Chem

Chemistry 311
Lecture Notes
Fall 2009
David E. Henderson
Chem 311
Chapter 2 & 3
How do we measure things
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
Chem 311 Fall 2009
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
8/3/2009
Chem 311 Fall 2009
2-
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
Chem 311 Fall 2009
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
Chem 311 Fall 2009
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.
8/3/2009
Chem 311 Fall 2009
2-
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 Chem 311 Fall 2009
8/3/2009
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
Chem 311 Fall 2009
Common error
1.234*(40.35-40.31)=0.04936
How many significant figures should we
have?
1.234*0.04= 0.04936
0.05
8/3/2009
Chem 311 Fall 2009
2-
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
Chem 311 Fall 2009
10
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.
8/3/2009
Measured value - real

The significant figures which denote the
uncertainty of the value are those in the
mantissa.
Chem 311 Fall 2009
11
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

9.1x10-2 < [H+] < 9.5x10-2
9.5x10-14
13.02 < pH < 13.04

9.1x10-14 < [H+] <
Note that the relative uncertainties are the

same.+/- 2/93 = 2% rel.unc.
8/3/2009
Chem 311 Fall 2009
12
2-
SIGNIFICANT FIGURES
The Correct Way
Uncertainty of sum or difference
y abc
s y sa sb2 sc2
2
40.34 ml +/- .01 ml

-1.03 ml +/- .01 ml
39.31 ml +/-0.014 ml
8/3/2009
0.0002 0.014
Chem 311 Fall 2009
13
SIGNIFICANT FIGURES
MULTIPLYING OR DIVIDING
UNCERTAINTY OF THE RESULT IS
THE SUM OF THE RELATIVE
UNCERTAINTIES.
a *b
y
c
2
8/3/2009
s s s
s y a b c
a b c 14
Chem 311 Fall 2009
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
.0047 2 + .034 2 + .0012 2 0. 034

.03 * 100% = 3% rel. unc.
8/3/2009
Chem 311 Fall 2009
15
2-
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
Chem 311 Fall 2009
16
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 0.01 0.014 ml

2
8/3/2009
Chem 311 Fall 2009
17
CALCULATION OF MAXIMUM
EXPERIMENTAL ERROR Examples.
Multiplication and division
Molarity of titrant = grams acid*(mol / gm)/vol. titrant
M 0.5675 .0001 gm
1 mol
1
1l
204.23 .02 gm 43.01 0.014 ml 1000 ml
M=.0646066 Mol/l
error
8/3/2009
0.0001gm 2
.02gm mol
0.014 ml 2
) (
)2 (
)
.5675gm
204.23 gm mol
43.01 ml
Chem 311 Fall 2009
18
2-
CALCULATION OF MAXIMUM
EXPERIMENTAL ERROR Examples.
error
0.0001gm 2
.02gm mol
0.014 ml 2
) (
)2 (
)
.5675gm
204.23 gm mol
43.01 ml
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
8/3/2009
Chem 311 Fall 2009
19
SIGNIFICANT FIGURES
Other Functions
Partial Derivative Calculus- how does y change
with small changes in the value
of the measured quantity a
y ax
sy
s
x a
y
a
8/3/2009
y log(a)
sa
sy
2.303a
y 10 a
sy
2.303sa
y
Chem 311 Fall 2009
20
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- H2O
Measure OH with buret
Moles OH = moles H
8/3/2009
Chem 311 Fall 2009
21
2-
Stoichiometry
Conservation of Mass
Mass of each atom is conserved in all Reactions
Types of Reactions
Acid Base = Protons Conserved

Redox = electrons conserved
Complexation = electron pairs conserved
Precipitation = Charge conserved
8/3/2009
Chem 311 Fall 2009
22
Stoichiometry
Conservation of Mass
C4H10 + O2 4 CO2 + 5 H2O
Moles CO2 = 4 x moles C4H10
Moles H2O = 5 x moles C4H10
2 x Moles H2O = 10 x moles C4H10
8/3/2009
Chem 311 Fall 2009
23
Stoichiometry Example
Anabuse
C10H20N2S4 + oxidizing agent 4 SO2

SO2 + H2O2 H2SO4
H2SO4 + 2 NaOH 2 H2O
Measure moles of NaOH (buret)
moles NaOHx
1molH 2 SO4
1molSO2
1molAnabuse
x
x
2molNaOH 1molH 2 SO4
4molSO2
mol Anabuse
Overall Ratio 1:8

8/3/2009
Chem 311 Fall 2009
24
2-
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
Chem 311 Fall 2009
25
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
Chem 311 Fall 2009
26
Relative resolution uncertainty of measurement

1.0000 gm weighed on the balance
0.0001/1.0000 * 1000 = 0.1 ppt
150.0000 gm weighed on same balance

0.0001/150.0000 * 1,000,000 = 0.7 ppm
0.0100 gm
0.0001/0.0100 * 1000 = 10 ppt
30.00 ml titrant
0.01/30.00 * 1000 = 0.3 ppt
8/3/2009
Chem 311 Fall 2009
27
2-
Conversion devices
Quantity being measured
Transducer
Sensor
Thermistor
Temperature probe
Intermediate conversion device
Serial Interface Box

12 Bit A/D
Readout Conversion
Data Logger on PC
Calibrated T readout
8/3/2009
Chem 311 Fall 2009
28
Linear Scales
Interpolation
Estimate position between smallest division
Bias in Interpolation reading

Look at Buret data from lab
8/3/2009
Chem 311 Fall 2009
29
Volume to Length Converters
Buret
Pipet
Graduate Cylinder
Sensitivity depends on inner diameter
length=V/pi r2
Sensitivity = 1/pi r2
smaller r= greater sensitivity = more length
change per unit volume - Examples
8/3/2009
Chem 311 Fall 2009
30
2-
10

Pipet Two 10 ml pipets one has 6 mm ID
and the other has 3 mm ID
Sensitivity = 1/pi r2
1/3.1415x32 = 0.0354
1/3.1415x 1.52 = 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
Chem 311 Fall 2009
31
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
Chem 311 Fall 2009
32
Digital Pipetters
Actual Data for 3 Fisher adjustable pipetters
Pipette
"20-200
Sample Size
8/3/2009
20
200-1000
200
1000-5000
200
1000
1000
5000
19.9
200.8 200.3
1004.1
998.4
5021.7
20
200.8 199.7
1004.1
997.6
5019.8
20
200.7 199.9
1003.1
998.3
5013.6
20
200.7 200.5
1003.5
998
5014.1
20
200.7 200.4
1003
997.2
5008.3
Average
19.98
200.74 200.2
1003.6
997.9
5015.5
SD
0.045
0.0548 0.344
0.5273
0.5
5.346494
CV
0.224
0.0273 0.172
0.0525
0.0501
0.106599
Chem 311 Fall 2009
33
2-
11
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 212 = 4096
Serial Interface 0-5 volts ==> 0 to 4096

Resolution 5 volts/4096 = 0.0012
volts/digit
8/3/2009
Chem 311 Fall 2009
34
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 vs 24 bit
8/3/2009
Chem 311 Fall 2009
35
A/D - Discrete Data

Convert continuous function to discrete
function
Stairstep
Potentiometric titration
Sequence of discrete points
Sampling signal at specific times
Discrete Calculus
8/3/2009
Chem 311 Fall 2009
36
2-
12
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
Chem 311 Fall 2009
37
Accuracy and Precision

Not Accurate and Not
Precise
Accurate but Not

Precise
Accurate and Precise
8/3/2009
Not Accurate but

Precise
Chem 311 Fall 2009
38
Accuracy
ACCURACY - This is how close the
experimental value is to the "true value"
if this is known
Only measurable if you know true
value of measured quantity
Estimate using statistics - more later
Measure using Standard Reference
Materials
8/3/2009
Chem 311 Fall 2009
39
2-
13
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
Chem 311 Fall 2009
40
Control Chart Example

Control Chart
Result
1.4
1.2
1
0.8
Data
0.6
0.4
-2 stdev
+ 2 stdev
0.2
0
0
10
Experiment Day
8/3/2009
Chem 311 Fall 2009
41
Identification and Separation

Identification - Not part of 208
Chem 211-2 Organic
Chem 312 Instrumental
Separation - We spend lots of time on this

Remove Interferences
Quantitation of Mixtures
Identification of Mixtures
8/3/2009
Chem 311 Fall 2009
42
2-
14
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
Chem 311 Fall 2009
43
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
Chem 311 Fall 2009
44
Solids - A Few Components

Are they the volatile components.
Headspace analysis, Solid Phase
microextraction, or purge and trap.
Non-volatile components
separation is necessary by boiling, soxhlet
extraction, sonication, microwave
digestion, supercritical fluid extraction.
8/3/2009
Chem 311 Fall 2009
45
2-
15
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
Chem 311 Fall 2009
46
Strange Techniques
8/3/2009
Soxhlet Extraction Solid Phase ExtractionSolid Phase MicroextractionHeadspace SamplingPurge and Trap analysisMicrowave methodsSonication MethodsChem 311 Fall 2009
47
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).
8/3/2009
Chem 311 Fall 2009
48
2-
16
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
Chem 311 Fall 2009
49
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
8/3/2009
Chem 311 Fall 2009
50
2-
17
Chem 311
Chapter 4 Lecture 1
Meaurement and Calculation
Accuracy and Precision
8/3/2009
Chem 311 Fall 2009
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
MNaOH = 0.8 g x 1 mol/204.23g /0.040 l = 0.09793 mol / liter
Uncertainty on value
2
0.0001 0.02 0.014

undertainty

0.0004
0.8 204.23 40
0.09793 0.0004 or 0.0979 0.0004

About 4 ppt uncertainty
8/3/2009
Chem 311 Fall 2009
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
8/3/2009
Chem 311 Fall 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
Chem 311 Fall 2009
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
Chem 311 Fall 2009
MEANS OF 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
8/3/2009
Chem 311 Fall 2009
calculations.
4-
Standard Deviation
Assume Gaussian Distribution of
Experimental Values
8/3/2009
Chem 311 Fall 2009
Standard Deviation
Standard Terms associated with statistical
treatment of data.
(x i )
Mean - arithmetic average. x n
n
Median - middle value of the set

Mode - most common value of the set
Chem 311 Fall 2009
8/3/2009
Standard Deviation
x x
Standard Deviation - sx (sample)

sx
or x (population >24)
x x
i
n 1
Variance = square of standard deviation
V = sx2 or V = x2
8/3/2009
Chem 311 Fall 2009
4-
Standard Deviation
Relative standard deviation.
s
% sx
ppt s
100 %
1000 ppt
% relative std. dev. = Coefficient of

Variation (CV)
8/3/2009
Chem 311 Fall 2009
10
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
Chem 311 Fall 2009
11
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.
8/3/2009
Chem 311 Fall 2009
12
4-
Deviation
99.8% within 3 std. dev.
As a general rule, virtually all experimental

Fall 2009
data should lie inChem
the311range
4 s.
8/3/2009
13
Relative Standard Deviation

Standard deviation is fine for
comparing things that are the same. It is like
absolute error and has the same units as the
quantity being measured.
Relative standard deviation useful

comparing the deviations of data of differing
magnitude to see which has the smallest
deviation.
8/3/2009
Chem 311 Fall 2009
14
Relative Standard Deviation

eg. One student reports 25.34% KHP in an
unknown with a standard deviation of 0.05
while another reports 64.34% KHP with a
standard deviation of 0.09.Which is a better
result? Calculate the relative standard
deviation for each. The smallest relative
standard deviation gets the best grade.
(2.0 ppt and 1.4 ppt)
8/3/2009
Chem 311 Fall 2009
15
4-
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.
= true value
tn = Students t for the particular C.L. and number of replicates
n = number of replicates
sx = std. dev.
x bar = mean value
8/3/2009
tn sx
n
Chem 311 Fall 2009
16
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
Chem 311 Fall 2009
17
Confidence Limits
The 99% confidence interval includes a larger
area than the 90% and hence the confidence
limits will be greater.
Note that the larger the value of n, the smaller
the confidence limit. This is due to the fact that
larger n not only increases the denominator but
also decreases Student's t.
t s
x n x
n
8/3/2009
Chem 311 Fall 2009
18
4-
Confidence Limits
Example mean = 144 ppm sx = 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.
8/3/2009
Chem 311 Fall 2009
tn s x
n
19
Types of Errors
Random Errors - Indeterminate errors
Have no definite cause
random errors in reading a buret
Result in decreased precision and wider

confidence limits
8/3/2009
Chem 311 Fall 2009
20
Types of Errors
Systemmatic Errors - Determinate errors
Have a definite cause
Balance not calibrated
Solution at wrong temperature for volumetric work
Result in measured values which fall outside

the confidence interval of the measurement
May be constant relative or absolute error

Vary Sample size to evaluate
8/3/2009
Chem 311 Fall 2009
21
4-
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
Chem 311 Fall 2009
22
Types of Errors
Personal Errors
Analyst makes a mistake
8/3/2009
Chem 311 Fall 2009
23
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
8/3/2009
Chem 311 Fall 2009
24
4-
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
Chem 311 Fall 2009
25
ELIMINATION OF
Undefinable error. These are of two types.
Errors due to learning to do the experiment
correctly or due to problems in start up. These
are common in student experiments.
You may discard all data prior to any point and
call that data practice.
Note this in the notebook with the data.
8/3/2009
Chem 311 Fall 2009
26
ELIMINATION OF
Undefinable error. These are of two types.
Errors due to deterioration of reagents, operator
patience, or equipment. These occur at the end
of a series of experiments.
You may discard all data after any given data set
and assume some unknown problem occurred.
You may NOT discard data in the middle of

a data set unless there is a definable error!!!
8/3/2009
Chem 311 Fall 2009
27
4-
ELIMINATION OF
Q-test.
measure of the gap between an outlying value
and its nearest neighbor compared to the total
range of the data.
xnearest
x
Qexp outlier
EXAMPLE
xhigh xlow
0.1013, 0.1014, 0.1016 , 0.1024,
0.1024 0.1016
Q
0.727
0.1024 0.1013
95% confidence Q4 =0.83 > 0.73
Cannot discard value
8/3/2009
Chem 311 Fall 2009
28
Significance Testing
Assume Gaussian Distribution
Assign Probability to two possible
outcomes
NULL HYPOTHESIS
Differences observed are due to random,
indeterminate errors
ALTERNATIVE HYPOTHESIS
Differences observed are real
8/3/2009
Chem 311 Fall 2009
29
NEVER PROVE ANYTHING IS TRUE
Prove NOT FALSE
Use alpha = the area of the distribution not

included usually 0.05
95% of area is included and 5% excluded
If p<0.05 then the Null Hypothesis can be

rejected. Differences not due to random errors
8/3/2009
Chem 311 Fall 2009
30
4-
10
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 give the same result?
Compare two data sets
F-Test
Compare standard deviations for data sets or methods
8/3/2009
Chem 311 Fall 2009
31
T-test example
Measured pH of natural and machine made
snow. Are they the same or different?
Various t-tests
Variance on each data set is the same
Variance on each data set is different
Data in two sets are paired or not
Excel Tools Data Analysis

8/3/2009
Chem 311 Fall 2009
32
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
8/3/2009
Chem 311 Fall 2009
33
4-
11
t-Test: Two-Sample Assuming Unequal Variances

Data for [H+] of Snow Samples
Natural
Machine
Mean
Variance
Observations
Hypothesized Mean Difference
1.47E-05
4.19E-11
7
0
1.51E-06
7.65E-12
11
df
t Stat
P(T<=t) one-tail
t Critical one-tail
P(T<=t) two-tail
7
5.086709
0.00071
1.894579
0.00142
t Critical two-tail
8/3/2009
2.364624
Chem 311 Fall 2009
34
What does this mean

