208Spring2002 33
SEQUENTIAL COMPLEXATION BY
SIMPLE LIGANDS
Ni(H
2
O)
4
2+
+4 Cl
-
<===> NiCl
4
2-
+
4H
2
O) REACTION OCCUR IN A SERIES OF
STEPS
Ni (H
2
O)
4
2+
+Cl
-
<===> Ni (H
2
O)
3
Cl
+
Ni (H
2
O)
3
Cl
+
+Cl
-
<===>Ni (H
2
O)
2
Cl
2
+
Ni (H
2
O)
2
Cl
2
+
+Cl
-
<===> Ni (H
2
O) Cl
3
-
Ni (H
2
O) Cl
3
-
+Cl
-
<===> NiCl
4
-2
K
1
=
[NiCl
+
]
[Ni
2+
][Cl
]
K
2
=
[NiCl
2
]
[NiCl
+
][Cl
]
K
3
=
[NiCl
3
]
[NiCl
2
][Cl
]
K
4
=
[NiCl
4
2
]
[NiCl
3
][Cl
]
12
8/15/2006 Chem208Spring2002 34
Overall Formation constants

1
=K
1
2
=K
1
K
2
3
=K
1
K
2
K
3
4
=K
1
K
2
K
3
K
4
WRITE A MASS BALANCE FOR NICKEL

C
Ni
=[Ni
2+
] +[NiCl
+
] +[NiCl
2
] +[NiCl
3-
] +[NiCl
4
2-
]
2
=
[NiCl
2
]
[Ni
2+
][Cl
]
2
8/15/2006 Chem208Spring2002 35
General Approach
Calculate the fraction of each form in an
equilibrium under specified conditions
will be used to identify fraction

Ni2+
=fraction of total Ni in the Ni
2+
form

HA
=fraction of weak acid (HA) in
undissociatedform.

A-
=fraction of weak acid (HA) in dissociated
form.
8/15/2006 Chem208Spring2002 36
Species present
Three techniques
Calculate fraction in each form- alpha
Log Concentration plots (computer)
Line diagrams (qualitative)
13
8/15/2006 Chem208Spring2002 37
Species present at a pH
Calculate alphas
HA <==> H
+
+A
-
C
HA
=[HA] +[A] Mass balance
Define fraction of total in specific form
HA
A
HA
HA
C
A
C
HA ] [

] [

= =
8/15/2006 Chem208Spring2002 38
Substitute values from K
a
C
HA
=[HA] +[A]
HA
A
HA
HA
C
A
C
HA ] [

] [

= =
] [
] [
] [
] ][ [
] [
'
' +
+
= =
H
HA K
A
K
A H
HA
a
a
8/15/2006 Chem208Spring2002 39
For monoproticacid - Exact equation
'
'
'
] [
] [
] [
a
a
A
a
HA
K H
K
K H
H
+
=
+
=
+
+
+
14
8/15/2006 Chem208Spring2002 40
For polyproticacid - Exact equation
' '
2
'
1
2 '
2
'
1
1 '
1
'
2
'
1
2
2
' '
2
'
1
2 '
2
'
1
1 '
1
' '
2
'
1
' '
2
'
1
2 '
2
'
1
1 '
1
.. ... ] [ ] [ ] [
] [
.. ... ] [ ] [ ] [
.. ] [
.. ... ] [ ] [ ] [
] [
an a a
n
a a
n
a
n
a a
n
A
an a a
n
a a
n
a
n
an a a
m n
Am
an a a
n
a a
n
a
n
n
HA
K K K H K K H K H
K K H
K K K H K K H K H
K K K H
K K K H K K H K H
H
+ + +
=
+ + +
=
+ + +
=
+ + +
+
+ + +
+
+ + +
+
8/15/2006 Chem208Spring2002 41
Each term in denominator represents one
form of the acid
' '
2
'
1
2 '
2
'
1
1 '
1
'
2
'
1
2
2
' '
2
'
1
2 '
2
'
1
1 '
1
' '
2
'
1
' '
2
'
1
2 '
2
'
1
1 '
1
.. ... ] [ ] [ ] [
] [
.. ... ] [ ] [ ] [
.. ] [
.. ... ] [ ] [ ] [
] [
an a a
n
a a
n
a
n
a a
n
A
an a a
n
a a
n
a
n
an a a
m n
Am
an a a
n
a a
n
a
n
n
HA
K K K H K K H K H
K K H
K K K H K K H K H
K K K H
K K K H K K H K H
H
+ + +
=
+ + +
=
+ + +
=
+ + +
+
+ + +
+
+ + +
+
8/15/2006 Chem208Spring2002 42
Plot vspH - Useful to see what is present
as function of pH
Sketch approximate alpha plots
pH=pK at 50% crossover
H2A
pK'1
A
2-
pK'2
0.0
0.2
0.4
0.6
0.8
1.0
0 2 4 6 8 10 12 14
p[H3O
+
]
a
lp
h
a
15
8/15/2006 Chem208Spring2002 43
Log Conc. Plot
Excel Spreadsheet
Depend on Conc.
8/15/2006 Chem208Spring2002 44
Advantages of alpha
Rigorous No assumptions
Define pH and do calculation
Concentration of total acid does not matter
Useful for simplifying equilibrium
calculations for non-acid/base chemistry
Useful in predicting titration curves
8/15/2006 Chem208Spring2002 45
Ladder Diagrams
Easy way to see what is going on
1. What component will vary in the system
Acids and bases pH
Complexation pLigand
Redox- Potential
Draw line to represent the possible values
As you adjust this parameter along the line, the
ratio of reactants and products must change.
16
8/15/2006 Chem208Spring2002 46
Drawing ladder diagrams
Set up pH line
Put in point for pH=pK for each acid
present (approximate only)
Label the major species present in each
section.
8/15/2006 Chem208Spring2002 47
Mix of HF
And NH
3
pH=pK
NH3=9.24
pH=pk
HF=3.17
NH4+
NH3
F-
HF
8/15/2006 Chem208Spring2002 48
At any pH
Look at the predominate species
pH=pK
NH3=9.24
pH=pk
HF=3.17
NH4+
NH3
F-
HF
17
8/15/2006 Chem208Spring2002 49
Ladder Diagrams for Any
Equilibrium
Redox line is E potential in Volts
Complexation line is ligand concentration
Great way to see what is present at a
specific set of conditions.
Figuring out what is present is hardest part.
8/15/2006 Chem208Spring2002 50
Ladder for complexation
Ligand conc. Is line
1
8/15/2006 Chem208Spring2002 1
Chem 208
Chapter 7
Titration
8/15/2006 Chem208Spring2002 2
Volumetric Analysis
Acid Base Reactions
ComplexationReactions
REDOX Reactions
Add reagent of known Concentration
Monitor Something that indicates progress
of reaction
8/15/2006 Chem208Spring2002 3
Volumetric Analysis
Equivalence Point - Theory
moles titrant=moles analyte
End Point - Practice
Indicator Changes Color
Signal reaches specified value
Analysis of output indicates Equivalence point
2
8/15/2006 Chem208Spring2002 4
Detecting Endpoints
Visual Indicators
Potentiometric Titrations
Accurate graph of data. Use compass to
find midpoint of graph.
Curvefit to theory
Derivatives of plot
Titrate to specific pH
Alkalinity methods for water analysis do
this
8/15/2006 Chem208Spring2002 5
Visual Indicators
http://inst.santafe.cc.fl.us/~chem/intbl.html
8/15/2006 Chem208Spring2002 6
End Points
End point is where the titration is stopped
Equivalence point is where acid=base
Not always equal
Indicator error ==>Do blank titration
add salt present at Equivalence point +
indicator and titrate
3
8/15/2006 Chem208Spring2002 7
Types of Titration Methods
Direct Titration
Acid Base and EDTA are examples
Back Titration
Add excess reagent and titrate excess
Vitamin C in Chem112
Add IO
3
I
2
reacts with Vit. C
TitrateUnreactedI
2
Displacement Titration
Ca and Mg EDTA
8/15/2006 Chem208Spring2002 8
Simple Titrations
No Equilibrium Chemistry to worry about
Strong Acid-Strong Base
Anything with really huge K
eq
Stoichiometry determines curve
8/15/2006 Chem208Spring2002 9
Simple Titrations
Moles titrant =C
t
V
t
Moles Analyte =C
o
V
o
Moles un-titrated =C
o
V
o
C
t
V
t
C
t o
t t o o
a
V V
V C V C
C
+
=
4
8/15/2006 Chem208Spring2002 10
Molarity vs volume
0.1 M HCl with 0.1 M NaOH
0
0.02
0.04
0.06
0.08
0.1
0.12
0 5 10 15 20
Series1
8/15/2006 Chem208Spring2002 11
pH vs Volume
Note rapid change at 15 ml
0
1
2
3
4
5
6
7
8
0 5 10 15 20
Series2
8/15/2006 Chem208Spring2002 12
End Points
At end of Titration Stoichiometry
produces very large changes in the relative
amount of analyte for small additions of
titrant.
5
8/15/2006 Chem208Spring2002 13
After the Endpoint
Stoichiometric addition of excess titrant
Relative excess changes rapidly at EP
Change becomes more gradual as you get
farther along.
t o
o o t t
t
V V
V C V C
C
+
=
8/15/2006 Chem208Spring2002 14
Simple Titration Curve
0
2
4
6
8
10
12
14
0 5 10 15 20 25 30
Series2
8/15/2006 Chem208Spring2002 15
Titration Calculations
Moles Analyte =moles Titrant (x
stoichiometry factor from reaction)
C
a
V
a
=C
t
V
t
(Stoichiometry factor)
1
Chapter 8
Activity
ACTIVITY
Describes the real, effective concentration of
the species in the solution (Their chemical
potential)
Not always same as concentration
Activity is not always easy to determine.
Some techniques actually measure activity
directly (eg.pH electrode) but unless they can
be calibrated with standards of known activity
this doesn't do any good.
Activity of HCl solutions
Activity vs Concentratino for HCl
0
100
200
300
400
500
600
700
800
0 5 10 15
Concentrati on (F)
A
c
tiv
ity
H
+
Activity
Activity vs Concentration for HCl
0
0.2
0.4
0.6
0.8
1
0 0.2 0.4 0.6 0.8 1
Concentrati on (F)
A
c
tiv
ity
H
+
Series1
2
ACTIVITY see Chapter 8
Activity coefficient
Finagle constant
x x x
C =
ACTIVITY
Problem of definition. What are the
conditions where f=1.00000 ? The value of
K
eq
will depend on how this is chosen.
Henryan System
Used for Solutions
f =1 at limit C ==>0
Chemistry at infinite dilution.
Graph values at low Conc. then extrapolate to
1.0 M
At infinite dilution, the solvent is unperturbed
by any solute particles and the each solute
particle feels no forces other than from the
solvent.
3
Concentration Effects
Concentration increases
Ions get close enough to be attracted and
repelled by other ions
All ions are solvated (Solvation number)
Very High concentration
Run out of solvent for hydration
Activity coefficients range widely
Raoultian System
f =1 at 1.00 mole fraction.
Thus a =[conc.] for a pure substance.
This reference system is useful for pure
liquids and solids.
Distillation
It is also used in some types of
chromatography.
Use Henryan System
Unless othewise specified
Note- Activities at std. State
all =1.00 mol/l
Values in K
eq
expression
Dimensionless
b
o
b
b
a
o
a
a
d
o
d
d
c
o
c
c
eq
K
4
Activity Coefficients
Function of Ionic Strength
Ionic strength depends on charges of ions
More highly charged ions make S>>[Conc]
Na
2
SO
4
[Na]=2x [SO
4
] but S=1/2([Na] +[SO
4
]x4)
S=1/2(2x [SO
4
] +4x [SO
4
] ) =3x [SO
4
]
+ + =
+
... ] [ ] [
2
1 2 2
B
z
A
z
z B z A S
Calculating Activity Coef.
DHLL - excellent but only under trivial
conditions S=0.0001 M or less
DHE (EDHE)- OK to S=0.01
still not terrible useful
Doesnt work for blood, urine, or sea water
Davies Equation - empirical - curve fit
Cut to the Chase
Ignore Activity Coefficients in most work
Cant get them accurately
Places limit on absolute accuracy of all
equilibrium calculations
Biochemists making buffers to match
physiological conditions
Use 1:1 salts for best accuracy
Use highly charged ions to get high ionic
strength at low conc.
5
Cut to the Chase
Makes Great excuse for why things
dont work quite right
Potentiometric titration data fit
Use activity coefficients as excuse for
some deviations from theory
Cut to the Chase
Activity effects on Non-polar species
Activity coefficients >1
Salting in and Salting out
Add a lot of salt to force something out of
solution -
Protein separations
Acetone extractions
1
8/15/2006 Chem208Spring2002 1
Chem 208
Chapter 9
SystemmaticEquilibrium
8/15/2006 Chem208Spring2002 2
Systemmatic Approach to
Equilibrium Problems
If exact answers are needed
Set of steps to take
Use Acid Base as example
8/15/2006 Chem208Spring2002 3
Acid Base Equilibria
Formal Approach to solving Equilibrium
Problems
Step 1. Write all chemical reactions and
their equilibrium constant expressions.
2
8/15/2006 Chem208Spring2002 4
Example - Benzoic Acid
2 H
2
O <===> H
3
O
+
+ OH
-
K
w
=1.00 X 10
-14
C
6
H
5
COOH + H
2
O <==> C
6
H
5
COO
-
+ H
3
O
+
Too Many Unknowns to solve
Need additional Equations
K
a
=
[H
+
][BA
]
[HBA]
= 6.3x10
5
8/15/2006 Chem208Spring2002 5
CHARGE BALANCE Solution is
electrically neutral
Sum [+ions] =Sum [- ions]
[H
+
] = [BA
-
] + [OH
-
]
[BA
-
] = [H
+
] - [OH
-
]
8/15/2006 Chem208Spring2002 6
MASS BALANCE - Conservation of
matter
C
HBa
=[HBA] +[BA
-
]
3
8/15/2006 Chem208Spring2002 7
Substitute from Charge Balance into Mass
Balance
[BA
-
] = [H
+
] - [OH
-
]
C
HBA
=[HBA] +[H
+
] - [OH
-
]
REARRANGE
[HBA] =C
HBA
- [ H
+
] +[OH
-
]
SUBSTITUTE INTO K
a
EXPRESSION
] [ ] [
] ][ [
+
+
+
=
OH H C
Ba H
K
HBa
a
8/15/2006 Chem208Spring2002 8
Exact solution
Substitute from
Charge Balance
[OH
-
] =K
w
/[H
+
]
( )
] [ ] [
] [ ] [ ] [
+
+ +
+

=
OH H C
OH H H
K
HBa
a
] [
] [
] [
] [ ] [
+
+
+
+ +
+

=
H
K
H C
H
K
H H
K
w
HBa
w
a
8/15/2006 Chem208Spring2002 9
Exact solution
Clean it up
You dont really want to do this. (cubic)
Solutions of very weak acids HCN
K
a
=6x10
-10
( )
] [
] [
] [
2
+
+
+
+

=
H
K
H C
K H
K
w
HBa
w
a
4
8/15/2006 Chem208Spring2002 10
Approximate solution
Ignore OH
OH <<H since this is an acid
If C
HBA
>>[H
+
]
] [
] ][ [
+
+
=
H C
Ba H
K
HBa
a
[ H
+
] = C
HA
K
a
8/15/2006 Chem208Spring2002 11
Solution
Solve on calculator
Works as long as water contribution [OH] is
negligible
] [
] ][ [
+
+
=
H C
Ba H
K
HBa
a
0 ] [ ] [
2
= +
+ +
a HA a
K C H K H
8/15/2006 Chem208Spring2002 12
Monoprotic base
Two ways to solve - Approximate
More Exact
0 ] [ ] [
2
= +

b b b
K C K OH OH
b b
C K OH =
] [
5
8/15/2006 Chem208Spring2002 13
Solving on a TI Calculator
TI-86
POLY
Order =2 Enter
A2=1
A1=K
a
A0=-C
a
xK
a
SOLVE
You get two answers- choose the reasonable
one (positive and not greater than C
a
)
8/15/2006 Chem208Spring2002 14
TI- 83
Set up Equation in Solver
Put in values for things you know
Put cursor on unknown and press SOLVE
Select bounds to eliminate negative answers
If You get two answers- choose the reasonable
one (positive and not greater than C
a
)
8/15/2006 Chem208Spring2002 15
TI- 82
Math
go down list of functions to Solve (last function on
list)
(H^2+K*H-C*K,H,guess .00000001)
numbers for K and C use small value for guess
Make sure result is positive
Make sure result is less than C
6
8/15/2006 Chem208Spring2002 16
Very complicated
Multiple species
Lots of Algebra
Identifying all Chemical Reactions often
hardest part
1
8/15/2006 Chem208Spring2002 1
Chem 208
Chapter 10
Acid-Base
8/15/2006 Chem208Spring2002 2
Calculation pH of anything
Four cases to address
Weak Acid
Acid or conjugate acid of weak base
Weak Base
Base of conjugate base of weak acid
Buffer
Acid plus its conjugate base
Amphiprotic - polyproticsystems
8/15/2006 Chem208Spring2002 3
Monoprotic acid or base
Acid
Base
0 ] [ ] [
2
= +

b b b
K C K OH OH
0 ] [ ] [
2
= +
+ +
a a a
K C H K H
2
8/15/2006 Chem208Spring2002 4
Buffers
Resist Change in pH
Added acid
Added base
dilution
Very important in all chemistry
Acid +conjugate base
8/15/2006 Chem208Spring2002 5
Exact Solution buffers
Example - Base Buffer
.200M + .100M
NH
3
+ NH
4
NO
3
NH
3
+H
2
0 <====>NH
4
+
+OH
-
NO
3
-
disassociated - no Rx
8/15/2006 Chem208Spring2002 6
Buffer Example
Change balance [NH
4
+
] +[H
+
] =[OH
-
] +[NO
3
-
]
Mass balance 0.200 +0.100 =[NH
3
] +[NH
4
+
]
By substitution from Charge Balance
[NH
4
+
] =[OH
-]
+[NO
3
-
]- [H
+
]
0.200 +0.100 =[NH
3
] +[NO
3
-
] +[OH
-
] - [H
+
]
[NO
3
-
] =0.100 Therefore subtract from both
sides
0.200 =[NH
3
] +[OH
-
] - [H
+
]
[NH
3
] = 0.200 - [OH
-]
+ [H
+
]
3
8/15/2006 Chem208Spring2002 7
Buffer Example
Mass Balance rearranged
[NH
3
]=0.200 +0.100 - [NH
4
+
]
substitute in expression for NH
3
0.100 =[NH
4+
] - [OH
-
] +[H
+
]
Rearranging we see that :
[NH
4
+
] = 0.100 + {[OH
-
] - [H
+
]}
and
[NH
3
] = 0.200 - {[OH
-
] - [H
+
]}
8/15/2006 Chem208Spring2002 8
Buffer Example
Substitute
Exact Solution
J ust like acid but with Concof Conjugate included
] [
] ][ [
3
4 '
NH
NH OH
K b
+
=
( ) { }
( ) ] [ ] [
] [ ] [ ] [
3
4 '
+
+

