Chemistry 311 Lecture Notes

Chemistry 311
Lecture Notes

Fall 2009

David E. Henderson
1
Chapter 2 & 3
How do we measure things
Chem 311
8/3/2009 Chem311Fall 2009 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.
8/3/2009 Chem311Fall 2009 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-
2
8/3/2009 Chem311Fall 2009 4
SIGNIFICANT FIGURES
KEEP ONLY THE FIRST UNCERTAIN
DIGIT IN A RESULT.
Assume that data is presented this way
by other responsible scientists.
8/3/2009 Chem311Fall 2009 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
8/3/2009 Chem311Fall 2009 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.
2-
3
8/3/2009 Chem311Fall 2009 7
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
8/3/2009 Chem311Fall 2009 8
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.
8/3/2009 Chem311Fall 2009 9
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
2-
4
8/3/2009 Chem311Fall 2009 10
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
8/3/2009 Chem311Fall 2009 11
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.
8/3/2009 Chem311Fall 2009 12
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.
2-
5
8/3/2009 Chem311Fall 2009 13
SIGNIFICANT FIGURES
The Correct Way
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 =
8/3/2009 Chem311Fall 2009 14
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
8/3/2009 Chem311Fall 2009 15
SIGNIFICANT FIGURES
EXAMPLE
21.1 * 0.029 * 83.2 =50.91008
Relative Uncertainty
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
2-
6
8/3/2009 Chem311Fall 2009 16
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.
8/3/2009 Chem311Fall 2009 17
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
8/3/2009 Chem311Fall 2009 18
CALCULATION OF MAXIMUM
EXPERIMENTAL ERROR -
Examples.
Multiplication and division
Molarity of titrant =grams acid*(mol / gm)/vol. titrant
M=.0646066 Mol/l
error = (
0.0001gm
.5675gm
)
2
+ (
.02gm/ mol
204.23 gm/ mol
)
2
+ (
0.014 ml
43.01 ml
)
2
ml 1000
l 1

ml 0.014 43.01
1
gm .02 204.23
mol 1
gm .0001 0.5675 M
=
2-
7
8/3/2009 Chem311Fall 2009 19
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
8/3/2009 Chem311Fall 2009 20
SIGNIFICANT FIGURES
Other Functions
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
=
=
8/3/2009 Chem311Fall 2009 21
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
2-
8
8/3/2009 Chem311Fall 2009 22
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
8/3/2009 Chem311Fall 2009 23
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
8/3/2009 Chem311Fall 2009 24
Stoichiometry Example
Anabuse
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
2-
9
8/3/2009 Chem311Fall 2009 25
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
8/3/2009 Chem311Fall 2009 26
Resolution
How finely can the quantity be measured
4 place balance - Resolution 0.0001 gm
Sartorius balance - Resolution 0.00001 gm
50 ml Buret - resolution 0.01 ml
Relative resolution depends on quantity
measured
8/3/2009 Chem311Fall 2009 27
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
2-
10
8/3/2009 Chem311Fall 2009 28
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
8/3/2009 Chem311Fall 2009 29
Linear Scales
Interpolation
Estimate position between smallest division
Bias in Interpolation reading
Look at Buret data from lab
8/3/2009 Chem311Fall 2009 30
Volume to Length Converters
Buret
Pipet
Graduate Cylinder
Sensitivity depends on inner diameter
length=V/pi r
2
Sensitivity =1/pi r
2
smaller r=greater sensitivity =more length
change per unit volume - Examples
2-
11
8/3/2009 Chem311Fall 2009 31
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
8/3/2009 Chem311Fall 2009 32
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
8/3/2009 Chem311Fall 2009 33
Digital Pipetters
Actual Data for 3 Fisher adjustable pipetters
0.106599 0.0501 0.0525 0.172 0.0273 0.224 CV
5.346494 0.5 0.5273 0.344 0.0548 0.045 SD
5015.5 997.9 1003.6 200.2 200.74 19.98 Average
5008.3 997.2 1003 200.4 200.7 20 5
5014.1 998 1003.5 200.5 200.7 20 4
5013.6 998.3 1003.1 199.9 200.7 20 3
5019.8 997.6 1004.1 199.7 200.8 20 2
5021.7 998.4 1004.1 200.3 200.8 19.9 1
5000 1000 1000 200 200 20 Sample Size
1000-5000 200-1000 "20-200 Pipette
2-
12
8/3/2009 Chem311Fall 2009 34
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
8/3/2009 Chem311Fall 2009 35
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
8/3/2009 Chem311Fall 2009 36
A/D - Discrete Data
Convert continuous function to discrete
function
Stairstep
Potentiometric titration
Sequence of discrete points
Sampling signal at specific times
Discrete Calculus
2-
13
8/3/2009 Chem311Fall 2009 37
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
8/3/2009 Chem311Fall 2009 38
Accuracy and Precision
Accurate and Precise Not Accurate but
Precise
Not Accurate and Not
Precise
Accurate but Not
Precise
8/3/2009 Chem311Fall 2009 39
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
2-
14
8/3/2009 Chem311Fall 2009 40
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.
8/3/2009 Chem311Fall 2009 41
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
8/3/2009 Chem311Fall 2009 42
Identification and Separation
Identification - Not part of 208
Chem211-2 Organic
Chem312 Instrumental
Separation - We spend lots of time on this
Remove Interferences
Quantitation of Mixtures
Identification of Mixtures
2-
15
8/3/2009 Chem311Fall 2009 43
Sample Preparation
See Chapter 28
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
8/3/2009 Chem311Fall 2009 44
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
8/3/2009 Chem311Fall 2009 45
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.
2-
16
8/3/2009 Chem311Fall 2009 46
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
8/3/2009 Chem311Fall 2009 47
Strange Techniques
Soxhlet Extraction -
Solid Phase Extraction-
Solid Phase Microextraction-
Headspace Sampling-
Purge and Trap analysis-
Microwave methods-
Sonication Methods-
8/3/2009 Chem311Fall 2009 48
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).
2-
17
8/3/2009 Chem311Fall 2009 49
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.
8/3/2009 Chem311Fall 2009 50
Using EXCEL
Excel powerful tool
Calculations
Graphing
Curve fitting
Learn to use it well
Format cells for clear data display
Learn to document your spreadsheet
Add equations and text to explain what you have done
Do your own!!
Include your name in the File Name
2-
1
8/3/2009 Chem311Fall 2009 1
Chem 311
Chapter 4 Lecture 1
Meaurement and Calculation
Accuracy and Precision
8/3/2009 Chem311Fall 2009 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 uncertainty
0004 . 0
40
014 . 0
23 . 204
02 . 0
8 . 0
0001 . 0
y undertaint
2 2 2
= |
.
|
\
|
+ |
.
|
\
|
+ |
.
|
\
|
=
8/3/2009 Chem311Fall 2009 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
4-
2
8/3/2009 Chem311Fall 2009 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
8/3/2009 Chem311Fall 2009 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.
8/3/2009 Chem311Fall 2009 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.
4-
3
8/3/2009 Chem311Fall 2009 7
Standard Deviation
Assume Gaussian Distribution of
Experimental Values
8/3/2009 Chem311Fall 2009 8
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 =

E
n
( x
i
)
n
8/3/2009 Chem311Fall 2009 9
Standard Deviation
Standard Deviation - s
x
(sample)
or o
x
(population >24)
Variance =square of standard deviation
V =s
x
2
or V =o
x
2
1
2
|
.
|
\
|