P<0.05 95% confidence that values are
different.
One Tail all 5% area under curve on one side
Use when reason to expect one value to be smaller
and the other larger
Two tail 2.5 % of area at each end of curve

More rigorous test
Use when no reason to predict the order of values
8/3/2009
Chem 311 Fall 2009
35
Stream pH
Average pH data
7
6.9
6.8
6.7
pH
6.6
Off Mt
6.5
On Mt
6.4
6.3
6.2
6.1
6
8/1/2004 9/20/200 11/9/200 12/29/20 2/17/200 4/8/2005 5/28/200 7/17/200 9/5/2005 10/25/20 12/14/20
4
4
04
5
5
5
05
05
Date
8/3/2009
Chem 311 Fall 2009
36
4-
12
t-Test: Paired Two Sample for Means

[H+] for two Streams Nov-May
Mean
Variance
Giant
1.35E-06
1.67E-12
Coleman
3.87E-07
8.53E-14
Observations
Pearson Correlation
Hypothesized Mean Difference
df
t Stat
8
0.929994
0
7
2.649175
P(T<=t) one-tail
t Critical one-tail
P(T<=t) two-tail
8/3/2009
t Critical two-tail
0.016491
1.894579
0.032981
Chem 311 Fall 2009
2.364624
37
4-
13
Chapter 4 Part 2
Calibration, Standardization
8/3/2009
Chem 311
Smeas = kCAnalyte + Sreagent

Calibration necessary for all measurements
What to use as STANDARDS
Primary Standards
8/3/2009
Known stoichiometry
Known purity
Stable for long periods
List in Appendix 2 text
Ex. KHP for Titration
Chem 311
Standard Solutions
Weigh Primary Std. Into Volumetric Flask
Dilute to mark only after all dissolved
Do not heat
Mix at least 10 times
8/3/2009
Chem 311
4 part 2-
Standard Solutions
Serial Dilution for Calibration or dilute
Pipet portion of stock solution into Vol.
Flask and dilute
CoVo=CdVd
Many ways to get to specific concentration
Errors not the same for each
problem 4-11
8/3/2009
Chem 311
Calibration Methods
Single Point
External Standard Method Calibration
Curve
Internal Standard Method
Standard Addition Method
8/3/2009
Chem 311
Single Point Calibration

k= Sstand/Cs
One Standard
Assumes response factor k is constant over
entire range
Assumes linear relationship
Ignores Blank due to reagents
Generally not a good idea

8/3/2009
Chem 311
4 part 2-
External Standard Method

Using a Standard Curve
Known concentrations vs Signal
Intercept need not be Zero
Identify non-linearity if present
8/3/2009
Chem 311

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
Chem 311

Chromatography
make calibration curve
run sample and compare
Limited by injection accuracy
2-4%
1-2%
0.5%
8/3/2009
manual
auto
valve injections
Chem 311
4 part 2-
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
Chem 311
10
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
Chem 311
11
ASSUMPTIONS
Values for the x axis are relatively free from error.
Concentration = independent variable
Values for the the y axis and may contain some

error.
Signal = what you measured
The standard least squares method assumes all

have the same error
more sophisticated approaches allow weighing factors
for cases where errors differ greatly.
8/3/2009
Chem 311
12
4 part 2-
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
Chem 311
13
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
Chem 311
14

Excel Getting values for Calculations
=Slope(yvalues, xvalues)
=Intercept(yvalues, xvalues)
=RSQ(yvalues,xvalues)
bo
b1
R2
All Values are Live and update changes
8/3/2009
Chem 311
15
4 part 2-

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
Chem 311
16
Does line go through ZERO

SEy >= y intercept
Y-intercept lies within 1 SD of 0
Repeat Calculation and select Force Zero
Intercept
SEy < y intercept

Use full equation from Least Squares
Be sure to use SEy not overall SE

8/3/2009
Chem 311
17
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.
R2 correlation coefficient 0.0 to +1.0
Variance or standard deviation of the fit
8/3/2009
Chem 311
18
4 part 2-
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
Chem 311
19
Excel Least Squares

A Closer look
0
1.1
2
3.98
5.2
Y values
0.1
1
2.1
4
5
Least Squares
Depe ndent
V a r ia b le
X values
y = 0.9624x + 0.0763
R2 = 0.9974
6
4
Y values
2
0
0
Linear (Y
values)
Independent Variable
8/3/2009
Chem 311
20
Residuals
X values
0
1.1
2
3.98
5.2
Y values Y calc
Residuals
0.1
0.0763
-0.0237
1 1.13494 0.13494
2.1
2.0011
-0.0989
4 3.906652 -0.09335
5 5.08078 0.08078
Residuals
Residuals
0.2
0.1
0
-0.1 0
Residuals
2
-0.2
8/3/2009
X values
Chem 311
21
4 part 2-
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
Chem 311
22
Non-linear data
X values
0
1.1
2
3.98
5.2
Y values Y calculateResiduals
-0.05
0.0669
0.1169
1.15 1.12257 -0.02743
2.14
1.9863
-0.1537
3.89 3.886506 -0.00349
4.99 5.05734 0.06734
Same R squared
as previous data
Non-linear Data
Y values
y = 0.9597x + 0.0669
R 2 = 0.9974
Y values
4
2
Linear (Y
values)
0
-2 0
X values
8/3/2009
Chem 311
23
Residuals
Note pattern in residuals
Residuals
Residuals
0.2
0.1
0
-0.1 0
Residuals
2
-0.2
X value
8/3/2009
Chem 311
24
4 part 2-
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
Chem 311
25
Calculating an Unknown
The value for an unknown result xc is calculated
from measured value yc
Xs
y s bo
b1
b1=slope bo=intercept
8/3/2009
Chem 311
26
Precision of Unknown
m = number of observations of unknown
n = number of calibration points
sx
sr
8/3/2009
sr
b1
( yx y)2
1 1

m n b12 ( xi x) 2
(y
y i ) 2
n2
( yi y i ) residual for point

Chem 311
27
4 part 2-
Equation hard to use. Errors occur if any
sum value is not taken with all digits
sx
sr
sr
b1
( y y)2
1 1
2 x
m n b1 ( xi x) 2
(y
y i ) 2
n2
8/3/2009
( yi y i ) residual for point

Chem 311
28
Precision of Regression
sx function has a minimum value when the mean
value for the unknown is close to the mean value
of the calibration curve.
(yxbar - 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
Chem 311
29
Confidence Limit for unknown
xtrue x ts x
8/3/2009
Chem 311
30
4 part 2-
10
Error in b1 and bo
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
Chem 311
31
95% CL best case

6
5
Y calues
4
Exp. Values
-b+m
+b-m
1
0
-1 0
X values
8/3/2009
Chem 311
32
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 + cx2
Calculation of Unknown gives 2 values
Quadratic Solution Required
8/3/2009
Chem 311
33
4 part 2-
11
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

8/3/2009
Chem 311
34
4 part 2-
12
Chapter 5
QA & Calibration
8/3/2009
Chem 311
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
Chem 311
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)
8/3/2009
Chem 311
5-
Range of Method
Limit of Detection
Limit of Quantitation
Limit of Linearity
Limit of response
Range statistics vary depending on many

factors R2=0.995 in text is rarely met
8/3/2009
Chem 311
Limit of Detection
8/3/2009
Chem 311
Limit of Detection (LOD)

Slod= S + z sigma
Z=3 gives 99.86% confidence that
substance is present
S=mC +b from Least Squares
Clod = (Slod b)/m
< 1% chance of false positive
8/3/2009
Chem 311
5-
Quantitation Limit
Sloq= S + 10 sigma
Gives quantitation with 10% rel. error
Cloq = (Sloq b)/m
1 sig fig quantitative result possible
8/3/2009
Chem 311
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
Chem 311
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

Interlaboratory Precision
8/3/2009
Chem 311
5-
Accuracy
8/3/2009
Run SRMs similar matrix

Use two independent methods
Spike sample with know amount - Recovery
Compare Standard Addition with
Calibration result
Chem 311
10
Recovery Study
Validate sample preparation/extraction etc
Run Sample
Run Sample with known addition
8/3/2009
Chem 311
11
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
8/3/2009
Chem 311
12
5-
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
Chem 311
13
Graphical treatment of std.

addition
Graph Signal vs Spike Concentration
X-Intercept= -CAVo/Vf
8/3/2009
Chem 311
14
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
8/3/2009
Chem 311
15
5-
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
Chem 311
16

Data from GC
experiment
20:0
16:0
7.0000
10.0000
6.0000
Area Ratio
Area Ratio
8.0000
y = 0.9287x + 0.0426
R2 = 0.9999
5.0000
y = 0.8809x + 0.0549
4.0000
3.0000
2.0000
R = 0.9999
6.0000
4.0000
2.0000
1.0000
0.0000
0.0000
0
8/3/2009
10
15
Mass Ratio
Mass Ratio
Chem 311
17
Selecting an Internal Standard

Chemically Similar to Analyte
HPLC Lab Theophylline for Caffeine
Isomers or close homologs
GC Lab C17 fatty acid
Once Internal Std and Sample are made

homogeneous all further steps need not be
quantitative (within limits of linearity and
detection)
8/3/2009
Chem 311
18
5-
Selecting an Internal Standard

Mass Spectrometry Isotopic labels
D, 13C, 15N, etc
All chemical properties same as Analyte
Mass Spec gives separate signals
Cocaine methods for Chem 312
8/3/2009
Chem 311
19
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
Chem 311
20
Control Chart
Monitor QA over time
Hg Analyzer
Run 100 ul of 1 ppm each day as first sample
Put result on graph
Monitor changes
8/3/2009
Chem 311
21
5-
Chem 311
Chapter 6
Chemical Equilibrium
8/3/2009
Chem 311
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
Chem 311
Equilibrium constant
Free Energy change is related to chemical
potential
G G o RT ln Q
c
c d
o o
Note difference from Eq 6.2
c d
Ratio of activities to Std. St. Q
a
b
a b
Dimensionless
o o
a b
8/3/2009
Chem 311
6-
At Equilibrium
G o RT ln K eq
c d
o o
K eq c a d b
a b
o o
a b
where a' s are equilibrium values
8/3/2009
Chem 311
Normally write using Concentrations
(6-2)
K eq
Ceqa Deqd
Aeqa Beqb
All values are dimensionless (ratio to std state)

Ignore Activity effects in most cases
(Chapter 8)
8/3/2009
Chem 311
Properties of Keq
Reverse Reaction Invert K
Cu + 4 NH3 Cu(NH3)4
Kf
[Cu ( NH 3 ) 4 ]
[Cu 2 ][ NH 3 ]4
Cu(NH3)4 Cu + 4 NH3
Ki
8/3/2009
1 [Cu 2 ][ NH 3 ]4
Kf
[Cu ( NH 3 ) 4 ]
Chem 311
6-
Precipitation Reactions
AgCl(s) Ag+ + ClKsp= [Ag+] [ Cl-]

Note solid does not appear in different phase
Only works for a few species
Silver halides
BaSO4
All others have complex chemistry which make

the simple treatment useless.
8/3/2009
Chem 311
Precipitation Reactions
PbI2(s) Pb+2 + 2I Ksp=[Pb+2 ][ 2I-]
Also need to consider
PbI+ , PbI3- , and PbI42 Simple equilibrium gives wrong answers
8/3/2009
Chem 311
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 + H2O <===> H3O+ + Cl acid + base
8/3/2009
conj.acid conj.base
Chem 311
6-
Look at half reactions
8/3/2009
Acid1 ==> H+ + Base1

Base2 + H+ ==> Acid2
Sum these to get
Acid 1 + Base2 ==> Base1 + Acid2
conjugate Acid-Base pairs (subscripts)
Chem 311
10
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 + H2O ==> Cl- + H3O+
Stronger acid+ stronger base==> weaker base + weaker acid
8/3/2009
Chem 311
11
Acid-Base Strength
Relative
In water - H3O+ is strongest acid
Anything above it reacts with
water to produce H3O+
HCl, HwSO4, HBr, HI, HClO4
Anything below water is weak acid

Farther below water the weaker it gets
None react completely
Equilibrium
8/3/2009
Chem 311
12
6-
Strong Acids
Above H3O+
HCl + H2O <===> Cl- + H3O+
Cl- very weak proton acceptor

HCl very strong proton donor.
Thus there is a complete proton transfer.
Ka >>> 100
8/3/2009
Chem 311
13
Weak acids
Acetic acid for example:
CH3COOH + H2O <===> CH3COO- + H3O+
Ka = 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
Chem 311
14
Strong bases
O2- + H2O <===> 2 OH>>>100 strong base
Kb
Anything which yields OH- ions in

solutions
NaOH, KOH, Ba2OH - must be soluble
Cu(OH)2 insoluble not strong base
8/3/2009
Chem 311
15
6-
Weak bases
NH3 + H2O <===> NH4+ + OH Kb= 1.7x10-5 weak base
Amines, conjugate bases of weak acids
phosphates, carbonates, acetate, etc.
8/3/2009
Chem 311
16
Acid and Conjugate Base

Strengths are related to the solvent.
CH3COOH + H2O <===> CH3COO- + H3O+
Ka
[H ][OAc ]
[HOAc]
CH3COO- + H2O <===> CH3COOH + OH-
Kb
8/3/2009
[OH ][HOAc]
[OAc ]
Chem 311
17
Add reactions multiply Ks

CH3COOH + H2O + <===> CH3COO- + H3O+
Ka
CH3COO- + H2O <===> CH3COOH + OH- Kb
8/3/2009
+ add two reactions

2 H2O <===> H3O+ + OHSolvent Autoprotolysis
Equilibrium constant Kw = Ka x Kb
Chem 311
18
6-
Acid- Base Pair Relationship

Ka x Kb = Kw
This relationship is true for any conjugate acid
base pair in water.
KaKb
= Ks
For any solvent
If you know the Ka you can always get the Kb.

