Professional Documents
Culture Documents
Lecture Notes
Fall 2009
David E. Henderson
Chem 311
Chapter 2 & 3
How do we measure things
8/3/2009
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
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2-
SIGNIFICANT FIGURES
KEEP ONLY THE FIRST UNCERTAIN
DIGIT IN A RESULT.
Assume that data is presented this way
by other responsible scientists.
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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
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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.
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2-
SIGNIFICANT FIGURES
MULTIPLYING OR DIVIDING
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.
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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
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2-
SIGNIFICANT FIGURES
DURING CALCULATIONS
ONE ADDITIONAL INSIGNIFICANT DIGIT
SHOULD BE CARRIED IN
INTERMEDIATE RESULTS.
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.
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11
12
2-
SIGNIFICANT FIGURES
The Correct Way
Uncertainty of sum or difference
y abc
s y sa sb2 sc2
2
0.0002 0.014
13
SIGNIFICANT FIGURES
MULTIPLYING OR DIVIDING
UNCERTAINTY OF THE RESULT IS
THE SUM OF THE RELATIVE
UNCERTAINTIES.
a *b
y
c
2
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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
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.
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16
CALCULATION OF
MAXIMUM EXPERIMENTAL
ERROR -Examples.
Addition and subtraction
Volume of titrant = Final buret reading initial reading
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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
M=.0646066 Mol/l
error
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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
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
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y log(a)
sa
sy
2.303a
y 10 a
sy
2.303sa
y
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
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21
2-
Stoichiometry
Conservation of Mass
Mass of each atom is conserved in all Reactions
Types of Reactions
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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
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23
Stoichiometry Example
Anabuse
1molH 2 SO4
1molSO2
1molAnabuse
x
x
2molNaOH 1molH 2 SO4
4molSO2
mol Anabuse
24
2-
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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
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26
0.0100 gm
0.0001/0.0100 * 1000 = 10 ppt
30.00 ml titrant
0.01/30.00 * 1000 = 0.3 ppt
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27
2-
Conversion devices
Quantity being measured
Transducer
Sensor
Thermistor
Temperature probe
Readout Conversion
Data Logger on PC
Calibrated T readout
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28
Linear Scales
Interpolation
Estimate position between smallest division
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29
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
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30
2-
10
31
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32
Digital Pipetters
Actual Data for 3 Fisher adjustable pipetters
Pipette
"20-200
Sample Size
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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
33
2-
11
34
35
Discrete Calculus
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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
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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
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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.
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40
Result
1.4
1.2
1
0.8
Data
0.6
0.4
-2 stdev
+ 2 stdev
0.2
0
0
10
Experiment Day
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41
42
2-
14
Sample Preparation
See Chapter 28
43
Solids
Is the particle size appropriate for
dissolution or extraction?
If not, mill, grind, chop, blend etc.
44
Non-volatile components
separation is necessary by boiling, soxhlet
extraction, sonication, microwave
digestion, supercritical fluid extraction.
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45
2-
15
46
Strange Techniques
8/3/2009
Soxhlet Extraction Solid Phase ExtractionSolid Phase MicroextractionHeadspace SamplingPurge and Trap analysisMicrowave methodsSonication MethodsChem 311 Fall 2009
47
48
2-
16
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49
Using EXCEL
Excel powerful tool
Calculations
Graphing
Curve fitting
Do your own!!
Include your name in the File Name
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50
2-
17
Chem 311
Chapter 4 Lecture 1
Meaurement and Calculation
Accuracy and Precision
8/3/2009
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
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
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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
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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.
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MEANS OF MULTIPLE
EXPERIMENTAL
DETERMINATIONS
4-
Standard Deviation
Assume Gaussian Distribution of
Experimental Values
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Standard Deviation
Standard Terms associated with statistical
treatment of data.
(x i )
Mean - arithmetic average. x n
n
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Standard Deviation
x x
or x (population >24)
x x
i
n 1
V = sx2 or V = x2
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4-
Standard Deviation
Relative standard deviation.
s
% sx
ppt s
100 %
1000 ppt
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.
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11
Significance of Standard
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.
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12
4-
Significance of Standard
Deviation
99.8% within 3 std. dev.
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13
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14
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.