=

n
x x
s
n
i
x
n
x x
n
i
x
|
.
|
\
|

=
2
o
4-
4
8/3/2009 Chem311Fall 2009 10
Standard Deviation
Relative standard deviation.
% relative std. dev. =Coefficient of
Variation (CV)
% s
x
=
s
x
x
- 100 %
ppt s
x
=
s
x
x
- 1000 ppt
8/3/2009 Chem311Fall 2009 11
Significance of Standard
Deviation
One standard deviation is the distance from
the mean in which 69.3% of all values are
expected to lie.
8/3/2009 Chem311Fall 2009 12
Deviation
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.
4-
5
8/3/2009 Chem311Fall 2009 13
Deviation
99.8% within 3 std. dev.
As a general rule, virtually all experimental
data should lie in the range 4 s.
8/3/2009 Chem311Fall 2009 14
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.
8/3/2009 Chem311Fall 2009 15
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)
4-
6
8/3/2009 Chem311Fall 2009 16
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
=
8/3/2009 Chem311Fall 2009 17
Confidence Limits
Assumes there is no determinate error
present.
Different confidence levels include different
areas of the Gaussian distribution.
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
8/3/2009 Chem311Fall 2009 18
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.
n
s t
x
x n
=
4-
7
8/3/2009 Chem311Fall 2009 19
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.
n
s t
x
x n
=
8/3/2009 Chem311Fall 2009 20
Types of Errors
Random Errors - Indeterminate errors
Have no definite cause
randomerrors in reading a buret
Result in decreased precision and wider
confidence limits
8/3/2009 Chem311Fall 2009 21
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
4-
8
8/3/2009 Chem311Fall 2009 22
Types of Errors
Sampling Errors
Sample not representative of whole
M&M experiment
Method error
Interferences
Measurement Errors
Temperature not 20 deg for Volumetric flask
8/3/2009 Chem311Fall 2009 23
Types of Errors
Personal Errors
Analyst makes a mistake
8/3/2009 Chem311Fall 2009 24
Finding Errors
Constant errors
Balance reads 0.002 g high
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
4-
9
8/3/2009 Chem311Fall 2009 25
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.
8/3/2009 Chem311Fall 2009 26
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.
8/3/2009 Chem311Fall 2009 27
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!!!
4-
10
8/3/2009 Chem311Fall 2009 28
ELIMINATION OF
Q-test.
measure of the gap 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
8/3/2009 Chem311Fall 2009 29
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
8/3/2009 Chem311Fall 2009 30
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
If p<0.05 then the Null Hypothesis can be
rejected. Differences not due to randomerrors
4-
11
8/3/2009 Chem311Fall 2009 31
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?
Compare two data sets
F-Test
Compare standard deviations for data sets or methods
8/3/2009 Chem311Fall 2009 32
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
8/3/2009 Chem311Fall 2009 33
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
4-
12
8/3/2009 Chem311Fall 2009 34
2.364624 t Critical two-tail
0.00142 P(T<=t) two-tail
1.894579 t Critical one-tail
0.00071 P(T<=t) one-tail
5.086709 t Stat
7 df
0 Hypothesized Mean Difference
11 7 Observations
7.65E-12 4.19E-11 Variance
1.51E-06 1.47E-05 Mean
Machine Natural
Data for [H+] of Snow Samples
t-Test: Two-Sample Assuming Unequal Variances
8/3/2009 Chem311Fall 2009 35
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
8/3/2009 Chem311Fall 2009 36
Stream pH
Average pH data
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7
8/1/2004 9/20/200
4
11/9/200
4
12/29/20
04
2/17/200
5
4/8/2005 5/28/200
5
7/17/200
5
9/5/2005 10/25/20
05
12/14/20
05
Date
p
H
Off Mt
On Mt
4-
13
8/3/2009 Chem311Fall 2009 37
2.364624 t Critical two-tail
0.032981 P(T<=t) two-tail
1.894579 t Critical one-tail
0.016491 P(T<=t) one-tail
2.649175 t Stat
7 df
0 Hypothesized Mean Difference
0.929994 Pearson Correlation
8 8 Observations
8.53E-14 1.67E-12 Variance
3.87E-07 1.35E-06 Mean
Coleman Giant
[H+] for two Streams Nov-May
t-Test: Paired Two Sample for Means
4-
1
8/3/2009 Chem311 1
Chapter 4 Part 2
Calibration, Standardization
8/3/2009 Chem311 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
8/3/2009 Chem311 3
Standard Solutions
Weigh Primary Std. Into Volumetric Flask
Dilute to mark only after all dissolved
Do not heat
Mix at least 10 times
4 part 2-
2
8/3/2009 Chem311 4
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
8/3/2009 Chem311 5
Calibration Methods
Single Point
External Standard Method Calibration
Curve
Internal Standard Method
Standard Addition Method
8/3/2009 Chem311 6
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
4 part 2-
3
8/3/2009 Chem311 7
External Standard Method
Using a Standard Curve
Known concentrations vsSignal
Intercept need not be Zero
Identify non-linearity if present
8/3/2009 Chem311 8
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.
8/3/2009 Chem311 9
Chromatography
make calibration curve
run sample and compare
Limited by injection accuracy
2-4% manual
1-2% auto
0.5% valve injections
4 part 2-
4
8/3/2009 Chem311 10
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
8/3/2009 Chem311 11
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
8/3/2009 Chem311 12
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.
4 part 2-
5
8/3/2009 Chem311 13
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
8/3/2009 Chem311 14
Computing Least Squares
Excel -
Make XY scatter Graph
Click on data in graph
Right click Select Add Trendline
Options -
display equation
display R squared
8/3/2009 Chem311 15
Excel Getting values for Calculations
=Slope(yvalues, xvalues) b
o
=Intercept(yvalues, xvalues) b
1
=RSQ(yvalues,xvalues) R
2
All Values are Live and update changes
4 part 2-
6
8/3/2009 Chem311 16
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
Results Dead and dont update changes
8/3/2009 Chem311 17
Does line go through ZERO
SE
y
>=y intercept
Y-intercept lies within 1 SD of 0
Repeat Calculation and select Force Zero
Intercept
SE
y
<y intercept
Use full equation fromLeast Squares
Be sure to use SE
y
not overall SE
8/3/2009 Chem311 18
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
4 part 2-
7
8/3/2009 Chem311 19
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.
8/3/2009 Chem311 20
Excel Least Squares
A Closer look
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
8/3/2009 Chem311 21
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
4 part 2-
8
8/3/2009 Chem311 22
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.
8/3/2009 Chem311 23
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
8/3/2009 Chem311 24
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
4 part 2-
9
8/3/2009 Chem311 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.
8/3/2009 Chem311 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
8/3/2009 Chem311 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
4 part 2-
10
8/3/2009 Chem311 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
8/3/2009 Chem311 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.
8/3/2009 Chem311 30
Confidence Limit for unknown
x true
ts x x
4 part 2-
11
8/3/2009 Chem311 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.
8/3/2009 Chem311 32
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
8/3/2009 Chem311 33
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: (See Text fopr
EXCEL Instructions)
y=a +bx+cx
2
Calculation of Unknown gives 2 values
Quadratic Solution Required
4 part 2-
12
8/3/2009 Chem311 34
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 Linear Range of the
Method
Dont drop points from the middle
4 part 2-
1
8/3/2009 Chem311 1
Chapter 5
QA & Calibration
8/3/2009 Chem311 2
QA/QC
Critical part of any analysis
Does the method work?
Instruments OK?
Reagents OK?
Is the analyst doing a good job
How much confidence in result?
8/3/2009 Chem311 3
Important terms
Sensitivity Slope of Calibration
Selectivity Do other things give a
response?
False Positive
PSA Test for Prostate Cancer
False Negative
Mammograms (X-Ray)
5-
2
8/3/2009 Chem311 4
Range of Method
Limit of Detection
Limit of Quantitation
Limit of Linearity
Limit of response
Range statistics vary depending on many
factors R
2
=0.995 in text is rarely met
8/3/2009 Chem311 5
Limit of Detection
8/3/2009 Chem311 6
Limit of Detection (LOD)
S
lod
=S +z sigma
Z=3 gives 99.86% confidence that
substance is present
S=mC +b from Least Squares
C
lod
=(S
lod
b)/m
<1% chance of false positive
5-
3
8/3/2009 Chem311 7
Quantitation Limit
S
loq
=S +10 sigma
Gives quantitation with 10% rel. error
C
loq
=(S
loq
b)/m
1 sigfig quantitative result possible
8/3/2009 Chem311 8
Limit of Linearity
Data deviates by 10% from Least SQ line
Usually Negative deviation so results from
calibration are high
No mathematical way to determine this
Interpolate residuals to find 10%
Requires enough points to get good value
Compare linear and quadratic fit
8/3/2009 Chem311 9
Precision
Repeated analysis of same homogeneous
material Instrument precision
Repeated sample preparation of same
sample Method precision
Precision between instruments and analysts
Method Ruggedness
Precision between labs
InterlaboratoryPrecision
5-
4
8/3/2009 Chem311 10
Accuracy
Run SRMs similar matrix
Use two independent methods
Spike sample with know amount - Recovery
Compare Standard Addition with
Calibration result
8/3/2009 Chem311 11
Recovery Study
Validate sample preparation/extraction etc
Run Sample
Run Sample with known addition
8/3/2009 Chem311 12
Standard Addition
Useful if matrix of sample has background
signal which cannot be accounted for in a
blank or calibration curve
Quick analysis if only one or two samples
are to be run
5-
5
8/3/2009 Chem311 13
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 signal for dilution.
8/3/2009 Chem311 14
Graphical treatment of std.
addition
Graph Signal vsSpike Concentration
X-Intercept=-C
A
V
o
/V
f
8/3/2009 Chem311 15
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
5-
6
8/3/2009 Chem311 16
Analysis Methods
eliminates injection error in Chromatography
- only integration error is left
Eliminates need for careful sample workup in
many methods (Chocolate Lab)
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
8/3/2009 Chem311 17
Data fromGC
experiment
16:0
y=0.8809x+0.0549
R
2
=0.9999
0.0000
2.0000
4.0000
6.0000
8.0000
10.0000
0 5 10 15
Mass Ratio
A
r
e
a