Also note that for a solvent of different Ks the
relationship between the Ka and Kb will be
different.
8/3/2009
Chem 311
19
Temperature Effects
Ka and Ks vary with temperature
Temperature (C )
0
25
50
100
37
8/3/2009
Kw
0.114 x 10-14
1.01x10-14
5.47x10-14
49x10-14
3.2 x10-14
Chem 311
20
What is a neutral solution?
8/3/2009
25 deg C Kw= 1.0x10-14 = [H3O+] [OH-]

[H3O+]= 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
Chem 311
21
6-
Solubility Equilibria
AgCl <==> Ag+ + Cl Ksp = [Ag+] [ Cl-]
Solubility = how many moles dissolve in 1 liter of
water
S=[Ag+] = [Cl-] = Ksp1/2
Al(OH)3 < Al3+ + 3 OH S= [Al3+ ] = 1/3 [OH-] so [OH-] = 3S
Ksp = [Al3+ ] [OH-] 3 = S x (3S)3 = 9S4
8/3/2009
Chem 311
22
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 complexing agents- must consider
fraction of metal in uncomplexed form
8/3/2009
Chem 311
23
Bottom Line
AgCl, AgBr, AgI and BaSO4 are the only
cases which work even tolerably well
Silver halides form higher complex ions
AgCl2-, AgCl3-2, AgCl4-3, AgCl(aq)
8/3/2009
Chem 311
24
6-
Coordination Complexes
Lewis Acid Base Theory
Acids = electron pair acceptor
H+, Metal ions Fe3+, Cu2+, etc.
Base = Ligand = electron pair donor

OH-, Cl-, :NH3, CN-, H2O, etc.
Acid + Base ==> Complex ion

8/3/2009
Chem 311
25
Coordination Complexes
Complex ions Co complex lab
All ions in water are hydrated
Cations =Lewis acids ==> accept electron pair
from :O on H2O
Fe3+ + 6 H2O <==> [Fe(H2O)6]3+
hexaaquo iron (III) ion
Formation of other complex ions by

Displacement
8/3/2009
Chem 311
26
SEQUENTIAL COMPLEXATION BY
SIMPLE LIGANDS
Ni(H2O)42+ + 4 Cl- <===> NiCl42- +
4H2O) REACTION OCCUR IN A SERIES OF
STEPS
[NiCl ]
K
Ni (H2O)42+ + Cl- <===> Ni (H2O)3 Cl+ 1
Ni (H2O)3 Cl+ + Cl- <===> Ni (H2O)2 Cl2+K 2

Ni (H2O)2 Cl2+ + Cl- <===> Ni (H2O) Cl3- K 3
Ni (H2O) Cl3- + Cl- <===> NiCl4-2
8/3/2009
Chem 311
K4
[Ni ][Cl ]
[NiCl 2 ]
[NiCl ][Cl ]
[NiCl 3 ]
[NiCl 2 ][Cl ]
[NiCl 4 2 ]
[NiCl 3 ][Cl ]
27
6-
Overall Formation constants

1= K1
2= K1K2
2
[NiCl 2 ]
2
3= K1K2K3
4=K1K2K3K4
[Ni ][Cl ]
WRITE A MASS BALANCE FOR NICKEL

CNi = [Ni 2+ ] + [NiCl+ ] + [NiCl2 ] + [NiCl3- ] + [NiCl42- ]
8/3/2009
Chem 311
28
General Approach Complicated

Equilibrium Systems
Calculate the fraction of each form in an
equilibrium under specified conditions
will be used to identify fraction
Ni2+ = fraction of total Ni in the Ni2+ form
HA = fraction of weak acid (HA) in
undissociated form.
A- = fraction of weak acid (HA) in dissociated
form.
8/3/2009
Chem 311
29
Species present
Three techniques
Calculate fraction in each form - alpha
Log Concentration plots (computer)
Line diagrams (qualitative)
8/3/2009
Chem 311
30
6-
10
Species present at a pH
Calculate alphas
HA <==> H+ + ACHA = [HA] + [A] Mass balance
Define fraction of total in specific form
HA
8/3/2009
[ HA]
C HA
[ A ]
C HA
Chem 311
31
Substitute values from Ka
CHA=[HA] + [A]
[ HA]
HA
C HA
[ HA]
8/3/2009
[ A ]
C HA
K ' [ HA]
[ H ][ A ]
[ A ] a
'
[H ]
Ka
Chem 311
32
For monoprotic acid - Exact equation
HA
A
8/3/2009
[H ]
[ H ] K a'
K a'
[ H ] K a'
Chem 311
33
6-
11
For polyprotic acid - Exact equation
HA
[ H ]n
'
[ H ]n K a' 1[ H ]n 1 K a' 1 K a' 2 [ H ]n 2 ...K a' 1 K a' 2 ..K an
Am
'
[ H ]n m K a' 1 K a' 2 ..K an
'
[ H ]n K a' 1[ H ]n 1 K a' 1 K a' 2 [ H ]n 2 ...K a' 1 K a' 2 ..K an
A2
[ H ]n 2 K a' 1 K a' 2
'
[ H ] K [ H ] K a' 1 K a' 2 [ H ]n 2 ...K a' 1 K a' 2 ..K an
n
'
a1
n 1
8/3/2009
Chem 311
34
HA
n 1
[H ] K [H ]
'
a1
[ H ]n
'
K a' 1 K a' 2 [ H ]n 2 ...K a' 1 K a' 2 ..K an
Am
'
[ H ]n m K a' 1 K a' 2 ..K an
n2
'
'
'
[ H ] K [ H ] K a1 K a 2 [ H ] ...K a' 1 K a' 2 ..K an
A2
[ H ]n 2 K a' 1 K a' 2
'
[ H ] K [ H ] K a' 1 K a' 2 [ H ]n 2 ...K a' 1 K a' 2 ..K an
'
a1
'
a1
n 1
n 1
Each term in denominator represents one

form of the acid
8/3/2009
Chem 311
35
Plot vs pH - Useful to see what is present
as function of pH
Sketch approximate alpha plots
pH=pK at 50% crossover
pK '1 pK '2
1.0
A2-
H2A
0.8
alpha
0.6
0.4
0.2
0.0
8/3/2009
Chem 311
p[H3O+]
10 12 14
36
6-
12
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
Chem 311
37
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
Chem 311
38
8/3/2009
Chem 311
39
6-
13
8/3/2009
Chem 311
40
6-
14
Chem 311
Chapter 8
Activity
Systematic Equilibrium
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
Chem 311
Activity of HCl solutions
Activity vs Concentration for HCl

Activity
0.8
0
10
Concentration (F)
15
Activity H+
Activity H+
Activity vs Concentratino for HCl

800
700
600
500
400
300
200
100
0
0.6
Series1
0.4
0.2
0
0
8/3/2009
Chem 311
0.2
0.4
0.6
Concentration (F)
0.8
8-
ACTIVITY see Chapter 8

Activity coefficient
Finagle constant
8/3/2009
x xC x
Chem 311
ACTIVITY
Problem of definition. What are the
conditions where f=1.00000 ? The value
of Keq will depend on how this is
chosen.
8/3/2009
Chem 311
Henryan System
Used for Solutions
f = 1 at limit C ==> 0
8/3/2009
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.
Chem 311
8-
Concentration Effects
Concentration increases
Ions get close enough to be attracted and
repelled by other ions
All ions are solvated (Solvation number)
Very High concentration

Run out of solvent for hydration
Activity coefficients range widely

8/3/2009
Chem 311
Activity Coefficients
Function of Ionic Strength
S
1
[ A z ]z A 2 [ B z ]z B 2 ...
2
Ionic strength depends on charges of ions

More highly charged ions make S>> [Conc]
Na2SO4 [Na]=2x [SO4] but S=1/2([Na] + [SO4]x4)
S=1/2(2x [SO4] + 4x [SO4] ) = 3x [SO4]
8/3/2009
Chem 311
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
8/3/2009
Chem 311
8-
Cut to the Chase

Ignore Activity Coefficients in most work
Cant get them accurately
Places limit on absolute accuracy of all
equilibrium calculations
Biochemists making buffers to match

physiological conditions
8/3/2009
Use 1:1 salts for best accuracy

Use highly charged ions to get high ionic
strength at low conc.
Large errors due to Chem
lack311of dissociation
10
Cut to the Chase

Makes Great excuse for why things dont
work quite right
Potentiometric titration data fit
Use activity coefficients as excuse for some
deviations from theory
8/3/2009
Chem 311
11
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
8/3/2009
Chem 311
12
8-
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
Chem 311
13
Acid Base Equilibria

Step 1. Write all chemical reactions and
their equilibrium constant expressions.
8/3/2009
Chem 311
14
Example - Benzoic Acid

2 H2O <===> H3O+ + OH
Kw = 1.00 X 10-14
C6H5COOH + H2O <==> C6H5COO- + H3O+
Ka
[H ][BA ]
5
6.3x10
[HBA]
Too Many Unknowns to solve

Need additional Equations
8/3/2009
Chem 311
15
8-

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
Chem 311
16

MASS BALANCE - Conservation of
matter
Multiple equations for complex systems
CHBa = [HBA] + [BA- ]
8/3/2009
Chem 311
17

Step 3 Use Algebra to simplify things and remove
unknowns
Substitute from Charge Balance into Mass Balance
[BA- ] = [H +] - [OH- ]
CHBA = [HBA] + [H +] - [OH- ] REARRANGE

[HBA] = CHBA - [ H+ ] + [OH- ]
SUBSTITUTE INTO Ka EXPRESSION
Ka
8/3/2009
[ H ][ BA ]
CHBa Chem
[ H311 ] [OH ]
18
8-
Exact solution
Substitute from
Ka
Charge Balance
[OH-] = Kw/[H+]
8/3/2009
[ H ] [ H ] [OH ]
C HBa [ H ] [OH ]
K
[ H ] [ H ] w
[
]
H
Ka
K
C HBa [ H ] w
[H ]
Chem 311
19
Exact solution
Clean it up
Ka
[ H
] Kw
K
C HBa [ H ] w
[H ]
You dont really want to do

this. (cubic)
[ H ]3 K a [ H ]2 K a [ H ]C HA K w [ H ] K w K a 0
Solutions of very weak acids
HCN
Ka= 6x10-10
8/3/2009
Chem 311
20

Approximate solution
Ignore OH
OH <<H since this is an acid
Ka
If CHBA>> [H+]
8/3/2009
[ H ][ Ba ]
C HBa [ H ]
[H] CHAKa
Chem 311
21
8-

Solution
Ka
[ H ][ Ba ]
C HBa [ H ]
[ H ]2 K a [ H ] C HA K a 0
Solve on calculator
Works as long as water contribution [OH] is
negligible
8/3/2009
Chem 311
22
Monoprotic base
Two ways to solve - Approximate
[OH ] K bCb
More Exact
[OH ]2 [OH ]K b Cb K b 0
8/3/2009
Chem 311
23
Solving on a TI Calculator
TI-86
POLY
Order =2 Enter
A2=1
A1=Ka
A0=-CaxKa
SOLVE
You get two answers- choose the reasonable

one (positive and not greater than Ca)
8/3/2009
Chem 311
24
8-
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 Ca)
8/3/2009
Chem 311
25
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
Chem 311
26
Very complicated
Acid base component
Coordination complex component
Multiple species
Lots of Algebra fun
Identifying all Chemical Reactions often
hardest part
8/3/2009
Chem 311
27
8-
Chem 311
Chapter 9
Monoprotic Acid-Base
8/3/2009
Chem 311
Calculation pH of anything
Six cases to address
Strong Acid/Base > 10-6 M
Strong Acid/Base <10-6 M
Weak Acid
Acid or conjugate acid of weak base
Weak Base
Base of conjugate base of weak acid
Buffer
Acid plus its conjugate base
Amphiprotic substance - polyprotic systems Chapter 10

8/3/2009
Chem 311
Strong Acid/Base
Completely dissociated
[H+] = CHA
[OH-] = CB
If Conc > 1x10-6 M then dissociation of

H2O can be ignored
8/3/2009
Chem 311
9-
Strong Acid/Base
If Conc. < 1x10-6 M then dissociation of
H2O must be included
Acids never > pH 7
Bases never < pH 7
Quadratic solution from Systematic
treatment
[H+]2 + CHA[H+] Kw = 0
8/3/2009
Chem 311
Monoprotic acid or base

Acid
[ H ]2 K a [ H ] C a K a 0
Base
[OH ]2 [OH ]K b Cb K b 0
8/3/2009
Chem 311
Buffers
Resist Change in pH
Added acid
Added base
dilution
Very important in all chemistry

Acid + conjugate base
8/3/2009
Chem 311
9-
Exact Solution buffers

Example - Base Buffer
.200M
+
.100M
NH3
+ NH4NO3
NH3 + H20 <====> NH4+ + OH NO3- disassociated - no Rx
8/3/2009
Chem 311
Buffer Example
Change balance [NH4+] + [H+] = [OH-] + [NO3-]
Mass balance 0.200 + 0.100 = [NH3] + [NH4+]
By substitution from Charge Balance

[NH4+] = [OH-] + [NO3-]- [H+]
0.200 + 0.100 = [NH3] + [NO3-] + [OH-] - [H+]
[NO3-] = 0.100 Therefore subtract from both
sides
0.200 = [NH3] + [OH-] - [H+]
[NH3] = 0.200 - [OH-] + [H+]
8/3/2009
Chem 311
Buffer Example
Mass Balance rearranged
[NH3]= 0.200 + 0.100 - [NH4+]
substitute in expression for NH3
0.100 = [NH4+] - [OH-] + [H+]

Rearranging we see that :
[NH4+] = 0.100 + {[OH-] - [H+]}

and
[NH3] = 0.200 - {[OH-] - [H+]}

8/3/2009
Chem 311
9-
Buffer Example
K 'b
[OH ][ NH 4 ]
[ NH 3 ]
Substitute
K 'b
[OH ] C NH 4 [OH ] [ H ]
C NH 3 [OH ] [ H ]
Exact Solution
Just like acid but with Conc of Conjugate included
8/3/2009
Chem 311
10
Acidic buffer
[ H ]C [ H ] [OH ]
K
'
a
C HA [ H ] [OH ]
Completely rigorous all monoprotic cases

Nasty cubic equation
Simplify
If Acidic H+>>OH- Drop OH term
If Basic OH>>H Drop H term
K a'
8/3/2009
Chem 311
[ H ] C A [ H ]
C HA [ H ]
11
Acidic buffer
Simplify
If Concentrations of Acid and Salt are high
C >> [H+]
C >> {[H+]-[OH-]}
Drop those terms

K a'
8/3/2009
[ H ]C A
C HA
Chem 311
12
9-
Acidic buffer
K a'
[ H ]C A
C HA
Take -log of equation

Henderson-Hasselback
8/3/2009
Chem 311
[ H ]C A
log K a' log
CHA
C
pK a log[ H ] log A
CHA
C

pK a pH log A
CHA
C
pH pK a log A
CHA 13
Solution to Ammonia example
Base/Acid
0.200
pH 9.244 log
9.545
0.100
NH3 is base (A-)
NH4+ 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.
8/3/2009
Chem 311
14
Buffers- Resist pH Change

Weak Acid-Conjugate Base pair
Henderson-Hasselbach Eq
C
pH pK a log A
C HA
Since CHA and CA apply to same

solution==> volumes cancel
Use ratio of moles salt/moles acid
8/3/2009
Chem 311
15
9-
Buffer Capacity
Maximum at pH = pK
Useable in range pH=
pK 2
8/3/2009
Chem 311
16
9-
Chem 311
Chapter 10
Poly-protic systems
8/3/2009
Chem 311
Polyprotic systems
Each proton is harder to remove
mostly due to the increased energy
required to separate the charges.
H3PO4 Ka1 = 7.5x10-3 Ka2 = 6.2x10-8

Ka3 = 4.8x10-13
Polyprotic bases.
PO4-3
polyamines such as ethylene diamine.
NH2-CH2-CH2-NH2 Kb1 = 9.1x10-5 Kb2

= 1.5x10-7
8/3/2009
Chem 311
Polyprotic systems
If K1 >K2 solve as monoprotic acid
(or base)
Use K1 and quadratic
[H+] from first Eq. Suppress other reactions

Additional dissociation = K2
8/3/2009
Chem 311
10-
Polyprotic systems
H3P04 + H20 <===>
K a1
H2P04-
H30+
[H ][H 2 PO ]
[H 3 PO4 ]
H2PO4-2 + H20 <===> HP042- + H30+

2
K a2
HP04-2 + H20
[H ][H PO 4 ]
[H 2 PO 4 ]
<===> P04-3+ H30+

3
K a3
8/3/2009
[H ][ PO 4 ]
[H PO
2
4
Chem 311
How much does K2 contribute

2
K a2
[H ][H PO4 ]
[H 2 PO4 ]
[ H ] [ H 2 PO4 ]
2
K2
[ H ][ H PO4 ]
[H ]
2
K 2 [ H PO4 ]
8/3/2009
Chem 311
Amphiprotic substances
Both acids and base reactions
Intermeditate forms
HC03 H2P04, HP042-
Amino acids (e.g.alanine)

weak acid - weak base salts
ammonium acetate
ammonium formate
8/3/2009
Chem 311
10-
NaH malate example

HMal- + H20 <===> H2Mal + 0H Kb2 = Kw/Ka1 =2.5x10-11
HMal- + H20 <===> Mal-2 + H30+

Ka2 =8.9x10-6
Which reaction has larger K?