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tn sx
n
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
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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
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18
4-
Confidence Limits
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tn s x
n
19
Types of Errors
Random Errors - Indeterminate errors
Have no definite cause
random errors in reading a buret
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20
Types of Errors
Systemmatic Errors - Determinate errors
Have a definite cause
Balance not calibrated
Solution at wrong temperature for volumetric work
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
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22
Types of Errors
Personal Errors
Analyst makes a mistake
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23
Finding Errors
Constant errors
Balance reads 0.002 g high
Proportional errors
Balance reads 0.1% high
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.
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25
ELIMINATION OF
QUESTIONABLE RESULTS
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.
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26
ELIMINATION OF
QUESTIONABLE RESULTS
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.
27
4-
ELIMINATION OF
QUESTIONABLE RESULTS
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
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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
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29
Significance Testing
NEVER PROVE ANYTHING IS TRUE
Prove NOT FALSE
30
4-
10
Significance Testing
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?
F-Test
Compare standard deviations for data sets or methods
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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
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
33
4-
11
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
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2.364624
Chem 311 Fall 2009
34
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
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4-
12
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
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
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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
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Chem 311
Calibration Methods
Single Point
External Standard Method Calibration
Curve
Internal Standard Method
Standard Addition Method
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Chem 311
Chem 311
4 part 2-
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Chem 311
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Chem 311
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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
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10
Chem 311
11
ASSUMPTIONS
Values for the x axis are relatively free from error.
Concentration = independent variable
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12
4 part 2-
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
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13
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14
bo
b1
R2
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4 part 2-
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4 part 2-
Chem 311
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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
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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
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X values
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21
4 part 2-
Residuals
Random distribution ==> Data is inherently
linear
Non-Random ==> Data not linear
Use a different model
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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
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23
Residuals
Note pattern in residuals
Residuals
Residuals
0.2
0.1
0
-0.1 0
Residuals
2
-0.2
X value
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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.
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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
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26
Precision of Unknown
m = number of observations of unknown
n = number of calibration points
sx
sr
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sr
b1
( yx y)2
1 1
m n b12 ( xi x) 2
(y
y i ) 2
n2
27
4 part 2-
Precision of Unknown
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
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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
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29
Precision of Unknown
Confidence Limit for unknown
xtrue x ts x
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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.
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31
4
Exp. Values
-b+m
+b-m
1
0
-1 0
X values
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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
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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
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4 part 2-
12
Chapter 5
QA & Calibration
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Chem 311
QA/QC
Critical part of any analysis
Does the method work?
Instruments OK?
Reagents OK?
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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)
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5-
Range of Method
Limit of Detection
Limit of Quantitation
Limit of Linearity
Limit of response
Chem 311
Limit of Detection
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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
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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
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
Chem 311
5-
Accuracy
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10
Recovery Study
Validate sample preparation/extraction etc
Run Sample
Run Sample with known addition
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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
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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
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5-
Analysis Methods
Internal Standard Method
eliminates injection error in Chromatography
- only integration error is left
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16
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
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10
15
Mass Ratio
Mass Ratio
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5-
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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-
Equilibrium constant
At Equilibrium
G o RT ln K eq
c d
o o
K eq c a d b
a b
o o
a b
8/3/2009
Chem 311
Equilibrium constant
Normally write using Concentrations
(6-2)
K eq
Ceqa Deqd
Aeqa Beqb
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
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-
8/3/2009
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
Chem 311
12
6-
Strong Acids
Above H3O+
HCl + H2O <===> Cl- + H3O+
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
Kb
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
Ka
[H ][OAc ]
[HOAc]
Kb
8/3/2009
[OH ][HOAc]
[OAc ]
Chem 311
17
Ka
8/3/2009
18
6-
KaKb
= Ks
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
8/3/2009
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
Solubility Equilibria
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.
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
Chem 311
26
SEQUENTIAL COMPLEXATION BY
SIMPLE LIGANDS
Ni(H2O)42+ + 4 Cl- <===> NiCl42- +
4H2O) REACTION OCCUR IN A SERIES OF
STEPS
[NiCl ]
K
Chem 311
K4
[Ni ][Cl ]
[NiCl 2 ]
[NiCl ][Cl ]
[NiCl 3 ]
[NiCl 2 ][Cl ]
[NiCl 4 2 ]
[NiCl 3 ][Cl ]
27
6-
2= K1K2
2
[NiCl 2 ]
2
3= K1K2K3
4=K1K2K3K4
[Ni ][Cl ]
8/3/2009
Chem 311
28
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
Species present at a pH
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
Species present at a pH
For monoprotic acid - Exact equation
HA
A
8/3/2009
[H ]
[ H ] K a'
K a'
[ H ] K a'
Chem 311
33
6-
11
Species present at a pH
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
Species present at a pH
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
Chem 311
35
Species present at a pH
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
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
0.8
0
10
Concentration (F)
15
Activity H+
Activity H+
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-
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
8-
Concentration Effects
Concentration increases
Ions get close enough to be attracted and
repelled by other ions
All ions are solvated (Solvation number)
Chem 311
Activity Coefficients
Function of Ionic Strength
S
1
[ A z ]z A 2 [ B z ]z B 2 ...