R
a
tio
20:0
y=0.9287x+0.0426
R
2
=0.9999
0.0000
1.0000
2.0000
3.0000
4.0000
5.0000
6.0000
7.0000
0 2 4 6 8
Mass Ratio
A
re
a
R
a
tio
8/3/2009 Chem311 18
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)
5-
7
8/3/2009 Chem311 19
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
8/3/2009 Chem311 20
Blanks
Complicated issue
Reagent Blank Add all reagents at their final
concentrations to the solvent
Method Blank Carry a matrix matched blank
sample through the entire method
Field Blank Create a blank when samples are
collected to include all environmental
conditions
8/3/2009 Chem311 21
Control Chart
Monitor QA over time
Hg Analyzer
Run 100 ul of 1 ppmeach day as first sample
Put result on graph
Monitor changes
5-
1
8/3/2009 Chem311 1
Chem 311
Chapter 6
Chemical Equilibrium
8/3/2009 Chem311 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
8/3/2009 Chem311 3
Equilibrium constant
Free Energy change is related to chemical
potential
Note difference from Eq6.2
Ratio of activities to Std. St.
Dimensionless
Q RT G G
o
ln + A = A
b
o
b
b
a
o
a
a
d
o
d
d
c
o
c
c
Q
(
=
o
o
o
o
o
o
o
o
6-
2
8/3/2009 Chem311 4
At Equilibrium
eq
o
K RT G ln = A
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
(
=
o
o
o
o
o
o
o
o
8/3/2009 Chem311 5
Normally write using Concentrations
(6-2)
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 =
8/3/2009 Chem311 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
+
= =
6-
3
8/3/2009 Chem311 7
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.
8/3/2009 Chem311 8
Precipitation Reactions
PbI
2(s)
Pb
+2
+2I
-
K
sp
=[Pb
+2
][ 2I
-
]
Also need to consider
PbI
+
, PbI
3
-
, and PbI
4
2-
Simple equilibriumgives wrong answers
8/3/2009 Chem311 9
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
6-
4
8/3/2009 Chem311 10
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)
8/3/2009 Chem311 11
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
8/3/2009 Chem311 12
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
6-
5
8/3/2009 Chem311 13
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
8/3/2009 Chem311 14
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).
8/3/2009 Chem311 15
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
6-
6
8/3/2009 Chem311 16
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.
8/3/2009 Chem311 17
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
]
8/3/2009 Chem311 18
Add reactions multiply Ks
CH
3
COOH +H
2
O +<===> CH
3
COO
-
+H
3
O
+
K
a
CH
3
COO
-
+ H
2
O <===> CH
3
COOH + OH
-
K
b
+ add two reactions
2 H
2
O <===>H
3
O
+
+ OH
-
Solvent Autoprotolysis
Equilibrium constant K
w
=K
a
x K
b
6-
7
8/3/2009 Chem311 19
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.
8/3/2009 Chem311 20
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
8/3/2009 Chem311 21
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
6-
8
8/3/2009 Chem311 22
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
8/3/2009 Chem311 23
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
8/3/2009 Chem311 24
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)
6-
9
8/3/2009 Chem311 25
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
8/3/2009 Chem311 26
Coordination Complexes
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
8/3/2009 Chem311 27
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
]
6-
10
8/3/2009 Chem311 28
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
8/3/2009 Chem311 29
General Approach Complicated
Equilibrium Systems
Calculate the fraction of each form in an
equilibrium under specified conditions
o will be used to identify fraction
o
Ni2+
=fraction of total Ni in the Ni
2+
form
o
HA
=fraction of weak acid (HA) in
undissociatedform.
o
A-
=fraction of weak acid (HA) in dissociated
form.
8/3/2009 Chem311 30
Species present
Three techniques
Calculate fraction in each form- alpha
Log Concentration plots (computer)
Line diagrams (qualitative)
6-
11
8/3/2009 Chem311 31
Species present at a pH
Calculate alphas
HA <==> H
+
+A
-
C
HA
=[HA] +[A] Mass balance
Define o fraction of total in specific form
HA
A
HA
HA
C
A
C
HA ] [

] [

= = o o
8/3/2009 Chem311 32
Substitute values from K
a
C
HA
=[HA] +[A]
HA
A
HA
HA
C
A
C
HA ] [

] [

= = o o
] [
] [
] [
] ][ [
] [
'
' +
+
= =
H
HA K
A
K
A H
HA
a
a
8/3/2009 Chem311 33
For monoproticacid - Exact equation
'
'
'
] [
] [
] [
a
a
A
a
HA
K H
K
K H
H
+
=
+
=
+
+
+
o
o
6-
12
8/3/2009 Chem311 34
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
+ + +
=
+ + +
=
+ + +
=
+ + +
+
+ + +
+
+ + +
+
o
o
o
8/3/2009 Chem311 35
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
+ + +
=
+ + +
=
+ + +
=
+ + +
+
+ + +
+
+ + +
+
o
o
o
8/3/2009 Chem311 36
Plot o 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
6-
13
8/3/2009 Chem311 37
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
8/3/2009 Chem311 38
Using Fraction Titrated
Avoids problem of figuring out
stoichiometry
Avoids need to recognize species present
Pick pH and calculate fraction titrated
Only way to do spreadsheets
8/3/2009 Chem311 39
6-
14
8/3/2009 Chem311 40
6-
1
Chem 311
Chapter 8
Activity
Systematic Equilibrium
8/3/2009 Chem311 2
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.
8/3/2009 Chem311 3
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
8-
2
8/3/2009 Chem311 4
ACTIVITY see Chapter 8
Activity coefficient
Finagle constant x x x
C o =
8/3/2009 Chem311 5
ACTIVITY
Problem of definition. What are the
conditions where f=1.00000 ? The value
of K
eq
will depend on how this is
chosen.
8/3/2009 Chem311 6
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.
8-
3
8/3/2009 Chem311 7
Concentration Effects
Concentration increases
Ions get close enough to be attracted and
repelled by other ions
All ions are solvated (Solvationnumber)
Very High concentration
Run out of solvent for hydration
Activity coefficients range widely
8/3/2009 Chem311 8
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
8/3/2009 Chem311 9
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
8-
4
8/3/2009 Chem311 10
Cut to the Chase
Ignore Activity Coefficients in most work
Cant get themaccurately
Places limit on absolute accuracy of all
equilibriumcalculations
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.
Large errors due to lack of dissociation
8/3/2009 Chem311 11
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 fromtheory
8/3/2009 Chem311 12
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
8-
5
8/3/2009 Chem311 13
Systematic Approach to
Equilibrium Problems
If exact answers are needed
Systematic Set of steps to take to solve a
problem Any problem
Use Acid Base as example
8/3/2009 Chem311 14
Acid Base Equilibria
Step 1. Write all chemical reactions and
their equilibrium constant expressions.
8/3/2009 Chem311 15
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-
6
8/3/2009 Chem311 16
Step 2 Write additional equations defining the system One
Charge Balance
CHARGE BALANCE Solution is electrically neutral
Sum [+ions] =Sum [- ions]
[H
+
] = [BA
-
] + [OH
-
]
rearrange
[BA
-
] = [H
+
] - [OH
-
]
Phosphoric acid /potassium phosphate equilibrium
8/3/2009 Chem311 17
MASS BALANCE - Conservation of
matter
Multiple equations for complex systems
C
HBa
=[HBA] +[BA
-
]
8/3/2009 Chem311 18
Step 3 Use Algebra to simplify things and remove
unknowns
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-
7
8/3/2009 Chem311 19
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
8/3/2009 Chem311 20
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
0 ] [ ] [ ] [ ] [
2 3
=
+ + + +
a w w HA a a
K K H K C H K H K H
8/3/2009 Chem311 21
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-
8
8/3/2009 Chem311 22
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
8/3/2009 Chem311 23
Monoprotic base
Two ways to solve - Approximate
More Exact
0 ] [ ] [
2
= +

b b b
K C K OH OH
b b
C K OH =
] [
8/3/2009 Chem311 24
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-
9
8/3/2009 Chem311 25
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
)
8/3/2009 Chem311 26
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
8/3/2009 Chem311 27
Very complicated
Acid base component
Coordination complex component
Multiple species
Lots of Algebra fun
Identifying all Chemical Reactions often
hardest part
8-
1
8/3/2009 Chem311 1
Chem 311
Chapter 9
MonoproticAcid-Base
8/3/2009 Chem311 2
Calculation pH of anything
Six cases to address
Strong Acid/Base >10
-6
M
Strong Acid/Base <10
-6
M
Weak Acid
Acid or conjugateacid of weak base
Weak Base
Baseof conjugatebaseof weak acid
Buffer
Acid plus its conjugatebase
Amphiproticsubstance- polyproticsystems Chapter 10
8/3/2009 Chem311 3
Strong Acid/Base
Completely dissociated
[H
+
] =C
HA
[OH
-
] =C
B
If Conc>1x10
-6
M then dissociation of
H
2
O can be ignored
9-
2
8/3/2009 Chem311 4
Strong Acid/Base
If Conc. <1x10
-6
M then dissociation of
H
2
O must be included
Acids never >pH 7
Bases never <pH 7
Quadratic solution from Systematic
treatment
[H
+
]
2
+C
HA
[H
+
] K
w
=0
8/3/2009 Chem311 5
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
8/3/2009 Chem311 6
Buffers
Resist Change in pH
Added acid
Added base
dilution
Very important in all chemistry
Acid +conjugate base
9-
3
8/3/2009 Chem311 7
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
8/3/2009 Chem311 8
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
+
]
8/3/2009 Chem311 9
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
+
]}
9-
4
8/3/2009 Chem311 10
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
8/3/2009 Chem311 11
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
8/3/2009 Chem311 12
Acidic buffer
Simplify
If Concentrations of Acid and Salt are high
C >>[H
+
]
C >>{[H
+
]-[OH
-
]}
Drop those terms
HA
A
a
C
C H
K

+
=
] [
'
9-
5
8/3/2009 Chem311 13
Acidic buffer
Take -log of equation
Henderson-Hasselback
HA
A
a
C
C H
K