+
=
H OH C
H OH C OH
K
NH
NH
b
8/15/2006 Chem208Spring2002 9
Acidic buffer
Completely rigorous all monoproticcases
Nasty cubic equation
Simplify
If Acidic H
+
>>OH
-
Drop OH term
If Basic OH>>H Drop H term
{ }
] [ ] [
] [ ] [ ] [
'
+
+ +
+
+
=

OH H C
OH H C H
K
HA
A
a
{ }
] [
] [ ] [
'
+
+ +
+
=

H C
H C H
K
HA
A
a
4
8/15/2006 Chem208Spring2002 10
Acidic buffer
Simplify
If Concentrations of Acid and Salt are high
C>>H
+
Drop those terms
HA
A
a
C
C H
K

+
=
] [
'
8/15/2006 Chem208Spring2002 11
Acidic buffer
Take -log of equation
Henderson-Hasselback
sometimes invert
and change sign of
ratio term. Watch out
HA
A
a
C
C H
K

+
=
] [
'
+
+
HA
A
a
HA
A
a
HA
A
a
C
C
pH pK
C
C
H pK
C
C H
K
log
log ] log[
] [
log log
'
8/15/2006 Chem208Spring2002 12
Solution to Ammonia example
Base/Acid
NH
3
is base
NH
4
+
is Acid
Things to check
If more base than Acid then pH should be
higher than pK. Good way to make sure you
get signs right.
545 . 9
100 . 0
200 . 0
log 244 . 9 =
+ = pH
5
8/15/2006 Chem208Spring2002 13
Using line diagram to check
Compare pH to pK
Look at diagram
Is the correct species the predominate one
In example NH
3
>NH
4
+
pH >pK
8/15/2006 Chem208Spring2002 14
Buffers- Resist pH Change
Weak Acid-Conjugate Base pair
Henderson-Hasselbach Eq
Since C
HA
and C
A
apply to same
solution==>volumes cancel
Use ratio of moles salt/moles acid
+ =

HA
A
a
C
C
pK pH log
8/15/2006 Chem208Spring2002 15
Buffer Capacity
Maximum at pH =pK
Useable in range pH=pK 2
6
8/15/2006 Chem208Spring2002 16
Two techniques
Calculate fraction in each form
Log Concentration plots
1
8/15/2006 Chem208Spring2003 1
Chem 208
Chapter 11
Poly-proticsystems
8/15/2006 Chem208Spring2003 2
Polyprotic systems
Each proton is harder to remove
mostly due to the increased energy
required to separate the charges.
H
3
PO
4
K
a1
=7.5x10
-3
K
a2
=6.2x10
-8
K
a3
=4.8x10
-13
Polyprotic bases.
PO
4
-3
polyamines such as ethylene diamine.
NH
2
-CH
2
-CH
2
-NH
2
K
b1
=9.1x10
-5
K
b2
=1.5x10
-7
8/15/2006 Chem208Spring2003 3
Polyprotic systems
If K
1
>K
2
solve as monoprotic acid
(or base)
Use K
1
and quadratic
[H
+
] from first Eq. Suppress other reactions
2
8/15/2006 Chem208Spring2003 4
Polyprotic systems
H
3
P0
4
+ H
2
0 <===> H
2
P0
4- +
H
3
0
+
H
2
PO
4
-2
+ H
2
0 <===> HP0
4
2-
+ H
3
0
+
HP0
4
-2
+ H
2
0 <===> P0
4
-3
+ H
3
0
+
K
a1
=
[ H + ] [ H
2
PO
4
]
[ H
3
PO
4
]
K
a2
=
[ H + ] [ H PO
2
4
]
[ H
2
PO

4
]
K
a3
=
[ H + ] [ PO
3
4
]
[ H PO
2
4
]
8/15/2006 Chem208Spring2003 5
How much does K
2
contribute
K
a2
=
[ H + ] [ H PO
2
4
]
[ H
2
PO

4
]
] [
] [
] ][ [
] [ ] [
2
4 2
2
4
2
4 2
+
+
+
=
=
PO H K
H
PO H H
K
PO H H
8/15/2006 Chem208Spring2003 6
Amphiprotic substances
Both acids and bases at the same time
HC0
3-
H
2
P0
4,
HP0
4
2-
Intermediate forms of polyprotics
Amino acids (e.g.alanine)
weak acid - weak base salts
ammonium acetate
ammonium formate
3
8/15/2006 Chem208Spring2003 7
NaH malate example
HMal
-
+H
2
0 <===>H
2
Mal +0H
-
K
b2
=K
w
/K
a1
=2.5x10
-11
HMal
-
+H
2
0 <===>Mal
-2
+H
3
0
+
K
a2
=8.9x10
-6
Which reaction has larger K?
Solution will be acidic
8/15/2006 Chem208Spring2003 8
NaH malate example
5 Unknowns [H
2
Mal] , [HMal
-
] ,[Mal
2-
], [H
+
] ,
and [0H
-
] Need 5 equations
Mass Balance
C
NaHMAl
=[H
2
Mal] +[HMal
-
] +[Mal
2-
]
Charge Balance
[Na
+
] +[H
+
] =[HMal
-
] +[0H
-
] +2[Mal
2-
]
8/15/2006 Chem208Spring2003 9
NaH malate example
5 Unknowns [H
2
Mal] , [HMal
-
] ,[Mal
2-
],
[H
+
] , and [0H
-
] Need 5 equations
K
a2
=[H
+
] [Mal
2-
] / [HMal
-
]
K
b2
=[0H
-
] [H
2
Mal] / [HMal
-
]
K
w
=[H
+
] [0H
-
]
4
8/15/2006 Chem208Spring2003 10
NaH malate example
[Na
+
] +[H
+
] =[HMal
-
] +[0H
-
] +2[Mal
2-
]
[Na
+
] =C
NaHMAl
C
NaHMal
+[H
+
] =[HMal
-
] +[OH
-
] +2[Mal
2-
]
Subtract Mass balance below
C
NaHMAl
=[H
2
Mal] +[HMal
-
] +[Mal
2-
]
8/15/2006 Chem208Spring2003 11
NaH malate example
[H
+
] =[Mal
2-
] - [H
2
Mal] +[0H
-
]
] [
] [
] [
] [
] ][ [
2 2
2
2
+
+
=
=
H
HMal K
Mal
HMal
Mal H
K
a
a
1
1
2
1
2
2
] ][ [
] [
] [
] [
] [
] ][ [
a
w
a
w
a
w
b
K
H HMal
H
K
K
HMal K
Mal H
K
K
HMal
OH Mal H
K
+
+
= =
= =
] [
] [
+
=
H
K
OH
w
8/15/2006 Chem208Spring2003 12
NaH malate example
[H
+
] =[Mal
2-
] - [H
2
Mal] +[0H
-
]
( )
w a a a a
w a
a
w a
a
w
a
a
w
a
a
K K HMal K K HMal K H
K HMal K
K
HMal
H
K HMal K
K
H HMal
H
K
K
H HMal
HMal K H
H
K
K
H HMal
H
HMal K
H
2 2 2 2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
] [ ] [ ] [
] [
] [
1 ] [
] [
] ][ [
] [
] ][ [
] [ ] [
] [
] ][ [
] [
] [
] [
+ = +
+ =
+
+ = +
+ =
+ =
+
+
+
+
+
+
+
+
+
5
8/15/2006 Chem208Spring2003 13
NaH malate example
Rigorous equation
=
+
] [H
[H
+
] =
K
a1
(K
a2
[HMal
] + K
w
)
K
a1
+ [HMal
]
8/15/2006 Chem208Spring2003 14
NaH malate example
Simplifying Assumptions
1. If [HMAl] >>K
A1
then K
a1
+[HMal
-
] = [HMal
-
]
[H
+
] =
K
a1
(K
a2
[HMal
] + K
w
)
K
a1
+ [HMal
]
[H
+
] =
K
a1
(K
a2
[HMal
] + K
w
)
[HMal
]
8/15/2006 Chem208Spring2003 15
NaH malate example
Simplifying Assumptions
If K
A2
[HMAl] >>Kw
then drop last term - conc. cancels
[H
+
] =
K
a1
(K
a2
[HMal
] + K
w
)
[HMal
]
[H
+
] = K
a1
K
a2
[H
+
] =
K
a1
K
a2
[HMal
]
[HMal
]
6
8/15/2006 Chem208Spring2003 16
NaH malate example
Equation independent of concentration
Works at high concentration only
Use for Amino Acids
[H
+
] = K
a1
K
a2
8/15/2006 Chem208Spring2003 17
NaH malate example
For NaMal
[H
+
] = (4.0x10
-4
x 8.9 x 10
-6
)
1/2
=5.97 x
10
-5
[H
+
] = K
a1
K
a2
8/15/2006 Chem208Spring2003 18
NaH malate example
What if assumptions dont work
0.0005 M NaMal
[HMAl] K
A1
Calculate approximate value using
simple form
7
8/15/2006 Chem208Spring2003 19
NaH malate example
[H
+
] = =5.97 x 10
-5

1
at this pH
[H
+
]
2
=2.56x10
-9
K1 [H
+
] =2.39x10
-8
K
1
K
2
=3.56x10
-9
770 . 0
10 10 . 3
10 39 . 2
8
8
1
= =
x
x
8/15/2006 Chem208Spring2003 20
NaH malate example
Calculate [Hmal
-
] =
0.0005x0.770=0.000385
Plug into rigorous equation
770 . 0
10 10 . 3
10 39 . 2
8
8
1
= =
x
x
( )
5
4
14 6 4
10 18 . 4
00038 . 0 10 4
10 00 . 1 00038 . 0 10 9 . 8 10 4
] [

+
=
+
+
= x
x
x x x x
H
8/15/2006 Chem208Spring2003 21
NaH malate example
Calculate new alpha at this pH
alpha=0.76
Repeat
Result OK
( )
5
4
14 6 4
10 25 . 4
000381 . 0 10 4
10 00 . 1 000381 . 0 10 9 . 8 10 4
] [

+
=
+
+
= x
x
x x x x
H
8
8/15/2006 Chem208Spring2003 22
Summary
Weak Acid
Weak Base
Buffer
Amphiprotic [H
+
] = K
a1
K
a2
0 ] [ ] [
2
= +
+ +
a HA a
K C H K H
0 ] [ ] [
2
= +

b b b
K C K OH OH
+ =
HA
A
a
C
C
pK pH log
8/15/2006 Chem208Spring2003 23
Summary
Have to be able to tell what is present
Stoichiometry
Recognize acids and bases
8/15/2006 Chem208Spring2003 24
Decision Tree for Titration
Problems
pH=1/2(pK1 +pK2)
Amphiprotic Substance
Buffer - HH Equation
Weak Acid +Conjugate Base
Quadratic solution using Kb1
Weak Base Only
What chemicals are Present?
Yes
What is limiting reagent?
Calculate concentration of all reactants and products
Quadratic solution using Ka1
No
Weak Acid Only
Is more that one reagent added
9
8/15/2006 Chem208Spring2003 25
Practice Problem
Calculate the pH of 25.0 ml of a 0.100 M
solution of phthalicacid after the addition
of 0, 10.0, 25.0, 35.0, 50.0 and 75.0 ml of
0.100 M NaOH
K
1
=1.12x10
-3
pK
1
=2.95
K
2
=3.90x10
-6
pK
2
=5.41
8/15/2006 Chem208Spring2003 26
Practice Problem
0 ml weak diproticacid only treat as
monoprotic
K
1
=1.12x10
-3
97 . 1 0106 . 0 ] [
0 10 12 . 1 10 . 0 ] [ 10 12 . 1 ] [
3 3 2
= =
= +
+
+ +
pH H
x x H x H
8/15/2006 Chem208Spring2003 27
Practice Problem
10.0 ml base
moles acid =25.0x0.1 =2.5 mmoles
moles base =10.0x0.1 =1.0 mmoles
acid reacted to NaHP=1.0 mmoles
H
2
P acid remaining =1.5 mmoles
Buffer solution - use HH ignore dilution
77 . 2
5 . 1
0 . 1
log 95 . 2 =
+ = pH
10
8/15/2006 Chem208Spring2003 28
Practice Problem
25.0 ml base
H
2
P acid reamining=0 mmoles
Amphiproticignore dilution
18 . 4
10 6 . 6 10 90 . 3 10 12 . 1 ] [
5 6 3
=
= =
+
pH
x x x x H
8/15/2006 Chem208Spring2003 29
Practice Problem
35.0 ml base
base remaining =1.0 mmole
NaHP reacted with base =1.0 mmoles
NaHP remaining =1.5 mmol
P
2-
formed =1.0 mmole buffer HH ignore
dilution
8/15/2006 Chem208Spring2003 30
Practice Problem
35.0 ml base
NaHP remaining =1.5 mmol
P
2-
formed =1.0 mmole buffer HH
23 . 5
5 . 1
0 . 1
log 41 . 5 =
+ = pH
11
8/15/2006 Chem208Spring2003 31
Practice Problem
50.0 ml base
moles acid (both protons) =25.0x0.1x2 =
5.0 mmoles
All acid converted to phthlateP
2-
Weak base - treat as monoprotic-with
dilution
97 . 8 03 . 5 10 24 . 9 ] [
0
10 90 . 3
10 1
75
25
1 . 0
10 90 . 3
10 1
] [ ] [
0 ] [ ] [
6
6
14
6
14
2
2 2
2
= = =
= +
= +

pH pOH x OH
x
x
x x
x
x
OH OH
K
K
C
K
K
OH OH
a
w
b
a
w
8/15/2006 Chem208Spring2003 32
Practice Problem
75.0 ml base
moles protons =25.0mlx0.1mM/mlx2 protons M=
5.0 mmoles
phthlateconc=2.5 mmoles/100 ml =0.025
Excess base=2.5 mmoles/100 ml =0.025 M
Strong base +weak base ==>strong base
[OH
-
]=0.025 M pOH=1.60 pH=12.40
1
8/15/2006 Chem208Spring2002 1
Chem 208
Chapter 12 -5 & 9
Titration Endpoints
8/15/2006 Chem208Spring2002 2
Weak Acid- Strong Base
Traditional way to calculate titration curves
Break into regions
Acid
Buffer
Conj. Base
strong base
Complex and not possible on computer
8/15/2006 Chem208Spring2002 3
Finding Endpoints
Indicators
Change color +/- 1 pH unit around pK
Equivalence point pH must be close to pK
Spectrophotometricand Fluorescence
Monitor pH - Lab titration
Plot data and analyze accurate graph
Take 1
st
and 2
nd
derivatives
GranPlots V
b
x[H
+
] vsV
b
EP at zero
Temperature
Pressure
2
8/15/2006 Chem208Spring2002 4
Derivatives
Slope at tangent between points
dpH/dV vs (V2-V1)/2
Second derivative is derivative of first der.
Points do not need to be equally spaced
Automatic titrations
Systemstops When 2
nd
Der. =zero
Smooth thedata
Titrate to final pH value
Alkalinity determination
8/15/2006 Chem208Spring2002 5
Derivative Spreadsheet
Vol pH Av Vol 1
st
Der Av Vol 2
nd
Der
8/15/2006 Chem208Spring2002 6
Alternative Approach
Introduce Fraction Titrated
acid moles
base moles
= =
a a
b b
V C
V C
3
8/15/2006 Chem208Spring2002 7
At any point in a titration the following
must be true:
Mass Balance Equation
And moles base added =moles acid

reacted
] [ ] [ ] [
2
2

+ + =
+
A HA A H
V V
V C
b a
a a
( ) ( )
a
a
a
HA
a b b
V OH H C C V OH H C ] [ ] [ 2 ] [ ] [
2
+ +
+ + = +

8/15/2006 Chem208Spring2002 8
Fraction titrated then becomes
If we define Fraction Dissociated for the

acid as
b
a
a HA
C
H OH
C
H OH
] [ ] [
1
] [ ] [
2 2
+
+
+ +
=

+ =
2
2
A HA
ad
F
8/15/2006 Chem208Spring2002 9
Fraction Titrated
b
w
a
w
ad
C
H K H
C
H K H
F
] /[ ] [
1
] /[ ] [
+ +
+ +
=
4
8/15/2006 Chem208Spring2002 10
Volume of base added
Related to fraction
titrated
Calculate theoretical
V
b
to get to any pH
b
a a
b
C
V C
V

=
acid moles
base moles
= =
a a
b b
V C
V C
8/15/2006 Chem208Spring2002 11
Can be extended to titration with a weak
base (carbonate) as well.
denominator corrected for the fraction
of the carbonate in various forms.
b
w
bd
a
w
ad
C
H K H
F
C
H K H
F
] /[ ] [
] /[ ] [
+ +
+ +
=
1 0
2 + =
bd
F
8/15/2006 Chem208Spring2002 12
Rigorous - No assumptions or
approximations
J .ChemEd. Vol 70, pp.-209-217, (1993)
b
w
bd
a
w
ad
C
H K H
F
C
H K H
F
] /[ ] [
] /[ ] [
+ +
+ +
=
b
a a
b
C
V C
V

=
5
8/15/2006 Chem208Spring2002 13
Diprotic Acid Titration
Program
Start with the pH of the solution and calculate
the Fraction Titrated at that pH. A single
equation can do this at all points.
Calculate volume of base required to reach
the fraction titrated
Excel Spreadsheet - calculates the theoretical
curve for a diprotic weak acid titrated with
NaOH
8/15/2006 Chem208Spring2002 14
Program
Enter your pH data with volume of titrant
Molarity of NaOH
K
1
and K
2
of your acid
Volume of diprotic acid
Concentration of acid solution - Calculate from
end point
8/15/2006 Chem208Spring2002 15
Program
Adjust parameters to get a good fit. Use
the sum of the squares of the deviations
to measure fit.
Molarity of acid- Adjust until endpoints
exactly coincide. Use molarity of acid
and mass used to calculate MW of acid
Mass/moles =MW
6
8/15/2006 Chem208Spring2002 16
Program
Ks these are susceptible to errors due
to temperature and ionic strength.
Adjust for best fit of lines. K
1
before 1st
EP and K
2
after 1st EP
8/15/2006 Chem208Spring2002 17
Program
DONT CHANGE THESE
Molarity of base (You standardized it)
Volume of acid (measured accurately)
If fit is bad at high pH, you can delete
some points if you want.
pH electrode also responds to sodium ions
Typical good fit has Sum of Squares <4
8/15/2006 Chem208Spring2002 18
Program
Original Data
Good Fit
Not at high pH
Sum
2
=81
Titration curve - Theory vs actual
0
2
4
6
8
10
12
14
-10 0 10 20 30 40
Vol NaOH (ml)
p
H
Actual
Theoretical
7
8/15/2006 Chem208Spring2002 19
Program
Delete last points
Exc Fit
Sum
2
=1.5
0
2
4
6
8
10
12
14
-5 0 5 10 15 20 25
Vol NaOH (ml)
p
H
Actual
Theoretical
8/15/2006 Chem208Spring2002 20
Program
Expand EP region - Drop last 3 points
6
6.5
7
7.5
8
8.5
9
9.5
10
10.5
11
16 16.5 17 17.5 18 18.5 19
Vol NaOH (ml)
p
H
Actual
Theoretical
Conc. Acid = 0.0314 Diprotic Acid
Conc. Base = 0.089914 K1= 0.000038
Vol. Acid = 25 K2= 0.000006
Sumof deviations sq.= 0.44676964
1
8/15/2006 Chem 208 Spring 2001 1
Chem 208
Chapter 13
Complexometric Titration
8/15/2006 Chem 208 Spring 2001 2
SEQUENTIAL COMPLEXATION
BY SIMPLE LIGANDS
Ni(H
2
O)
4
2+
+ 4 Cl
-
<===> NiCl
4
2-
+ 4H
2
O)
REACTION OCCUR IN A SERIES OF STEPS
Ni (H
2
O)
4
2+
+ Cl
-
<===> Ni (H
2
O)
3
Cl
+
Ni (H
2
O)
3
Cl
+
+ Cl
-
<===> Ni (H
2
O)
2
Cl
2
+
Ni (H
2
O)
2
Cl
2
+
+ Cl
-
<===> Ni (H
2
O) Cl
3
-
Ni (H
2
O) Cl
3
-
+ Cl
-
<===> NiCl
4
-2
K
1
=
[NiCl
+
]
[Ni
2 +
][Cl
]
K
2
=
[NiCl
2
]
[NiCl
+
][Cl
]
K
3
=
[NiCl
3