2
Chem 311
8/3/2009
Chem 311
8-
8/3/2009
10
8/3/2009
Chem 311
11
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
8/3/2009
Chem 311
14
Kw = 1.00 X 10-14
Ka
[H ][BA ]
5
6.3x10
[HBA]
Chem 311
15
8-
[BA- ] = [H +] - [OH- ]
Phosphoric acid /potassium phosphate equilibrium
8/3/2009
Chem 311
16
8/3/2009
Chem 311
17
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 ]
[ 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
Ka
If CHBA>> [H+]
8/3/2009
[ H ][ Ba ]
C HBa [ H ]
[H] CHAKa
Chem 311
21
8-
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
Chem 311
24
8-
Solving on a TI Calculator
TI- 83
8/3/2009
Chem 311
25
Solving on a TI Calculator
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
Chem 311
26
Solubility Equilibria
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
Chem 311
Strong Acid/Base
Completely dissociated
[H+] = CHA
[OH-] = CB
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
[ 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
8/3/2009
Chem 311
9-
.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+]
8/3/2009
Chem 311
Buffer Example
Mass Balance rearranged
[NH3]= 0.200 + 0.100 - [NH4+]
substitute in expression for NH3
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 ]
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-]}
[ H ]C A
C HA
Chem 311
12
9-
Acidic buffer
K a'
[ H ]C A
C HA
8/3/2009
Chem 311
[ H ]C A
CHA
C
pK a log[ H ] log A
CHA
C
pK a pH log A
CHA
C
pH pK a log A
CHA 13
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
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.
Chem 311
Polyprotic systems
If K1 >K2 solve as monoprotic acid
(or base)
Use K1 and quadratic
8/3/2009
Chem 311
10-
Polyprotic systems
H3P04 + H20 <===>
K a1
H2P04-
H30+
[H ][H 2 PO ]
[H 3 PO4 ]
K a2
HP04-2 + H20
[H ][H PO 4 ]
[H 2 PO 4 ]
K a3
8/3/2009
[H ][ PO 4 ]
[H PO
2
4
Chem 311
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-
Chem 311
10-
Chem 311
Chem 311
Chem 311
10-
Chem 311
10
[ 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
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-
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
[H ]
K a1 [HMal ]
Simplifying Assumptions
1. If [HMAl] >> KA1
then Ka1 + [HMal-] = [HMal-]
[H ]
K a1 (K a2 [HMal ] K w )
[HMal ]
8/3/2009
Chem 311
14
K a1 (K a2 [HMal ] K w )
[HMal ]
Simplifying Assumptions
If KA2 [HMAl] >> Kw
[H ]
K a1 K a2 [HMal ]
[HMal ]
[H ]
8/3/2009
K a1 K a2
Chem 311
15
10-
[H ]
K a1 K a2
Chem 311
16
[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
Chem 311
18
10-
3.56x10-9
8/3/2009
2.39 x10 8
0.770
3.10 x10 8
Chem 311
19
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
8/3/2009
Chem 311
20
Repeat
[H ]
Result OK
8/3/2009
Chem 311
21
10-
Summary
Weak Acid
[ H ]2 K a [ H ] C HA K a 0
2
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
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
Ct
8/3/2009
Chem 311
CtVt CoVo
Vo Vt
6
11-
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
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
10
Visual Indicators
http://inst.santafe.cc.fl.us/~chem/intbl.html
8/3/2009
Chem 311
11
End Points
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
Yes
What is limiting reagent?