Solution will be acidic
8/3/2009
Chem 311
NaH malate example

5 Unknowns [H2Mal] , [HMal-] ,[Mal2-], [H+] ,
and [0H-] Need 5 equations
Mass Balance
CNaHMAl = [H2Mal] + [HMal-] + [Mal2-]
Charge Balance
[Na+] + [H+] = [HMal-] + [0H-] + 2[Mal2-]
8/3/2009
Chem 311
NaH malate example

5 Unknowns [H2Mal] , [HMal-] ,[Mal2-],
[H+] , and [0H-] Need 5 equations
Ka2 = [H+] [Mal2-] / [HMal-]
Kb2 = [0H-] [H2Mal] / [HMal-]
Kw = [H+] [0H-]
8/3/2009
Chem 311
10-
NaH malate example

[Na+] + [H+] = [HMal-] + [0H-] + 2[Mal2-]
[Na+] = CNaHMAl
CNaHMal + [H+] = [HMal-] + [OH-] +2[Mal2-]
Subtract Mass balance below
CNaHMAl = [H2Mal] + [HMal-] + [Mal2-]

8/3/2009
Chem 311
10
NaH malate example

[H+] = [Mal2-] - [H2Mal] + [0H-]
Ka2
[ H ][ Mal 2 ]
[ HMal ]
[ Mal 2 ]
[OH ]
Kw
[H ]
K a 2 [ HMal ]
[ H Mal ][OH ] K w
[H ]
Kb2 2
[ HMal ]
K a1
[ H 2 Mal ]
8/3/2009
K w [ HMal ] [ HMal ][ H ]
K
K a1
K a1 w
[H ]
Chem 311
11
NaH malate example

K [ HMal ] [ HMal ][ H ] K w
Ka2
[H ]
[H ]
a 2 2-
[ H+] ]= [Mal
[H
] - [H2Mal] + [0H ]
[ H ]2 K a 2 [ HMal ]
[ H ]2
8/3/2009
[ HMal ][ H ]2
Kw
Ka2
[ HMal ][ H ]2
K a 2 [ HMal ] K w
Ka2
[ HMal ]
K a 2 [ HMal ] K w
[ H ]2 1
K a 2
[ H ]2 K a 2 [ HMal ]
K311
Chem
a 2 K a 2 [ HMal ] K a 2 K w
12
10-
NaH malate example

Rigorous equation
[H ]
K a1 (K a2 [HMal ] K w )
[H ]
K a1 [HMal ]
Problem
Dont know equilibrium concentration till we
solve equation
8/3/2009
Chem 311
13
NaH malate example

[H ]
K a1 [HMal ]
Simplifying Assumptions
1. If [HMAl] >> KA1
then Ka1 + [HMal-] = [HMal-]
[H ]
[HMal ]
8/3/2009
Chem 311
14
NaH malate example

[H ]
[HMal ]
Simplifying Assumptions
If KA2 [HMAl] >> Kw
[H ]
K a1 K a2 [HMal ]
[HMal ]
then drop last term - conc. cancels
[H ]
8/3/2009
K a1 K a2
Chem 311
15
10-
NaH malate example
[H ]
K a1 K a2
Equation independent of concentration

Works at high concentration only
Use for Amino Acids
8/3/2009
Chem 311
16
NaH malate example
[H ]
K a1 K a2
For NaHMal
[H+] = (4.0x10-4 x 8.9 x 10-6 )1/2 = 5.97 x
10-5
8/3/2009
Chem 311
17
NaH malate example

What if assumptions dont work
0.0005 M NaHMal
[HMAl] KA1
Calculate approximate value using

simple form
8/3/2009
Chem 311
18
10-
NaH malate example

[H+] = = 5.97 x 10-5
1 at this pH
[H+]2 = 2.56x10-9
K1 [H+] = 2.39x10-8
K1K2
3.56x10-9
8/3/2009
2.39 x10 8
0.770
3.10 x10 8
Chem 311
19
NaH malate example

1
2.39 x10 8
0.770
3.10 x10 8
Calculate [Hmal-] =
0.0005x0.770=0.000385
8.9 x10
Plug into
equation
4 rigorous
6
[H ]
4 x10
x0.00038 1.00 x10 14

4.18 x10 5
4 x10 0.00038
4
8/3/2009
Chem 311
20
NaH malate example

Calculate new alpha at this pH
alpha=0.76
Repeat
[H ]
4 x10 4 8.9 x10 6 x0.000381 1.00 x10 14

4.25 x10 5
4 x10 4 0.000381
Result OK
8/3/2009
Chem 311
21
10-
Summary
Weak Acid
[ H ]2 K a [ H ] C HA K a 0
2
Weak Base [OH ] [OH ]K b Cb K b 0

C
pH pK a log A
C HA
Buffer
Amphiprotic
8/3/2009
[H ]
K a1 K a2
Chem 311
22
Summary
Have to be able to tell what is present
Stoichiometry
Recognize acids and bases
8/3/2009
Chem 311
23
Fractional forms
See previous lecture notes
Alpha calculations
8/3/2009
Chem 311
24
10-
Chem 311
Chapter 11
Titration
8/3/2009
Chem 311
Simple Titrations
No Equilibrium Chemistry to worry about
Strong Acid-Strong Base
Anything with really huge Keq
Stoichiometry determines curve
8/3/2009
Chem 311
Simple Titrations
Moles titrant = CtVt
Moles Analyte = CoVo
C
Moles un-titrated = CoVo CtVt
Ca
8/3/2009
CoVo CtVt
Vo Vt
Chem 311
11-
Molarity vs volume
0.1 M HCl with 0.1 M NaOH
0.12
0.1
0.08
0.06
Series1
0.04
0.02
0
0
8/3/2009
Chem 311
10
15
20
pH vs Volume
Note rapid change at 15 ml
8
7
6
5
4
Series2
3
2
1
8/3/2009
Chem 311
0
0
5
10
15
20
After the Endpoint

Stoichiometric addition of excess titrant
Relative excess changes rapidly at EP
Change becomes more gradual as you get
farther along.
Ct
8/3/2009
Chem 311
CtVt CoVo
Vo Vt
6
11-
Simple Titration Curve

14
12
10
8
Series2
6
4
2
0
0
8/3/2009
10
15
20
25
30
Chem 311
Titration Calculations
Moles Analyte = moles Titrant (x
stoichiometry factor from reaction)
CaVa = CtVt (Stoichiometry factor)
8/3/2009
Chem 311
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
8/3/2009
Chem 311
11-
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
8/3/2009
Alkalinity methods for water analysis do

Chem 311
this
10
Visual Indicators
http://inst.santafe.cc.fl.us/~chem/intbl.html
8/3/2009
Chem 311
11
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
8/3/2009
Chem 311
12
11-
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
Chem 311
13
Decision Tree for Weak

Acid/Base Titration Problems
Is more that one reagent added
Yes
What is limiting reagent?
Calculate concentration of all reactants and products
No
Weak Acid Only
What chemicals are Present?
Quadratic solution using Ka1
Amphiprotic Substance
Weak Acid + Conjugate Base
Weak Base Only
pH=1/2(pK1 + pK2)
Buffer - HH Equation
Quadratic solution using Kb1
8/3/2009
Chem 311
14
Practice Problem
Calculate the pH of 25.0 ml of a 0.100 M
solution of phthalic acid after the addition
of 0, 10.0, 25.0, 35.0, 50.0 and 75.0 ml of
0.100 M NaOH
K1=1.12x10-3 pK1 = 2.95
K2=3.90x10-6 pK2 = 5.41
8/3/2009
Chem 311
15
11-
Practice Problem
0 ml weak diprotic acid only treat as
monoprotic
K1=1.12x10-3
[ H ]2 1.12 x10 3 [ H ] 0.10 x1.12 x10 3 0

[ H ] 0.0106
8/3/2009
pH 1.97
Chem 311
16
Practice Problem
8/3/2009
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
H2P acid remaining = 1.5 mmoles
Buffer solution - use HH ignore dilution
1.0
pH 2.95 log 2.77
1.5 17
Chem 311
Practice Problem
8/3/2009
25.0 ml base
H2P acid remaining = 0 mmoles
Amphiprotic ignore dilution
[ H ] 1.12 x10 3 x3.90 x10 6 6.6 x10 5
Chem 311
18
pH 4.18
11-
Practice Problem
8/3/2009
35.0 ml base
base remaining = 1.0 mmole
NaHP reacted with base = 1.0 mmoles
NaHP remaining = 1.5 mmol
P2- formed = 1.0 mmole buffer HH ignore
dilution
Chem 311
19
Practice Problem
35.0 ml base
NaHP remaining = 1.5 mmol
P2- formed = 1.0 mmole buffer HH
1.0
pH 5.41 log 5.23
1.5
8/3/2009
Chem 311
20
Practice Problem
50.0 ml base
moles acid (both protons) = 25.0x0.1x2 =
5.0 mmoles
All acid converted to phthlate P2 Weak base - treat as monoprotic -with
dilution [OH ]2 [OH ] K w C K w 0
Ka2
Ka2
1x10
25 1x10 14
0.1x x
0
6
3
.
90
x
10
75
3.90 x10 6 21
Chem 311
6
[OH ] 9.24 x10
pOH 5.03 pH 8.97
[OH ]2 [OH ]
8/3/2009
14
11-
Practice Problem
75.0 ml base
moles protons = 25.0mlx0.1mM/mlx2 protons M=
5.0 mmoles
phthlate conc = 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
Chem 311
22
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
Chem 311
23
8/3/2009
Chem 311
24
11-
8/3/2009
Chem 311
25
11-
Chem 311
Chapter 12
Complexometric Titration
8/3/2009
Chem 311
SEQUENTIAL COMPLEXATION
BY SIMPLE LIGANDS
Ni(H2O)42+ + 4 Cl- <===> NiCl42- + 4H2O)
REACTION OCCUR IN A SERIES OF STEPS
Ni (H2O)42+ + Cl- <===> Ni (H2O)3 Cl+
K1
Ni (H2O)3 Cl+ + Cl- <===> Ni (H2O)2 Cl2+

Ni (H2O)2 Cl2+ + Cl- <===> Ni (H2O) Cl3 Ni (H2O) Cl3- + Cl- <===> NiCl4-2
8/3/2009
K4
[NiCl ]
2
[Ni ][Cl ]
K2
K3
[NiCl 2 ]
[NiCl ][Cl ]
[NiCl 3 ]
[NiCl 2 ][Cl ]
[NiCl 4 2 ]
[NiCl 3 ][Cl ]
Chem 311
Overall Formation constants

1= K1
2= K1K2 3= K1K2K3
4=K1K2K3K4
[NiCl ]
[Ni ][Cl ]
WRITE A MASS BALANCE FOR NICKEL

CNi = [Ni 2+ ] + [NiCl+ ] + [NiCl2 ] + [NiCl3- ] + [NiCl42- ]
Rearrange K's to get mass balance in terms
of [Ni ] and [Cl ]
8/3/2009
Chem 311
12-
Rearrange Betas
[NiCl+ ] = K1 [Ni2+][Cl- ]
[NiCl2 ] = K1 K2 [Ni2+][Cl- ]2 = 2 [Ni2+][Cl- ]2
[NiCl3- ] = 3 [Ni2+][Cl- ]3
[NiCl42-] = 4 [Ni2+][Cl- ]4
Substitute into Mass Balance
CNi = [Ni2+] + b1 [Ni2+][Cl-] + b2 [Ni2+][Cl-]2+ b3
[Ni2+][Cl-]3+ b4 [Ni2+][Cl-]4
8/3/2009
Chem 311
Define alpha metal

2
[Ni ]
C Ni
Ni
Ni
[Ni ]
2
[Ni ] + 1 [Ni ][Cl ] + 2 [Ni ][Cl ] + 3 [Ni ][Cl ] + 4 [Ni ][Cl ]
Ni
1
1 +1[Cl] +2[Cl]2+ 3[Cl]3+ 4[Cl]4
1
2
1 + 1 [L ] +2 [L ] + 3 [L] +... n [L]
8/3/2009
Chem 311
Multidentate Ligands for

Complex Formation
Chelate Effect - Formation of rings
5 or 6 member rings are best.
N
O
O
Cu
Ni
N
8/3/2009
Chem 311
12-
Multidentate Ligands for

Complex Formation
Entropy effects enhance stability of
multidentate ligands.
M(H2O)6 + 6 L <===> ML6 + 6H2O
7 <===> 7 entropy change small
M(H2O)6 + L <===> ML + 6H2O

2 <===> 7 large increase in entropy
greater entropy change for the latter reaction.
8/3/2009
Chem 311
Complexation Titration
For titrations, 1:1 complexes best
Intermediate equilibria make titration
breaks less sharp for higher No. of ligands.
14
ML6
pM
ML
8/3/2009
Chem 3110
8
EP
Volume Titrant
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
8/3/2009
Chem 311
metal
12-
ACIDS
EDTA - Ethylenediamine
tetraacetic acid.
H
O
H
This has 6 Lewis base sites, 4

oxygen and 2 amine electron
pairs.
designated as H4Y and its anion as
Y4 All six complexation sites are able
to bind to a single metal ion for
many metals.
8/3/2009
O
N
N
H
O
H
O
O
Chem 311
10
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.
8/3/2009
This is for 10 coordinate metals such as the

Chem 311
actinides.
11
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.
H
O
H
O
O
N
N
H
8/3/2009
Chem 311
12
O
O
12-
LIGANDS
Use EDTA as an example:
H4Y Primary standard but relatively
insoluble
Na2H2Y .2H2O Not a true primary standard
but soluble to more than 0.05 m/l .
At low pH most EDTA is in the form H2Y28/3/2009
Chem 311
13
LIGANDS
At low pH most EDTA is in the form H2Y2-pK '1
-pK '2
-pK '4
-pK '3
-2
log alpha
-4
-6
H2Y2-
HY3-
-8
-10
Y4-
H3Y-
-12
H4Y
-14
0
8/3/2009
10
12
14
]
p[H3O311
Chem
14
LIGANDS
M+n + H2Y-2 <===> MYn-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+.
8/3/2009
Chem 311
15
12-
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
16
Control pH to Control [Y]
8/3/2009
Chem 311
17
Conditional Formation
Constants
M + Y-4 <===> MY
[CaY]
Kf
[Ca 2 ][Y4 ]
[Y4- ] = 4 CEDTA
K 'f K f 4
[CaY]
[Ca 2 ]C EDTA
NOTE The conditional formation applies in this

case at only the pH used to calculate 4 for
EDTA.
8/3/2009
Chem 311
18
12-
EXAMPLE
Using conditional formation constants to
predict feasibility of titration.
50.00 ml of 0.0100 m Cu2+ is titrated with
0.0100 m EDTA at pH 3.00
Kf = 6.3x1018
Note this is a large POSITIVE exponent.
8/3/2009
Chem 311
19
EXAMPLE
Calculate 4 at pH = 3.00
4 = 2.5 x 10-11
Calculate Kf' = 6.3 x 1018 * 2.5 x 10-11
= 1.6 x 108
8/3/2009
Kf= [CuEDTA]
[Cu]CEDTA
Chem 311
20
Example
Initial pCu = -log 0.0100 = 2.00
Before equivalence point:
[Cu 2 ]
moles Cu untitrated
total volume
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
8/3/2009
Chem 311
21
12-
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
22
Example
At equivalence point. Since Kf' is so large
we know the reaction will lie far to the
right.
Cu+2 + Y-4 ====> CuY-2
[CuEDTA] C0 V0
=
V0 + VT
0.0100 * 0.0500
0.0500 + 0.0500
=0.0050M
[Cu2+ ] = C EDTA
8/3/2009
Chem 311
23
Example
8/3/2009
Kf' = 1.6x108
[CuEDTA]
[Cu2+ ]C EDTA
[Cu2+ ] = 5.5x10-6
Chem 311
0.00500
[Cu2+ ]2
pCu = 5.26
24
12-
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 = CT VT - C0 V0
V0 + VT
8/3/2009
Chem 311
25
Example
C EDTA= 0.0100 * 0.0600 - 0.0100 * 0.0500
0.0500 + 0.0600
= 0.000910
[CuY2- ] = C0 V0
= 0.000500
V 0 + VT
0.110
= 0.00450
8/3/2009
Chem 311
26
Example
[Cu2+ ] = 0.00450
= 3.1x10-8
0.000910 * 1.6x108
pCu = 7.51
Kf' determines sharpness of titration
break.For Cu+2
pH = 2.00 Kf' = 2.3x105
pH = 3.00 Kf' = 2.1x1010 larger Kf' gives
sharper break.
8/3/2009
Chem 311
27
12-
How large a Kf is Needed

Rule of thumb - 1 ppt of sample untitrated
at the end point and a consequent 1 ppt
excess of titrant.
For CY m/l EDTA we can do the following
calculation:
K 'f
0.999 xCm 0
[ MY ]
106
[ M ]C EDTA 0.001xCm 0 x 0.001xCY CY
CY 0.01
8/3/2009
Chem 311
28
K 'f 108
Auxillary Ligands
Problems
High pH required and metal not soluble
Two metals with similar Kf and we want to
titrate just one
Add auxillary ligand - MASKING AGENT
8/3/2009
Chem 311
29
Auxillary Ligands
[M] = m Cm where m refers to the
auxiliary ligand.
Kf
[ MY ]
[ MY ]
[ M ] 4CY mCm 4CY
K 'f' m 4 K f
8/3/2009
Chem 311
30
12-
10
EXAMPLE
Titration of Cd2+ with EDTA at pH 9.24 in
a NH3
NH4+ buffer which also serves to mask Cd
to prevent precipitation.
Both NH3 and NH4+ are 0.100 m/l
Kf for CdEDTA = 3.16x1016
Cd2+ + 5 NH3 <===> Cd(NH3)52+
8/3/2009
Chem 311
31
Example
Cd2+ + 5 NH3 <===> Cd(NH3)52+
K1 = 398 K2 = 316 K3 = 24.5 K4 = 7.59
K5 = 2.09
1
m
1 K1[ L] K1 K 2 [ L]2 K1 K 2 K 3[ L]3 ...