]
[NiCl
2
][Cl
]
K
4
=
[NiCl
4
2
]
[NiCl
3
][Cl
]
8/15/2006 Chem 208 Spring 2001 3
Overall Formation constants

1
= K
1
2
= K
1
K
2
3
= K
1
K
2
K
3
4
=K
1
K
2
K
3
K
4
WRITE A MASS BALANCE FOR NICKEL

C
Ni
= [Ni
2+
] + [NiCl
+
] + [NiCl
2
] + [NiCl
3-
] + [NiCl
4
2-
]
Rearrange K's to get mass balance in terms
of [Ni ] and [Cl ]
2
=
[NiCl
2
]
[Ni
2 +
][Cl
]
2
2
8/15/2006 Chem 208 Spring 2001 4
Rearrange Betas
[NiCl
+
] = K
1
[Ni
2+
][Cl
-
]
[NiCl
2
] = K
1
K
2
[Ni
2+
][Cl
-
]
2
=
2
[Ni
2+
][Cl
-
]
2
[NiCl
3
-
] =
3
[Ni
2+
][Cl
-
]
3
[NiCl
4
2-
] =
4
[Ni
2+
][Cl
-
]
4
Substitute into Mass Balance
C
Ni
= [Ni
2+
] + b
1
[Ni
2+
][Cl
-
] + b
2
[Ni
2+
][Cl
-
]
2
+ b
3
[Ni
2+
][Cl
-
]
3
+ b
4
[Ni
2+
][Cl
-
]
4
8/15/2006 Chem 208 Spring 2001 5
Define alpha metal
Ni
=
[Ni
2 +
]
C
Ni
Ni
=
[Ni
2 +
]
[Ni
2 +
] +
1
[Ni
2 +
][Cl
] +
2
[Ni
2 +
][Cl
]
2
+
3
[Ni
2 +
][Cl
]
3
+
4
[Ni
2 +
][Cl
]
4
Ni
=
1
1+
1
[Cl
] +
2
[Cl
]
2
+
3
[Cl
]
3
+
4
[Cl
]
4
M
=
1
1 +
1
[L
] +
2
[L
]
2
+
3
[L
]
3
+...+
n
[L
]
n
8/15/2006 Chem 208 Spring 2001 6
Multidentate Ligands for
Complex Formation
Chelate Effect - Formation of rings
5 or 6 member rings are best.
N
N
Ni
N
N
O
O
O
O
Cu
3
8/15/2006 Chem 208 Spring 2001 7
Multidentate Ligands for
Complex Formation
Entropy effects enhance stability of
multidentate ligands.
M(H
2
O)
6
+ 6 L <===> ML
6
+ 6H
2
O
7 <===> 7 entropy change small
M(H
2
O)
6
+ L <===> ML + 6H
2
O
2 <===> 7 large increase in entropy
greater entropy change for the latter reaction.
8/15/2006 Chem 208 Spring 2001 8
Complexation Titration
For titrations, 1:1 complexes best
Intermediate equilibria make titration
breaks less sharp for higher No. of ligands.
Volume Titrant
pM
7
0
14
EP
ML6
ML
8/15/2006 Chem 208 Spring 2001 9
AMINO POLYCARBOXYLIC
ACIDS
Most common and useful polydentate
ligands.
They are used extensively for titrations
Tie up metal for a variety of applications
Mayonnaise
Available with 4, 6, 8 and 10 coordination
sites
Match to Maximum Coordination No. of
metal
4
8/15/2006 Chem 208 Spring 2001 10
ACIDS
EDTA - Ethylenediamine
tetraacetic acid.
This has 6 Lewis base sites, 4
oxygen and 2 amine electron
pairs.
designated as H
4
Y and its anion as
Y
4-
All six complexation sites are able
to bind to a single metal ion for
many metals.
N
N
O
O
O
O
O
O
H
H
H
O
O
H
8/15/2006 Chem 208 Spring 2001 11
ACIDS
NTA - Nitrilotriacetic acid.
Tetradentate - this is best for small metals.
DETPA - Diethylenetriamine pentacetic
acid.
Best for 8 coordinate metals such as the 3rd
Transition series and the Lanthanides.
TTHA - Triethylenetetramine hexacetic
acid.
This is for 10 coordinate metals such as the
actinides.
8/15/2006 Chem 208 Spring 2001 12
ACID-BASE CHEMISTRY OF
LIGANDS
Virtually all ligands are Bronsted bases,
strong or weak
For weak base ligands, the pH of the solution
determines how much of the total concentration
of the ligand is free to form the complex.
N
N
O
O
O
O
O
O
H
H
H
O
O
H
5
8/15/2006 Chem 208 Spring 2001 13
LIGANDS
Use EDTA as an example:
H
4
Y Primary standard but relatively
insoluble
Na
2
H
2
Y
.
2H
2
O Not a true primary standard
but soluble to more than 0.05 m/l .
At low pH most EDTA is in the form H
2
Y
2-
8/15/2006 Chem 208 Spring 2001 14
LIGANDS
At low pH most EDTA is in the form H
2
Y
2-
H4Y
H3Y
-
-pK'1
H2Y
2-
-pK'2
HY
3-
Y
4-
-pK'3 -pK'4
-14
-12
-10
-8
-6
-4
-2
0
0 2 4 6 8 10 12 14
p[H3O
+
]
lo
g

a
lp
h
a
8/15/2006 Chem 208 Spring 2001 15
LIGANDS
M
+n
+ H
2
Y
-2
<===> MY
n-2
+ 2 H
+
Normally titrations must be buffered to
keep pH constant.
A further complication lies in the fact that
the reaction of this form with metal liberates
H
+
.
6
8/15/2006 Chem 208 Spring 2001 16
Conditional Formation
Constants
Titrations. These are useful for describing
the formation constant under a specific set
of experimental conditions.
pH
Auxillary Ligands
Titrations always buffered to hold pH
constant
8/15/2006 Chem 208 Spring 2001 17
Conditional Formation
Constants
M + Y
-4
<===> MY
[Y
4-
] =
4
C
EDTA
NOTE The conditional formation applies in this
case at only the pH used to calculate
4
for
EDTA.
K
f
=
[CaY]
[Ca
2 +
][Y
4
]
K
'
f
= K
f
4
=
[CaY]
[Ca
2 +
]C
EDTA
8/15/2006 Chem 208 Spring 2001 18
EXAMPLE
Using conditional formation constants to
predict feasibility of titration.
50.00 ml of 0.0100 m Cu
2+
is titrated with
0.0100 m EDTA at pH 3.00
K
f
= 6.3x10
18
Note this is a large POSITIVE exponent.
7
8/15/2006 Chem 208 Spring 2001 19
EXAMPLE
Calculate
4
at pH = 3.00

4
= 2.5 x 10
-11
Calculate K
f
' = 6.3 x 10
18
* 2.5 x 10
-11
= 1.6 x 10
8
K
f
= [CuEDTA]
[Cu]C
EDTA
8/15/2006 Chem 208 Spring 2001 20
Example
Initial pCu = -log 0.0100 = 2.00
Before equivalence point:
moles CuEDTA dissociated = moles EDTA not reacted

During most of the titration the excess Cu
suppresses the second term until close to the
endpoint. Common ion effect
volume total
untitrated Cu moles
] [
2
+
Cu
8/15/2006 Chem 208 Spring 2001 21
Example
40.00 ml of titrant added:
[Cu
+2
] = 0.0500 * 0.0100 - 0.0400 * 0.0100 = 0.0011
0.0400 + 0.0500
pCu = 2.96
8
8/15/2006 Chem 208 Spring 2001 22
Example
At equivalence point. Since K
f
' is so large
we know the reaction will lie far to the
right.
Cu
+2
+ Y
-4
====> CuY
-2
[CuEDTA] C
0
V
0
= 0.0100 * 0.0500 =
V
0
+ V
T
0.0500 + 0.0500
=0.0050M
[Cu
2+
] = C
EDTA
8/15/2006 Chem 208 Spring 2001 23
Example
K
f
' = 1.6x10
8
= [CuEDTA] = 0.00500
[Cu2+ ]C
EDTA
[Cu
2+
]
2
[Cu
2+
] = 5.5x10
-6
pCu = 5.26
8/15/2006 Chem 208 Spring 2001 24
Example
After the Equivalence point-
EDTA is now in stoichiometric excess
60.00 ml of titrant:
moles EDTA = stoichiometric excess +
dissociation of CuEDTA
C
EDTA
= C
T
V
T
- C
0
V
0
V
0
+ V
T
9
8/15/2006 Chem 208 Spring 2001 25
Example
C
EDTA
= 0.0100 * 0.0600 - 0.0100 * 0.0500
0.0500 + 0.0600
= 0.000910
[CuY
2-
] = C
0
V
0
= 0.000500
V
0
+ V
T
0.110
= 0.00450
8/15/2006 Chem 208 Spring 2001 26
Example
[Cu
2+
] = 0.00450 = 3.1x10
-8
0.000910 * 1.6x10
8
pCu = 7.51
K
f
' determines sharpness of titration
break.For Cu
+2
pH = 2.00 K
f
' = 2.3x10
5
pH = 3.00 K
f
' = 2.1x10
10
larger K
f
' gives
sharper break.
8/15/2006 Chem 208 Spring 2001 27
How large a K
f
is Needed
Rule of thumb - 1 ppt of sample untitrated
at the end point and a consequent 1 ppt
excess of titrant.
For C
Y
m/l EDTA we can do the following
calculation:
8 '
6
0
0 '
10
01 . 0
10
001 . 0 001 . 0
999 . 0
] [
] [
= = =
f
Y
Y Y m
m
EDTA
f
K
C
C xC x xC
xC
C M
MY
K
10
8/15/2006 Chem 208 Spring 2001 28
Auxillary Ligands
Problems
High pH required and metal not soluble
Two metals with similar K
f
and we want to
titrate just one
Add auxillary ligand - MASKING AGENT
8/15/2006 Chem 208 Spring 2001 29
Auxillary Ligands
[M] =
m
C
m
where
m
refers to the
auxiliary ligand.
f m f
Y m m Y
f
K K
C C
MY
C M
MY
K
4
' '
4 4
] [
] [
] [

=
= =
8/15/2006 Chem 208 Spring 2001 30
EXAMPLE
Titration of Cd
2+
with EDTA at pH 9.24 in
a NH
3
NH
4
+
buffer which also serves to mask Cd
to prevent precipitation.
Both NH
3
and NH
4
+
are 0.100 m/l
K
f
for CdEDTA = 3.16x10
16
Cd
2+
+ 5 NH
3
<===> Cd(NH
3
)
5
2+
11
8/15/2006 Chem 208 Spring 2001 31
Example
Cd
2+
+ 5 NH
3
<===> Cd(NH
3
)
5
2+
K
1
= 398 K
2
= 316 K
3
= 24.5 K
4
= 7.59
K
5
= 2.09

4
= 0.087
K
f
' = 3.16x10
16
* 1.4x10
-4
* 0.087 = 3.8x10
11
4
3
3 2 1
2
2 1 1
10 4 . 1
479 2291 3018 1258 8 . 39 1
1
... ] [ ] [ ] [ 1
1
=
+ + + + +
=
+ + + +
=
x
L K K K L K K L K
m
m
8/15/2006 Chem 208 Spring 2001 32

Example
Titration should work well.
But note that the titration break is not as
great as in the absence of NH
3
.
You could not do it without NH
3
due to
precipitation of Cd(OH)
2
.
8/15/2006 Chem 208 Spring 2001 33
End Point Detection
Colored Complexes.
CuEDTA is bright blue and can be monitored
with a spectrometer.
a colored complex with a substantially lower
K
f
' can be used to monitor the endpoint of a
colorless complex. eg. Cu and Fe(III) can be
titrated together.
12
8/15/2006 Chem 208 Spring 2001 34
End Point Detection
Abs.
Vol. Titrant
Cu titration with EDTA
8/15/2006 Chem 208 Spring 2001 35
End Point Detection
Abs.
Vol. Titrant
Cu and Fe titration with EDTA
Fe E.P.
Cu E.P.
8/15/2006 Chem 208 Spring 2001 36
End Point Detection
Electrochemical.
Endpoints have been monitored by change in
pH
mercury electrode
ion selective electrodes.
13
8/15/2006 Chem 208 Spring 2001 37
End Point Detection
Indicators.
metallochromic indicators
Serendipity science.
1945 - a group of Swiss chemists headed by
Swartzenbach accidentally synthesized the
compound Murexide (Roman purple) which
proved to be an indicator for metals.
8/15/2006 Chem 208 Spring 2001 38
Murexide
N
N O
O
N
O
N
N
-O
O
O
8/15/2006 Chem 208 Spring 2001 39
EBT
This lead to the development of Eriochrom
Black T
note nitro and sulfito groups
N
O
O
S
O
O
O
OH
N
N
HO
14
8/15/2006 Chem 208 Spring 2001 40
Calmagite
N,N,O,O tetradentate complexes
S
O
O
O
OH
N
N
HO
8/15/2006 Chem 208 Spring 2001 41
Indicator equilibria
pK = 6.3 pK = 11.5
H2In- <========> HIn-2 <========> In-3
| | |
red blue yellow-orange
In
-3
+ M <====> MIn RED
with Mg, Ca, Zn, Cd, Hg,
Al, Ga, In, Pb, Fe, Ti, Co, Ni, Cu, Pt, Rare earth
8/15/2006 Chem 208 Spring 2001 42
Indicator reaction
K
fEDTA
>> K
fIn
Before endpoint there is excess metal so the
indicator is present as M-In complex. Solutions
are red.
At end point, EDTA strips the metal from
the indicator complex and the solution
becomes blue. This only works well if the
reaction is fast.
15
8/15/2006 Chem 208 Spring 2001 43
Indicator reaction
For slow reactions the best approach is a
back titration
excess EDTA is added and allowed to react.
Then the solution is back titrated with a
standard Mg
+2
solution to the reverse endpoint.
8/15/2006 Chem 208 Spring 2001 44
Ca titration - most common
Direct EDTA Titration work
The addition of Mg to the titrant normally
makes the Ca determination better
Ca binds more tightly to EDTA than Mg, the
indicator should be stripped of Ca near the
endpoint and be present in the MgIn complex
form.
This then gives a sharp endpoint just after the
Ca equivalence point.
8/15/2006 Chem 208 Spring 2001 45
Back Titration
Used when complex formation is slow
Add excess EDTA
Titrate with Mg
2+
or Mn
2+
16
8/15/2006 Chem 208 Spring 2001 46
Displacement Titration
Useful when indicator is lacking for specific
metal titration.
Add excess MgY complex (standardized
amount)
Allow the metal of interest to displace the
Mg from the complex
Titrate the Mg released with EDTA
8/15/2006 Chem 208 Spring 2001 47
Back titration Example
A 50.0 ml solution containing Ni
2+
and Zn
2+
was
treated with 25.0 ml of 0.0452 M EDTA to bind all
the metal. The excess EDTA was titrated with
0.0123 M Mg
2+
and required 12.4 ml to reach the
endpoint. An excess of the reagent 2,3-dimetcapto-
2-propanol was then added to the solution to
displace the EDTA from the Zn
2+
. The solution was
then titrated with additional Mg
2+
and 29.2 ml of
the titrant was required to reach the new endpoint.
Calculate the molarity of Ni
2+
and Zn
2+
in the
original solution.
8/15/2006 Chem 208 Spring 2001 48
Back titration Example
Moles EDTA added= 1.130 mMol
Moles Mg to titrate excess=0.152 mMol
Moles Zn + Ni= EDTA-Mg = 0.978 mMol
Moles Mg after Zn displaced = moles Zn=
0.359 mMol
Moles Ni= total - moles Zn =0.619 mMol
17
8/15/2006 Chem 208 Spring 2001 49
Example 2
A mixture of Mn
2+
, Mg
2+
, and Zn
2+
was analyzed as follows: A 25.00
ml sample was treated with 0.25 gm of hydroxylamine hydrochloride ,
a reducing agent to keep the Mn
2+
from being oxidized. The solution
was buffered at pH 10.0 by the addition of 10 ml of ammonia-
ammonium buffer. A few drops of Eriochrom Black T indicator were
added and the sample was diluted to 100 ml. It was warmed to 40
o
C
and titrated with 39.98 ml of 0.04500 M EDTA to the blue endpoint.
At this point 2.5 gm of NaF were added to displace Mg
2+
from its
EDTA complex. The solution was titrated with 10.26 ml of 0.02065
M Mn
2+
to a red endpoint, indicating all EDTA was now complexed.
After this endpoint is reached, 5 ml of 15 % (w/w) aqueous KCN was
added to displace the Zn
2+
from its EDTA complex. It required 15.47
ml of the standard Mn
2+
solution to reach the third endpoint. Calculate
the number of milligrams of each metal in the initial 25 ml sample of
unknown.
8/15/2006 Chem 208 Spring 2001 50
Example 2
Moles EDTA = total moles metal = 1.799 mMol
moles Mn to 2nd EP = moles Mg=0.211 mMol
moles Mn to third EP = moles Zn=0.319 mMol
Moles Mn in initial = total - (Mg + Zn)=1.269
mMol
8/15/2006 Chem 208 Spring 2001 51
Applications
Spot Tests
Test Strips
Flow Injection Analysis
Immunoassays
1
8/15/2006 Chem 208 Spring 2001 1
Chem 208
Chapter 14-15
Potentiometry
8/15/2006 Chem 208 Spring 2001 2
Electrochemical Cell
2 Half Cells
Salt Bridge
Potential = E
V
Salt Bridge
OX RED
e-
Pt
Ce (IV) 1.0
m/l
Fe (II) 1.0
m/l
Pt
8/15/2006 Chem 208 Spring 2001 3
Chemical Analysis
.A variety of chemical analyses involve
electrochemical cells.
Potentiometry 208
voltametry 312
amperometry
chronopotentiometry
coulometry
others
2
8/15/2006 Chem 208 Spring 2001 4
Electron Energy Levels
Electron Flow in Metals
Band Theory
Electrons Move in Conducting Band
Electron Levels in Metal Electrodes
Fermi Level
If Fermi Level > Empty Orbital of Ion
Reduction occurs
Vice Versa
8/15/2006 Chem 208 Spring 2001 5
Electron Transfer
Fermi level of
electrons in electrodes
shown
Fe
2+
filled orbital
Ce
4+
empty orbital
8/15/2006 Chem 208 Spring 2001 6
Electron Transfer
Potential is difference
in energy of two
electrodes.
Measure with no
electron flow
Thermodynamic
potential
G = -nFE
3
8/15/2006 Chem 208 Spring 2001 7
Measuring Potential
Operational amplifiers
measure potential while allowing little current
flow.
The input of the Op Amp appears to the cell as
a simple resistor with resistance ranging from
10
10
to possibly as high as 10
15
ohms depending
on quality and cost. This value is often called
the input impedance of the op amp.
We can calculate the rate of the reaction
occurring for any given cell EMF and input
impedance
8/15/2006 Chem 208 Spring 2001 8
Potentiometer Circuit
Salt Bridge
Voltage Follower
Amp
Voltmeter
8/15/2006 Chem 208 Spring 2001 9
Measuring Potential
Ohm's Law E = iR.
E= EMF volts
R = impedance or resistance ohms
i = current amps
For R=10
12
, if the cell EMF is 1.00 volts
then i = 1x10
-12
amps.
For n=1, the application of Faraday's law
(moles reacted = it/nF t in seconds) yields a
rate of reaction of 1.04x10
-17
moles/sec or
about 600000 atoms/sec reacting. This is really
slow.
4
8/15/2006 Chem 208 Spring 2001 10
Example of a Cell
This cell is constructed without a salt bridge. It uses a
porous membrane. The [H
+
] is the same on both sides of
the membrane, hence no concentration potential develops.
Porous
membrane
Ag electrode
with AgCl
Pt
electrode
with H2
bubbler
1.0 M
HCl
V
8/15/2006 Chem 208 Spring 2001 11
Schematic Description of Cell
Pt, H
2
(1 atm.)/ H
+
(a = 1.0)/AgCl/Ag
Anode Cathode
OX RED
LEFT side is the ANODE-OXIDATION
The cell reaction will have as reactants the
REDUCED form of the material in the
LEFT half cell and the OXIDIZED form of
the material in the RIGHT half cell.
8/15/2006 Chem 208 Spring 2001 12
Half Reactions
H
2
==> 2H
+
+ 2 e
-
Oxidation
AgCl + e
-
==> Ag + Cl
-
Reduction
H
2
+2AgCl
==>
2H
+
+ 2 Ag + 2Cl
-
Redox Couple
Conjugate oxidation- reduction pair
E=0.2222 volts at STP
5
8/15/2006 Chem 208 Spring 2001 13
Conventions for the
Description of Electrochemical
Cells.
Pt / Fe
+2
(1.0 M) // Ce
+4
(1.0 M) / Pt
Fe
2+
=====> Fe
3+
+ e
-
and Ce
4+
+ e
-
====> Ce
3+
All concentrations and partial pressures are
listed in parenthesis.
Phase boundaries are shown with a single '/
Gaseous reactants are normally show in the
same phase as the electrode material. eg.
Pt, H
2
(1.0. atm) / H
+
(1.0 M)
8/15/2006 Chem 208 Spring 2001 14
Conventions for the
Description of Electrochemical
Cells.
Every phase boundary has a potential
difference associated with it.
Salt bridge or phase boundaries assumed to
generate no potential is designated by the
double '//'.
These are constructed from a tube filled with a solution of an inert
electrolyte. Normally the tube is closed with glass frits or are made from
an electrolyte in agar to minimize the mixing of the bridge solution with
the cell solutions. In many commercial batteries, porous membranes
(paper or cardboard are often used) are employed in place of a traditional
salt bridge.
8/15/2006 Chem 208 Spring 2001 15
Half Reactions
Cant measure just One half
Need a Standard Reference
Normal hydrogen Electrode
This is made from a Pt electrode in 1 M H
+
in 1
atm H
2
. The Pt electrode must be carefully coated
with Pt black. The potential of this HALF CELL is
defined as exactly 0.00000 volts at STP. The
operation of this electrode is also hazardous due to
the use of hydrogen.
6
8/15/2006 Chem 208 Spring 2001 16
1953 IUPAC Conventions
1. Standard Potential E
o
all reactants and
products at 1.0 M activity.
2. Std.Potential for a 1/2 cell is the cell
EMF of that 1/2 cell coupled with the NHE.
3. The sign of the Std. Potential is the sign
of the 1/2 cell electrode where the NHE is
operating as the ANODE in the reaction.
(Galvanic cells thus have + Std.Potentials
and Electrolytic cells have - Std.Potentials.
8/15/2006 Chem 208 Spring 2001 17
Tables of Std.Potentials
Appendix H- Standard Potentials E
o
Formal Potentials - Ignore Activity E
o
Oxidized form on left will transfer electrons