+
=
] [
'
|
|
.
|
\
|
+ =
|
|
.
|
\
|
=
|
|
.
|
\
|
=
|
|
.
|
\
|
=
+
+
HA
A
a
HA
A
a
HA
A
a
HA
A
a
C
C
pK pH
C
C
pH pK
C
C
H pK
C
C H
K
log
log
log ] log[
] [
log log
'
8/3/2009 Chem311 14
Solution to Ammonia example
Base/Acid
NH
3
is base (A-)
NH
4
+
is Acid (HA)
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
8/3/2009 Chem311 15
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
9-
6
8/3/2009 Chem311 16
Buffer Capacity
Maximum at pH =pK
Useable in range pH=
pK 2
9-
1
8/3/2009 Chem311 1
Chem 311
Chapter 10
Poly-proticsystems
8/3/2009 Chem311 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
8/3/2009 Chem311 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
Additional dissociation =K
2
10-
2
8/3/2009 Chem311 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
]
8/3/2009 Chem311 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
8/3/2009 Chem311 6
Amphiprotic substances
Both acids and base reactions
Intermeditate forms
HC0
3-
H
2
P0
4,
HP0
4
2-
Amino acids (e.g.alanine)
weak acid - weak base salts
ammonium acetate
ammonium formate
10-
3
8/3/2009 Chem311 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
8/3/2009 Chem311 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-
]
8/3/2009 Chem311 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
-
]
10-
4
8/3/2009 Chem311 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-
]
8/3/2009 Chem311 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
8/3/2009 Chem311 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 ] [
] [
] ][ [
] [
] ][ [
] [ ] [
] [
] ][ [
] [
] [
] [
+ = +
+ =
|
|
.
|
\
|
+
+ = +
+ =
+ =
+
+
+
+
+
+
+
+
+
10-
5
8/3/2009 Chem311 13
NaH malate example
Rigorous equation
Problem
Dont know equilibrium concentration till we
solve equation
=
+
] [H
[H
+
] =
K
a1
(K
a2
[HMal
] + K
w
)
K
a1
+ [HMal
]
8/3/2009 Chem311 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
]
8/3/2009 Chem311 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
]
10-
6
8/3/2009 Chem311 16
NaH malate example
Equation independent of concentration
Works at high concentration only
Use for Amino Acids
[H
+
] = K
a1
K
a2
8/3/2009 Chem311 17
NaH malate example
For NaHMal
[H
+
] = (4.0x10
-4
x 8.9 x 10
-6
)
1/2
=5.97 x
10
-5
[H
+
] = K
a1
K
a2
8/3/2009 Chem311 18
NaH malate example
What if assumptions dont work
0.0005 M NaHMal
[HMAl] ~ K
A1
Calculate approximate value using
simple form
10-
7
8/3/2009 Chem311 19
NaH malate example
[H
+
] = =5.97 x 10
-5
o
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
o
8/3/2009 Chem311 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
o
( )
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
8/3/2009 Chem311 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
10-
8
8/3/2009 Chem311 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
8/3/2009 Chem311 23
Summary
Have to be able to tell what is present
Stoichiometry
Recognize acids and bases
8/3/2009 Chem311 24
Fractional forms
See previous lecture notes
Alpha calculations
10-
1
8/3/2009 Chem311 1
Chem 311
Chapter 11
Titration
8/3/2009 Chem311 2
Simple Titrations
No Equilibrium Chemistry to worry about
Strong Acid-Strong Base
Anything with really huge K
eq
Stoichiometry determines curve
8/3/2009 Chem311 3
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
+
=
11-
2
8/3/2009 Chem311 4
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
8/3/2009 Chem311 5
pH vs Volume
Note rapid change at 15 ml
0
1
2
3
4
5
6
7
8
0 5 10 15 20
Series2
8/3/2009 Chem311 6
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
+
=
11-
3
8/3/2009 Chem311 7
Simple Titration Curve
0
2
4
6
8
10
12
14
0 5 10 15 20 25 30
Series2
8/3/2009 Chem311 8
Titration Calculations
Moles Analyte =moles Titrant (x
stoichiometry factor from reaction)
C
a
V
a
=C
t
V
t
(Stoichiometry factor)
8/3/2009 Chem311 9
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
11-
4
8/3/2009 Chem311 10
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
8/3/2009 Chem311 11
Visual Indicators
http://inst.santafe.cc.fl.us/~chem/intbl.html
8/3/2009 Chem311 12
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
11-
5
8/3/2009 Chem311 13
End Points
At end of Titration Stoichiometry
produces very large changes in the relative
amount of analyte for small additions of
titrant.
8/3/2009 Chem311 14
Decision Tree for Weak
Acid/Base 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
8/3/2009 Chem311 15
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
11-
6
8/3/2009 Chem311 16
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
8/3/2009 Chem311 17
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
8/3/2009 Chem311 18
Practice Problem
25.0 ml base
H
2
P acid remaining =0 mmoles
Amphiproticignore dilution
18 . 4
10 6 . 6 10 90 . 3 10 12 . 1 ] [
5 6 3
=
= =
+
pH
x x x x H
11-
7
8/3/2009 Chem311 19
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
8/3/2009 Chem311 20
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
8/3/2009 Chem311 21
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
11-
8
8/3/2009 Chem311 22
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
8/3/2009 Chem311 23
Using Fraction Titrated
Avoids problem of figuring out
stoichiometry
Avoids need to recognize species present
Pick pH and calculate fraction titrated
Only way to do spreadsheets
8/3/2009 Chem311 24
11-
9
8/3/2009 Chem311 25
11-
1
8/3/2009 Chem 311 1
Chem 311
Chapter 12
Complexometric Titration
8/3/2009 Chem 311 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
]
8/3/2009 Chem 311 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
12-
2
8/3/2009 Chem 311 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
8/3/2009 Chem 311 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
1
1 +
1
[L
] +
2
[L
]
2
+
3
[L
]
3
+...
n
[L
]
n
8/3/2009 Chem 311 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
12-
3
8/3/2009 Chem 311 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.
8/3/2009 Chem 311 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
8/3/2009 Chem 311 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
12-
4
8/3/2009 Chem 311 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
8/3/2009 Chem 311 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.
8/3/2009 Chem 311 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
12-
5
8/3/2009 Chem 311 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-
8/3/2009 Chem 311 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
8/3/2009 Chem 311 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
+
.
12-
6
8/3/2009 Chem 311 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
8/3/2009 Chem 311 17
Control pH to Control [Y]
8/3/2009 Chem 311 18
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
12-
7
8/3/2009 Chem 311 19
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.
8/3/2009 Chem 311 20
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
8/3/2009 Chem 311 21
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
12-
8
8/3/2009 Chem 311 22
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
8/3/2009 Chem 311 23
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
8/3/2009 Chem 311 24
Example
K
f
' = 1.6x10
8
= [CuEDTA] = 0.00500
[Cu2+ ]C
EDTA
[Cu
2+
]
2
[Cu
2+
] = 5.5x10
-6
pCu = 5.26
12-
9
8/3/2009 Chem 311 25
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
8/3/2009 Chem 311 26
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
8/3/2009 Chem 311 27
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.
12-
10
8/3/2009 Chem 311 28
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
8/3/2009 Chem 311 29
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
8/3/2009 Chem 311 30
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
] [
] [
] [

12-
11
8/3/2009 Chem 311 31
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+
8/3/2009 Chem 311 32
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
8/3/2009 Chem 311 33

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
.
12-
12
8/3/2009 Chem 311 34
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.
8/3/2009 Chem 311 35
End Point Detection
Abs.
Vol. Titrant
Cu titration with EDTA
8/3/2009 Chem 311 36
End Point Detection
Abs.
Vol. Titrant
Cu and Fe titration with EDTA
Fe E.P.
Cu E.P.
12-
13
8/3/2009 Chem 311 37
End Point Detection
Electrochemical.
Endpoints have been monitored by change in
pH
mercury electrode
ion selective electrodes.
8/3/2009 Chem 311 38

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.
8/3/2009 Chem 311 39
Murexide
N
N O
O
N
O
N
N
-O
O
O
12-
14
8/3/2009 Chem 311 40
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
8/3/2009 Chem 311 41
Calmagite
N,N,O,O tetradentate complexes
S
O
O
O
OH
N
N
HO
8/3/2009 Chem 311 42
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
12-
15
8/3/2009 Chem 311 43
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.
8/3/2009 Chem 311 44
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.
8/3/2009 Chem 311 45
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.
12-
16
8/3/2009 Chem 311 46
Back Titration
Used when complex formation is slow
Add excess EDTA
Titrate with Mg
2+
or Mn
2+
8/3/2009 Chem 311 47
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
8/3/2009 Chem 311 48
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.
12-
17
8/3/2009 Chem 311 49
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
8/3/2009 Chem 311 50
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.
8/3/2009 Chem 311 51
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
12-
18
8/3/2009 Chem 311 52
Applications
Spot Tests
Test Strips
Flow Injection Analysis
Immunoassays
12-
1
8/3/2009 Chem 311 1
Chem 311
Chapter 14
Electrochemistry
8/3/2009 Chem 311 2
Oxidation - Reduction
Electrons transferred coulomb (q)
1 coulomb = 1 amp for 1 sec
Electricity
q= nF coulombs = number of electrons x F
F= 96,500 electrons/coulomb
G= -nFE = -Eq
8/3/2009 Chem 311 3
Ohms Law
E=IR
Voltage = current(amps)x resistance (ohms)
14-
2
8/3/2009 Chem 311 4
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
8/3/2009 Chem 311 5
Chemical Analysis
A variety of chemical analyses involve
electrochemical cells.
Potentiometry 311
voltametry 312
amperometry
chronopotentiometry
coulometry
others
8/3/2009 Chem 311 6
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
14-
3
8/3/2009 Chem 311 7
Electron Transfer
Fermi level of
electrons in electrodes
shown
Fe
2+
filled orbital
Ce
4+
empty orbital
8/3/2009 Chem 311 8
Electron Transfer
Potential is difference
in energy of two
electrodes.
Measure with no
electron flow
Thermodynamic
potential
G = -nFE
8/3/2009 Chem 311 9
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
14-
4
8/3/2009 Chem 311 10
Potentiometer Circuit
Salt Bridge
Voltage Follower
Amp
Voltmeter
8/3/2009 Chem 311 11
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.
8/3/2009 Chem 311 12
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
14-
5
8/3/2009 Chem 311 13
Line Notation 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.
8/3/2009 Chem 311 14
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
8/3/2009 Chem 311 15
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)
14-
6
8/3/2009 Chem 311 16
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.
8/3/2009 Chem 311 17
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.
8/3/2009 Chem 311 18
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.
14-
7
8/3/2009 Chem 311 19
Tables of Std.Potentials
Appendix - 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)
8/3/2009 Chem 311 20
8/3/2009 Chem 311 21
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