1
1.4 x10 4
1 39.8 1258 3018 2291 479
4 = 0.087
Kf' = 3.16x1016 * 1.4x10-4 * 0.087 = 3.8x1011
8/3/2009
Chem 311
32
Example
Titration should work well.
But note that the titration break is not as
great as in the absence of NH3.
You could not do it without NH3 due to
precipitation of Cd(OH)2.
8/3/2009
Chem 311
33
12-
11
End Point Detection

Colored Complexes.
CuEDTA is bright blue and can be monitored
with a spectrometer.
a colored complex with a substantially lower
Kf' 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
34
End Point Detection
Cu titration with EDTA
Abs.
Vol. Titrant
8/3/2009
Chem 311
35
End Point Detection
Cu and Fe titration with EDTA

Cu E.P.
Abs.
Fe E.P.
Vol. Titrant
8/3/2009
Chem 311
36
12-
12
End Point Detection

Electrochemical.
Endpoints have been monitored by change in
pH
mercury electrode
ion selective electrodes.
8/3/2009
Chem 311
37
End Point Detection

Indicators.
metallochromic indicators
Serendipity science.
1945 - a group of Swiss chemists headed by
Swartzenbach accidentally synthesized the
compound Murexide (Roman purple) which
proved to be an indicator for metals.
8/3/2009
Chem 311
38
Murexide
-O
N
O
O
N
N
O
O
8/3/2009
Chem 311
39
12-
13
EBT
This lead to the development of Eriochrom
Black T
note nitro and sulfito groups
O
N
N
O
HO
8/3/2009
OH
O ChemO311
40
Calmagite
N,N,O,O tetradentate complexes
N
N
O
HO
S
O
8/3/2009
OH
O
Chem 311
41
Indicator equilibria
pK = 6.3
pK = 11.5
H2In- <========> HIn-2 <========> In-3
|
|
|
red
blue
yellow-orange
In-3 + M <====> MIn RED
with Mg, Ca, Zn, Cd, Hg,

Al,
Ga, In, Pb, Fe, Ti, Co, Ni, Cu, Pt, Rare earth
8/3/2009
Chem 311
42
12-
14
Indicator reaction
KfEDTA >> KfIn
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
43
Indicator reaction
For slow reactions the best approach is a
back titration
excess EDTA is added and allowed to react.
Then the solution is back titrated with a
standard Mg+2 solution to the reverse endpoint.
8/3/2009
Chem 311
44
Ca titration - most common

Direct EDTA Titration work
The addition of Mg to the titrant normally
makes the Ca determination better
Ca binds more tightly to EDTA than Mg, the
indicator should be stripped of Ca near the
endpoint and be present in the MgIn complex
form.
This then gives a sharp endpoint just after the
Ca equivalence point.
8/3/2009
Chem 311
45
12-
15
Back Titration
Used when complex formation is slow
Add excess EDTA
Titrate with Mg2+ or Mn2+
8/3/2009
Chem 311
46
Displacement Titration
Useful when indicator is lacking for specific
metal titration.
Add excess MgY complex (standardized
amount)
Allow the metal of interest to displace the
Mg from the complex
Titrate the Mg released with EDTA
8/3/2009
Chem 311
47
Back titration Example

A 50.0 ml solution containing Ni2+ and Zn2+ 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 Mg2+ and required 12.4 ml to reach the
endpoint. An excess of the reagent 2,3-dimetcapto2-propanol was then added to the solution to
displace the EDTA from the Zn2+. The solution was
then titrated with additional Mg2+ and 29.2 ml of
the titrant was required to reach the new endpoint.
Calculate the molarity of Ni2+ and Zn2+ in the
original solution.
8/3/2009
Chem 311
48
12-
16
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
49
Example 2
A mixture of Mn2+, Mg2+, and Zn2+ was analyzed as follows: A 25.00
ml sample was treated with 0.25 gm of hydroxylamine hydrochloride ,
a reducing agent to keep the Mn2+ from being oxidized. The solution
was buffered at pH 10.0 by the addition of 10 ml of ammoniaammonium buffer. A few drops of Eriochrom Black T indicator were
added and the sample was diluted to 100 ml. It was warmed to 40oC
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 Mg2+ from its
EDTA complex. The solution was titrated with 10.26 ml of 0.02065
M Mn2+ 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 Zn2+ from its EDTA complex. It required 15.47
ml of the standard Mn2+ 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
50
Example 2
8/3/2009
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
Chem 311
51
12-
17
Applications
8/3/2009
Spot Tests
Test Strips
Flow Injection Analysis
Immunoassays
Chem 311
52
12-
18
Chem 311
Chapter 14
Electrochemistry
8/3/2009
Chem 311
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
Ohms Law
E=IR
Voltage = current(amps)x resistance (ohms)
8/3/2009
Chem 311
14-
Electrochemical Cell
2 Half Cells
Salt Bridge
Potential = E
eV
Salt Bridge
Pt
Pt
OX
8/3/2009
Chem 311
Ce (IV) 1.0
m/l
RED
Fe (II) 1.0
m/l
Chemical Analysis
A variety of chemical analyses involve
electrochemical cells.
8/3/2009
Potentiometry 311
voltametry 312
amperometry
chronopotentiometry
coulometry
others
Chem 311
Electron Energy Levels

Electron Flow in Metals
Band Theory
Electrons Move in Conducting Band
Electron Levels in Metal Electrodes

Fermi Level
If Fermi Level > Empty Orbital of Ion
Reduction occurs
Vice Versa
8/3/2009
Chem 311
14-
Electron Transfer
Fermi level of
electrons in electrodes
shown
Fe2+ filled orbital
Ce4+ empty orbital
8/3/2009
Chem 311
Electron Transfer
Potential is difference
in energy of two
electrodes.
Measure with no
electron flow
Thermodynamic
potential
G = -nFE
8/3/2009
Chem 311
Measuring Potential
Operational amplifiers
8/3/2009
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
1010 to possibly as high as 1015 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
Chem 311
14-
Potentiometer Circuit
Voltmeter
Voltage Follower
Amp
Salt Bridge
8/3/2009
Chem 311
10
Measuring Potential
Ohm's Law E = IR.
E= EMF volts
R = impedance or resistance ohms
I = current amps
For R=1012, if the cell EMF is 1.00 volts

then I = 1x10-12 amps.
8/3/2009
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
Chem 311
slow.
11
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.
V
Porous
membrane
8/3/2009
Pt
electrode
with H2
bubbler
1.0 M
HCl
Chem 311
Ag electrode
with AgCl
12
14-
Line Notation of Cell

Pt, H2 (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
13
Half Reactions
H2 ==> 2H+ + 2 eOxidation
AgCl + e ==> Ag + ClReduction
8/3/2009
H2 + 2AgCl ==> 2H+ + 2 Ag + 2ClRedox Couple

Conjugate oxidation- reduction pair
E=0.2222 volts at STP
Chem 311
14
Conventions for the

Description of Electrochemical
Cells.
+2
+4
Pt / Fe (1.0 M) // Ce (1.0 M) / Pt
Fe2+ =====> Fe3+ + e- and Ce4+ + e- ====> Ce3+
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, H2 (1.0. atm) / H+ (1.0 M)
8/3/2009
Chem 311
15
14-
Conventions for the

Description of Electrochemical
Cells.
Every phase boundary has a potential

difference associated with it.
Salt bridge or phase boundaries assumed to
generate no potential is designated by the
double '//'.
These are constructed from a tube filled with a solution of an inert

electrolyte. Normally the tube is closed with glass frits or are made from
an electrolyte in agar to minimize the mixing of the bridge solution with
the cell solutions. In many commercial batteries, porous membranes
(paper or cardboard are often used) are employed in place of a traditional
8/3/2009 salt bridge.
Chem 311
16
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 H2. 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
17
1953 IUPAC Conventions

1. Standard Potential Eo all reactants and
products at 1.0 M activity.
2. Std.Potential for a 1/2 cell is the cell
EMF of that 1/2 cell coupled with the NHE.
3. The sign of the Std. Potential is the sign
of the 1/2 cell electrode where the NHE is
operating as the ANODE in the reaction.
(Galvanic cells thus have + Std.Potentials
and Electrolytic cells have - Std.Potentials.
8/3/2009
Chem 311
18
14-
Tables of Std.Potentials
Appendix - Standard Potentials Eo
Formal Potentials - Ignore Activity Eo
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
19
8/3/2009
Chem 311
20
Nerst Equation
Nerst eq. applies equally well to 1/2 RX and
cell RX.
E Eo
8/3/2009
ac ad
0.05916
log Ca Db
n
a A aB
Chem 311
21
14-
Calculate EMF of CELL RX

From Cell Schematic (Shorthand Notation)
Fe/Fe2+(1.0) // Sn4+(1.0), Sn2+ (1.0) /Pt
Fe + Sn4+ =====> Fe2+ + Sn2+
Fe2+ + 2e ====> Fe
Eo Fe2+,Fe = - .447
reduction
Sn4+ + 2e ===>Sn2+ Eo Sn4+ +,Sn2+ = + .151
More negative potential will be oxidation
reverse reaction will occur
8/3/2009
Chem 311
22
Calculate EMF of CELL RX
Ecell = EFe,Fe2+ - ESn2+,Sn4+

0.151 (-0.447) = 0.598 volts
Ecell = EoCathode- EoAnode
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
23
Calculate E1/2 using Nerst At non-standard conditions
Fe3+(.200)//Mn04-(.010),
+
, H (1.0)/Pt
look for 1/2 Rx in table
Fe3+====> Fe2+ + eE Fe3+,Fe2+ = 0.771
EFe3+,Fe2+ = 0.771
Mn04- + 8H+ + 5e- ====> Mn2+ + 4H20
E Mn04-,Mn2+ = 1.51
Pt/Fe2+(.100),
Mn2+(1.0x10-4)
8/3/2009
Chem 311
24
14-

As oxidation
EFe3+,Fe2+ = Eo - 0.0591 log [Fe2+]
1
[Fe3+]
E = 0.771 - 0.0591 log 0.100 = 0.771 + 0.0178
1
0.200
= 0.789 volts
8/3/2009
Chem 311
25

EMn04 ,Mn2+ = 1.51 - .0591 log [Mn2+]H20
5
[Mn04-][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
26

Balanced RX
5Fe2+ + Mn04- + 8H+ + 5e- 5Fe3+ + Mn2 +
4 H20 + 5e-
Ecoll = 1.536 -(0.789) = + 0.747

Note Potential of 5 Fe2+ =
as for 1
8/3/2009
Chem 311
5Fe3+ same
27
14-
Methods of Analysis
Direct Potentiometry
Nerst Eq. activity measurements
Calibration Curve conc. Measurements
Standard Addition
Potentiometric Titration
8/3/2009
Chem 311
28
14-
10
Chem 311
Chapter 15
Potentiometry
8/3/2009
Chem 311
Reference Electrodes
Ag/AgCl reference electrode is also
popular. 0.2222 v
Most common
No Hg
8/3/2009
Chem 311
Reference Electrodes
Saturated Calomel electrode SCE
Hg,Hg2Cl2/ Hg2+2 (sat.), KCl (sat.)// E =
+0.2415 volts
8/3/2009
Chem 311
15-
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
Junction Potentials
K+ and Cl
Best choice
8/3/2009
Chem 311
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 23 mV.
Not a problem in titrations and std. add
calibration curves can be a problem matrix
See Rainwater example in text
8/3/2009
Chem 311
15-
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
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
MEMBRANE INDICATOR
ELECTRODES
8/3/2009
Glass electrode (pH) is good example.

If only one type of charged ion can move
Emem = - RT ln a2
nF
a1
activity. of solutions on two sides of
membrane
Chem 311
15-
MEMBRANE INDICATOR
ELECTRODES
If a1 is a constant
Emem = K - RT ln a2
nF
Glass membrane
Si02 Li20 or Na20 Ca0
8/3/2009
Chem 311
10
Glass Membrane Electrode

Need
solutions for
electrical
contact with
membrane
Dry Glass
External
Solution
8/3/2009
Internal
Solution
Chem 311
11
Hydrated gel
pH electrode
Volt
meter
Total Potential
includes potential
of electrode on
both sides of the
membrane.
pHunk = pHs +
.0591
Eunk-Es
8/3/2009
Reference
Solution
External
Electrode
Internal Solution
and Electrode
Chem 311
Porous Junction
12
15-
Glass Electrodes
Glass electrodes with Na20 Al203 B203
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
13
Ion SELECTIVE Electrodes

If electrode responds to several different
ions (most electrodes do)
relative response described by selectivity
coefficient kx,y
relative response to ion x in presence of ion y
ns
0.05916
log [ X ] k x , y [Y ] n y
E constant
y
nx
8/3/2009
Chem 311
14
Ion SELECTIVE Electrodes

ns
0.05916
log [ X ] k x , y [Y ] n y
E constant
y
nx
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.
8/3/2009
Chem 311
15
15-
Example of Selectivity
Na+ glass electrode 11% Na20 18% Al203 71%
Si02
Specific example for sodium electrode and

potassium interference
KK/Na = 2800
Na+ response is = 2800 x K+ response
KK/Na = 1/KK/Na = 1/2800 = 3.57x10-4
8/3/2009
Chem 311
16
Example of Selectivity
For interference < 1% of concentration
[X] > 0.01 * Kx,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
17
Liquid Ion Exchange

Membranes
Immiscible organic phase
interposed between
internal reference solution
and external solution.
Organic phase contains
some substance which will
transport one ion
preferentially across
membrane. Porous to one
type of ion.
8/3/2009
Chem 311
18
15-
Liquid Ion Exchange

Membranes
Ca didecylphosphate neutral (org. soluble)
overall E = Q + .0591 log [ Ca2+]
2
Note N = 2 sensitivity only 1/2 that for N = 1
8/3/2009
Chem 311
19
Liquid Ion Exchange

Membranes
Other Liquid exchanges
Macrocyclics
penicillin
Aureomycin
Valinomycin
Crown Ethers K+ 16 crown 5

Alkyl phosphates Ca2+
Bidentate ligands (transition metals)
8/3/2009
Chem 311
O
O
20
Liquid Ion Exchange

Membranes
Ion Pair Formation
Tetraalkyl ammonium salts
Alkyl amines
1,10 phenanthroline Fe (III) salts
8/3/2009
Chem 311
21
15-
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
8/3/2009
KCl
filling
solution
Chem 311 Organic ion

Exchanger
Tungsten wire
22
Treated Tip
Solid State Membranes

LaF3 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- KA = 6.8x10-4
pH must be controlled (5.5 typical)
8/3/2009
Chem 311
23

also F- + Mn+ MF metal complexes
add NTA or EDTA to free F Other Solid State Electrodes
AgS
+ AgCl AgBr AgI AgSCN
Rapid and selective
8/3/2009
(Ag+, S2-, Cl-.)
Chem 311
24
15-

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
25

Shape of
curve
E
Slope= 0.0592/ z
Solubility limit
Log_Conc entration
8/3/2009
Chem 311
26
Heterogenous membranes
Silicone polymer + precipitate
of some insoluble salt.
PVC
Coated wire electrodes
Tungsten wire - treated to
become pH electrode
8/3/2009
Chem 311
27
15-
Gas Sensing Electrodes

Gas permeable membrane
Ammonia Electrode
NH3 + H+ <===> NH4+

pH electrode or NH4+ electrode
Also available for CO2, S02,
H2S & N02
8/3/2009
Chem 311
28
Enzyme Electrodes
ISE, pH, or gas electrode
Urease <===>NH3
Decarboxylazes===>H+
pH Electrode
Enzyme in
acrylamide gel
2 enzyme sandwich 8/3/2009
cholesterol electrode
Chem 311
29
Enzyme Electrodes
Nerve gas electrode (& phosphate
pesticides) - Cholinesterase inhibitors
enzyme film
add substrate to sample
monitor reduced activity
Vitamin D electrode etc.