Calculate concentration of all reactants and products
No
Weak Acid Only
Amphiprotic Substance
pH=1/2(pK1 + pK2)
Buffer - HH Equation
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
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
moles acid = 25.0x0.1 = 2.5 mmoles
moles base = 25.0x0.1 = 2.5 mmoles
acid reacted to NaHP= 2.5 mmoles
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
moles acid = 25.0x0.1 = 2.5 mmoles
moles base = 35.0x0.1 = 3.5 mmoles
acid reacted to NaHP= 2.5 mmoles
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
moles base = 50.0x0.1 = 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
moles base = 75.0x0.1 = 7.5 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
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
K4
[NiCl ]
2
[Ni ][Cl ]
K2
K3
[NiCl 2 ]
[NiCl ][Cl ]
[NiCl 3 ]
[NiCl 2 ][Cl ]
[NiCl 4 2 ]
[NiCl 3 ][Cl ]
Chem 311
2= K1K2 3= K1K2K3
4=K1K2K3K4
[NiCl ]
[Ni ][Cl ]
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
[Ni ]
C Ni
Ni
Ni
[Ni ]
2
Ni
1
1 +1[Cl] +2[Cl]2+ 3[Cl]3+ 4[Cl]4
1
2
8/3/2009
Chem 311
O
O
Cu
Ni
N
8/3/2009
Chem 311
12-
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
12-
AMINO POLYCARBOXYLIC
ACIDS
EDTA - Ethylenediamine
tetraacetic acid.
H
O
H
O
N
N
H
O
H
O
O
Chem 311
10
AMINO POLYCARBOXYLIC
ACIDS
NTA - Nitrilotriacetic acid.
Tetradentate - this is best for small metals.
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-
ACID-BASE CHEMISTRY OF
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
ACID-BASE CHEMISTRY OF
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
ACID-BASE CHEMISTRY OF
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
Chem 311
16
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
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
Chem 311
21
12-
Example
40.00 ml of titrant added:
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-
0.999 xCm 0
[ MY ]
106
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
8/3/2009
Chem 311
29
Auxillary Ligands
[M] = m Cm where m refers to the
auxiliary ligand.
Kf
[ MY ]
[ MY ]
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
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
8/3/2009
Chem 311
34
Abs.
Vol. Titrant
8/3/2009
Chem 311
35
Fe E.P.
Vol. Titrant
8/3/2009
Chem 311
36
12-
12
8/3/2009
Chem 311
37
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
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.
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
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
Chem 311
48
12-
16
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
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
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
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
8/3/2009
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-
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
14
Pt / Fe (1.0 M) // Ce (1.0 M) / Pt
Chem 311
15
14-
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
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-
8/3/2009
Chem 311
22
8/3/2009
Chem 311
23
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-
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
5
[Mn04-][H+]8
= 1.51 -.0591 log 1.0 x 10-4
5
0.010x(1.0)8
8/3/2009
Chem 311
26
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.
Chem 311
15-
Indicator Electrodes
Electrodes of the 1st kind (Probe the
primary reactant metal)
Metal electrodes don't work very well
Contamination
Interferences
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
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
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
y
nx
8/3/2009
Chem 311
14
y
nx
Chem 311
15
15-
Example of Selectivity
Na+ glass electrode 11% Na20 18% Al203 71%
Si02
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
Chem 311
18
15-
2
Note N = 2 sensitivity only 1/2 that for N = 1
8/3/2009
Chem 311
19
Chem 311
O
O
20
8/3/2009
Chem 311
21
15-
KCl
filling
solution
Tungsten wire
22
Treated Tip
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
8/3/2009
Chem 311
24
15-
Chem 311
25
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-
8/3/2009
Chem 311
28
Enzyme Electrodes
ISE, pH, or gas electrode
Urease <===>NH3
Decarboxylazes===>H+
pH Electrode
Enzyme in
acrylamide gel
cholesterol electrode
Chem 311
29
Enzyme Electrodes
Nerve gas electrode (& phosphate
pesticides) - Cholinesterase inhibitors
enzyme film
add substrate to sample
monitor reduced activity
Chem 311
30
15-
10
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
8/3/2009
Chem 311
35
8/3/2009
Chem 311
36
15-
12
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
Methods of Application of
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
Methods of Application of
ISE's
If addition causes little volume change Ej
will stay the same
Std. allows compensation for unknown
matrix
8/3/2009
n
Es = Q + .0591 log VsCs+VxCx
n
Vs+Vx
Chem 311
40
Methods of Application of
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)
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
Methods of Application of
ISE's
8/3/2009
Chem 311
-Cx
42
Unk.