14-
8
8/3/2009 Chem 311 22
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 will occur
8/3/2009 Chem 311 23
Calculate EMF of CELL R
X
E
cell
= E
Fe,Fe2+
- E
Sn
2+
,Sn
4+
0.151 (-0.447) = 0.598 volts
E
cell
= 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.
8/3/2009 Chem 311 24
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+
= 0.771
E
Fe3+,Fe2+
= 0.771
Mn0
4
-
+ 8H
+
+ 5e- ====> Mn
2+
+ 4H
2
0
E
Mn04-,Mn2+
= 1.51
14-
9
8/3/2009 Chem 311 25
Calculate E
1/2
using Nerst -
As oxidation
E
Fe
3+,
Fe
2+ =
E
o
- 0.0591 log [Fe
2+
]
1 [Fe
3+
]
E = 0.771 - 0.0591 log 0.100 = 0.771 + 0.0178
1 0.200
= 0.789 volts
8/3/2009 Chem 311 26
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
8/3/2009 Chem 311 27
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 -(0.789) = + 0.747
Note Potential of 5 Fe
2+
= 5Fe
3+
same
as for 1
14-
10
8/3/2009 Chem 311 28
Methods of Analysis
Direct Potentiometry
Nerst Eq. activity measurements
Calibration Curve conc. Measurements
Standard Addition
Potentiometric Titration
14-
1
8/3/2009 Chem 311 1
Chem 311
Chapter 15
Potentiometry
8/3/2009 Chem 311 2
Reference Electrodes
Ag/AgCl reference electrode is also
popular. 0.2222 v
Most common
No Hg
8/3/2009 Chem 311 3
Reference Electrodes
Saturated Calomel electrode SCE
Hg,Hg
2
Cl
2
/ Hg
2
+2
(sat.), KCl (sat.)// E =
+0.2415 volts
15-
2
8/3/2009 Chem 311 4
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 -
8/3/2009 Chem 311 5
Junction Potentials
K+ and Cl
Best choice
8/3/2009 Chem 311 6
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
See Rainwater example in text
15-
3
8/3/2009 Chem 311 7
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
8/3/2009 Chem 311 8
Indicator Electrodes
Electrodes of the 2nd kind (probe a
secondary reactant)
Ag/AgCl - potentiometric titration of Cl
Quinone/hydroquinone pH electrode
8/3/2009 Chem 311 9
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
15-
4
8/3/2009 Chem 311 10
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
8/3/2009 Chem 311 11
Glass Membrane Electrode
Need
solutions for
electrical
contact with
membrane
Dry Glass
Hydrated gel
External
Solution
Internal
Solution
8/3/2009 Chem 311 12
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
15-
5
8/3/2009 Chem 311 13
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)
8/3/2009 Chem 311 14
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
,
|
8/3/2009 Chem 311 15
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
,
|
15-
6
8/3/2009 Chem 311 16
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
8/3/2009 Chem 311 17
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
8/3/2009 Chem 311 18
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.
15-
7
8/3/2009 Chem 311 19
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
8/3/2009 Chem 311 20
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
8/3/2009 Chem 311 21
Liquid Ion Exchange
Membranes
Ion Pair Formation
Tetraalkyl ammonium salts
Alkyl amines
1,10 phenanthroline Fe (III) salts
15-
8
8/3/2009 Chem 311 22
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
8/3/2009 Chem 311 23
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)
8/3/2009 Chem 311 24
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
-
.)
15-
9
8/3/2009 Chem 311 25
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
8/3/2009 Chem 311 26
Shape of
curve
Slope=0.0592/z
Solubility limit
Log_Concentration
E
8/3/2009 Chem 311 27
Heterogenous membranes
Silicone polymer + precipitate
of some insoluble salt.
PVC
Coated wire electrodes
Tungsten wire - treated to
become pH electrode
15-
10
8/3/2009 Chem 311 28
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
8/3/2009 Chem 311 29
Enzyme Electrodes
pH Electrode
Enzyme in
acrylamide gel
ISE, pH, or gas electrode
Urease <===>NH
3
Decarboxylazes===>H
+
2 enzyme sandwich -
cholesterol electrode
8/3/2009 Chem 311 30
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
15-
11
8/3/2009 Chem 311 31
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 - sensor array
spoiled food, ripe fruit, mine detection
8/3/2009 Chem 311 32
Semiconductor Electronics
Silicon (4 e) doped with
Al 3 e p-type
P 5 e n-type
8/3/2009 Chem 311 33
Diode
Allows electron flow only np
15-
12
8/3/2009 Chem 311 34
FET
2 PN Junctions
8/3/2009 Chem 311 35
Analysis using FET
8/3/2009 Chem 311 36
FET and MOS-FET
Sensors for Solution
Sensors for vapors
Made in arrays 10x10 etc
Each FET with different Chemical sensor
Multiple sensitivities
15-
13
8/3/2009 Chem 311 37
Problems with ISE's
Slow response (minutes) for Liquid
Junction
Short life ISE's months
Enzymes days or hours
High resistance - good pH meters required
8/3/2009 Chem 311 38
Methods of Application of
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
8/3/2009 Chem 311 39
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.
15-
14
8/3/2009 Chem 311 40
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
8/3/2009 Chem 311 41
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
8/3/2009 Chem311 42
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
15-
15
8/3/2009 Chem311 43
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.
8/3/2009 Chem311 44
Clinical Analyzers
Electrochem on a Chip
Disposable
ISEs
8/3/2009 Chem311 45
Resurgence of ISEs
New Techniques
Ppb level analysis
Pb in drinking water < 5 ppb
Add EDTA to filling
Free metal very low
Prevent transfer to analyte
15-
1
8/3/2009 Chem311 1
Chem 311
Chapter 18
Fundamentals of
Spectrometry
8/3/2009 Chem311 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
8/3/2009 Chem311 3
q =Refractive index = speed in vac
speed in medium
q>1.00
function of wavelength
light of different energy moves at different rates
in mediumother than vacuum.
Fiber Optics separate wavelengths of a pulse
18-
2
8/3/2009 Chem311 4
u (wave number) =waves/cm=cm
-1
= 1/
in vacuum
Frequency (v ) =waves passing fixed
point/sec.
period (p) time per wave crest v =1/p
all are related v =c

E =h v =hc/ =hcu
8/3/2009 Chem311 5
Electromagnetic spectrum
8/3/2009 Chem311 6
Interaction of matter and
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
18-
3
8/3/2009 Chem311 7
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.)
8/3/2009 Chem311 8
Molecular Energy Levels
E H A orbitals
Bonding and Antibonding orbitals
CAChe and Spartan calculate these
8/3/2009 Chem311 9
Molecular Energy Levels
Computation chemistry
Compute energy of all MOs
Predict UV-Vis spectra
Predict IR Spectra
Predict NMR Spectra
18-
4
8/3/2009 Chem311 10
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.
8/3/2009 Chem311 11
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
8/3/2009 Chem311 12
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
18-
5
8/3/2009 Chem311 13
Absorption
log(P
0
/P) =k/2.303 Cb =c bC or (abC)
P = Transmittance (T)
P
o
-log T =A Absorbance - A is dimensionless
A =abc C in gm/l
A= cbc C in moles/l
bC =cm*mol/1000 cm
3
=mol/1000 cm
2
a units cm
2
/gm c unit =cm
2
/mol
(old literature often dm
2
/gm)
8/3/2009 Chem311 14
Limitations on Beers Law
Light must be monochromatic
Parallel
Enter at a right angle.
8/3/2009 Chem311 15
Accuracy
Deviations from Beers Law
Instrumental Deviations
Non-monochromatic light
value of c or a not constant across bandwidth of
spectrometer.
Negative deviation at high conc.
Concentration error
lower sensitivity.
Need more standards.
A
Conc.
18-
6
8/3/2009 Chem311 16
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)
8/3/2009 Chem311 17
Bandwidth effects
8/3/2009 Chem311 18
Bandwidth effects
18-
7
8/3/2009 Chem311 19
Chemical Deviations
Equilibria - acid base pH control
Activity coef.
Temperature
Solvent effects - concentration changes
dielectric constant of solution
Refractive Index change due to Conc.
8/3/2009 Chem311 20
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
8/3/2009 Chem311 21
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)
18-
8
8/3/2009 Chem311 22
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)