Nick Schmit with Gary Rechnitz
8/3/2009
Chem 311
30
15-
10
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 semiconductor circuits (Metal Oxide
semiconductors) now used as sensors.
Mechanical nose - sensor array

spoiled food, ripe fruit, mine detection
8/3/2009
Chem 311
31
Semiconductor Electronics
Silicon (4 e) doped with
Al 3 e p-type
P 5 e n-type
8/3/2009
Chem 311
32
Diode
Allows electron flow only np
8/3/2009
Chem 311
33
15-
11
FET
2 PN Junctions
8/3/2009
Chem 311
34
Analysis using FET
8/3/2009
Chem 311
35
FET and MOS-FET
Sensors for Solution

Sensors for vapors
Made in arrays 10x10 etc
Each FET with different Chemical sensor
Multiple sensitivities
8/3/2009
Chem 311
36
15-
12
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
37
Methods of Application of
ISE's
Calibration Curves
Standards and samples of similar ionic strength
Keep Ejunct 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
38
ISE's
Standard Addition
Measure sample
E = Q - .0591 log Cx
n
Measure sample + Std.
E = Q - .0591 log (Cx+Cs)
n
Assumes linear portion of response.
8/3/2009
Chem 311
39
15-
13
ISE's
If addition causes little volume change Ej
will stay the same
Std. allows compensation for unknown
matrix
8/3/2009
eg. ionic strength and Ej effects minimized

Ex = Q + .0591 log Cx
n
Es = Q + .0591 log VsCs+VxCx
n
Vs+Vx
Chem 311
40
ISE's
n( E x E s )
V C Vx C x
log C x log s s
Vs Vx
0.05916
n(E E)
Take inverse log

rearrange
x s
Cx(Vs Vs)
100.0592
VsCs VxCx
Cx
8/3/2009
Cs Vs
n(E E )0.0592
(Vx Vs ) 10
Chem 311
Vx
41
ISE's
Graphical treatment of std. addition

Log C on X axis
Put Cx at 0
Plot Standards
Extrapolate to
Y=0
Potential
8/3/2009
Chem 311
-Cx
42
Unk.
Unk+S1
Unk+S2
15-
14
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
Chem 311
43
Clinical Analyzers
Electrochem on a Chip
Disposable
ISEs
8/3/2009
Chem 311
44
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
8/3/2009
Chem 311
45
15-
15
Chem 311
Chapter 18
Fundamentals of
Spectrometry
8/3/2009
Chem 311
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
8/3/2009
vacuum 3.00 x 108 m/sec

slower in all other media - slowed by electronic
interactions
Chem 311
= Refractive index = speed in vac
speed in medium
>1.00
function of wavelength
light of different energy moves at different rates
in medium other than vacuum.
Fiber Optics separate wavelengths of a pulse
8/3/2009
Chem 311
18-
(wave number) = waves/cm = cm-1 = 1/
in vacuum
Frequency ( ) = waves passing fixed

point/sec.
period (p) time per wave crest = 1/p
all are related
= c
E = h = hc/ = hc
8/3/2009
Chem 311
Electromagnetic spectrum
8/3/2009
Chem 311
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
Rotational Transition
Vibrational Transition
Electronic Transition
8/3/2009
Chem 311
18-
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.
8/3/2009
Studied for molecules (hard to exited enough to

emit.)
Chem 311
Molecular Energy Levels

orbitals
Bonding and Antibonding orbitals
CAChe and Spartan calculate these
8/3/2009
Chem 311
Molecular Energy Levels
8/3/2009
Computation chemistry
Compute energy of all MOs
Predict UV-Vis spectra
Predict IR Spectra
Predict NMR Spectra
Chem 311
18-
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
Chem 311
10
Absorption
dx
P(o)
P(x)
if each thin segment (dx) absorbs some

fraction of all of incident light (Px)
dP = -k Px C dx
8/3/2009
Chem 311
11
Absorption
dP = -k Px C dx
k= constant for absorbing species
cross section area of absorbing region of molecule (0rbital or
conjugated system
Probability of transition occurring (0 => 1 )
PX =Incident radiation power

C = Concentration
-dP/Px = kC dx
Integrate over entire path length x= 0 to b
8/3/2009
Chem 311
12
18-
Absorption
log(P0/P) = k/2.303 Cb = bC
or (abC)
P = Transmittance (T)
Po
-log T = A Absorbance - A is dimensionless
A = abc
A= bc
C in gm/l
C in moles/l
bC = cm*mol/1000 cm3 = mol/1000 cm2

a units cm2/gm
unit = cm2/mol
(old literature often dm2/gm)
8/3/2009
Chem 311
13
Limitations on Beers Law

Light must be monochromatic
Parallel
Enter at a right angle.
8/3/2009
Chem 311
14
Accuracy
Deviations from Beers Law
Instrumental Deviations
Non-monochromatic light
value of or a not constant across bandwidth of
spectrometer.
8/3/2009
Negative deviation at high conc.

Concentration error
lower sensitivity.
A
Need more standards.
Chem 311
15
Conc.
18-

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
Chem 311
16
Bandwidth effects
8/3/2009
Chem 311
17
Bandwidth effects
8/3/2009
Chem 311
18
18-

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
Chem 311
19

Diagnostic Tool for Deviations
Plot A vs path length.
Beers Law - straight line (deviations must

be chemical)
Stray Light - negative deviations
Reflections inside instrument
Higher orders from a grating
slit diffraction around entrance slit
8/3/2009
Chem 311
20
Concentration Errors- Precision

Assume error is a constant value of
Transmittance (T)
A=abC A= -log T
C = - log(T)/ab
take derivative of C with respect to T
dC/dT = -0.4343/T(ab)
8/3/2009
Chem 311
21
18-

Want relative concentration error dC/C so
divide by
0.4343
dC C
C= -log(T)/ab
T
log(T)
dT
ab term cancels
dC
dT 0.4343
x
C
T
log(T)
dC/C has a minimum at T=0.368 (36.8%)
8/3/2009
Chem 311
22
Error as f(%T)
Working Range
very short
factor of 5-10
error
36.8%T
%T
8/3/2009
Chem 311
23
8/3/2009
Chem 311
24
18-
Chem 311
Chapter 19
Applications of
Spectrometry
8/3/2009
Chem 311
Applications of
Spectrophotometry
Direct determination of a chromophoric
compound
anything that absorbs strongly.
Absorptivities range from 0 to 500,000 , wide
range of sensitivities.
8/3/2009
Chem 311
Applications of
Spectrophotometry
Form a chromophore with non-absorbing species
metals react with ligands to form colored complexes large number of analytical methods developed to use
this
organic derivatives - 2,4-dinitrophenyl hydrozones
azo coupling- make azo dye -acid rain nitrate
vanillate ion in lab
breath-a-lizer alcohol detn.
8/3/2009
Chem 311
19-
Methods of Quantitation
Direct Use of Beers Law
Least Precise and Accurate (one point
calibration) assumes blank=0.00
Known concentrations vs Abs. Least Squares

Intercept need not be Zero
Identify non-linearity if present
Subtract Blank or Zero instrument with Blank
8/3/2009
Chem 311
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
Cunk
8/3/2009
C stdVstd
Aunk std
(Vstd Vunk ) Vunk
Aunk
Chem 311
Standard Addition
Useful if matrix of sample has background
absorbance which cannot be accounted for
in a blank or calibration curve
Quick if only one or two samples are to be
run
8/3/2009
Chem 311
19-
Extensions to Beers Law

Multi-component Systems
A 1 = 1bC1 + 2bC2 + 3bC3 +.....
Total abs. = sum of absorbencies of individual
absorbing species.
Measure at several wavelengths solve

simultaneous equations.
Calc. conc. of all species.
8/3/2009
Chem 311
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 ML2
.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
Chem 311
Spectrophotometry to study
reaction stoichiometry
Used for - metal complexes, enzyme
substrate complexes, etc.
Continuous variation - Jobs Method
Use where ratio is close to 1:1
Make series of solutions where Total moles of
two reactants constant.
Plot Mole ratio vs A
8/3/2009
Chem 311
19-
Jobs Method
8/3/2009
Chem 311
10
Scatchard Plot
Measure Equilibrium Constant
Biochemists
8/3/2009
Chem 311
11
Scathcard Plot
8/3/2009
Chem 311
12
19-
Scatchard Plot
Slope = - K
K=4.0x109
8/3/2009
Chem 311
13
Instrumentation for Optical

Spectroscopy
Source of
monochromatic
radiation
8/3/2009
Sample
Detector
Readout
Chem 311
14
Sources of Radiation
Black body radiators Tungsten lamp 2870oK - 1.5 micron peak
visible only - 330 nm minimum
8/3/2009
Chem 311
15
19-
Monochromators
Filters
Glass 30-50 nm band width 5-20% T at max
Interference Filters 10-20 nm bandwidth 40% T
Prisms - Quartz for UV-VIS

Gratings - parallel lines on glass
Most common dispersion device
Fourier Transform instruments - Nondispersive (IRs) - Details in Chem 312

8/3/2009
Chem 311
16
Dispersion by a Grating
8/3/2009
Chem 311
17
Wavelength Selectors
Definition of effective bandwidth
8/3/2009
Chem 311
18
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
Chem 311
19
Solvents - must be transparent

UV cutoff
Solvent
Acetone
Acetonitrile
Benzene
Carbon disulfide
Chloroform
Dichloromethane
Ether
Ethyl Acetate
Hexane
Methanol
Water
8/3/2009
UV Cutoff
330
210
280
380
245
233
220
260
210
210
200
Chem 311
20
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.
8/3/2009
Chem 311
21
19-
Detectors
Solid State Detectors
Photodiodes
Change Couple Devices.
Photodiodes - pn junction conduct in

reverse direction due to photon flux.
8/3/2009
Chem 311
22
Detectors
Linear photodiode Arrays
512 diodes - detect 512 wavelengths at once complete spectrum not scanned. HP
Spectrometer
.
8/3/2009
Good visibility sensitivity

Rapid response
high linearity
Chem 311
23
Instrument Designs
Photodiode array spectrophotometer

8/3/2009
Chem 311
24
19-
Fiber Optic Dip Probes

No cell required
Mirror reflects
8/3/2009
Chem 311
25
Flow Injection Analysis

Inject pulse of sample with valve
Mix with reagents
Pass through detector cell
8/3/2009
Chem 311
26
FIA
Calibration Data
Autoanalyzers
Common in Clinical
8/3/2009
Chem 311
27
19-
ELISA
8/3/2009
Chem 311
28
ELISA
Requires specific antibody for each analyte
Antibody bound to substrate
Magnetic particles
Solid supports
Etc
y = -0.6456Ln(x) - 0.8658
2
R = 0.9886
2.000
logit % B/Bo
1.500
1.000
0.500
0.000
-0.500
8/3/2009
-1.000
0.01
Chem
311
0.1
Conc. (ug/L)
29 1
19-
10
Chem 311
Chapter 23
Separations
General Chromatography
8/3/2009
Chem 311
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, Coprecipitation.
8/3/2009
Chem 311
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
8/3/2009
Chem 311
23-
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
SEPARATIONS AND
EXTRACTION
Separation necessary when measurement
technique not adequately selective
Contact between two immiscible phases
Separation of phases
8/3/2009
Chem 311
Phase Combinations
8/3/2009
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.
Chem 311
23-
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
HPLC
Thin layer G-L

Thin layer L-L or L-S
8/3/2009
Chem 311
Solvent Extraction
Equilibrium of solute Z between two
immiscible phases:
Z(Aq) <===> Z(org)
Keq = Kp = [Z]or
[Z]Aq
Partition Coefficient
Activity coef, rarely known - use conc.
8/3/2009
Chem 311
Solvent Extraction
Multiple equilibria often occur
major advantage of extraction methods.
M+ + L- <====> MLAq
+X
+H+
||
||
||
V
V
V
MX+
HL(Aq
ML(org) + B <===> ML . B(org)
||
V
HL(0rg)
8/3/2009
Chem 311
23-
Solvent Extraction
Most useful information - total conc of all
forms.
D = Cor =
[ML]or + [ML.B]or
Caq
[MX+] + [M+] + [ML]Aq
Distribution Ratio
8/3/2009
Chem 311
10
Fraction Extracted
Calculation of fraction of total in each
phase.
Start with all solute in Aq - phase.
q = fraction remaining in Aq phase = CAq
Co
Cor = (CoVAq - CAqVAq)/Vor = (Co-CAq) VAq
Vor
D = Cor = (CoVAq - CAq VAq)/Vor=Vaq x Co - CAq
CAq
CAq
Vor
CAq
8/3/2009
Chem 311
11
Fraction Extracted
Co Vor
1
xD
Caq Vaq
Co = initial conc.
Caq
Co
1
1
Vor
1
V
RD
1
D
Vaq
VR= Vor/Vaq
fraction in org. phase = p
8/3/2009
Chem 311
p 1 q
VR D
1 VR D
12
23-
Ps and qs
Special case VR = 1
q= 1
p=
D
1+D
1+D
% Extracted = 100p
8/3/2009
Chem 311
13
Simple extraction
p = fraction in organic phase D VR
1+DVR
q = fraction in aqueous phase
1
1+DVR
Extract efficiency depends on D & VR

For small values of D it becomes necessary to
increase VR to get complete extraction. Practical
limits to VR about 1000
8/3/2009
Chem 311
14
Repeated - Stepwise Extraction

Use several organic portions to extract
aqueous phase (Vanilla Experiment).
p1 = DVR
1+DVR
p2 = DVR
1+DVR
8/3/2009
q1 =
1
1 + DVR
1
= DVR
= pq
1+DVR (1+DVR)2
q2 = 1
* 1
= 1
= q2
1+DVR 1+DVR (1+DVR)2
Chem 311
15
23-
Repeated - Stepwise Extraction