to the reduced form of anything with lower
potential it on the right (similar to acid base
half reaction table)
8/15/2006 Chem 208 Spring 2001 18
7
8/15/2006 Chem 208 Spring 2001 19
Nerst Equation
Nerst eq. applies equally well to 1/2 RX and
cell RX.
b
B
a
A
d
D
c
C o
a a
a a
n
E E log
05916 . 0
=
8/15/2006 Chem 208 Spring 2001 20
Calculate EMF of CELL R
X
From Cell Schematic (Shorthand Notation)
Fe/Fe
2+
(1.0) // Sn
4+
(1.0), Sn
2+
(1.0) /Pt
Fe + Sn
4+ =====>
Fe
2+
+ Sn
2+
Fe
2+
+ 2e ====> Fe E
o
Fe2+,Fe
= - .447
reduction
Sn
4+
+ 2e ===>Sn
2+
E
o
Sn4+ +,Sn2+
= + .151
More negative potential will be oxidation
reverse reaction
8/15/2006 Chem 208 Spring 2001 21
X
Fe ===> Fe
2+
+ 2e oxidation
E
Fe
,
Fe2+ =
+.447 Oxidation Potential
E
Fe
,
Fe
2+ = +
.447 E
Sn
4+,
Sn
2+ =
+.151
G
cell
= G
Fe,Fe2+
+ G
Sn4+
,
Sn2+
nFE
cell
= nFE
Fe,Fe2+
+ nFE
Sn
4+
,Sn
2+
since nF is the same for all terms it cancels.
E
cell
= E
Fe,Fe2+
+ E
Sn
4+
,Sn
2+
8
8/15/2006 Chem 208 Spring 2001 22
X
E
cell
= E
Fe,Fe2+
+ E
Sn
4+
,Sn
2+
0.447 + 0.151 = 0.598 volts
E
cell
=E
red
+ E
ox
= E
o
Cathode
- E
o
Anode
NOTE E does not depend on how much
we use. (intensive property) 1 mole, 2
moles etc. Measured under conditions of
No Reaction.
G does depend on how much (extensive).
8/15/2006 Chem 208 Spring 2001 23
Calculate E
1/2
using Nerst -
At non-standard conditions
Pt/Fe
2+
(.100), Fe
3+
(.200)//Mn0
4
-(
.010)
,
Mn
2+
(1.0x10
-4
)
,
H
+
(1.0)/Pt
look for 1/2 Rx in table
Fe
3+
====> Fe
2+
+ e
-
E
Fe3+,Fe2+
= .771
E
Fe2+,Fe3+
= -.771
Mn0
4
-
+ 8H
+
+ 5e- ====> Mn
2+
+ 4H
2
0
E
Mn04-,Mn2+
= 1.51
8/15/2006 Chem 208 Spring 2001 24

Calculate E
1/2
using Nerst -
As oxidation
E
Fe
2+,
Fe
3+ =
E
o
- 0.0591 log [Fe
3+
]
1 [Fe
2+
]
E = -.771 - 0.0591 log 0.200 = -0.771 -.0178
1 0.100
= -0.789 volts
9
8/15/2006 Chem 208 Spring 2001 25
Calculate E
1/2
using Nerst -
E
Mn04 ,Mn2+
= 1.51 - .0591 log [Mn
2+
]H
2
0
5 [Mn0
4
-
][H
+
]
8
= 1.51 -.0591 log 1.0 x 10
-4
5 0.010x(1.0)
8
potential very sensitive to [H
+
]
= 1.51 + .026 = 1.536
8/15/2006 Chem 208 Spring 2001 26
Calculate E
1/2
using Nerst -
Balanced RX
5Fe
2+
+ Mn0
4-
+ 8H
+
+ 5e
-
5Fe
3+
+ Mn
2
+
4 H
2
0 + 5e
-
E
coll
= 1.536 + (-.789) = + 0.747
Note Potential of 5 Fe
2+
= 5Fe
3+
same
as for 1
8/15/2006 Chem 208 Spring 2001 27
Methods of Analysis
Direct Potentiometry
Nerst Eq. activity measurements
Calibration Curve conc. Measurements
Standard Addition
Potentiometric Titration
10
8/15/2006 Chem 208 Spring 2001 28
Reference Electrodes
Saturated Calomel electrode SCE
Hg,Hg
2
Cl
2
/ Hg
2
+2
(sat.), KCl (sat.)// E =
+0.2415 volts
Ag/AgCl reference electrode is also
popular. 0.2222 v
8/15/2006 Chem 208 Spring 2001 29
Junction Potentials
Can be a major problem 1-3 mV
Both positive and negative ions must be
equally mobile through junction.
Leaky electrodes - allow free passage of
solution
Salt Bridge -
K
+
and Cl
-
ions have same size
8/15/2006 Chem 208 Spring 2001 30
Errors
Error due to 1 mv inaccuracy =
4% rel conc. n = 1
8 % n = 2
etc.
Accuracy of direct potentiometry limited by
junction pot. Typical salt bridge can have up to 2-
3 mV.
Not a problem in titrations and std. add
calibration curves can be a problem - matrix
11
8/15/2006 Chem 208 Spring 2001 31
Indicator Electrodes
Electrodes of the 1st kind (Probe the
primary reactant metal)
Metal electrodes-
don't work very well
Contamination
Interferences
Rarely used for Direct Analysis
8/15/2006 Chem 208 Spring 2001 32
Indicator Electrodes
Electrodes of the 2nd kind (probe a
secondary reactant)
Ag/AgCl - potentiometric titration of Cl
Quinone/hydroquinone pH electrode
8/15/2006 Chem 208 Spring 2001 33
MEMBRANE INDICATOR
ELECTRODES
Glass electrode (pH) is good example.
If only one type of charged ion can move
E
mem
= - RT ln a
2
nF a
1
activity. of solutions on two sides of
membrane
12
8/15/2006 Chem 208 Spring 2001 34
MEMBRANE INDICATOR
ELECTRODES
If a
1
is a constant
E
mem
= K - RT ln a
2
nF
Glass membrane
Si0
2
Li
2
0 or Na
2
0 Ca0
8/15/2006 Chem 208 Spring 2001 35
Glass Membrane Electrode
Need
solutions for
electrical
contact with
membrane
Dry Glass
Hydrated gel
External
Solution
Internal
Solution
8/15/2006 Chem 208 Spring 2001 36
pH electrode
Volt
meter
Reference
Solution
External
Electrode
Internal Solution
and Electrode
Porous J unction
Total Potential
includes potential
of electrode on
both sides of the
membrane.
pH
unk
= pH
s
+ E
unk
-
E
s
.0591
13
8/15/2006 Chem 208 Spring 2001 37
Glass Electrodes
Glass electrodes with Na
2
0 Al
2
0
3
B
2
0
3
respond to Na
+
, Li
+
, & K
+
to some extent.
Useful as selective electrodes for these ions
(also respond to H
+
so [H
+
] must be kept
low)
8/15/2006 Chem 208 Spring 2001 38
Semiconductor pH electrodes
FET - Converts potential to current
Instrument must supply voltage
Cover semiconductor material with
something to generate chemically related
potential
Next generation of electrodes
8/15/2006 Chem 208 Spring 2001 39
Ion SELECTIVE Electrodes
If electrode responds to several different
ions (most electrodes do)
relative response described by selectivity
coefficient k
x,y
relative response to ion x in presence of ion y
y
n
n
y x
x
y
s
Y k X
n
E ] [ ] [ log
05916 . 0
constant
,
14
8/15/2006 Chem 208 Spring 2001 40
Ion SELECTIVE Electrodes
is measure of how closely the slope
follows the Nerst type behavior (Nerstian).
Should be approx = 1
Use sign on n - Positive ions give positive
slope, negative ions give negative slope.
y
n
n
y x
x
y
s
Y k X
n
E ] [ ] [ log
05916 . 0
constant
,
8/15/2006 Chem 208 Spring 2001 41

Example of Selectivity
Na
+
glass electrode 11% Na
2
0 18% Al
2
0
3
71%
Si0
2
Specific example for sodium electrode and
potassium interference
K
K/Na
= 2800
Na
+
response is = 2800 x K
+
response
K
K/Na
= 1/K
K/Na
= 1/2800 = 3.57x10
-4
8/15/2006 Chem 208 Spring 2001 42
Example of Selectivity
For interference < 1% of concentration
[X] > 0.01 * K
x,y
[Y]
[K
+
] would have to be 2800 x[Na
+
] to give
an equal potential
1% error if K
+
= 28 x Na
+
useful to establish working conditions
15
8/15/2006 Chem 208 Spring 2001 43
Liquid Ion Exchange
Membranes
Immiscible organic phase interposed
between internal reference solution and
external solution.
Organic phase contains some substance
which will transport one ion preferentially
across membrane. Porous to one type of ion.
8/15/2006 Chem 208 Spring 2001 44
Liquid Ion Exchange
Membranes
Ca didecylphosphate neutral (org. soluble)
overall E = Q + .0591 log [ Ca
2+
]
2
Note N = 2 sensitivity only 1/2 that for N = 1
8/15/2006 Chem 208 Spring 2001 45
Liquid Ion Exchange
Membranes
Other Liquid exchanges
Macrocyclics
penicillin
Aureomycin
Valinomycin
Crown Ethers K
+
16 crown 5
Alkyl phosphates Ca
2+
Bidentate ligands (transition metals)
O
O
O
O
O
16
8/15/2006 Chem 208 Spring 2001 46
Liquid Ion Exchange
Membranes
Ion Pair Formation
Tetraalkyl ammonium salts
Alkyl amines
1,10 phenanthroline Fe (III) salts
8/15/2006 Chem 208 Spring 2001 47
Liquid Ion Exchange
Membranes
Liquid exchangers somewhat limited
Generally work only down to 10
-3
or 10
-4
M
Limited by solubility of exchanger and of
organic solvent in water.
Slow response - depends on viscosity and
diffusion rate
2-20 micron capillary
measurements in
single living cells
Organic ion
Exchanger
KCl
filling
solution
Tungsten wire
Treated Tip
8/15/2006 Chem 208 Spring 2001 48
Solid State Membranes
LaF
3
single crystal (doped with Eu)
very conductive - F
-
ions mobile
Linear potential from 1 M to 1x10
-5 (
1x10
-7)
0H
-
interferes pH must not be too high.
But no response to HF
HF <===> H
+
+ F
-
K
A
= 6.8x10
-4
pH must be controlled (5.5 typical)
17
8/15/2006 Chem 208 Spring 2001 49
also F
-
+ M
n+
MF metal complexes
add NTA or EDTA to free F
-
Other Solid State Electrodes
AgS + AgCl AgBr AgI AgSCN
Rapid and selective (Ag
+
, S
2-
, Cl
-
.)
8/15/2006 Chem 208 Spring 2001 50
Linear response limited by Ksp (solubility
of exchange crystal)
10
-10
10
-12
M solutions often give good
values
lower solubility ===>lower limit of
linearity
I<Br<Cl
8/15/2006 Chem 208 Spring 2001 51
Shape of
curve
Slope=0.0592/z
Solubility limit
Log_Concentration
E
18
8/15/2006 Chem 208 Spring 2001 52
Heterogenous membranes
Silicone polymer + precipitate of some
insoluble salt.
PVC
Coated wire electrodes
Tungsten wire - treated to become pH
electrode
8/15/2006 Chem 208 Spring 2001 53
Gas Sensing Electrodes
Gas permeable membrane
Ammonia Electrode
NH
3
+ H
+
<===> NH
4+
pH electrode or NH
4
+ electrode
Also available for CO
2
, S0
2,
H
2
S & N0
2
8/15/2006 Chem 208 Spring 2001 54
Enzyme Electrodes
pH Electrode
Enzyme in
acrylamide gel
ISE, pH, or gas electrode
Urease <===>NH
3
Decarboxylazes===>H
+
2 enzyme sandwich -
cholesterol electrode
19
8/15/2006 Chem 208 Spring 2001 55
Enzyme Electrodes
Nerve gas electrode (& phosphate
pesticides) - Cholinesterase inhibitors
enzyme film
add substrate to sample
monitor reduced activity
Vitamin D electrode etc.
Nick Schmit with Gary Rechnitz
8/15/2006 Chem 208 Spring 2001 56
State Of The Art
Electrochemical sensors are often fabricated
like integrated circuits with multiple sensors
in a very small space.
Field Effect Transistors and other semi-
conductor circuits (Metal Oxide
semiconductors) now used as sensors.
Mechanical nose spoiled food, ripe fruit
mine detection
8/15/2006 Chem 208 Spring 2001 57
Problems with ISE's
Slow response (minutes) for Liquid
Junction
Short lifeISE's months
Enzymes days or hours
High resistance - good pH meters required
20
8/15/2006 Chem 208 Spring 2001 58
ISE's
Calibration Curves
Standards and samples of similar ionic strength
Keep E
junct
similar
Interfering ions minimized
N=1 59 mV/log C
N=2 28 mV/log C
N=3 20 mV/log C
8/15/2006 Chem 208 Spring 2001 59
ISE's
Standard Addition
Measure sample
E = Q - .0591 log C
x
n
Measure sample + Std.
E = Q - .0591 log (C
x
+C
s
)
n
Assumes linear portion of response.
8/15/2006 Chem 208 Spring 2001 60
ISE's
If addition causes little volume change E
j
will stay the same
Std. allows compensation for unknown
matrix
eg. ionic strength and E
j
effects minimized
E
x
= Q + .0591 log C
x
n
E
s
= Q + .0591 log
V
s
C
s+
V
x
C
x
n V
s
+V
x
21
8/15/2006 Chem 208 Spring 2001 61
ISE's
Take inverse log
rearrange
x s
x x s s
x
s x
V V
C V C V
C
E E n
+
+
=
log log
05916 . 0
) (
10
n(E
x
E
s
)
0.0592
=
C
x
( V
s
+V
s
)
V
s
C
s
+V
x
C
x
C
x
=
C
s
V
s
(V
x
+ V
s
) 10
n(E
x
E
s
)/ 0.0592
V
x
8/15/2006 Chem208 Spring 2001 62
ISE's
Graphical treatment of std. addition
Log C on X axis
Put C
x
at 0
Plot Standards
Extrapolate to
Y=0
-Cx Unk. Unk+S1 Unk+S2
Potential
8/15/2006 Chem208 Spring 2001 63
ISE's
Three approaches to Std. Add.
1. Add micro amounts of standard and ignore
dilution.
2. Add standard and correct for dilution.
3. Dilute unknown and standard additions to
constant volume.
Std. Add. Eliminates Many Matrix Effects
Multiple Std. Additions verify linearity
22
8/15/2006 Chem208 Spring 2001 64
ISE's
Potentiometric Titration
Very precise and accurate
but not applicable at such low conc. as direct
potentiometry
1st and 2nd Derivatives used for end point
determination.
8/15/2006 Chem208 Spring 2001 65
Clinical Analyzers
Electrochem on a Chip
Disposable
ISEs
8/15/2006 Chem208 Spring 2001 66
Resurgence of ISEs
New Techniques
Ppb level analysis
Pb in drinking water < 5 ppb
1
8/15/2006 Chem 208 Spring 2006 1
Chem 208
Chapter 16
Redox Titrations
8/15/2006 Chem 208 Spring 2006 2
Oxidation States
Free elements always 0
Monoatomic ions = charge on ion
Group 1 elements always +1
O always -2 except when bonding to other O
(HOOH O=-1)
H always +1 except when bound to metal
F always -1
Group 1 metals always +1
Sum of all Ox states = charge of ion or
molecule
8/15/2006 Chem 208 Spring 2006 3
Examples
H
2
SO
4
HClO
4
H
3
PO
4
Na
2
S
2
O
3
2
8/15/2006 Chem 208 Spring 2006 4
REVIEW BALANCING
EQUATIONS (Z 164-178)
1. Break overall reaction into OX and RED half reactions
2. Balance all elements other than O and H
3. Balance O by adding H
2
O
4. Balance H by adding H
+
5. Balance charge by adding e
-
6. If Basic, add OH
-
to both sides to remove all H
+
7. Multiply each half reaction by a factor to make e
-
the
same in both.
8. Add half reactions to get overall reaction
9. Cancel everything that appears on both sides to simplify.
8/15/2006 Chem 208 Spring 2006 5
Example
MnO
4
-
+ H
2
C
2
O
4
==> Mn
2+
+ CO
2(g)
Half reactions
MnO
4
-
==> Mn
2+
H
2
C
2
O
4
==> CO
2(g)
8/15/2006 Chem 208 Spring 2006 6
Example
MnO
4
-
==> Mn
2+
Mn balanced
Balance O with H
2
O
MnO
4
-
==> Mn
2+
+ 4 H
2
O
Balance H with H
+
MnO
4
-
+ 8H
+
==> Mn
2+
+ 4 H
2
O
Balance charge with electrons
MnO
4
-
+ 8H
+
+ 5e
-
==> Mn
2+
+ 4 H
2
O
Done
3
8/15/2006 Chem 208 Spring 2006 7
Example
H
2
C
2
O
4
==> CO
2(g)
Balance Cs
H
2
C
2
O
4
==> 2 CO
2(g)
O Balanced
Balance H with H
+
H
2
C
2
O
4
==> 2 CO
2(g)
+ 2H
+
Balance charge with electrons
H
2
C
2
O
4
==> 2 CO
2(g)
+ 2H
+
+ 2e
-
Done
8/15/2006 Chem 208 Spring 2006 8
Example
Add two half reactions
electrons must cancel
5x(H
2
C
2
O
4
==> 2 CO
2(g)
+ 2H
+
+ 2e
-
)
2x(MnO
4
-
+ 8H
+
+ 5e
-
==> Mn
2+
+ 4 H
2
O)
5H
2
C
2
O
4
==> 10 CO
2(g)
+ 10H
+
+ 10e
-
2MnO
4
-
+ 16H
+
+ 10e
-
==>2Mn
2+
+ 8 H
2
O
2MnO
4
-
+ 5H
2
C
2
O
4
+ 16H
+
+ 10e
-
==>2Mn
2+
+
8 H
2
O + 10 CO
2(g)
+ 10H
+
+ 10e
-
Clean up- eliminate common terms
2MnO
4
-
+ 5H
2
C
2
O
4
+ 6H
+
==>2Mn
2+
+ 8 H
2
O +
10 CO
2(g)
+
8/15/2006 Chem 208 Spring 2006 9
In base solution
Follow rules for acid solution
Add OH
-
to both sides to neutralize H
+
On side with H
+
combine with OH
-
to make
H
2
O
4
8/15/2006 Chem 208 Spring 2006 10
Last Example
I
-
+ IO
3
-
==> I
3
-
Half reactions
I
-
==> I
3
-
IO
3
-
==> I
3
-
Only one product gets used in both
Same is possible when one reactant goes to
two products -
disproportionation
8/15/2006 Chem 208 Spring 2006 11
Last Example
3 I
-
==> I
3
-
+ 2e
-
IO
3
-
==> I
3
-
+3 H
2
O
IO
3
-
+ 6H
+
+ 4e
-
==> I
3
-
+3 H
2
O
6 I
-
==> 2 I
3
-
+ 4e
-
( x 2)
Add
6 I
-
+ IO
3
-
+ 6H
+
+ 4e
-
==> 2 I
3
-
+ I
3
-
+3 H
2
O
+4e
-
6 I
-
+ IO
3
-
+ 6H
+
==> 3 I
3
-
+3 H
2
O
8/15/2006 Chem 208 Spring 2006 12
Oxidizing Agents - Titration
KMnO
4
- titrant for Fe, Sn, and Oxalate
Ce
4+
- Very stable in strong sulfuric acid,
Titrations of Fe and organics
K
2
Cr
2
O
7
- little used now due to hazards
and waste disposal of Cr
I
2
- add KI to increase solubility - Vit C,
wide range of applications
Karl Fischer Detn of H
2
O
5
8/15/2006 Chem 208 Spring 2006 13
Reducing Agents - Titration
Not too many of these
Fe
2+
not very stable Fe(NH
3
)
2
(SO
4
)
2
Thiosulfate - S
2
O
3
2-
===> S
4
O
6
2-
+ 2e
-
Iodine in some cases
Many common REDOX procedures use a reducing agent
to reduce the sample and then use an oxidizing agent titrant
for analysis rather than a reducing agent titrant.
reduce Fe
3+
to Fe
2+
(Zn reductor) then titrate with Ce.
8/15/2006 Chem 208 Spring 2006 14
Titration Curves
Classic Example
Fe
2+
+ Ce
4+
====> Fe
3+
+ Ce
31
E
o
cell
= 2.12 volts
K
eq
= 7 x 10
35
very favorable reaction
Titration should be possible even at low
concentrations. (Kinetically limited)
At any point in titration - reaction proceeds
until E
Fe
= E
Ce
8/15/2006 Chem 208 Spring 2006 15
Titration Curves
Initial Potential
(Fe
3+
) = 0
E = -
Therefore there will always be traces of
Fe
2+
=
+
+
] [
] [
log 0592 . 0
3
2
Fe
Fe
E E
o
6
8/15/2006 Chem 208 Spring 2006 16
Titration Curves
After First Addition of Titrant
Both
E = .68 - .0592 log [Fe2+]
[Fe3+]
and
E = 1.44 - .0592 log [Ce3+]
[Ce4+]
must hold.
8/15/2006 Chem 208 Spring 2006 17
Titration Curves
[Fe2+] = C
o
V
o
-C
T
V
T
V
o
+V
T
[Fe
3+
] = C
t
V
T
V
o
+V
T
V
o
+ V
T
cancels in Ratio
E = .68 -.0592 log C