8/3/2009 Chem311 23
Error as f(%T)
Working Range
very short
factor of 5-10
%T
error
36.8%T
8/3/2009 Chem311 24
18-
1
8/3/2009 Chem311 1
Chem 311
Chapter 19
Applications of
Spectrometry
8/3/2009 Chem311 2
Applications of
Spectrophotometry
Direct determination of a chromophoric
compound
anything that absorbs strongly.
Absorptivities range from0 to 500,000 , wide
range of sensitivities.
8/3/2009 Chem311 3
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.
19-
2
8/3/2009 Chem311 4
Methods of Quantitation
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
Subtract Blank or Zero instrument with Blank
8/3/2009 Chem311 5
Methods of Quantitation
Standard Addition Method
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

) (
8/3/2009 Chem311 6
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
19-
3
8/3/2009 Chem311 7
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.
8/3/2009 Chem311 8
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
8/3/2009 Chem311 9
Spectrophotometry to study
reaction stoichiometry
Used for - metal complexes, enzyme
substrate complexes, etc.
Continuous variation - J obs Method
Use where ratio is close to 1:1
Make series of solutions where Total moles of
two reactants constant.
Plot Mole ratio vsA
19-
4
8/3/2009 Chem311 10
Jobs Method
8/3/2009 Chem311 11
Scatchard Plot
Measure Equilibrium Constant
Biochemists
8/3/2009 Chem311 12
Scathcard Plot
19-
5
8/3/2009 Chem311 13
Scatchard Plot
Slope =- K
K=4.0x10
9
8/3/2009 Chem311 14
Instrumentation for Optical
Spectroscopy
Detector
Source of
monochromatic
radiation
Sample Readout
8/3/2009 Chem311 15
Sources of Radiation
Black body radiators -
Tungsten lamp 2870
o
K - 1.5 micron peak
visible only - 330 nmminimum
19-
6
8/3/2009 Chem311 16
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
8/3/2009 Chem311 17
Dispersion by a Grating
8/3/2009 Chem311 18
Definition of effective bandwidth
Wavelength Selectors
19-
7
8/3/2009 Chem311 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
8/3/2009 Chem311 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
8/3/2009 Chem311 21
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
19-
8
8/3/2009 Chem311 22
Detectors
Solid State Detectors
Photodiodes
Change Couple Devices.
Photodiodes - pnjunction conduct in
reverse direction due to photon flux.
8/3/2009 Chem311 23
Detectors
Linear photodiode Arrays
512 diodes - detect 512 wavelengths at once -
complete spectrumnot scanned. HP
Spectrometer
Good visibility sensitivity
Rapid response
high linearity
.
8/3/2009 Chem311 24
Instrument Designs
Photodiode array spectrophotometer
19-
9
8/3/2009 Chem311 25
Fiber Optic Dip Probes
No cell required
Mirror reflects
8/3/2009 Chem311 26
Flow Injection Analysis
Inject pulse of sample with valve
Mix with reagents
Pass through detector cell
8/3/2009 Chem311 27
FIA
Calibration Data
Autoanalyzers
Common in Clinical
19-
10
8/3/2009 Chem311 28
ELISA
8/3/2009 Chem311 29
ELISA
Requires specific antibody for each analyte
Antibody bound to substrate
Magnetic particles
Solid supports
Etc
y=-0.6456Ln(x) - 0.8658
R
2
=0.9886
-1.000
-0.500
0.000
0.500
1.000
1.500
2.000
0.01 0.1 1
Conc. (ug/L)
l
o
g
i
t

%

B
/
B
o
19-
1
8/3/2009 Chem 311 1
Chem 311
Chapter 23
Separations
General Chromatography
8/3/2009 Chem 311 2
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.
8/3/2009 Chem 311 3
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
23-
2
8/3/2009 Chem 311 4
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
8/3/2009 Chem 311 5
SEPARATIONS AND
EXTRACTION
Separation necessary when measurement
technique not adequately selective
Contact between two immiscible phases
Separation of phases
8/3/2009 Chem 311 6
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.
23-
3
8/3/2009 Chem 311 7
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
8/3/2009 Chem 311 8
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.
8/3/2009 Chem 311 9
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)
23-
4
8/3/2009 Chem 311 10
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
8/3/2009 Chem 311 11
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
8/3/2009 Chem 311 12
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
23-
5
8/3/2009 Chem 311 13
Ps and qs
Special case V
R
= 1
q = 1 p = D
1+D 1+D
% Extracted = 100p
8/3/2009 Chem 311 14
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
8/3/2009 Chem 311 15
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
23-
6
8/3/2009 Chem 311 16
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
8/3/2009 Chem 311 17
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
8/3/2009 Chem 311 18
Example
Extraction of Acetanilide from water into eth
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
p
T
= 1 - q
aq
or c
V
V D
e q
23-
7
8/3/2009 Chem 311 19
Multiple extractions
In general N = 5 is optimum gets within a factor of
5 of max. possible efficiency.
8/3/2009 Chem 311 20
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
8/3/2009 Chem 311 21
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
23-
8
8/3/2009 Chem 311 22
Vanilla Experiment
Vanillin is neutral molecule
Partitions into organic phase with high D
Add Base
De-protonated form of vanillin is anion
Soluble in aqueous phase
anion form not soluble in organic phase
Very efficient separation
Selective for neutral molecules with acidic
functional group
8/3/2009 Chem 311 23
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 o =
8/3/2009 Chem 311 24
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.
23-
9
8/3/2009 Chem 311 25
Simplest Metal Ext. Case
b
n
M
n+
+ nL
-
<===> ML
n(Aq]
+H+ K
pML
K
A
ML
N(or)
HL
(Aq)
K
pHL
HL
(or)
8/3/2009 Chem 311 26
Distribution
D = [ML]
or
[M+]
Aq
+ [ML]
Aq
b
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]
8/3/2009 Chem 311 27
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
23-
10
8/3/2009 Chem 311 28
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
(
(
=
+
+
] [
]
] [ ] [ |
8/3/2009 Chem 311 29
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
8/3/2009 Chem 311 30
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
23-
11
8/3/2009 Chem 311 31
Metal Distribution
Multidentate
ligands give
sharper pH
extraction
breaks
n=1
n=2
n=3
pH
% Ext.
8/3/2009 Chem 311 32
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
8/3/2009 Chem 311 33
A little History
Solvent Extraction Research - Alternative to
Precipitation
Craig Counter Current Extraction
Chromatography - Continuous Extraction
23-
12
8/3/2009 Chem 311 34
CHROMATOGRAPHY AND
SEPARATIONS
Problems with difficult separations-
Counter current extraction was slow.
Tswett Russian/Italian worked in Russian
Poland
Russian Japanese war and WW I problems
Made enemies of leaders of field before publication
Finally published in 1906 German Botanical
Chromatography
Color-writing or tsvet-writing
8/3/2009 Chem 311 35
CHROMATOGRAPHY AND
SEPARATIONS
Lederer's work in 1930's
Germany - also lost.
AJP Martin - Silica gel
has surface layer of bound
water.
Packed column with silica and
passed CHCl
3
through (mobile
phase) worked for fatty acids.
8/3/2009 Chem 311 36
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
23-
13
8/3/2009 Chem 311 37
Elution chromatography - most
common
Solute placed at top of column.
Eluted until each component reaches the
end (detector).
8/3/2009 Chem 311 38
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
8/3/2009 Chem 311 39
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
Vel = length/t
o
10 cm length must contain 1.00 ml of mobile phase
Vol mobile phase = t
o
x Flow Rate
23-
14
8/3/2009 Chem 311 40
Elution chromatography
Total column volume 4.6 mm diameter
x 10 cm
V
t
= pr
2
h = 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 F = V
s
/V
m
= .66/1.0 = 0.66
8/3/2009 Chem 311 41
time ratio in phases
K = Cs partition ratio
C
m
= K F Thermodynamic basis for
retention
mobile
stationary
mobile
stationary
mobile
stationary
V
V
x
C
C
T
T
=
8/3/2009 Chem 311 42
Capacity factor k' - Describes Retention
Most common for HPLC
time in stationary phase/time in mobile phase
k' varies from 0 to large numbers

ln k' = D H - D S - ln F
RT R
o
r
o
o r
o
o r
t
t
t
t t
V
V V
k
'
' =
=
23-
15
8/3/2009 Chem 311 43
ln k' = D H - D S - ln F
RT R
Vant Hoff Plots
Plots of ln k vs 1/T linear
show contribution of entropy and enthalpy
8/3/2009 Chem 311 44

Separation Factor
Relative Retention = t
r2
/ t
r1
= k
2
/k
1
8/3/2009 Chem 311 45
Band spreading in
chromatography
Inject Sample as a very narrow band -
square wave pulse of sample
Three Causes of Band Spreading