Unk+S1
Unk+S2
15-
14
Methods of Application of
ISE's
Potentiometric Titration
Very precise and accurate
but not applicable at such low conc. as direct
potentiometry
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
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
8/3/2009
Electromagnetic Radiation
= 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-
Electromagnetic Radiation
(wave number) = waves/cm = cm-1 = 1/
in vacuum
E = h = hc/ = hc
8/3/2009
Chem 311
Electromagnetic spectrum
8/3/2009
Chem 311
Rotational Transition
Vibrational Transition
Electronic Transition
8/3/2009
Chem 311
18-
____1s
Emission occurs when e- state drops.
Studied for atoms. Chemland
8/3/2009
Chem 311
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
Chem 311
10
Quantitative aspects of
Absorption
dx
P(o)
P(x)
dP = -k Px C dx
8/3/2009
Chem 311
11
Quantitative aspects of
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 )
-dP/Px = kC dx
Integrate over entire path length x= 0 to b
8/3/2009
Chem 311
12
18-
Quantitative aspects of
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
Chem 311
13
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
15
Conc.
18-
8/3/2009
Chem 311
16
Bandwidth effects
8/3/2009
Chem 311
17
Bandwidth effects
8/3/2009
Chem 311
18
18-
Activity coef.
Temperature
Chem 311
19
Chem 311
20
Chem 311
21
18-
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
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-
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
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
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
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
Chem 311
21
19-
Detectors
Solid State Detectors
Photodiodes
Change Couple Devices.
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
Chem 311
23
Instrument Designs
Chem 311
24
19-
8/3/2009
Chem 311
25
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
Chem 311
A little History
To get a multi-stage separation required
much time - weeks - years.
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
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
23-
8/3/2009
Chem 311
Solvent Extraction
Equilibrium of solute Z between two
immiscible phases:
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
8/3/2009
Chem 311
14
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-
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
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
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
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
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-
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.
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
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.
Chem 311
36
23-
12
8/3/2009
Chem 311
37
8/3/2009
Chem 311
38
8/3/2009
Chem 311
39
23-
13
Elution chromatography
8/3/2009
Chem 311
40
Elution chromatography
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
Elution chromatography
Capacity factor k'
- Describes Retention
k'
Vr Vo t r t o t ' r
Vo
to
to
RT
R
8/3/2009
Chem 311
42
23-
14
Elution chromatography
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 -
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
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
1e-1
10
100
54
23-
18
Reduced parameters
v = dpv/Dm
h = H/dp
Chem 311
55
Chem 311
56
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
59
Deviant Chromatographic
Behavior
Antilangmuir
Anti-langmuir case
sample acts
as
stationary
phase
Langmuir
8/3/2009
Saturate
sites
Langmuir case
Chem 311
60
23-
20
Deviant Chromatographic
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
Resolution in Chromatography
R
( 1 )
1
4
k'
k'
8/3/2009
k'
1 k'
separation factor
Chem 311
65
Resolution in Chromatography
3 Ways to control separation
Change relative affinity of solutes for stationary
phase Column Selectivity ()
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
Chem 311
67
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
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
Chem 311
72
23-
24
8/3/2009
Chem 311
73
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
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
Elution
Different solvent used - solute partitions into solvent
Solute elutes from column
Chem 311
80
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
1000
10-100
meters
10,000100,000
8/3/2009
0.05 1 mm
ID
Chem 311
Types of Columns
Capillary columns
Chem 311
24-
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
Polarity
Application
General separation
by polarity
Ethylene glycol
esters
DEGS di-ethylene
glycol succinate
Very polar
O
O
8/3/2009
O
O
Chem
311
O
5
n
Chem 311
24-
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
8/3/2009
Chem 311
10
Rr
Rs
Amp
V
Rr
8/3/2009
Rs
Reference
Chem 311
Column
Sample
Column
DC Power
Supply
11
Chem 311
12
24-
Flame
Electrometer
Sample +
Hydrogen
8/3/2009
Chem 311
13
Electrode
200 volt DC
Flame
Electrometer
Sample +
Hydrogen
Chem 311
14
Electrometer
Sample
Beta
source
Chem 311
15
24-
Electrometer
Power
Supply - DC
or Pulsed
+
Sample
Beta
source
8/3/2009
Chem 311
16
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.
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
MS-MS
Fragment a Fragment
Improved S/N ratio
8/3/2009
Chem 311
20
Detector Summary
Detector
FID
ECD
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
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
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
GC-IR
IR spectrum of peaks
8/3/2009
Chem 311
27
24-
Analysis Methods
External Standard Method
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
Internal Standard Method
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
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
Administrator, 11/13/2006
25-
Columns
Small particles
More pressure
Harder to pack.