o
V
o
-C
t
V
T
C
T
V
T
8/15/2006 Chem 208 Spring 2006 18
Titration Curves
E = .68 -.0592 log C
o
V
o
-C
t
V
T
C
T
V
T
ratio moles remaining = fraction titrated
moles titrated
[Ce
3+
] = C
T
V
T
[Ce
4+
] = ?
V
o
+V
T
solve K
eq
expression or Nerst Eq.
7
8/15/2006 Chem 208 Spring 2006 19
Titration Points
10% titrated
E = .68 - .0592 log 100-10 = .68 - .0591 log 9
10
E - .624
50% E = .68 - .0592 log 100-50
50
E =.68 Special point E
50%
= E
o
90% E = .68 - .0592 log 100-90= .736
90
8/15/2006 Chem 208 Spring 2006 20

Titration Points
At Equivalence Point
E
ep
= .68 - .0592 log [Fe
2+
]
[Fe
3+
]
E
ep
= 1.44 - .0592 log [Ce
3+
]
[Ce
4+
]
2 Eep = .68 + 1.44 - .0592 log [Ce

3+
] - 0.0592 log [Fe
2+
]
[Ce4
+
] [Fe
3+
]
E
ep
= 2.12 - .0592 log [Ce
3+
][Fe
2+
]
2 2 [Ce
4+
][Fe
3+
]
This is not the Nerst Equation!!
8/15/2006 Chem 208 Spring 2006 21

Titration Points
At Equivalence Point
E
ep
= 2.12 - .0592 log [Ce
3+
][Fe
2+
]
2 2 [Ce
4+
][Fe
3+
]
[Ce3+] = [Fe3+] and [Fe2+] = [Ce4+]
Eep = 2.12/2 = 1.06
8
8/15/2006 Chem 208 Spring 2006 22
Titration Points
After the Eq. Point E = E
Ce
E = 1.44 - 0.0592 log [Ce3+]
[Ce4+]
[Ce
3+
] = C
o
V
o
[Ce
4+
] = C
T
V
T
- C
o
V
o
V
o
+V
T
V
o
+ V
T
Again volumes cancel.
E = 1.44 - .0592 log CoVo
C
T
V
T
- C
o
V
o
8/15/2006 Chem 208 Spring 2006 23
Titration Points
After the Eq. Point E = E
Ce
Since C
o
V
o
= 100% of sample
let X = % titrated
E = 1.44 - .0592 log l00
X-100
8/15/2006 Chem 208 Spring 2006 24
Titration Points
110% titrated
E = 1.44 - .0592 log 100 = 1.44 - .0592 log 10
110-100
E - 1.381
200%
E = 1.44 - .0592 log 100
200-100 log 1 = 0
E = 1.44
9
8/15/2006 Chem 208 Spring 2006 25
SUMMARY OF REDOX
TITRATION CURVE
Titration begins beyond E
o
for reactant
50% titrated E = E
o
for reactant
200% titrated E = E
o
2
Titration curve INDEPENDENT OF
CONC.
But Kinetics are problem at low conc.
E
ep
=
n
1
E
o
1
+ n
2
E
o
2
n
1
+ n
2
8/15/2006 Chem 208 Spring 2006 26
Hydrogen ion dependence
Fe
2+
+ Cr
2
0
7
====> Fe
3+
+ Cr
3+
Fe
3+
+ e
- ====>
Fe
2+
E
o
= 0.68 V
Cr
2
0
7
-
+ 14H
+
+ 6e
-
====>2Cr
3+
+ 7H
2
0
E
o
= 1.33 V
6 Fe
2+
+ Cr
2
0
7
-2
+ 14H
+
==> 6 Fe
3+
+ 2Cr
3+
+ 7H
2
0
8/15/2006 Chem 208 Spring 2006 27
6 Fe
2+
+ Cr
2
0
7
-2
+ 14H
+
==> 6 Fe
3+
+ 2Cr
3+
+ 7H
2
0
At E eq pt
E
ep
= E
Fe
- .0592 log [Fe
2+
]
1 [Fe
3+
]
E
ep
= E
Cr
- .0592 log [Cr
3+
]
2
6 [Cr
2
0
7
][H
+
]
14
multiply x6 to clear
10
8/15/2006 Chem 208 Spring 2006 28
6 E
ep
= 6 E
o
Cr207
- 6 x .0592 log [Cr
3+
]
2
6 [Cr
2
0
7
][H
+
]
14
7 E
ep
= E
o
Fe
+ 6 E
o
Cr207
- 6 .0592 log [Fe
2+
] [Cr
3+
]
2
6 [Cr
2
0
7
][H
+
]
14
[Fe
3+
]
[Fe
2+
] = 6 [Cr
2
0
7
-2
] unreacted
[Fe
3+
] = 3 [Cr
3+
] 6:2 = 3:1
[Cr
2
0
7
] = X [Fe
3+
] = 3Y
[Fe
2+
] = 6X [Cr
3+
] = Y
8/15/2006 Chem 208 Spring 2006 29
E
ep
= E
Fe
+
6E
Cr 20 7
- .0592 log 6x Y
2
7 7 X3Y [H
+
]
14
E
ep
= .68 + 6 x 1.33 - .0592 log 2[Cr
3+
]
7 7 [H
+
]
14
= 1.24 - .0592 log 2 [Cr

3+
] + .0592 log [H
+
]
14
7 7
8/15/2006 Chem 208 Spring 2006 30
Multicomponent systems
E
o
differ by at least 0.20 v
Titration of Mixtures
Titrations with multiple Ox states
V
2+
===> V
3+
-0.255 V
V
3+
===> VO
2+
+0.337 V
VO
2+
===> VO
2
+
+1.001 V
Three break titration curve
11
8/15/2006 Chem 208 Spring 2006 31
End Point Detection
Potentiometric
Pt wire (inert) electrode vs Ref. Electrode
Responds to potential of Redox couples
Colored samples
Redox Indicators
Fe phenanthroline - reversible Redox
e
-
withdraw or denote shift E
o
I
8/15/2006 Chem 208 Spring 2006 32
Iodine - Thiosulfate
Sodium thiosulfate titrant (Na
2
S
2
O
3
)
Standardize against Primary Std. KIO
3
React Cu wire to form Cu
2+
, add excess I
-
Reduces to CuI and I
3
-
Titrate iodine released with thiosulfate
Add excess I
3
-
or IO
3
-
as oxidizing agents
Titrate unreacted oxidant
Vitamin C analysis
8/15/2006 Chem 208 Spring 2006 33
Karl Fischer titration
Analysis of Water
Use Redox which involves water
Set up so water is limiting reagent
SO
2
+ I
2
+ H
2
O ==> SO
3
+ 2H
+
+ 2I
-
Solvent is alcohol
Base is added (pyridine, imidazole,
dimethanolamine)
12
8/15/2006 Chem 208 Spring 2006 34
Indicators in Iodine systems
Color of I
3
-
dark brown can be used
Starch indicator - clear to black
8/15/2006 Chem 208 Spring 2006 35
Electrochemical Generation
Titrants made by Electrode Reaction
Apply volatge
Electrolytic cell
I
2
, Br
2
work very well
Ag
+
for halide titrations
Electrogravimetry (ugh)
Coulometric Titration - measure current and
time
1
8/15/2006 Chem208Spring2001 1
Chem 208
Chapter 10
Analysis by Absorption of
Light
Spectrometry
8/15/2006 Chem208Spring2001 2
Electromagnetic Radiation
Electronic Vector
Magnetic Vector at 90 deg
Characteristics
Wavelength () crest to crest distance
10
-9
NM ==>100 Km
Speed (c) speed of light
vacuum 3.00 x 10
8
m/sec
slower in all other media - slowed by electronic
interactions
8/15/2006 Chem208Spring2001 3
=Refractive index = speed in vac
speed in medium
>1.00
function of wavelength
light of different energy moves at different rates
in mediumother than vacuum.
Fiber Optics separate wavelengths of a pulse
2
8/15/2006 Chem208Spring2001 4
(wave number) =waves/cm=cm
-1
= 1/
in vacuum
Frequency ( ) =waves passing fixed
point/sec.
period (p) time per wave crest =1/p
all are related =c

E =h =hc/ =hc
8/15/2006 Chem208Spring2001 5
Electromagnetic spectrum
8/15/2006 Chem208Spring2001 6
Interaction of matter and
Radiation can interact with matter in a wide
variety of ways.
Most interaction depends on size of particle
relative to wavelength.
Particles <1/2 are transparent
AM radio around buildings
KBr pellets- particles become transparent to IR
3
8/15/2006 Chem208Spring2001 7
Electronic vector Interactions
X-rays- ionize atoms eject inner shell e
-
UV-VIS - energy charges in valence e
-
IR - molecular vibrations
microwave - molecular rotations
Electronic Transition
Vibrational Transition
Rotational Transition
8/15/2006 Chem208Spring2001 8
Identification of Compounds
Compare Actual spectrum to Library
IR and NMR most specific for organics
UV-Vis not very specific
Broad bands for solutions due to collisional
broadening
8/15/2006 Chem208Spring2001 9
Magnetic vector Interactions
( in magnetic field)
microwave - electron spin changes
radio wave - nuclear spin changes
4
8/15/2006 Chem208Spring2001 10
Energy Levels Diagrams
_____3s ____ 3p ____ 3d