2
Injector
+
2
Column
+
2
Detector
23-
16
8/3/2009 Chem 311 46
Band spreading in
chromatography
Broadening in column
As it moves through column, it begins to
spread -
o = std. dev. of peak profile
Number of theoretical plates N
N= t
r
2
/ o
2
peak width at base t
w
= 4 o
8/3/2009 Chem 311 47
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
8/3/2009 Chem 311 48
Band spreading in
chromatography
HETP = L/N
Plate Theory ===> van Deemter Equation
H= A + B/v + Cv
where v= mobile phase velocity
23-
17
8/3/2009 Chem 311 49
Band spreading in
chromatography
H= A + B/v + Cv
Three Components
A-Eddy diffusion
B-Longitudinal
diffusion
C-Rate of mass
transfer between
phases
8/3/2009 Chem 311 50
Band spreading in
chromatography
Longitudinal Diffusion of solutes
D 10
-5
to 10
-7
function of MW
8/3/2009 Chem 311 51
Band spreading in
chromatography
Mass Transport between phases
23-
18
8/3/2009 Chem 311 52
Band spreading in
chromatography
Multiple Flow Paths - Eddy Diffusion
Not present in Open Tubular Columns
8/3/2009 Chem 311 53
HETP 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
8/3/2009 Chem 311 54
HPLC vs GC
Differences due to
diffusion rates in liquids vs gases
particle sizes
LC
GC
1E-5 1e-4 1e-3 1e-2 1e-1 1 10 100
velocity cm/sec
H
23-
19
8/3/2009 Chem 311 55
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
-5
to 10
-7
D
m
10
-2
to 10
-1
8/3/2009 Chem 311 56
Very High Flow
Prev. Discussion Assumes Laminar Flow
Turbulent Flow Conditions
Faster Mass Transport
HETP Graph comes back down
Not as far as Laminar minimum
Useful for high speed sample prep
8/3/2009 Chem 311 57
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
23-
20
8/3/2009 Chem 311 58
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
8/3/2009 Chem 311 59
Deviant Chromatographic
Behavior
Non-linear
partition
isotherms
Ideal behavior
Anti-Langmuir
Isotherm
Langmuir
Isotherm
Conc
Stationary
phase
Conc mobile phase
8/3/2009 Chem 311 60
Behavior
Anti-
langmuir
sample acts
as
stationary
phase
Langmuir
Saturate
sites
Anti-langmuir case
Langmuir case
23-
21
8/3/2009 Chem 311 61
Behavior
Anti-Langmuir
8/3/2009 Chem 311 62
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
8/3/2009 Chem 311 63
Resolution in Chromatography
R= Dt
r
/4o = Dt
r
/W
av
= (t
r2
- t
r1
)/0.5(W
1
+ W
2
)
most common equation for determining R
from chromatogram
23-
22
8/3/2009 Chem 311 64
Resolution
8/3/2009 Chem 311 65
R =
1
4

(o 1)
o
k'
1 + k'
N
o =
k'
2
k'
1
= separation factor
8/3/2009 Chem 311 66
3 Ways to control separation
Change relative affinity of solutes for stationary
phase Column Selectivity ()
change stationary phase
change form of solute ( charge or size)
change temperature
Change pH
Change overall retention by changing k'
Change temperature in GC
change mobile phase in LC
Change N
flow rate
change column - particle size or length
23-
23
8/3/2009 Chem 311 67
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
8/3/2009 Chem 311 68
General Elution Problem
Single set of
conditions good
for only 6-8
components
8/3/2009 Chem 311 69
Capillary GC
of FAMES
Isothermal GC at
150 degrees
Note Tr separation
between even Cs
almost doubles
23-
24
8/3/2009 Chem 311 70
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
8/3/2009 Chem 311 71
Capillary GC of FAMES
Temperature Programmed run
8/3/2009 Chem 311 72
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
23-
25
8/3/2009 Chem 311 73
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
8/3/2009 Chem 311 74
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 quaternary gradients possible
Chocolate lab - Methanol, acetonitrile better than
either one alone
DryLab Software for optimization
8/3/2009 Chem 311 75
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.
23-
26
8/3/2009 Chem 311 76
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)
8/3/2009 Chem 311 77
Problems with programmed
elution
Time to return to initial conditions
GC - cool column oven
LC- equilibrate column
SFC - density gradient - re-equilibrate
8/3/2009 Chem 311 78
Frontal Chromatography
Mixture used
as mobile
phase
Great if you
want Solute 1
only
23-
27
8/3/2009 Chem 311 79
Displacement
Chromatograph
y Great for Preperative
Separations
Gram Quantities on
standard columns
Must have displacer
Must determine
isotherms
8/3/2009 Chem 311 80
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
Solute elutes from column
Affinity Columns - specific biological interaction -
antibody-antigen
Pre-concentration- pass large volume through
Sample Cleanup remove interferences
8/3/2009 Chem 311 81
Solid Phase Extraction
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
23-
1
8/3/2009 Chem311 1
Chem 311
Chapter 24
Gas Chromatography
8/3/2009 Chem311 2
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
8/3/2009 Chem311 3
Types of Columns
Capillary columns
steel, glass, fused silica.
.067 - 1 mmID
1000 -100 plates/ft
Columns may be very long due to low pressure
required
Typical 10 meters - 60 meters (GC Lab 30 meter)
30,00010
6
plate possible
100,000 typical
24-
2
8/3/2009 Chem311 4
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
8/3/2009 Chem311 5
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
8/3/2009 Chem311 6
Solid Supports for GC
Adsorption on solid surface - Strongest
stationary phase interaction
Useful for permanent gases
GSC silica, graphite, firebrick
1000 m
2
/gmsurface
2-5 gmin a column
As vapor pressure decreases need less
retention.
Minimizing Solid Support Interactions
24-
3
8/3/2009 Chem311 7
Solid Supports for GC
Inertsupports with low surface area
0.2 - 1 m
2
/gmsurface
Diatomaceous earths - high surface area
silica
Acid wash remove metals
Calcine reduce surface
silanize bond surface hydroxyl groups
Coat with liquid phase 1000 available
8/3/2009 Chem311 8
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
8/3/2009 Chem311 9
Split/Splitless
Injector
Constant Pressure
Split vent electrical valve
Expansion of solvent
Inert liner
quartz
silanizedglass
24-
4
8/3/2009 Chem311 10
Injection
Syringe Injection 0.1 to 10 ul +/- 2-3%
Gas samples can be up to 2 ml
Valve injection gases
SPME other desorptiontechniques
Headspace analysis
Purge and Trap
Cold Trapping
8/3/2009 Chem311 11
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
8/3/2009 Chem311 12
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
24-
5
8/3/2009 Chem311 13
Ionization Detectors- Flame
ionization
Sample +
Hydrogen
200 volt DC
Electrometer Flame
Electrode
8/3/2009 Chem311 14
Ionization Detectors- Flame
ionization
Used for C-H compounds.
Gramresponse 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
8/3/2009 Chem311 15
ECD Electron Capture Detector
b e
-
+carrier establish background standing
current
Beta
source
Sample
Power
Supply - DC
or Pulsed
Electrometer
-
+
24-
6
8/3/2009 Chem311 16
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
-
+
8/3/2009 Chem311 17
halogenated samples best pesticides PCB's
CHC1
3
1 x 10
-15
gm/sec
peaks for 11-
14
to 10
-12
gm
Linear range 300-10000 depends on
design
Sensitivity high due to high ionization Eff.
100%
Beta
source
Sample
Power
Supply - DC
or Pulsed
Electrometer
-
+
8/3/2009 Chem311 18
GC-MS
EI or CI ionization
EI sensitivity like FID
EI M +e
-
M
+.
+2 e
-
M
+
. D
+
+ D.
Fragments Daughter ions
Structural Information
Library Search
24-
7
8/3/2009 Chem311 19
GC-MS
CI Chemical Ionization
CH
4
+e
-
CH
5
+
CH
5
+
+M MH
+
+CH
4
High efficiency ionization
Few Fragment ions
Much higher sensitivity
No Library Search possible
8/3/2009 Chem311 20
GC-MS
Selected Ion Monitoring
Find characteristic ion in each spectrum
Produce chromatogramof specific ion
Total Ion Chromatogram
All ions in each spectrumsummed
MS-MS
Fragment a Fragment
Improved S/N ratio
8/3/2009 Chem311 21
Detector Summary
Non-destructive 10,000 10
-9
gm/sec
10
-7
gmin peak
TCD
Identification
and
quantitation
100,000 10
-12
to 10
-
15
gm/sec
Mass
Spectrometer
S FPD
N or P NPD
Cl and F
molecules
300-10,000 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
24-
8
8/3/2009 Chem311 22
Photo ionization
7-12 eV H
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
8/3/2009 Chem311 23
ThermonicDetector
N or P specific FID
alkali metal FID
Bead of CeI placed in flame of FID
Enhances the sensitivity to N and P
8/3/2009 Chem311 24
Flame photometric
Sulfur specific
Flame with photodetector
Useful foanalysis of fuels
24-
9
8/3/2009 Chem311 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
8/3/2009 Chem311 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
8/3/2009 Chem311 27
Identification of Peaks
Mass Spectrometry
Molecular weight of molecules
Fragmentation pattern for Identification
Library search 300,000 spectra available
Trace analysis sensitivity
GC-IR
IR spectrumof peaks
24-
10
8/3/2009 Chem311 28
Analysis Methods
make calibration curve
run sample and compare
Limited by injection accuracy
2-4% manual
1-2% auto
0.5% valve injections
8/3/2009 Chem311 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 vsmass ratio
8/3/2009 Chem311 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)
24-
1
8/3/2009 Chem 311 1
Chem 311
Chapter 25
HPLC
8/3/2009 Chem 311 2
A little History
8/3/2009 Chem 311 3
Types of particles
A4
25-
Slide 3
A4 LC/GC April 2006 p 10
Administrator, 11/13/2006
25-
2
8/3/2009 Chem 311 4
Columns
Small particles
More pressure
Harder to pack.
Pressure required increases
P=k/d
p
2
Small particles reduce C term in HETP
8/3/2009 Chem 311 5
A5
8/3/2009 Chem 311 6
Effect of particle size
Xanthines
A6
25-
Slide 5
A5 LC/GC December 2005 p 1251
Slide 6
A6 LC/GC April 2006 p 10
25-
3
8/3/2009 Chem 311 7
Columns
Smaller particles ==>faster separation
1.5 ml/min x 8 min=12 ml
1.5 ml/min x 1.6 min =2.4 ml
Same separation in 20% of time using
20% of the solvent
8/3/2009 Chem 311 8
8/3/2009 Chem 311 9
9 components in < 30 sec
Alkylphenones
25-
4
8/3/2009 Chem 311 10
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
8/3/2009 Chem 311 11
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
8/3/2009 Chem 311 12