Third ext.
p3 = p*q2 = DVR
* 1
=
D VR
1+DVR
(1+DVR)2
(1+DVR)3
q3 = 1
. 1
=
1
= q3
1+DVR (1+DVR)2
(1+DVR)3
org phase
pqN-1
Aq phase
qN
total solute if all organic layers combined = (1-qN)
pT = p1 + p2 + p3 = p + pq + pq2 = total fraction
Total quantity = pT CoVAq = (1-qN) CoVAq
8/3/2009
Chem 311
16
Example
Extraction of Acetanilide from water into ether
D = 3.0
Single Ext VR = 1
ptotal =
= 3/4 = 0.75
1+D
Final Conc = 0.75*Co = 0.75
Single Ext VR = 10 ptotal = D =
1+D
ptotal =3*10/(1+3*10) = 0.97

Final Conc = 0.97 * Co/10 = 0.097
8/3/2009
Chem 311
17
Example
Extraction of Acetanilide from water into eth
5 ext. VR = 1
pt = 1- (1 /1+D) 5 = 1-(1/4)5
ptotal = .999
Final conc = 0.999*Co/5 = 0.1998 = 0.2
much more efficient to do multiple extractions

Max possible extraction efficiency
limit where Vaq divided into infinite no. of portions
8/3/2009
pT = 1 - q
Dc
Chem 311
q e
Vor
Vaq
18
23-
Multiple extractions
In general N = 5 is optimum gets within a factor of
5 of max. possible efficiency.
8/3/2009
Chem 311
19
Separation of 2 solutes
Separation factor depends on ratio of
DcA/DcB
Ratio must be very large to obtain high
purity 106.
Optimum conditions for separation
VR = (DcA/DcB)1/2
8/3/2009
Chem 311
20
Solvent Extraction
Extraction of Organic Molecules
Acids, bases, and neutral molecules
8/3/2009
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
Chem 311
21
23-
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
22
Solvent Extraction
Extraction of Organic Molecules
Dc HA K p'
Text uses only case for monoprotic but
equation applies to all. Use alpha for neutral
form.
8/3/2009
Chem 311
23
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.
8/3/2009
Chem 311
24
23-
Simplest Metal Ext. Case
bn
Mn+ + nL- <===> MLn(Aq]
+H+
KpML
KA
MLN(or)
HL(Aq)
KpHL
HL(or)
8/3/2009
Chem 311
25
Distribution
D=
[ML]or
[M+]Aq + [ML]Aq
bn = [MLn]Aq
[M+N][L]n
KpML= [MLn]or
[ML]Aq
8/3/2009
KA= [H+][L-]
[HL](Aq)
KpHL = [HL]or
[HL]
Chem 311
26
Distribution
[MLn]or = KP[MLn]Aq [MLn]Aq = Bn[Mn+][L-]n
[MLn]or = KPBn[Mn+][L-]n
[HL]A = [HL]or
KPHL
[L-] = KA[HL]or
[H+ ]KPHL
8/3/2009
[L-] = KA[HL]A
[H+]
Chem 311
27
23-
Distribution
K HL]
[ MLn ]or K pML n [ M n ] a or
[ H ]K pHL
D = [MLn]or
[M+]Aq + [MLn]Aq
assume complex not water soluble [Mln]Aq =0
Generally [MLn]Aq is very small except acetyl
acetonates and water soluble ligands with
additional charged sites
8/3/2009
Chem 311
28
Metal Distribution
Dc
pML
n ( K a ) n [ HL ] n or
(K
) n [H
pHL
D c ( constant
8/3/2009
] naq
) n [H
] aq
Chem 311
29
Metal Distribution
Variables to control
pH
Organic Solvent (Changes Kp)
Dc
pML
n ( K a ) [ HL ]
(K
pHL
D c ( constant
8/3/2009
Chem 311
) n [H
) n [H
n
or
] naq
] aq
30
23-
10
Metal Distribution
Multidentate
ligands give
sharper pH
extraction
breaks
n=2
n=3
n=1
% Ext.
pH
8/3/2009
Chem 311
31
Metal Extraction
Control of pH allows separation of species
with different B's and different KPML'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
8/3/2009
MLn[org] + B <====>
increase extraction
MLn . B[org]
Chem 311
32
A little History
Solvent Extraction Research - Alternative to
Precipitation
Craig Counter Current Extraction
Chromatography - Continuous Extraction
8/3/2009
Chem 311
33
23-
11
CHROMATOGRAPHY AND
SEPARATIONS
Problems with difficult separationsCounter 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
34
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 CHCl3 through (mobile
phase) worked for fatty acids.
8/3/2009
Chem 311
35
CHROMATOGRAPHY AND
SEPARATIONS
Then invented Gas chromatography and the
common detectors for GC
Martin and Synge - Nobel Prize 1952
Developed extension of counter current math to
describe chromatography.
Not best treatment (ignores kinetics) but worth
some time.
Three types of chromatography - Elution,

Frontal, and Displacement
8/3/2009
Chem 311
36
23-
12
Elution chromatography - most

common
Solute placed at top of column.
Eluted until each component reaches the
end (detector).
8/3/2009
Chem 311
37

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
38

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/to
10 cm length must contain 1.00 ml of mobile phase

Vol mobile phase = to x Flow Rate
8/3/2009
Chem 311
39
23-
13
Elution chromatography
Total column volume 4.6 mm diameter

x 10 cm
2
Vt = pr h = 1.66 ml.
Mobile phase volume
Vm = 1.00 ml
Stationary phase volume
Vs = Vt- Vm = 1.66-1.00 = 0.66 ml
Phase Ratio F = Vs/Vm = .66/1.0 = 0.66
8/3/2009
Chem 311
40
time ratio in phases Tstationary
Tmobile
Cstationary Vstationary
x
Cmobile
Vmobile
K = Cs partition ratio
Cm
=KF
Thermodynamic basis for
retention
8/3/2009
Chem 311
41
Capacity factor k'
- Describes Retention
Most common for HPLC

time in stationary phase/time in mobile phase
k'
Vr Vo t r t o t ' r
Vo
to
to
k' varies from 0 to large numbers

ln k' = D H - D S - ln F
RT
R
8/3/2009
Chem 311
42
23-
14
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
43
Separation Factor
Relative Retention = tr2 / tr1 = k2/k1
8/3/2009
Chem 311
44
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
8/3/2009
Chem 311
45
23-
15
Band spreading in
chromatography
Broadening in column
As it moves through column, it begins to
spread -
= std. dev. of peak profile

Number of theoretical plates N
N= tr2/ 2
peak width at base tw = 4
8/3/2009
Chem 311
46
Band spreading in
chromatography
N= 16(Vr/Vw)2 = 5.54(Vr/V1/2)2
Vr = tr Vw = tw
use any units time, length, volume
8/3/2009
Chem 311
47
Band spreading in
chromatography
HETP = L/N
Plate Theory ===> van Deemter Equation
H= A + B/v + Cv
where v= mobile phase velocity
8/3/2009
Chem 311
48
23-
16
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
49
Band spreading in
chromatography
Longitudinal Diffusion of solutes
D 10-5 to 10-7 function of MW
8/3/2009
Chem 311
50
Band spreading in
chromatography
Mass Transport between phases
8/3/2009
Chem 311
51
23-
17
Band spreading in
chromatography
Multiple Flow Paths - Eddy Diffusion
Not present in Open Tubular Columns
8/3/2009
Chem 311
52
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
53
HPLC vs GC
Differences due to
diffusion rates in liquids vs gases
particle sizes
GC
H
LC
1E-5
8/3/2009
1e-4
1e-3
1e-2
Chem 311
veloc ity c m/ sec
1e-1
10
100
54
23-
18
Reduced parameters
v = dpv/Dm
h = H/dp
All curves superimpose with h minimum at

about 2dp = 2 particle diameters is the
minimum length of column required for a
theoretical plate. v optimum = 1
Dm for liquids 10-5 to 10-7
Dm 10-2 to 10-1
8/3/2009
Chem 311
55
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
56
Time required for separation

Plates/second
Optimum k = 2.0
12
Log tr
10
8
1 year
LC
6
1 day
GC
1 hour
8/3/2009
1 min
Chem 311
2
57
Log N
23-
19
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
58
Deviant Chromatographic
Behavior
Non-linear
partition
isotherms
Anti-Langmuir
Isotherm
Ideal behavior
Conc
Stationary
phase
Langmuir
Isotherm
8/3/2009
Chem 311
Conc mobile phase
59
Behavior
Antilangmuir
Anti-langmuir case
sample acts
as
stationary
phase
Langmuir
8/3/2009
Saturate
sites
Langmuir case
Chem 311
60
23-
20
Behavior
Anti-Langmuir
8/3/2009
Chem 311
61
Extra-column Volume
Broadens the injected band before it reaches
the column
Broadens the post column bands before the
detector
For good peak shape
Vcol >>> Vinlet + Vdetector+ Vconnections
8/3/2009
Chem 311
62
Resolution in Chromatography
R= Dtr/4 = Dtr/Wav
= (tr2 - tr1)/0.5(W1 + W2)
most common equation for determining R
from chromatogram
8/3/2009
Chem 311
63
23-
21
Resolution
8/3/2009
Chem 311
64
R
( 1 )
1
4
k'
k'
8/3/2009
k'
1 k'
separation factor
Chem 311
65
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
8/3/2009
Chem 311
66
23-
22
Types of Columns
Packed columns
open tubular
coating on walls
Packed column
For open tubular columns there is no flow

paths term He = 0
8/3/2009
Chem 311
67
General Elution Problem

Single set of
conditions good
for only 6-8
components
8/3/2009
Chem 311
68
Chem 311
69
Capillary GC
of FAMES
Isothermal GC at
150 degrees
Note Tr separation
between even Cs
almost doubles
8/3/2009
23-
23
Capillary GC of FAMES
Chart Title
5.45
6.98
10.04
16.24
22.02
31.68
45.58
8/3/2009
y = 0.079x - 0.2364
R2 = 0.9963
2.0000
1.5000
Log Tr
C number Tr
12
14
16
18
20
22
24
Log tr
1.0000
Linear (Log tr)
0.5000
0.0000
0
10
20
30
Carbon Number
Chem 311
70
Capillary GC of FAMES
Temperature Programmed run
8/3/2009
Chem 311
71

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
8/3/2009
Chem 311
72
23-
24

Initial conditions k' very large most peaks held
at start of column
Special case in capillary column GC cryofocusing.
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
73

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
8/3/2009
DryLab Software for

optimization
Chem
311
74
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.
8/3/2009
Chem 311
75
23-
25
Sample Capacity
Isocratic
Max sample proportional to tr/2N1/2
Programmed
Max sample proportion to tro/2N1/2
tro = retention time at initial conditions (very
large)
8/3/2009
Chem 311
76
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
77
Frontal Chromatography
Mixture used
as mobile
phase
Great if you
want Solute 1
only
8/3/2009
Chem 311
78
23-
26
Displacement
Chromatograph
Great for y
Preperative
Separations
Gram Quantities on
standard columns
Must have displacer
Must determine
isotherms
8/3/2009
Chem 311
79
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
80
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
8/3/2009
Chem 311
81
23-
27
Chem 311
Chapter 24
Gas Chromatography
8/3/2009
Chem 311
Types of Columns
Column type
Typical
length
Typical
diameter
Preparative
packed
Packed
Analytical
Fused silica
capillary
2-4 meters
to in OD 100
Typical
number of
plates
1-10 meters 1/8 in OD
1000
10-100
meters
10,000100,000
8/3/2009
0.05 1 mm
ID
Chem 311
Types of Columns
Capillary columns
steel, glass, fused silica.

.067 - 1 mm ID
1000 -100 plates/ft
Columns may be very long due to low pressure
required
Typical 10 meters - 60 meters (GC Lab 30 meter)
30,000 106 plate possible

100,000 typical
8/3/2009
Chem 311
24-
Silicone Phases for GC

Functional Group
Methyl
Polarity
Non-polar
Application
Boiling point
separations
Phenyl
Slightly polar and pi
5% - 100% of silicon
bonding
General application
with more polar
molecules being
retained longer for
same BP
Separation by
polarity
Works when others
dont
Cyano
Very polar
5% - 100% of silicon
C3F7
Moderately polar
8/3/2009
Temp limits 200Chem 311

300oC
Liquid Phases for GC

Polymer
Polarity
Application
Poly-ethylene glycol Very polar

Carbowax
HO- [CH2-CH2-0 ]nH
MWs to 20,000
General separation
by polarity
Ethylene glycol
esters
DEGS di-ethylene
glycol succinate
Fatty acids and other

polar moleucles
Very polar
O
O
8/3/2009
O
O
Chem
311
O
5
n
Solid Supports for GC

Adsorption on solid surface - Strongest
stationary phase interaction
Useful for permanent gases
GSC silica, graphite, firebrick
1000 m2/gm surface
2-5 gm in a column
As vapor pressure decreases need less

retention.
Minimizing Solid Support Interactions
8/3/2009
Chem 311
24-
Solid Supports for GC

Inertsupports with low surface area
0.2 - 1 m2/gm surface
Diatomaceous earths - high surface area

silica
Acid wash
remove metals
Calcine
reduce surface
silanize
bond surface hydroxyl groups
Coat with liquid phase
1000 available
8/3/2009
Chem 311
Instrumentation for GC
Carrier Gas Supply
Flow Controller
Pressure Controller
Pressure Programming
Injector
Flash Vaporization
On Column
Split- Capillary
8/3/2009
Column Oven
Low Thermal Mass
40 deg/min dT
Chem 311
Detector
Readout
Computer
Integrator
Recorder
Split/Splitless
Injector
8/3/2009
Constant Pressure
Split vent electrical valve
Expansion of solvent
Inert liner
quartz
silanized glass
Chem 311
24-
Injection
Syringe Injection 0.1 to 10 ul +/- 2-3%

Gas samples can be up to 2 ml
Valve injection gases
SPME other desorption techniques
Headspace analysis
Purge and Trap
Cold Trapping
8/3/2009
Chem 311
10
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
Rr
Rs
Amp
V
Rr
8/3/2009
Rs
Reference
Chem 311
Column
Sample
Column
DC Power
Supply
11
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 104
Detects everything except carrier gas
8/3/2009
Chem 311
12
24-
Ionization Detectors- Flame

ionization
Electrode
200 volt DC
Flame
Electrometer
Sample +
Hydrogen
8/3/2009
Chem 311
13
Ionization Detectors- Flame

ionization
Used for C-H compounds.
Electrode
200 volt DC
Flame
Gram response approximately constant

C=0 & C-X lower response
Formaldehyde H2CO
Electrometer
Sample +
Hydrogen
very low response
current 10-14 to 10-7 amps.