E ____ 2s ____ 2p
____1s
Emission occurs when e
-
state drops.
Studied for atoms. Chemland
Absorption occurs when photon of exactly the
energy of the transition strikes the atoms.
Studied for molecules (hard to exited enough to
emit.)
8/15/2006 Chem208Spring2001 11
Molecular Energy Levels
orbitals
Bonding and Antibonding orbitals
CAChe and Spartan calculate these
8/15/2006 Chem208Spring2001 12
Molecular Energy Levels
Computation chemistry
Compute energy of all MOs
Predict UV-Vis spectra
Predict IR Spectra
Predict NMR Spectra
5
8/15/2006 Chem208Spring2001 13
Instrumentation for Optical
Spectroscopy
Detector
Source of
monochromatic
radiation
Sample Readout
8/15/2006 Chem208Spring2001 14
Sources of Radiation
Black body radiators -
Tungsten lamp 2870
o
K - 1.5 micron peak
visible only - 330 nmminimum
Quartz - Iodide - 3600
o
K more UV & VIS
Discharge Lamps
H
2
or D
2
165 to 360 Nm.
D
2
lasts longer and brighter than H
2
.
8/15/2006 Chem208Spring2001 15
Monochromators
Filters
Glass 30-50 nmband width 5-20% T at max
Interference Filters 10-20 nmbandwidth 40% T
Prisms - Quartz for UV-VIS
Gratings - parallel lines on glass
Most common dispersion device
Fourier Transform instruments - Non-
dispersive (IRs) - Details in Chem312
6
8/15/2006 Chem208Spring2001 16
Dispersion by a Grating
8/15/2006 Chem208Spring2001 17
Illumination of Monochromator Exit
Slit by Radiation
2
8/15/2006 Chem208Spring2001 18
Definition of effective bandwidth
Wavelength Selectors
7
8/15/2006 Chem208Spring2001 19
Cells
Glass or Plastic - Vis only
Quartz - UV-VIS-NIR $60-100 each
Flat parallel windows best
Cylindrical cells must always be in the same
position
(mark on spec 20 cells)
Flow Cells
Fiber Optic Probes
8/15/2006 Chem208Spring2001 20
Solvents - must be transparent
UV cutoff
Solvent UV Cutoff
Acetone 330
Acetonitrile 210
Benzene 280
Carbon disulfide 380
Chloroform 245
Dichloromethane 233
Ether 220
Ethyl Acetate 260
Hexane 210
Methanol 210
Water 200
8/15/2006 Chem208Spring2001 21
Detectors
Eye - Colorimetry
No numerical Readout
Most sensitive to Green
Photovoltaic Cells
No power supply needed
Portable
Response like eye
8
8/15/2006 Chem208Spring2001 22
Detectors
Photo-tube
Photo emissive surface
Work function -photon energy needed to eject
e
-'
s
photo cathodes designed for various regions of
the spectrum.
each photon produces 1 or more e
-
some thermal e
-
also produced
shot noise
dark current function of temp.
A
8/15/2006 Chem208Spring2001 23
Detectors
Solid State Detectors
Photodiodes
Change Couple Devices.
Photodiodes - pnjunction conduct in
reverse direction due to photon flux.
8/15/2006 Chem208Spring2001 24
Detectors
Linear photodiode Arrays
512 diodes - detect 512 wavelengths at once -
complete spectrumnot scanned. HP
Spectrometer
Good visibility sensitivity
Rapid response
high linearity
.
9
8/15/2006 Chem208Spring2001 25
Instrumentation for Optical
Spectroscopy
Spectrometer - Record light through sample at
given wavelength.
Spectronic20, ST 320
Spectrophotometer - Ratioof 2 beams in space
PE 330
Spectrophotometer - Ratio of 2 beams in time HP
Diode Array Ocean Optics spectrometer
Detector
Source of
monochromatic
radiation
Sample Readout
8/15/2006 Chem208Spring2001 26
Instrument Designs
Single Beam
Dual Beam-In-Space
Dual Beam-In-Time
8/15/2006 Chem208Spring2001 27
Instrument Designs
Photodiode array spectrophotometer
10
8/15/2006 Chem208Spring2001 28
Quantitative aspects of
Absorption
Beer's Law Beer-Lambert Law
(Harry Gray's version)
The taller the glass, the darker the brew, the less
amount of light comes through.
Absorption depends on probability of a
photon striking and being absorbed by a
molecule.
8/15/2006 Chem208Spring2001 29
Absorption
if each thin segment (dx) absorbs some
fraction of all of incident light (P
x
)
dP =-k P
x
C dx
dx
P(x)
P(o)
P
8/15/2006 Chem208Spring2001 30
Absorption
dP =-k P
x
C dx
k=constant for absorbing species
cross section areaof absorbing region of molecule(0rbital or
conjugated system
Probability of transition occurring (0 => 1 )
P
X
=Incident radiation power
C =Concentration
-dP/P
x
=kC dx
Integrate over entire path length x=0 to b
11
8/15/2006 Chem208Spring2001 31
Absorption
log(P
0
/P) =k/2.303 Cb = bC or (abC)
P = Transmittance (T)
P
o
-log T =A Absorbance - A is dimensionless
A =abc C in gm/l
A= bc C in moles/l
bC =cm*mol/1000 cm
3
=mol/1000 cm
2
a units cm
2
/gm unit =cm
2
/mol
(old literature often dm
2
/gm)
8/15/2006 Chem208Spring2001 32
Limitations on Beers Law
Light must be monochromatic
Parallel
Enter at a right angle.
8/15/2006 Chem208Spring2001 33
Extensions to Beers Law
Multi-component Systems
A
1
=
1
bC
1
+
2
bC
2
+
3
bC
3
+.....
Total abs. =sumof absorbencies of individual
absorbing species.
Measure at several wavelengths solve
simultaneous equations.
Calc. conc. of all species.
12
8/15/2006 Chem208Spring2001 34
Accuracy
Deviations from Beers Law
Instrumental Deviations
Non-monochromatic light
value of or a not constant across bandwidth of
spectrometer.
Negative deviation at high conc.
Concentration error
lower sensitivity.
Need more standards.
A
Conc.
8/15/2006 Chem208Spring2001 35
Wide slits give lower A values -
Value measured on ST320 (7nm band) or
Spec 20 (20 nm band) will be less than for
HP diode array (1 nm band) which may be
less than PE 330/Hitachi (0.1-10 nm band)
8/15/2006 Chem208Spring2001 36
Bandwidth effects
13
8/15/2006 Chem208Spring2001 37
Bandwidth effects
8/15/2006 Chem208Spring2001 38
Chemical Deviations
Equilibria - acid base pH control
Activity coef.
Temperature
Solvent effects - concentration changes
dielectric constant of solution
Refractive Index change due to Conc.
8/15/2006 Chem208Spring2001 39
Diagnostic Tool for Deviations
Plot A vspath length.
Beers Law - straight line (deviations must
be chemical)
Stray Light - negative deviations
Reflections inside instrument
Higher orders froma grating
slit diffraction around entrance slit
14
8/15/2006 Chem208Spring2001 40
Concentration Errors- Precision
Assume error is a constant value of
Transmittance (T)
A=abC A=-log T
C =- log(T)/ab
take derivative of C with respect to T
dC/dT =-0.4343/T(ab)
8/15/2006 Chem208Spring2001 41
Want relative concentration error dC/C so
divide by
C=-log(T)/ab
abterm cancels
dC/C has a minimum at T=0.368 (36.8%)
dC/ C
dT
=
0.4343
T log(T)
dC
C
=
dT
T
x
0.4343
log(T)

8/15/2006 Chem208Spring2001 42
Error as f(%T)
Working Range
very short
factor of 5-10
%T
error
36.8%T
15
8/15/2006 Chem208Spring2001 43
Applications of
Spectrophotometry
Direct determination of a chromophoric
compound
anything that absorbs strongly.
Absorptivities range from0 to 500,000 , wide
range of sensitivities.
WORST
1. use tabulated absorbance
2 measure absorbance froma single standard
BEST 3. prepare calibration curve
8/15/2006 Chem208Spring2001 44
Applications of
Spectrophotometry
Forma chromophore with non-absorbing species
metals react with ligands to formcolored complexes -
large number of analytical methods developed to use
this
organic derivatives - 2,4-dinitrophenyl hydrozones
azocoupling- make azodye -acid rain nitrate
vanillateion in lab
breath-a-lizer alcohol detn.
8/15/2006 Chem208Spring2001 45
Direct Use of Beers Law
Least Precise and Accurate (one point
calibration) assumes blank=0.00
Known concentrations vsAbs. Least Squares
Intercept need not be Zero
Identify non-linearity if present
16
8/15/2006 Chem208Spring2001 46
Prepare solutions by adding known amounts of
analyte to the unknown
one or more different additions
For one addition
unk unk std
unk
std unk
std std
unk
V V V
A
A
V C
C
+
=
+
) (
8/15/2006 Chem208Spring2001 47
Standard Addition
Useful if matrix of sample has background
absorbance which cannot be accounted for
in a blank or calibration curve
Quick if only one or two samples are to be
run
8/15/2006 Chem208Spring2001 48
Spectrophotometric
Titration
A-Titrant only absorbs
.B-Product of Reaction
absorbs
C- Sample only absorbs
D-Two successive
absorbing species are
formed eg. ML then ML
2
.E- Colored analyte is
converted to colorless
product by colored titrant.
(brominate a red dye)
F- Similar to D but second form
absorbs less
17
8/15/2006 Chem208Spring2001 49
Spectrophotometry to study
reaction stoichiometry
Used for - metal complexes, enzyme
substrate complexes, etc.
J obs Method - Continuous variation
Use where ratio is close to 1:1
Make series of solutions where Total moles of
two reactants constant.
Plot Mole ratio vsA
8/15/2006 Chem208Spring2001 50
Jobs Method
Mole
F ti
0 0.5 1.0
Abs.
1:1
1:2
8/15/2006 Chem208Spring2001 51
Mole-Ratio method
Use where ratio is large
Like a titration. Treat constant molar conc.
of metal with varying molar amounts of
ligand. (Plot A vsMoles ligand/moles
metal)
0 1 2 3 4
Moles L/moles M
Abs.
18
8/15/2006 Chem208Spring2001 52
Slope Ratio Method
Use where binding is weak and large
excesses of reagent are required to force
complete reaction.
xM +yL =====> M
x
L
y
8/15/2006 Chem208Spring2001 53
Slope Ratio Method
Add small amounts of metal to large excess of
ligand - drives reaction to completion even if K
f
is
small. Measure slope of graph of Abs vsconc.
Slope =eb/x
Add small amounts of ligand with large constant
excess of metal - excess metal drives reaction to
completion.
Slope =eb/y
(eb/x)/(eb/y) =y/x
1
8/15/2006 Chem208Spring2001 1
Chem 208
Separations
General Chromatography
8/15/2006 Chem208Spring2001 2
SEPARATIONS AND
EXTRACTION
Separation necessary when measurement
technique not adequately selective
Size Separation -
Filtration
Dialysis
Size Exclusion Chromatography
Density
Centrifugation
Change in State
Distillation
Precipitation Recrystallization
Sublimation
8/15/2006 Chem208Spring2001 3
Relative Volatility
RV=P
o
A
/P
o
B
Substance with greater vapor pressure is
more volatile
Volatility related to
Molecular weight
Intermolecular attraction
Mole Fraction of each component
2
8/15/2006 Chem208Spring2001 4
Relative Volatility
P
total
=P
A
+P
B
=X
A
P
o
A
+X
B
P
o
B
Vapor over a boiling mixture contains more
of the volatile component that the liquid
Show this in phase diagram
8/15/2006 Chem208Spring2001 5
Gas-Liquid
BULK SEPARATIONS
Distillation - Phase Diagram
Mole Fraction A B
Vapor
Composition
Liquid
Composition
Vapor
Liquid
Temp
.
1 Plate
2nd
plate
3rd
8/15/2006 Chem208Spring2001 6
Distilation
Collect volatile fraction - get more A
Repeat the distillation to improve purity
Distillation columns do this without need
for multiple pot stills
3
8/15/2006 Chem208Spring2001 7
Theoretical Plates
Each stage in distillation is called a "plate".
If we analyze distillate and know phase
diagram we can determine the number of
effective plates involved to produce the
observed separation.
Theoretical plate
1 plate = 1 discrete thermodynamic
equilibration
8/15/2006 Chem208Spring2001 8
Non-ideal solutions
High Boiling Azeotrope
Composition of vapor
reaches constant value
Independent of starting
composition
Preparation of Conc.
Acids
High boiling azeotrop
8/15/2006 Chem208Spring2001 9
Non-ideal solutions
Low Boiling Azeotrope
Composition of liquid
reaches constant value
Independent of starting
composition
lowboiling azeotrope
4
8/15/2006 Chem208Spring2001 10
Purifying azeotropic mixtures
Add third component
Ethanol-water 95% azeotrope 190 proof
Add benzene get 100% Ethanol
Dont ever drink it - contains benzene
8/15/2006 Chem208Spring2001 11
A little History
Precipitation - Liq - Solid Sep.
Quantitative Analysis
Qualitative Analysis
Purification of synthetic products
Precipitation often does not produce a
very pure product - Inclusion and
Occlusion, Co-precipitation.
8/15/2006 Chem208Spring2001 12
A little History
To get a multi-stage separation required
much time - weeks - years.
Rare earth separation
precipitation
L S
L SL S
L SL SL S
1000 stages Ph.D. Thesis
5
8/15/2006 Chem208Spring2001 13
A little History
Solvent Extraction Research
Alternative to Alternative to Precipitation
Easier to remove contamination
Craig Counter Current Extraction
Multistage solvent extraction
Chromatography - Continuous
Extraction
8/15/2006 Chem208Spring2001 14
SEPARATIONS AND
EXTRACTION
Separation necessary when
measurement technique not
adequately selective
Contact between two immiscible phases
Separation of phases
8/15/2006 Chem208Spring2001 15
Phase Combinations
16 possible phase combinations
G L S SCF
G * * *
L * * *
S * *
SCF *
Each represents 1 or more separation
methods depending on how phases are
brought together.
6
8/15/2006 Chem208Spring2001 16
Mechanisms for phase contact
Bulk separations - recognizable
volumes of each phase.
Solvent Extraction
Distillation
Thin - layer separations - 1 phase
present as a 2 dimensional layer.
GC Thin layer G-L
HPLC Thin layer L-L or L-S
8/15/2006 Chem208Spring2001 17
Solvent Extraction
Equilibrium of solute Z between two
immiscible phases:
Z
(Aq)
<===> Z
(org)
K
eq
=K
p
=[Z]
or
[Z]
Aq
Partition Coefficient
Activity coef, rarely known - use conc.
8/15/2006 Chem208Spring2001 18
Solvent Extraction
Multiple equilibria often occur
major advantage of extraction methods.
M
+
+ L
-
<====>ML
Aq
+X +H+
|| || ||
V V V
MX
+
HL
(Aq
ML
(org)
+ B <===> ML
.
B
(org)
||
V
HL
(0rg)
7
8/15/2006 Chem208Spring2001 19
Solvent Extraction
Most useful information - total conc of
all forms.
D =C
or
= [ML]
or
+[ML
.
B]
or
C
aq
[MX
+
] +[M
+
] +[ML]
Aq
Distribution Ratio
8/15/2006 Chem208Spring2001 20
Fraction Extracted
Calculation of fraction of total in
each phase.
Start with all solute in Aq - phase.
q =fraction remaining in Aq phase =C
Aq
C
o
C
or
=(C
o
V
Aq
- C
Aq
V
Aq
)/V
or
=(C
o
-C
Aq
)
V
Aq
V
or
D =C
or
=(C
o
V
Aq
- C
Aq
V
Aq
)/V
or
=V
aq
x C
o
- C
Aq
C
Aq
C
Aq
V
or
C
Aq
8/15/2006 Chem208Spring2001 21
Fraction Extracted
C
o
=initial conc.
V
R
=V
or
/V
aq
fraction in org. phase =p
xD
V
V
C
C
aq
or
aq
o
+ = 1
D V
D
V
V
C
C
q
R
aq
or
o
aq
+
=
+
= =
1
1
1
1
D V
D V
q p
R
R
+
= =
1
1
8
8/15/2006 Chem208Spring2001 22
Ps and qs
Special case V
R
=1
q = 1 p = D
1+D 1+D
% Extracted =100p
8/15/2006 Chem208Spring2001 23
Simple extraction
p =fraction in organic phase D V
R
1+DV
R
q =fraction in aqueous phase 1
1+DV
R
Extract efficiency depends on D & V
R
For small values of D it becomes necessary
to increase V
R
to get complete extraction.
Practical limits to V
R
about 1000
8/15/2006 Chem208Spring2001 24
Repeated - Stepwise
Extraction
Use several organic portions to extract
aqueous phase (Vanilla Experiment).
p
1
=DV
R
q
1
= 1
1+DV
R
1 +DV
R
p
2
=DV
R
* 1 = DV
R
=pq
1+DV
R
1+DV
R
(1+DV
R
)
2
q
2
= 1 * 1 = 1 =q
2
1+DV
R
1+DV
R
(1+DV
R
)
2
9
8/15/2006 Chem208Spring2001 25
Repeated - Stepwise
Extraction
Third ext.
p
3
=p*q
2
= DV
R
* 1 = D V
R
1+DV
R
(1+DV
R
)
2
(1+DV
R
)
3
q
3
= 1 . 1 = 1 =q
3
1+DV
R
(1+DV
R
)
2
(1+DV
R
)
3
org phase pq
N-1
Aq phase q
N
total solute if all organic layers combined =(1-q
N
)
p
T
=p
1
+p
2
+p
3
=p +pq +pq
2
=total fraction
Total quantity =p
T
C
o
V
Aq
=(1-q
N
) C
o
V
Aq
8/15/2006 Chem208Spring2001 26
Example
Extraction of Acetanilide from water into ether
D =3.0
Single Ext V
R
=1 p
total
= D =3/4 = 0.75
1+D
Final Conc =0.75*C
o
=0.75
Single Ext V
R
=10 p
total
= D =
1+D
p
total
=3*10/(1+3*10) = 0.97
Final Conc =0.97 * C
o
/10 =0.097
8/15/2006 Chem208Spring2001 27
Example
Extraction of Acetanilide from water into ether
5 ext. V
R
=1
p
t
=1- (1 /1+D)
5
=1-(1/4)
5
p
total
=.999
Final conc =0.999*C
o
/5 =0.1998 =0.2
much more efficient to do multiple
extractions
Max possible extraction efficiency
limit where V
aq
divided into infinite no. of portions
aq
or c
V
V D
e q
10
8/15/2006 Chem208Spring2001 28
Multiple extractions
In general N =5 is optimum gets within a
factor of 5 of max. possible efficiency.
8/15/2006 Chem208Spring2001 29
Separation of 2 solutes
Separation factor depends on ratio of
D
cA
/D
cB
Ratio must be very large to obtain high
purity 10
6
.
Optimum conditions for separation
V
R
=(D
cA
/D
cB
)
1/2
8/15/2006 Chem208Spring2001 30
Solvent Extraction
Extraction of Organic Molecules
Acids, bases, and neutral molecules
Only Neutral form partitions into Org.
Acids extract at low pH
Bases extract at high pH
Neutrals unaffected by pH
Used for separation of chemical classes
11
8/15/2006 Chem208Spring2001 31
Vanilla Experiment
Vanillin is neutral molecule
Partitions into organic phase with high D
Add Base
Deprotonatedformof vanillin is anion
Soluble in aqueous phase
anion formnot soluble in organic phase
Very efficient separation
Selective for neutral molecules with acidic
functional group
8/15/2006 Chem208Spring2001 32
Solvent Extraction
Extraction of Organic Molecules
Text uses only case for monoprotic but
equation applies to all. Use alpha for
neutral form.
'
p HA c
K D =
8/15/2006 Chem208Spring2001 33
Solvent Extraction
Metal separations-common use of Ext.
Extraction has several advantages over
precipitation.
Less contamination (entrapment or
adsorption on precipitate is a problem).
More variables in Equilibrium. - exploited
to give separation.
Organic extractants - fine tune for specific
separation.
12
8/15/2006 Chem208Spring2001 34
Simplest Metal Ext. Case

n
M
n+
+nL
-
<===> ML
n(Aq]
+H+ b K
pML
b K
A
ML
N(or)
HL
(Aq)
b K
pHL
HL
(or)
8/15/2006 Chem208Spring2001 35
Distribution
D = [ML]
or
[M+]
Aq
+[ML]
Aq
n
= [ML
n
]
Aq
K
A
=[H
+
][L
-
]
[M+
N
][L]
n
[HL]
(Aq)
K
pML
=
[ML
n
]
or
K
pHL
=[HL]
or
[ML]
Aq
[HL]
8/15/2006 Chem208Spring2001 36
Distribution
[ML
n
]
or
=K
P
[ML
n
]
Aq
[ML
n
]
Aq
=B
n
[M
n+
][L
-
]
n
[ML
n
]
or
=K
P
B
n
[M
n+
][L
-
]
n
[L
-]
=K
A
[HL]
A
[H
+
]
[HL]
A
=[HL]
or
[L
-]
=K
A
[HL]
or
K
PHL
[H
+
]K
PHL
13
8/15/2006 Chem208Spring2001 37
Distribution
D = [MLn
]
or
[M
+
]
Aq
+[MLn]
Aq
assume complex not water soluble [Mln]
Aq
=0
Generally [ML
n
]
Aq
is very small except
acetyl acetonates and water soluble
ligands with additional charged sites
n
p
or a n
n p or n
HL
ML
K H
HL K
M K ML
=
+
+
] [
]
] [ ] [
8/15/2006 Chem208Spring2001 38
Metal Distribution
D
c
=
K
pML

n
(K
a
)
n
[HL ]
n
or
(K
pHL
)
n
[H
+
]
n
aq
D
c
= (constant )
n
[H
+
]
n
aq
8/15/2006 Chem208Spring2001 39
Metal Distribution
Variables to control
pH
Organic Solvent (Changes K
p
)
D
c
=
K
pML