Polar Force
Normal phase LC
Silica or Aluminum solid absorbent
Si0
2
Al
2
0
3
25-
5
8/3/2009 Chem 311 13
Polar Force Normal Phase
8/3/2009 Chem 311 14
Dispersive Forces -Reverse
Phase London forces & Van der Waals
Induced dipoles
Reversed phase LC
R =C
18
H
37
C
8
H
17
C
3
H
6
NH
2
C
4
H
9
8/3/2009 Chem 311 15
Reverse Phase
Elutropic Series Weak solvent
Water lowest eluting power
Acetonitrile
Methanol
THF
CH
2
Cl
2
Great eluting power
Non-aqueous reverse phase
25-
6
8/3/2009 Chem 311 16
Reverse Phase
Solvent Triangle
ACN, MeOH, THF
Binary and ternary mixtures
Lab MeOH + ACN better than either alone
Computer Optimization Map space for best
separations
Drylab Software Run 4 Chromatograms
Predict most effects of parameter changes
8/3/2009 Chem 311 17
Reverse Phase
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
8/3/2009 Chem 311 18
HPLC Instrument
Solvent Supply
Pump
Constant flow
Gradient or Mixture
Injector
Valve
Column
Readout
Computer
Integrator
Strip Chart
Detector
25-
7
8/3/2009 Chem 311 19
Pumps
Single piston- cheap - need pulse
damper
Dual piston - most common
8/3/2009 Chem 311 20
Pumps
Screw piston- small bore column
Air pressure - dissolved bubbles a
problem
Isocratic and Gradient Systems
Solvent Mixing pumps
3 o4 4 Solvent inlets
Valves proportion in solvent
Mixes in pump body
8/3/2009 Chem 311 21
Solvent Preparation
Bubbles due to cavitations
Bubbles in detector
Helium Sparge (LDC in lab)
Vacuum Degasser (Thermo - LC/MS)
Intelligent pumps (Hitachi in lab)
Filter mobile phases to eliminate particles
Cause damage to check valves
25-
8
8/3/2009 Chem 311 22
HPLC Instrument
Injector Valve
8/3/2009 Chem 311 23
Capillary Columns
Capillary HPLC
1mm ID column loosely packed with large
particles
fused silica capillary coated with liq. phase
typical separation 0.1 - 24 hrs.
10
6
10
7
plates
8/3/2009 Chem 311 24
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 -
25-
9
8/3/2009 Chem 311 25
AbsorptionUV - Vis
Flow Cell in Spectrometer
Diode Array for full spectrum
10-
12
gm/sec sensitivity best case
Absorptivity100 - 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
Linear range 3000+
8/3/2009 Chem 311 26
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
8/3/2009 Chem 311 27
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
25-
10
8/3/2009 Chem 311 28
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
8/3/2009 Chem 311 29
Other Detectors
Evaporative Light
Scattering
Sample sprayed
into chamber
Solute particles
scatter light
Universal
detector - non-
volatile solutes
8/3/2009 Chem 311 30
Other Detectors
LC-MS
Same advantages as GC-MS
Identification
Very high sensitivity
Very high selectivity
Electrospray (ESI)
Polar molecules better
APCI
Non-polar molecules better
25-
11
8/3/2009 Chem 311 31
Comparison of Detectors
8/3/2009 Chem 311 32
Peak integration
Drop perpendicular
Most common method
Simple
Small peak + error
Large Peak error
8/3/2009 Chem 311 33
Peak integration
Valley Method
Always produces errors
Errors can be large
Useful for
Multiple peaks
Complex baseline
25-
12
8/3/2009 Chem 311 34
Peak integration
Skim Method
Exponential Skim
Linear Skim
R<1
Small peak < 5% of large
8/3/2009 Chem 311 35
Peak integration
Gaussian Skim
More Complicated
Less error than other skim methods
8/3/2009 Chem 311 36
Peak integration
Errors depend
Relative Peak areas
Resolution
Method used
Improve Resolution!!!
25-
13
8/3/2009 Chem 311 37
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
+
8/3/2009 Chem 311 38
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
8/3/2009 Chem 311 39
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
25-
14
8/3/2009 Chem 311 40
Ion Chromatograph
8/3/2009 Chem 311 41
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.
8/3/2009 Chem 311 42
Ion chromatography
Cr(VI) (Erin Brockovich)
Toxic form of Cr
ICP Total
Cr
3+
good
25-
15
8/3/2009 Chem 311 43
Ion chromatography
Cation Analysis
Soils
Water
8/3/2009 Chem 311 44
Ion chromatography
Anions
Water pollution
Soil Analysis
8/3/2009 Chem 311 45
Stearic Exclusion Ch. 26
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
25-
16
8/3/2009 Chem 311 46
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
8/3/2009 Chem 311 47
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
8/3/2009 Chem 311 48
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
25-
17
8/3/2009 Chem 311 49
Steric Exclusion
Calibrate V
R
vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
25-
1
8/3/2009 Chem311 1
Chem 311
Chapter 26
IC, GPC and CE
8/3/2009 Chem311 2
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
+
8/3/2009 Chem311 3
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
26-
2
8/3/2009 Chem311 4
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
8/3/2009 Chem311 5
Ion Chromatograph
8/3/2009 Chem311 6
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.
26-
3
8/3/2009 Chem311 7
Ion chromatography
Cr(VI) (Erin Brockovich)
Toxic formof Cr
ICP Total
Cr
3+
good
8/3/2009 Chem311 8
Ion chromatography
Cation Analysis
Soils
Water
8/3/2009 Chem311 9
Ion chromatography
Anions
Water pollution
Soil Analysis
26-
4
8/3/2009 Chem311 10
Reagent Free IC
Electrochemical Generation of mobile
phase
8/3/2009 Chem311 11
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
8/3/2009 Chem311 12
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
26-
5
8/3/2009 Chem311 13
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
8/3/2009 Chem311 14
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
8/3/2009 Chem311 15
Steric Exclusion
Calibrate V
R
vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
26-
6
8/3/2009 Chem311 16
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
8/3/2009 Chem311 17
Capillary Electrophoresis
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
8/3/2009 Chem311 18
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.
26-
7
8/3/2009 Chem311 19
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.
8/3/2009 Chem311 20
Electroosmoticflow
V
eof
=u
eof E
k layer thic double * charge wall *
viscosity * 4
* cons dielectric
k Zeta
pi
Zeta
u
eof
8/3/2009 Chem311 21
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
26-
8
8/3/2009 Chem311 22
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
8/3/2009 Chem311 23
Instrument Diagram
8/3/2009 Chem311 24
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
26-
9
8/3/2009 Chem311 25
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 - add absorbing species to buffer
and look at vacancies.
8/3/2009 Chem311 26
Example of CE
DNA fragments
8/3/2009 Chem311 27
MECC - Micelles
Micelles trap neutral organics and form
dynamic stationary phase.
Micelles migrate with or against the flow
depending on charge
26-
10
8/3/2009 Chem311 28
MECC - Micelles
Typical surfactants used
8/3/2009 Chem311 29
MECC -
Micelles
Example of MECC
8/3/2009 Chem311 30
CyclodextrinCE
Cyclodextrinsare basket shaped molecules
which can trap small molecules inside them.
26-
11
8/3/2009 Chem311 31
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
8/3/2009 Chem311 32
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.
8/3/2009 Chem311 33
CEC
Flat flow profile less
band broadening then
HPLC
26-
12
8/3/2009 Chem311 34
CEC
Column packingsbeing developed with
ODS bonded to silica but sufficient
silica surface to pump buffers.
Chiral separations-
Cyclodextrinand other additives
8/3/2009 Chem311 35
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
8/3/2009 Chem311 36
CE on a Chip
State of the art 96
parallel CE channels on a
chip
Detection
Laser fluorescence
MS
Separation in a few seconds
26-

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Chemistry 311

Rule of Thumb Keep the same

95% confidence is most commonly used.

WRITE A MASS BALANCE FOR NICKEL

WRITE A MASS BALANCE FOR NICKEL

moles CuEDTA dissociated = moles EDTA not reacted

8/3/2009 Chem 311 33

8/3/2009 Chem 311 38

Oxidized form on left will transfer electrons

k' varies from 0 to large numbers

8/3/2009 Chem 311 44

8/3/2009 Chem 311 12

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