10-12 gm/sec detection
Total peak 0.1 to 1ng
linear range 106

ionization efficiency .1%
8/3/2009
Chem 311
14
ECD Electron Capture Detector

Power
Supply - DC
or Pulsed
Electrometer
Sample
Beta
source
b e- + carrier establish background standing

current
8/3/2009
Chem 311
15
24-

Standing Current 10-9 to 10-10 amps
Sample + esamplereduces standing current
negative peaks
sensitivity depends on electron affinity
varies by 104
Electrometer
Power
Supply - DC
or Pulsed
+
Sample
Beta
source
8/3/2009
Chem 311
16

halogenated samples best pesticides PCB's
CHC13 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%
Power
Supply - DC
or Pulsed
8/3/2009
Chem 311
Sample
Electrometer
17
Beta
source
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
8/3/2009
Chem 311
18
24-
GC-MS
CI Chemical Ionization
CH4 + e- CH5+
CH5+ + M MH+ + CH4
High efficiency ionization
Few Fragment ions
Much higher sensitivity
No Library Search possible
8/3/2009
Chem 311
19
GC-MS
Selected Ion Monitoring
Find characteristic ion in each spectrum
Produce chromatogram of specific ion
Total Ion Chromatogram

All ions in each spectrum summed
MS-MS
Fragment a Fragment
Improved S/N ratio
8/3/2009
Chem 311
20
Detector Summary
Detector
FID
ECD
Best case limit

Linear range
of detection
10-12 gm/sec
106
10-11 gm in peak
Applicability
10-15 gm/sec
300-10,000
10-14 gm in peak
Cl and F
molecules
NPD
N or P
FPD
Mass
Spectrometer
10-12 to 1015gm/sec
TCD
10-9 gm/sec
10,000
10-7 gm in peak Chem 311
8/3/2009
All
hydrocarbons
100,000
Identification
and
quantitation
Non-destructive
21
24-
Photo ionization
7-12 eV H2 Lyman line - far UV

Ionization Eff 1%
Less background noise than FID
More sensitive by 10 - 50 then FID
Somewhat selective depending on
wavelength
C1-C4 He N2 not detected
8/3/2009
Chem 311
22
Thermonic Detector
8/3/2009
N or P specific FID
alkali metal FID
Bead of CeI placed in flame of FID
Enhances the sensitivity to N and P
Chem 311
23
Flame photometric
Sulfur specific
Flame with photodetector
Useful fo analysis of fuels
8/3/2009
Chem 311
24
24-
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
Chem 311
25
Identification of Peaks
Plot Log tr vs Carbon Number
Linear graph for homologous series
Log plots on two different columns
Identify
compound
log Tr
Col A
8/3/2009
Chem 311
26
log Tr Col B
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 spectrum of peaks
8/3/2009
Chem 311
27
24-
Analysis Methods
make calibration curve
run sample and compare
Limited by injection accuracy
2-4%
1-2%
0.5%
manual
auto
valve injections
8/3/2009
Chem 311
28
Analysis Methods
eliminates injection error - only integration
error is left
add standard to each sample use it to correct for
variation in injection and in sample workup etc.
Single point - determine peak ratio with one
standard mixture
Calibration curve - plot area ratio vs mass ratio
8/3/2009
Chem 311
29

Data from GC experiment
C22:1
C16:0
y = 0.6548x - 0.0804
R2 = 0.9989
5
4
Series1
20
Series1
15
10
Linear
(Series1)
y = 1.0405x 0.3135
R2 = 0.9991
Linear
(Series1)
5
0
0
0
8/3/2009
Chem 311
10
20
30
24-
10
Chem 311
Chapter 25
HPLC
8/3/2009
Chem 311
A little History
8/3/2009
Chem 311
Types of particles
8/3/2009
Chem 311
A4
25-
Slide 3
A4
LC/GC April 2006 p 10
Administrator, 11/13/2006
25-
Columns
Small particles
More pressure
Harder to pack.
Pressure required increases

P=k/dp2
Small particles reduce C term in HETP
8/3/2009
Chem 311
A5
8/3/2009
Chem 311
Effect of particle size

Xanthines
8/3/2009
Chem 311
A6
25-
Slide 5
A5
LC/GC December 2005 p 1251
Slide 6
A6
LC/GC April 2006 p 10
25-
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
8/3/2009
Chem 311
9 components in < 30 sec

Alkylphenones
8/3/2009
Chem 311
25-
Columns
Short Columns
3 and 5 cm length Fast HPLC
Narrow Bore 1 mm and 2 mm ID
low flow rate for LC-MS
Use less solvent
Increased sensitivity
8/3/2009
Chem 311
10
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
11
Polar Force
Normal phase LC
Silica or Aluminum solid absorbent
Si02
Al203
8/3/2009
Chem 311
12
25-
Polar Force Normal Phase
8/3/2009
Chem 311
13
Dispersive Forces -Reverse

Phase London forces & Van der Waals
8/3/2009
Induced dipoles
Reversed phase LC
R = C18 H37
C8 H17
C3H6NH2
C4H9
Chem 311
14
Reverse Phase
Elutropic Series Weak solvent
Water lowest eluting power

Acetonitrile
Methanol
THF
CH2Cl2 Great eluting power
Non-aqueous reverse phase
8/3/2009
Chem 311
15
25-
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
16
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
17
HPLC Instrument
Solvent Supply
Pump
Constant flow
Gradient or Mixture
Injector
Valve
Column
Detector
Readout
Computer
Integrator
Strip Chart
8/3/2009
Chem 311
18
25-
Pumps
Single piston - cheap - need pulse
damper
Dual piston - most common
8/3/2009
Chem 311
19
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
20
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
8/3/2009
Chem 311
21
25-
HPLC Instrument
Injector Valve
8/3/2009
Chem 311
22
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.
8/3/2009
106
107 plates
Chem 311
23
Detectors
Refractive index - Difference
between solvent and sample solution.
Universal sensitivity
10-9gm/sec best case RI = 1x10-7
Very temp. sensitive 10-4RI/oC
0.001 C stability req.
Linear Range 3000

peaks may be + or 8/3/2009
Chem 311
24
25-
Absorption UV - Vis
Flow Cell in Spectrometer
Diode Array for full spectrum
10-12gm/sec sensitivity best case
Absorptivity 100 - 500,000/mole
8l flow cells with 1 cm path length
3l flow cell with 0.5 cm path
0.1 l flow cell with 0.2 cm path
0.03l flow cell with 0.10 cm
Linear range 3000+

8/3/2009
Chem 311
25
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
26
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
8/3/2009
Chem 311
27
25-
Sample
Electrochemical
Cell
Voltage
source
Amperometry
biological studies A
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
8/3/2009
Chem 311
28
Other Detectors
Evaporative Light
Scattering
Sample sprayed
into chamber
Solute particles
scatter light
Universal
detector - nonvolatile solutes
8/3/2009
Chem 311
29
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
8/3/2009
Chem 311
30
25-
10
Comparison of Detectors
8/3/2009
Chem 311
31
Peak integration
Drop perpendicular
Most common method

Simple
Small peak + error
Large Peak error
8/3/2009
Chem 311
32
Peak integration
Valley Method
Always produces errors
Errors can be large
Useful for
Multiple peaks
Complex baseline
8/3/2009
Chem 311
33
25-
11
Peak integration
Skim Method
Exponential Skim
Linear Skim
R<1
Small peak < 5% of large
8/3/2009
Chem 311
34
Peak integration
Gaussian Skim
More Complicated
Less error than other skim methods
8/3/2009
Chem 311
35
Peak integration
Errors depend
Relative Peak areas
Resolution
Method used
Improve Resolution!!!
8/3/2009
Chem 311
36
25-
12
Ionic forces
Ion exchange LC
Polymer Resin -
Styrene- DVB
PVP-DVB
Bonded Silica
Anion exchange
Cation Exchange
R SO3 H
O
R
8/3/2009
OH
Chem 311
37
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
38
Ion chromatography
DIONEX
2 Columns - Separator and Ion suppressor
Conductivity Detector
for anions - suppressor is cation

exchange
mobile phase
Na0H or Na2CO3 in
H20
8/3/2009
NaCl ==> HCl

NaF ==> HF
Chem 311
Mobile phase H2O
or H2CO3
39
25-
13
Ion Chromatograph
8/3/2009
Chem 311
40
Ion chromatography
Sample Range ppb
Cations
to100%
alkali metals, NH4+ ,etc

transition metals + ligand post columns
spectrophotometric
Anions NO3-, SO42- halides, S04, PO4, NO3

inositol phosphates, etc.
8/3/2009
Chem 311
41
Ion chromatography
Cr(VI) (Erin Brockovich)
Toxic form of Cr
ICP Total
Cr3+ good
8/3/2009
Chem 311
42
25-
14
Ion chromatography
Cation Analysis
Soils
Water
8/3/2009
Chem 311
43
Ion chromatography
Anions
Water pollution
Soil Analysis
8/3/2009
Chem 311
44
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
8/3/2009
Chem 311
45
25-
15
Stearic Exclusion
Molecular sieves Zeolite clays
4, 5, 13 holes
Sephadex - dextran polymer

holes for molecules 100 - 5x105 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
46
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
47
Stearic Exclusion
Advantages
Vo =Vol. of mobile
phase not in pores
Vi = volume in all
pores
All peaks elute
between Vo
and Vo + Vi =
total void volume
8/3/2009
log M.W.
Chem 311
Vo small
molecules
Vo large
log Retention volume
48
25-
16
Steric Exclusion
Calibrate VR vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
8/3/2009
Chem 311
49
25-
17
Chem 311
Chapter 26
IC, GPC and CE
8/3/2009
Chem 311
Ionic forces
Ion exchange LC
Polymer Resin -
Styrene- DVB
PVP-DVB
Bonded Silica
Anion exchange
Cation Exchange
R SO3 H
O
R
8/3/2009
Chem 311
OH
2
Ion Exchange
Uses
Water purification
Amino acid analyzers
Rare earth (RE) separations
8/3/2009
30 ft columns in series
citrate complexing agent
pH gradient
citrate RE -- Resin RE competition
Chem 311
26-
Ion chromatography
DIONEX
2 Columns - Separator and Ion suppressor
Conductivity Detector
for anions - suppressor is cation

exchange
mobile phase
Na0H or Na2CO3 in
H20
8/3/2009
NaCl ==> HCl

NaF ==> HF
Chem 311
Mobile phase H2O
or H2CO3
Ion Chromatograph
8/3/2009
Chem 311
Ion chromatography
Sample Range ppb
Cations
to100%
alkali metals, NH4+ ,etc

transition metals + ligand post columns
spectrophotometric
Anions NO3-, SO42- halides, S04, PO4, NO3

inositol phosphates, etc.
8/3/2009
Chem 311
26-
Ion chromatography
Cr(VI) (Erin Brockovich)
Toxic form of Cr
ICP Total
Cr3+ good
8/3/2009
Chem 311
Ion chromatography
Cation Analysis
Soils
Water
8/3/2009
Chem 311
Ion chromatography
Anions
Water pollution
Soil Analysis
8/3/2009
Chem 311
26-
Reagent Free IC
Electrochemical Generation of mobile
phase
8/3/2009
Chem 311
10
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
Chem 311
11
Stearic Exclusion
Molecular sieves Zeolite clays
4, 5, 13 holes
Sephadex - dextran polymer

holes for molecules 100 - 5x105 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
12
26-
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
13
Stearic Exclusion
Advantages
Vo =Vol. of mobile
phase not in pores
Vi = volume in all
pores
All peaks elute
between Vo
and Vo + Vi =
total void volume
8/3/2009
log M.W.
Chem 311
Vo small
molecules
Vo large
log Retention volume
14
Steric Exclusion
Calibrate VR vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
8/3/2009
Chem 311
15
26-
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 Vep=uepE
uep= charge/6* pi* viscosity* radius or ion
8/3/2009
Chem 311
16
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
Chem 311
17
Fused silica tube
treated with base to get
free silanol groups
At high pH, surface is
negative Layer of
positive buffer ions
forms to counter the
charge.
8/3/2009
Chem 311
18
26-
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
Chem 311
19
Electroosmotic flow
Veof = ueof E
dielectric cons * Zeta
4 pi * viscosity
Zeta k * wall charge * double layer thick
ueof
8/3/2009
Chem 311
20
Ion Velocity
V=Vep + Veof
Cations both positive
V> Veof
Anions Vep negative

V<Veof
Neutral Species V=Veof

8/3/2009
Chem 311
21
26-
Separation
Depends on Electrophoretic mobility

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
Chem 311
22
Instrument Diagram
8/3/2009
Chem 311
23
Instrument
Injection
Pressure - siphon effect or gas pressure
Electrophoretic injection place capillary in
sample and apply voltage to draw sample in.
Mechanically more complex than HPLC or GC
and not as easy to reproduce.
Columns - Silica - surface treated

8/3/2009
Chem 311
24
26-
Instrument
Detection note MWs in 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 electroactive solutes

10-18 to 10-19 moles
Vacancy detection - add absorbing species to buffer

and look at vacancies.
8/3/2009
Chem 311
25
Example of CE
DNA fragments
8/3/2009
Chem 311
26
MECC - Micelles
Micelles trap neutral organics and form
dynamic stationary phase.
Micelles migrate with or against the flow
depending on charge
8/3/2009
Chem 311
27
26-
MECC - Micelles
Typical surfactants used
8/3/2009
Chem 311
28
MECC Micelles
Example of MECC
8/3/2009
Chem 311
29
Cyclodextrin CE
Cyclodextrins are basket shaped molecules
which can trap small molecules inside them.
8/3/2009
Chem 311
30
26-
10
Cyclodextrin CE
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
Chem 311
31
Capillary ElectroChromatography
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
Chem 311
32
CEC
Flat flow profile less
band broadening then
HPLC
8/3/2009
Chem 311
33
26-
11
CEC
Column packings being developed with
ODS bonded to silica but sufficient
silica surface to pump buffers.
Chiral separations Cyclodextrin and other additives
8/3/2009
Chem 311
34
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
Chem 311
35
CE on a Chip
State of the art 96
parallel CE channels on a
chip
Detection
Laser fluorescence
MS
Separation in a few seconds
8/3/2009
Chem 311
36
26-
12

Chem

Diunggah oleh

Informasi Dokumen

Hak Cipta

Format Tersedia

Bagikan dokumen Ini

Bagikan atau Tanam Dokumen

Opsi Berbagi

Apakah menurut Anda dokumen ini bermanfaat?

Apakah konten ini tidak pantas?

Hak Cipta:

Format Tersedia

Chem

Diunggah oleh

Hak Cipta:

Format Tersedia

Chemistry 311

RECORDING DATA - USE OF

REALS (Things you measure)

Chem 311 Fall 2009

Chem 311 Fall 2009

Chem 311 Fall 2009

Round off only once

Chem 311 Fall 2009

Chem 311 Fall 2009

Rule of Thumb Keep the same

Chem 311 Fall 2009

Chem 311 Fall 2009

EXCEPTION.The calculation of sums

Chem 311 Fall 2009

Measured value - real

LOGS - General rule

pH 13.03 [H+] = 9.3x10-14

1.02 < pH < 1.04

13.02 < pH < 13.04

Note that the relative uncertainties are the

Chem 311 Fall 2009

40.34 ml +/- .01 ml

Chem 311 Fall 2009

.0047 2 + .034 2 + .0012 2 0. 034

Chem 311 Fall 2009

Chem 311 Fall 2009

V= 43.24 0.01 ml - 0.230.01 ml =

error 0.01 0.01 0.014 ml

Chem 311 Fall 2009

204.23 .02 gm 43.01 0.014 ml 1000 ml

error = 0.00038 = 0.0004 relative error

Chem 311 Fall 2009

Chem 311 Fall 2009

Chem 311 Fall 2009

Acid Base = Protons Conserved

Chem 311 Fall 2009

Chem 311 Fall 2009

C10H20N2S4 + oxidizing agent 4 SO2

Overall Ratio 1:8

Chem 311 Fall 2009

A Look Inside What We Do

Chem 311 Fall 2009

Chem 311 Fall 2009

Relative resolution uncertainty of measurement

150.0000 gm weighed on same balance

Chem 311 Fall 2009

Intermediate conversion device

Serial Interface Box

Chem 311 Fall 2009

Bias in Interpolation reading

Chem 311 Fall 2009

Volume to Length Converters

Chem 311 Fall 2009

Volume to Length Converters

Chem 311 Fall 2009

Volume to Length Converters

Class A Pipets 1 ppt

Chem 311 Fall 2009

Chem 311 Fall 2009

Challenge of Digital Data

A/D converters electrical signal to number

Serial Interface 0-5 volts ==> 0 to 4096

Chem 311 Fall 2009

Thermistor Temperature probe