n
(K
a
)
n
[HL ]
n
or
(K
pHL
)
n
[H
+
]
n
aq
D
c
= (constant )
n
[H
+
]
n
aq
14
8/15/2006 Chem208Spring2001 40
Metal Distribution
Multidentate
ligandsgive
sharper pH
extraction
breaks
n=1
n=2
n=3
pH
% Ext.
8/15/2006 Chem208Spring2001 41
Metal Extraction
Control of pH allows separation of
species with different B's and different
K
PML
's.
Extraction Eq. also may be effected by
auxiliary ligands (masking agents)
which change conditional 's
i.e. Reduce extraction of masked ion.
Use of Adduct forming organic phases
also has an effect
ML
n[org]
+B <====> ML
n
.
B
[org]
increase extraction
8/15/2006 Chem208Spring2001 42
Solid Phase Extraction
Absorb solute on the surface of particles
Need lots of surface area
Selective absorption of solute of interest
Elution
Different solvent used - solute partitions into solvent
Soluteelutes fromcolumn
Affinity Columns - specific biological interaction -
antibody-antigen
Pre-concentration- pass large volume through
Sample Cleanup remove interferences
15
8/15/2006 Chem208Spring2001 43
Applications
Vanillin lab type - hold analyte and wash out
junk
Put SPE in Sample loop of HPLC
Flush out contaminants
Drug and metabolites in plasma
96 Well plates do 96 samples at once
8/15/2006 Chem208Spring2001 44
A little History
Solvent Extraction Research -
Alternative to Precipitation
Craig Counter Current Extraction
Chromatography - Continuous
Extraction
8/15/2006 Chem208Spring2001 45
CHROMATOGRAPHY AND
SEPARATIONS
Problems with difficult separations-
Counter current extraction was slow.
Goal- Continuous extraction - apparatus
without discrete stationery solvent units.
Tswett - lost in Russian Lit.
Lederer's work in 1930's Germany - also
lost.
AJ P Martin - Silica gel has surface layer
of bound water.
Packed column with silica and passed CHCl
3
through (mobile phase) worked for fatty acids.
16
8/15/2006 Chem208Spring2001 46
CHROMATOGRAPHY AND
SEPARATIONS
Then invented Gas chromatography
and the common detectors for GC
Martin and Synge - Nobel Prize 1952
Developed extension of counter current
math to describe chromatography.
Not best treatment (ignores kinetics) but
worth some time.
Three types of chromatography -
Elution, Frontal, and Displacement
8/15/2006 Chem208Spring2001 47
Elution chromatography -
most common
Solute placed at top of column.
Eluted until each component reaches
the end (detector).
8/15/2006 Chem208Spring2001 48
most common
Mobile phase velocity is the fastest
possible rate of movement through the
columns.
solutes retarded - based on time spent
in stationary phase.
TLC - Elute till solvent reaches top and
look for position of peaks
17
8/15/2006 Chem208Spring2001 49
most common
Change elution data to linear velocity in
the column
10 cm HPLC column x4.6 mm ID with
2.0 ml/min flow
if mobile phase moves through in 0.50
minutes - linear velocity =20 cm/min
10 cm length must contain 1.00 ml of
mobile phase
8/15/2006 Chem208Spring2001 50
Elution chromatography
Total column volume 4.6 mm diameter
x 10 cm
V
t
= r2h =1.66 ml.
Mobile phase volume
V
m
=1.00 ml
Stationary phase volume
V
s
=V
t
- V
m
=1.66-1.00 =0.66
ml
Phase Ratio = V
s
/V
m
=.66/1.0 =0.66
8/15/2006 Chem208Spring2001 51
time ratio in phases t
s
= C
s
V
s
=K Vs
t
m
C
m
V
m
V
m
=K
K =Cs partition ratio
C
m
Thermodynamic basis for retention
18
8/15/2006 Chem208Spring2001 52
Capacity factor k' - Describes
Retention
Most common for HPLC
time in stationary phase/time in mobile
phase
k'=V
R
- V
m
=t
r
-t
o
V
m
t
o
k' varies from 0 to large no.'s
ln k' = H - S - ln
RT R
8/15/2006 Chem208Spring2001 53
ln k' = H - S - ln
RT R
Plots of ln k vs 1/T linear
show contribution of entropy and enthlapy
8/15/2006 Chem208Spring2001 54
Band spreading in
chromatography
Inject Sample as a very narrow band -
square wave pulse of sample
As it moves through column, it begins to
spread - gaussian band profile
= std. dev. of peak profile
Number of theoretical plates N
= t
r
2
/
2
peak width at base t
w
=4
19
8/15/2006 Chem208Spring2001 55
Band spreading in
chromatography
N=16(V
r
/V
w
)
2
=5.54(V
r
/V
1/2
)
2
V
r
=t
r
V
w
=t
w
use any units time, length, volume
HETP =L/N
Plate Theory ===>van Deemter
Equation
H=A +B/v +Cv
where v=mobile phase velocity
8/15/2006 Chem208Spring2001 56
Band spreading in
chromatography
H=A +B/v +Cv
Three Components
A-Eddy diffusion
B-Longitudinal
diffusion
C-Rate of mass
transfer between
phases
velocity
H
8/15/2006 Chem208Spring2001 57
Band spreading in
chromatography
Problems with the Plate Theory
Assumes linear partition between phases
Assumes rapid equilibrium
Ignores molecular diffusion
Treats column as series of discrete plates
Rate Theory (J . C. Giddings)
H=H
d,m
+H
d,s
+H
s
+H
m
+H
e
all terms are flow rate dependent
20
8/15/2006 Chem208Spring2001 58
Rate Theory - optimum
conditions
k=1
small particle diameter
thin layer of stationary phase
high diffusion coefficients
( high temperature) low viscosity
Slope of rising portion of van Deemter
plot is function of particle diameter,
small particles flatten the graph allowing
efficient operation at high flow
8/15/2006 Chem208Spring2001 59
HETP
Result same
as
VanDeemter
velocity
H
H flow
H mass
transport
H diffusion
8/15/2006 Chem208Spring2001 60
HPLC vs GC
Differences due to
diffusion rates in liquids vsgases
particle sizes
LC
GC
1E-5 1e-4 1e-3 1e-2 1e-1 1 10 100
velocity cm/sec
H
21
8/15/2006 Chem208Spring2001 61
Reduced parameters
v =d
p
v/D
m
h =H/d
p
All curves superimpose with h minimum
at about 2d
p
=2 particle diameters is
the minimum length of column required
for a theoretical plate. v optimum =1
D
m
for liquids 10
-6
to 10
-7
D
m
10
-2
to 10
-1
8/15/2006 Chem208Spring2001 62
Very High Flow
Prev. Discussion Assumes Laminar Flow
Turbulent FlowConditions
Faster Mass Transport
HETP Graph comes back down
Not as far as Laminar minimum
Useful for high speed sample prep
8/15/2006 Chem208Spring2001 63
Types of Columns
Packed columns
For open tubular columns there is no
flow paths term H
e
=0
Packed column
open tubular
coating on walls
22
8/15/2006 Chem208Spring2001 64
Deviant Chromatographic
Behavior
Non-linear
partition
isotherms
Ideal behavior
Anti-Langmuir
Isotherm
Langmuir
Isotherm
Conc
Stationary
phase
Conc mobile phase
8/15/2006 Chem208Spring2001 65
Behavior
Anti-
langmuir
sample
acts as
stationary
phase
Langmuir
Saturate
sites
Anti-langmuir case
Langmuir case
8/15/2006 Chem208Spring2001 66
Behavior
Anti-Langmuir
23
8/15/2006 Chem208Spring2001 67
Extra-column Volume
Broadens the injected band before it
reaches the column
Broadens the post column bands before
the detector
For good peak shape
V
col
>>>V
inlet
+V
detector
+V
connections
8/15/2006 Chem208Spring2001 68
Resolution in Chromatography
R=t
r
/4 = t
r
/W
av
=(t
r2
- t
r1
)/0.5(W
1
+W
2
)
most common equation for determining
R from chromatogram
8/15/2006 Chem208Spring2001 69
Resolution
24
8/15/2006 Chem208Spring2001 70
R =
1
4

( 1)
k'
1+ k'
N
=
k'
2
k'
1
= separation factor
8/15/2006 Chem208Spring2001 71
3 Ways to control separation
Change affinity of solute for stationary
phase Column Selectivity
change stationary phase
change form of solute ( charge or size)
change temperature
Change retention by changing k'
Change temperature in GC
change mobile phase in LC
Change N
flow rate
change column - particle size or length
8/15/2006 Chem208Spring2001 72
Time required for separation
Plates/second
Optimum k =
2.0 12
10
8
6
4
2
0
2 3 4 5 6 7 8
Log N
Log tr
GC
LC
1 year
1 day
1 hour
1 min
25
8/15/2006 Chem208Spring2001 73
LC vs GC
LC less plates/sec due to slower mobile
phase velocity
LC applicable to much wider range of
compounds
GC always method of choice where
both work equally well. Volatile and low
MW. compounds
8/15/2006 Chem208Spring2001 74
General Elution Problem
Single set of
conditions
good for only 6-
8 components
8/15/2006 Chem208Spring2001 75
Capillary GC
of FAMES
Isothermal GC at
150 degrees
Note Tr separation
between even Cs
almost doubles
26
8/15/2006 Chem208Spring2001 76
Capillary GC of FAMES
Chart Title
y=0.079x- 0.2364
R
2
=0.9963
0.0000
0.5000
1.0000
1.5000
2.0000
0 10 20 30
Carbon Number
L
o
g

T
r
Logtr
Linear (Logtr)
C number Tr
12 5.45
14 6.98
16 10.04
18 16.24
20 22.02
22 31.68
24 45.58
8/15/2006 Chem208Spring2001 77
Capillary GC of FAMES
Temperature Programmed run
8/15/2006 Chem208Spring2001 78
Elution parameters must be varied to
separate complex mixtures
Temperature -
GC- column equilibrates quickly.
LC columns equilibrate very slowly, temp rarely
used
Mobile phase -
GC Mobile phase is inert and doesnt effect the
separation.
LC- most commonly varied parameter
27
8/15/2006 Chem208Spring2001 79
Initial conditions k' very large most peaks
held at start of column
Special case in capillary column GC cryo-
focusing.
If solvent is a liquid at initial temperature, the
peaks actually focus into a sharper band than
the injection band
Gradually reduce k' - peaks start to move
Temperature programming - linear ramps,
sometimes multi-stage ramps
8/15/2006 Chem208Spring2001 80
Initial conditions k' very large most peaks
held at start of column
Solvent programming -
solvent strength vs % composition is
exponential curve so programs are often
exponential in shape to produce linear
change in eluting power of mobile phase.
Vary polarity, pH, salt content
Ternary and quarternary gradients possible
Chocolate lab - Methanol, acetonitrile better than
either one alone
DryLabSoftware for optimization
8/15/2006 Chem208Spring2001 81
Gradient Elution
Peaks can be all about the same shape
No limit to number of components
N/t approximately constant in
programmed run. N is meaningless as
a measure of quality.
28
8/15/2006 Chem208Spring2001 82
Sample Capacity
Isocratic
Max sample proportional to t
r
/2N
1/2
Programmed
Max sample proportion to t
ro
/2N
1/2
t
ro
=retention time at initial conditions
(very large)
8/15/2006 Chem208Spring2001 83
Problems with programmed
elution
Time to return to initial conditions
GC - cool column oven
LC- equilibrate column
SFC - density gradient - re-
equilibrate
8/15/2006 Chem208Spring2001 84
Frontal Chromatography
Mixture used
as mobile
phase
Great if you
want Solute 1
only
29
8/15/2006 Chem208Spring2001 85
Displacement
Chromatograph
y Great for Preperative
Separations
GramQuantities on
standard columns
Must have displacer
Must determine
isotherms
8/15/2006 Chem208Spring2001 86
Gas-Solid Adsorption
Surface atoms dont have enough bonds
Form strong interactions with molecules or
ions
Equilibrium process - non-ideal
Adsorption Isotherms - Describe the
equilibrium - often non-linear
8/15/2006 Chem208Spring2001 87
Ideal and non-ideal isotherms
right graph- sites become filled and no more
abosrbed
left graph - molecules absorb on absorbed
molecules
Pressure of A
quantity
adsorbed
ideal
non-ideal
Pressure of A
quantity
adsorbed
ideal
non-ideal
non-ideal
ideal
30
8/15/2006 Chem208Spring2001 88
Solute Absorption
Absorb solute on the surface of particles
Need lots of surface area
Selective absorption of solute of interest
Elution
Different solvent used - solute partitions into solvent
Solute elutes fromcolumn
Affinity Columns - specific biological
interaction - antibody-antigen
1
8/15/2006 Chem208Spring2001 1
Chem 208
Chapter 24
Gas Chromatography
8/15/2006 Chem208Spring2001 2
4 factors influence retention
Vapor pressure f (Temp)
Heat of vaporization f (Temp)
Solubility (8m) in stationary phase f
(T)
Adsorption on solid surface
8/15/2006 Chem208Spring2001 3
Surface atoms dont have enough bonds
Form strong interactions with molecules or
ions
Equilibrium process - non-ideal
Adsorption Isotherms - Describe the
equilibrium - often non-linear
2
8/15/2006 Chem208Spring2001 4
Ideal and non-ideal isotherms
right graph- sites become filled and no more
abosrbed
left graph - molecules absorb on absorbed
molecules
Pressure of A
quantity
adsorbed
ideal
non-ideal
Pressure of A
quantity
adsorbed
ideal
non-ideal
non-ideal
ideal
8/15/2006 Chem208Spring2001 5
Solid Supports for GC
Adsorption on solid surface - Strongest
stationary phase interaction
Useful for permanent gases
GSC silica, graphite, firebrick
1000 m
2
/gm surface
2-5 gm in a column
As vapor pressure decreases need less
retention.
Minimizing Solid Support Interactions
8/15/2006 Chem208Spring2001 6
Solid Supports for GC
Inertsupports with low surface area
0.2 - 1 m
2
/gm surface
Diatomaceous earths - high surface
area silica
Acid wash remove metals
Calcine reduce surface
silanize bond surface hydroxyl
groups
Coat with liquid phase 1000 available
3
8/15/2006 Chem208Spring2001 7
Silicone Phases for GC
Temp limits 200-
300
o
C
Works when others
dont
Moderately polar C
3
F
7
Separation by
polarity
Very polar Cyano
5% - 100% of silicon
General application
with more polar
molecules being
retained longer for
same BP
Slightly polar and pi
bonding
Phenyl
5% - 100% of silicon
Boiling point
separations
Non-polar Methyl
Application Polarity Functional Group
8/15/2006 Chem208Spring2001 8
Liquid Phases for GC
O
O
O
O
O
O
O
O
n