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A5
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A6
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Slide 5
A5
Administrator, 11/13/2006
Slide 6
A6
Administrator, 11/13/2006
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
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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
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Chem 311
10
LIQUID
CHROMATOGRAPHY
Forces which lead to retention
Ionic force + and - ions
Size Exclusion
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11
Polar Force
Normal phase LC
Silica or Aluminum solid absorbent
Si02
Al203
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12
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13
8/3/2009
Induced dipoles
Reversed phase LC
R = C18 H37
C8 H17
C3H6NH2
C4H9
Chem 311
14
Reverse Phase
Elutropic Series Weak solvent
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15
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Reverse Phase
Solvent Triangle
ACN, MeOH, THF
Binary and ternary mixtures
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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
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17
HPLC Instrument
Solvent Supply
Pump
Constant flow
Gradient or Mixture
Injector
Valve
Column
Detector
Readout
Computer
Integrator
Strip Chart
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18
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Pumps
Single piston - cheap - need pulse
damper
Dual piston - most common
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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
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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)
Chem 311
21
25-
HPLC Instrument
Injector Valve
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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.
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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.
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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
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
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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
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27
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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
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28
Other Detectors
Evaporative Light
Scattering
Sample sprayed
into chamber
Solute particles
scatter light
Universal
detector - nonvolatile solutes
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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
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30
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10
Comparison of Detectors
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31
Peak integration
Drop perpendicular
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32
Peak integration
Valley Method
Always produces errors
Errors can be large
Useful for
Multiple peaks
Complex baseline
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33
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11
Peak integration
Skim Method
Exponential Skim
Linear Skim
R<1
Small peak < 5% of large
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34
Peak integration
Gaussian Skim
More Complicated
Less error than other skim methods
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35
Peak integration
Errors depend
Relative Peak areas
Resolution
Method used
Improve Resolution!!!
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36
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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
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38
Ion chromatography
DIONEX
2 Columns - Separator and Ion suppressor
Conductivity Detector
39
25-
13
Ion Chromatograph
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40
Ion chromatography
Sample Range ppb
Cations
to100%
Chem 311
41
Ion chromatography
Cr(VI) (Erin Brockovich)
Toxic form of Cr
ICP Total
Cr3+ good
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42
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14
Ion chromatography
Cation Analysis
Soils
Water
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43
Ion chromatography
Anions
Water pollution
Soil Analysis
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44
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45
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15
Stearic Exclusion
Molecular sieves Zeolite clays
4, 5, 13 holes
Polystyrene beads
MW 200 - 50 million
organic mobile phases
non-polar samples
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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
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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
48
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16
Steric Exclusion
Calibrate VR vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
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17
Chem 311
Chapter 26
IC, GPC and CE
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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
Ion Chromatograph
8/3/2009
Chem 311
Ion chromatography
Sample Range ppb
Cations
to100%
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
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Chem 311
26-
Reagent Free IC
Electrochemical Generation of mobile
phase
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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
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11
Stearic Exclusion
Molecular sieves Zeolite clays
4, 5, 13 holes
Polystyrene beads
MW 200 - 50 million
organic mobile phases
non-polar samples
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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
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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
14
Steric Exclusion
Calibrate VR vs log M.W. and determine
unknowns.
No equilibrium so no eq. band
broadening
Low separation factor.
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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
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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
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17
Capillary Electrophoresis
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.
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26-
Capillary Electrophoresis
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.
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19
Capillary Electrophoresis
Electroosmotic flow
Veof = ueof E
dielectric cons * Zeta
4 pi * viscosity
Zeta k * wall charge * double layer thick
ueof
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20
Ion Velocity
V=Vep + Veof
Cations both positive
V> Veof
Chem 311
21
26-
Separation
Chem 311
22
Instrument Diagram
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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.
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
Chem 311
25
Example of CE
DNA fragments
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26
MECC - Micelles
Micelles trap neutral organics and form
dynamic stationary phase.
Micelles migrate with or against the flow
depending on charge
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27
26-
MECC - Micelles
Typical surfactants used
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28
MECC Micelles
Example of MECC
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29
Cyclodextrin CE
Cyclodextrins are basket shaped molecules
which can trap small molecules inside them.
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30
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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
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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.
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32
CEC
Flat flow profile less
band broadening then
HPLC
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33
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11
CEC
Column packings being developed with
ODS bonded to silica but sufficient
silica surface to pump buffers.
Chiral separations Cyclodextrin and other additives
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34
8/3/2009
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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
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36
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12