Fatty acids and other
polar moleucles
Very polar Ethylene glycol
esters
DEGS di-ethylene
glycol succinate
General separation
by polarity
Very polar Poly-ethylene glycol
Carbowax
HO- [CH
2
-CH
2
-0 ]
n
H
MWsto 20,000
Application Polarity Polymer
8/15/2006 Chem208Spring2001 9
Types of Columns
10,000-
100,000
0.05 1 mm
ID
10-100
meters
Fused silica
capillary
1000 1/8 in OD 1-10 meters Packed
Analytical
100 to in OD 2-4 meters Preparative
packed
Typical
number of
plates
Typical
diameter
Typical
length
Column type
4
8/15/2006 Chem208Spring2001 10
Types of Columns
Capillary columns
steel, glass, fused silica.
.067 - 1 mm ID
1000 -100 plates/ft
Columns may be very long due to low
pressure required
Typical 10 meters - 60 meters (GC Lab 30
meter)
30,000 10
6
plate possible
100,000 typical
8/15/2006 Chem208Spring2001 11
Instrumentation for GC
Carrier Gas Supply
FlowController
Pressure Controller
Pressure Programming
Injector
Flash Vaporization
On Column
Split- Capillary
Column Oven
LowThermal Mass
40 deg/min dT
Detector Readout
Computer
Integrator
Recorder
8/15/2006 Chem208Spring2001 12
Split/Splitless
Injector
Constant Pressure
Split vent electrical valve
Expansion of solvent
Inert liner
quartz
silanizedglass
5
8/15/2006 Chem208Spring2001 13
Injection
Syringe Injection 0.1 to 10 ul +/- 2-3%
Gas samples can be up to 2 ml
Valve injection gases
SPME
Headspace analysis
Purge and Trap
8/15/2006 Chem208Spring2001 14
Detectors -Thermal conductivity
detector
Hot filament - Resistance is f(Temp)
Filament temp is varied with thermal
conductivity of gas in cell
He high TC
Org low TC
V
Amp
DC Power
Supply
Rs
Rs
Rr
Rr
Reference
Column
Sample
Column
8/15/2006 Chem208Spring2001 15
Detectors -Thermal conductivity
detector
Wheatstone Bridge
If resistances are matched V=O volts
sample reduces thermal cond.
increases temp ==>inc. resist
Voltage appears at V
Detection limit .1 to 1 microgram
injected - 10
-9
gm/sec
Linear Range 10
4
Detects everything except carrier gas
6
8/15/2006 Chem208Spring2001 16
Ionization Detectors- Flame
ionization
Sample +
Hydrogen
200 volt DC
Electrometer Flame
Electrode
8/15/2006 Chem208Spring2001 17
Ionization Detectors- Flame
ionization
Used for C-H compounds.
Gram response approximately constant
C=0 & C-X lower response
Formaldehyde H
2
CO very low response
current 10
-14
to 10
-7
amps.
10
-12
gm/sec detection
Total peak 0.1 to 1ng
linear range 10
6
ionization efficiency .1%
Sample +
Hydrogen
200 volt DC
Electrometer Flame
Electrode
8/15/2006 Chem208Spring2001 18
ECD Electron Capture
Detector
e
-
+carrier establish background
standing current
Beta
source
Sample
Power
Supply - DC
or Pulsed
Electrometer
-
+
7
8/15/2006 Chem208Spring2001 19
Detector
Standing Current 10
-9
to 10
-10
amps
Sample +e
-
sample
-
reduces standing current
negative peaks
sensitivity depends on electron affinity
varies by 10
4
Beta
source
Sample
Power
Supply - DC
or Pulsed
Electrometer
-
+
8/15/2006 Chem208Spring2001 20
Detector
halogenated samples best pesticides
PCB's
CHC1
3
1 x 10
-15
gm/sec
peaks for 11-
14
to 10
-12
gm
Linear range 300-100000 depends on
design
Sensitivity high due to high ionization
Eff. 100%
Beta
source
Sample
Power
Supply - DC
or Pulsed
Electrometer
-
+
8/15/2006 Chem208Spring2001 21
Detector Summary
Non-destructive 1000 10
-9
gm/sec
10
-7
gmin peak
TCD
Identification
and
quantitation
1000 10
-12
to 10
-
15
gm/sec
Mass
Spectrometer
S FPD
N or P NPD
Cl and F
molecules
100-1000 10
-15
gm/sec
10
-14
gmin peak
ECD
All
hydrocarbons
10
6
10
-12
gm/sec
10
-11
gmin peak
FID
Applicability Linear range Best case limit
of detection
Detector
8
8/15/2006 Chem208Spring2001 22
Photo ionization
7-12 eVH
2
Lyman line - far UV
Ionization Eff 1%
Less background noise than FID
More sensitive by 10 - 50 then FID
Somewhat selective depending on
wavelength
C
1-
C
4
He N
2
not detected
8/15/2006 Chem208Spring2001 23
Thermonic Detector
N or P specific FID
alkali metal FID
Bead of CeI placed in flame of FID
Enhances the sensitivity to N and P
8/15/2006 Chem208Spring2001 24
Flame photometric
Sulfur specific
Flame with photodetector
Useful fo analysis of fuels
9
8/15/2006 Chem208Spring2001 25
Microwave Emission
all elements excited in microwave
plasma.
Select light from one of interest.
1x10
-12
gm peaks for some elements.
Multiple channels - several elements
empirical formulas
Halogen analysis in water samples
Selenium in garlic
8/15/2006 Chem208Spring2001 26
Identification of Peaks
Plot Log t
r
vsCarbon Number
Linear graph for homologous series
Log plots on two different columns
log Tr
Col A
log Tr Col B
Identify
compound
8/15/2006 Chem208Spring2001 27
Identification of Peaks
Mass Spectrometry
Molecular weight of molecules
Fragmentation pattern for Identifcation
Library search 300,000 spectra available
Trace analysis sensitivity
GC-IR
IR spectrumof peaks
10
8/15/2006 Chem208Spring2001 28
Analysis Methods
make calibration curve
run sample and compare
Limited by injection accuracy
2-4% manual
1-2% auto
0.5% valve injections
8/15/2006 Chem208Spring2001 29
Analysis Methods
eliminates injection error - only integration
error is left
add standard to each sample use it to
correct for variation in injection and in
sample workup etc.
Single point - determine peak ratio with one
standard mixture
Calibration curve - plot area ratio vs mass ratio
8/15/2006 Chem208Spring2001 30
Data from GC experiment
C22:1
y =0.6548x - 0.0804
R
2
=0.9989
0
1
2
3
4
5
0 2 4 6 8
Series1
Linear
(Series1)
C16:0 y =1.0405x -
0.3135
R
2
=0.9991
0
5
10
15
20
0 10 20
Series1
Linear
(Series1)
11
8/15/2006 Chem208Spring2001 31
Analysis Methods
Standard Addition Methods
O
p
=KC
O
p
=output height or area
O
u
=Kcu
0
s
=K
(
V
u
C
u
+V
s
C
s
) .
V
u
+V
s
C
s
=V
u
C
u
+V
s
C
s
V
u
+V
s
1
8/15/2006 Chem208Spring2001 1
Chem 208
Chapter 25
HPLC
8/15/2006 Chem208Spring2001 2
LIQUID
CHROMATOGRAPHY
Forces which lead to retention
Ionic force -
+and - ions
Polar Force -
Dipole - Dipole attraction
Dispersive Forces -
London forces & Van der Waals
Size Exclusion
8/15/2006 Chem208Spring2001 3
Polar Force
Normal phase LC
Silica or Aluminum solid absorbent
Si0
2
Al
2
0
3
O
O
Si
OH
O
O
Si
O H
OH
O
O
O
SI
O
SI
O
SI
O
SI
O
SI
O
SI
O
O
O O
Si
O
Si
Si
O H
OH
OH
OH
O
O
Si
O H OH
O H
O H
2
8/15/2006 Chem208Spring2001 4
Polar Force
Elutropic series Solvents
Hexane low eluting power
CH
2
Cl
ether
alcohol high eluting power
Elution controlled by relative polarity of
mobile phase vs stationary phase.
8/15/2006 Chem208Spring2001 5
Dispersive Forces -
Induced dipoles
Reversed phase LC
R =C
18
H
37
C
8
H
17
C
3
H
6
NH
2
C
4
H
9
O
Si
O
Si
O
Si
O
Si
O
Si
O
Si
O
R
R R
R R R
R
R
R
R R R
8/15/2006 Chem208Spring2001 6
Dispersive Forces -
Elutropic Series Weak solvent
Water lowest eluting power
Acetonitrile
Methanol
THF
CH
2
Cl
2
Great eluting power
Non-aqueous reverse phase
3
8/15/2006 Chem208Spring2001 7
Dispersive Forces -
Solvent Triangle
ACN, MeOH, THF
Binary and ternary mixtures
Lab MeOH+ACN better than either alone
Computer Optimization Map space for
best separations
DrylabSoftware Run 4 Chromatograms
Predict most effects of parameter changes
8/15/2006 Chem208Spring2001 8
Dispersive Forces -
Advantages over Normal Phase
samples in aqueous solution
lower cost mobile phases easy disposal
rapid equil. of mobile phase
Disadvantages
columns more expensive - not much
Less efficient columns than normal phase
8/15/2006 Chem208Spring2001 9
Ionic forces
Ion exchange LC
Polymer Resin -
Styrene- DVB
PVP-DVB
Bonded Silica
Anion exchange
Cation Exchange
R
SO
3 H
R OH
O
N
R
R
R
+
4
8/15/2006 Chem208Spring2001 10
Ion Exchange
Uses
Water purification
Amino acid analyzers
Rare earth (RE) separations
30 ft columns in series
citrate complexing agent
pH gradient
citrate RE -- Resin RE competition
8/15/2006 Chem208Spring2001 11
Ion chromatography
DIONEX
2 Columns - Separator and Ion suppressor
Conductivity Detector
for anions - suppressor is cation
exchange
mobile phase Na0H or Na
2
CO
3
in
H
2
0
NaCl ==> HCl
NaF ==> HF
Mobile phase H
2
O or H
2
CO
3
8/15/2006 Chem208Spring2001 12
Ion Chromatograph
Detector
Ion exchange
analyzer column
Stripper columns
Selector
Valves
Regeneration
solvent
Regeneration
waste
5
8/15/2006 Chem208Spring2001 13
Ion chromatography
Sample Range ppb to100%
Cations
alkali metals, NH
4+
,etc
transition metals +ligand post columns
spectrophotometric
Anions -
NO
3
-
, SO
4
2-
halides, S0
4
, PO
4
, NO
3
inositol phosphates, etc.
8/15/2006 Chem208Spring2001 14
Stearic Exclusion
Volume available to each
species depends on its size
All species elute
"unretained" but void
volume varies with
molecular size.
Void volume =available
cross section x length
8/15/2006 Chem208Spring2001 15
Stearic Exclusion
Molecular sieves -
Zeolite clays
4, 5, 13 holes
Sephadex - dextran polymer
holes for molecules 100 - 5x10
5
M.W.
not rigid enough for high pressure
Polystyrene beads
MW 200 - 50 million
organic mobile phases
non-polar samples
6
8/15/2006 Chem208Spring2001 16
Stearic Exclusion
Molecular sieves -
Glass beads
MW 300 - 1,200,000
Polymer M.W. distributions
silica gels.
MW 400 - 8,000,000
C8 bonded silica and others
8/15/2006 Chem208Spring2001 17
Stearic Exclusion
Advantages
V
o
=Vol. of mobile
phase not in pores
V
i
=volume in all
pores
All peaks elute
between V
o
and V
o
+V
i
=
total void volume
log M.W.
log Retention volume
Vo large
Vo small
molecules
8/15/2006 Chem208Spring2001 18
Steric Exclusion
Calibrate V
R
vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
7
8/15/2006 Chem208Spring2001 19
HPLC Instrument
Solvent Supply
Pump
Constant flow
Gradient or Mixture
Injector
Valve
Column
Readout
Computer
Integrator
Strip Chart
Detector
8/15/2006 Chem208Spring2001 20
Pumps
Single piston- cheap - need pulse
damper
Dual piston - most common
Screw piston- small bore column
Gravity flow - LPLC only
Air pressure - dissolved bubbles a
problem
Isocratic and Gradient Systems
Solvent Mixing pumps
8/15/2006 Chem208Spring2001 21
Solvent Preparation
Bubbles due to cavitations
Bubbles in detector
HeliumSparge(LDC in lab)
VacuumDegasser (Thermo - LC/MS)
Intelligent pumps (Hitachi in lab)
Filter mobile phases to eliminate particles
Cause damage to check valves
8
8/15/2006 Chem208Spring2001 22
Columns
Large particle for Low Pressure LC
HPLC small particle
reduces diffusion distance
increases efficiency.
Evolution of HPLC
75 37 10 5 3 2 1 0.5
small particles - more pressure
Harder to pack.
8/15/2006 Chem208Spring2001 23
Columns
Typical columns of Analytical Work
4 or 4.6 mm ID x 10 - 25cm length
As particle size decreases. pressure
required increases P=k/dp^2
Below 2 microns flow causes significant
heating of the column.(Boil solvent in
extreme cases)
5000 to 20,000 plates typical.
8/15/2006 Chem208Spring2001 24
Columns
Smaller particles ==>faster separation
for a given capacity factor K =2
10 micron particles 25 cm column V
o
=
2ml
optimum flow 1ml/min
V
r
=6ml t
r
=6 min
3 micron particles
3 cm column V
o
=0.1ml
2ml/min optimum flow rate
V
r
=.3ml t
r
=9 sec.
Same separation in 2.5% of time using
5% f th l t
9
8/15/2006 Chem208Spring2001 25
Columns
Short Columns
3 and 5 cm length Fast HPLC
Narrow Bore 1 mm and 2 mm ID
low flow rate for LC-MS
Use less solvent
Increased sensitivity
8/15/2006 Chem208Spring2001 26
Capillary Columns
Capillary HPLC
1mm ID column loosely packed with large
particles
fused silica capillary coated with liq. phase
typical separation 1 - 24 hrs.
10
6
10
7
plates
8/15/2006 Chem208Spring2001 27
Detectors
Refractive index - Difference
between solvent and sample solution.
Universal sensitivity
10
-9
gm/sec best case RI =1x10
-7
Very temp. sensitive 10
-4
RI/o
C
0.001 C stability req.
Linear Range 3000
peaks may be +or -
10
8/15/2006 Chem208Spring2001 28
AbsorptionUV - Vis
Flow Cell in Spectrometer
Diode Array for full spectrum
10-
12
gm/sec sensitivity
depends on absorbtivity
100 - 500,000/mole
8l flow cells with 1 cm path length
3l flow cell with 0.5 cm path
0.1 l flow cell with 0.2 cm path
0.03l flow cell with 0.10 cm
Not as critical as RI - easy to use
8/15/2006 Chem208Spring2001 29
Detector Volume
Volume of detector and Injector and
connecting tubes <<<Void volume
Inj. & Det must have low volume 10
microliter detector volume for 25 cm 10
micron column
3 microliter volume for high speed
column
<0.1 ml detector volume now available
for capillary columns
8/15/2006 Chem208Spring2001 30
Fluorescence photometry
Sample absorbs light in UV and emits at
longer wavelength
Sensitivity 10
-15
gm/sec depends on
fluorescence of sample
very selective - most things dont
fluoresce
narrow linear range
Also used for Chemiluminescence
11
8/15/2006 Chem208Spring2001 31
Electrochemical
Amperometry -
biological studies -
neuroscience
measure current flow at fixed voltage
current due to oxidation or reduction of
molecules in solution
10
-13
moles injected 10
-15
moles/sec
may have very small cell vol. 0.1-3 l
used for capillary LC
Sample
A
Voltage
source
Cell
8/15/2006 Chem208Spring2001 32
Other Detectors
LC-MS
Same advantages as GC-MS
Identification
Very high sensitivity
Very high selectivity
Evaporative Light Scattering
Sample sprayed into chamber
Solute particles scatter light
Universal detector - non-volatile solutes
8/15/2006 Chem208Spring2001 33
Peak integration
Drop perpendicular
Most common method
Simple
Small peak +error
Large Peak error
12
8/15/2006 Chem208Spring2001 34
Peak integration
Valley Method
Always produces errors
Errors can be large
Useful for
Multipepeaks
Complex baseline
8/15/2006 Chem208Spring2001 35
Peak integration
Skim Method
Exponential Skim
Linear Skim
R<1
Small peak <5% of large
8/15/2006 Chem208Spring2001 36
Peak integration
Gaussian Skim
More Complicated
Less error than other skimmethods
13
8/15/2006 Chem208Spring2001 37
Peak integration
Errors depend
Relative Peak areas
Resolution
Method used
Improve Resolution!!!
1
8/15/2006 Chem208Spring2001 1
Chem 208
Capillary Electrophoresis
Chapter 26
8/15/2006 Chem208Spring2001 2
Traditional Electrophoresis
ion migration separation
Cations migrate toward the negative
electrode and Anions toward the positive
electrode
Neutral molecules do not migrate in an
electric field.
Rate of ion migration depends on size and
charge V
ep
=u
ep
E
u
ep
=charge/6* pi* viscosity* radius or ion
8/15/2006 Chem208Spring2001 3
Adds flow of buffer through the capillary
Buffer migrates toward the Negative
Electrode
Cations move faster
Anions swim upstream and may or may
not elute
Neutrals go with the flow
2
8/15/2006 Chem208Spring2001 4
Fused silica tube
treated with base to get
free silanol groups
At high pH, surface is
negative Layer of
positive buffer ions
forms to counter the
charge.
8/15/2006 Chem208Spring2001 5
Excess cations in the
area near the walls
move toward the
cathode DRAG THE
SOLVENT WITH
THEM
At low pH, no charge
on silica and it doesnt
work well.
8/15/2006 Chem208Spring2001 6
Electroosmoticflow
V
eof
=u
eof E
k layer thic double * charge wall *
viscosity * 4
* cons dielectric
k Zeta
pi
Zeta
u
eof
=
=
3
8/15/2006 Chem208Spring2001 7
Ion Velocity
V=V
ep
+V
eof
Cations both positive
V>V
eof
Anions V
ep
negative
V<V
eof
Neutral Species V=V
eof
8/15/2006 Chem208Spring2001 8
Separation
Depends on Electrophoreticmobility
Different Size
Different charge
Manipulate pH, ionic strength, dielectric
constant to change charge and shape
Additives to adduct with solutes
Cyclodextrins
Micelles
8/15/2006 Chem208Spring2001 9
Instrument Diagram
4
8/15/2006 Chem208Spring2001 10
Instrument
Injection
Pressure - siphon effect or gas pressure
Electrophoreticinjection place capillary in
sample and apply voltage to draw sample in.
Mechanically more complex than HPLC or GC
and not as easy to reproduce.
Columns - Silica - surface treated
8/15/2006 Chem208Spring2001 11
Instrument
Detection note MWsin 100s to 100,000s
Fluorescence (Laser induced LIF) very sensitive
10
-18
to 10
-20
moles injected
UV-Vis through column - remove polyimide coating
not very sensitive
10
-13
to 10
-16
moles
Mass Spec Electrospray works great
10
-16
to 10
-17
moles
Electrochemical works well for electroactivesolutes
10
-18
to 10
-19
moles
Vacancy detection - like acid rain add absorbing
species to buffer and look at vacancies.
8/15/2006 Chem208Spring2001 12
Example of CE
DNA fragments
5
8/15/2006 Chem208Spring2001 13
MECC - Micelles
Micelles trap neutral organics and form
dynamic stationary phase.
Micelles migrate with or against the flow
depending on charge
8/15/2006 Chem208Spring2001 14
MECC - Micelles
Typical surfactants used
8/15/2006 Chem208Spring2001 15
MECC -
Micelles
Example of MECC
6
8/15/2006 Chem208Spring2001 16
CyclodextrinCE
Cyclodextrinsare basket shaped molecules
which can trap small molecules inside them.
8/15/2006 Chem208Spring2001 17
CyclodextrinCE
CDs available with many functional groups
Added to mobile phase to increase
separation dynamic mobile phases.
CDs are chiral so chiral separations are
possible
8/15/2006 Chem208Spring2001 18
Capillary Electro-
Chromatography
Silica particles packed in column extend
the Electro-osmotic effect across entire
column
Packing pumps mobile phase through
column - no pressure drop.
7
8/15/2006 Chem208Spring2001 19
CEC
Flat flow profile less
band broadening then
HPLC
8/15/2006 Chem208Spring2001 20
CEC
Column packingsbeing developed with
ODS bonded to silica but sufficient
silica surface to pump buffers.
Chiral separations-
Cyclodextrinand other additives
8/15/2006 Chem208Spring2001 21
Comparison of HPLC and
CEC
HPLC column length limited by high
pressure drop for small particle columns.
CEC has no such limitations.
CEC and CE devices can be built on microchips
8
8/15/2006 Chem208Spring2001 22
CE on a Chip
State of the art 96
parallel CE channels on a
chip
Detection
Laser fluorescence
MS
Separation in a few seconds
1
Chapter 28
Sampling
Sampling
Cant Analyze everything
Get Representative Sample
M&M lab results
Is target population homogeneous?
If yes random sampling
If no how do we get a representative
sample?
Mix
Sample different parts
Sampling
Grab Sample
What you did to get a chocolate or
fluoride sample
Composite Sample
Buy chocolate from many lots, mix them
together
In situ Sampling
Put a probe in the production line
2
How Much Sample
N= number of particles
P=probability of finding one
S=std. Dev. Of sampling
For M&Ms 27% or sample was RED
P=0.27 for s=.10 n= 270
Need 270 M&Ms to get 10% std. Dev.
2
1 1
s
x
p
p
n

=
Trace Components
P=0.14 1.4 % extraneous candy
S=0.10
N=7042 particles (M&Ms)
About 8 liters of candy
Good argument for grinding up
samples
Make homogeneous
Increase number of particles/unit weight
How many samples?
Sampling error related to square root
of number of samples
Error can be reduced by taking more
samples
4x more samples gives the error
3
Sampling Plan
Physically Obtain Sample
Gases Bottles and Bags pump or syringe
Solids shovels, corers, etc
Liquids Bottles, drum thief,
Sampling device must not add or remove
analyte
Teflon, Stainless Steel, borosilicate glass
Sampling Plan
Preserve Sample until analysis
Cool
Acidify
Storage Container Critical
Plastic for trace metals
Glass for trace organics
Teflon but very expensive
Cleaning 50% nitric acid soak, etc.
Sampling Plan
Problems with Preserving Samples
Microbial degradation
Adsorption of analytes on surfaces
Contamination by storage medium
Glass Sodium, Potassium, Aluminum, Calcium Boron,
Silicon
Plastic Plasticizers (primarily phthalates), UV
inhibitors, polymerization catalysts (Al)
Sample/Matrix/Analyte/Container
Dependent
4
Sampling Plan
Sample Preparation
Solids most difficult
Reduce particle Size
Grinders, mills, etc. must not contaminate
Get homogeneous Sample
Cone and quarter
Dissolving
Acid
Base
Fusion
Microwave Digestion
Recovery
R=C
A
/C
oA
What fraction of original concentration
is present at analysis?
Test with Spiked Samples Chocolate
Lab
Add known amount
R=C
R
-C
A
/C
oR
Difference between Spiked-Unspiked/amount
of spike

Bai Giang Chung PDF

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Bai Giang Chung PDF

Uploaded by

Copyright:

Available Formats

Chemistry 208

Rule of Thumb Keep the same

.03 * 100% =3% rel. unc.

.03 * 100% =3% rel. unc.

95% confidence is most commonly used.

WRITE A MASS BALANCE FOR NICKEL

And moles base added =moles acid

If we define Fraction Dissociated for the

WRITE A MASS BALANCE FOR NICKEL

moles CuEDTA dissociated = moles EDTA not reacted

8/15/2006 Chem 208 Spring 2001 32

Oxidized form on left will transfer electrons

8/15/2006 Chem 208 Spring 2001 24

8/15/2006 Chem 208 Spring 2001 41

E = .68 -.0592 log C

8/15/2006 Chem 208 Spring 2006 20

2 Eep = .68 + 1.44 - .0592 log [Ce

8/15/2006 Chem 208 Spring 2006 21

= 1.24 - .0592 log 2 [Cr

You might also like